D6-2 Bio 03-Hirfan10

Project no.
(GOCE) 003985
Acronym: EuroDemo
Project title: European Platform for Demonstration of Efficient Soil and Groundwater
Remediation
Instrument: Coordination Action
Thematic Priority: Global change and ecosystems
D6-2 Status report on technological reliability for demonstrated soil and

groundwater management technologies with special focus on the situation
in Europe (update on the bioremediation part only)
Due date of deliverable: November 2005

Actual submission date: November 2005, revision February 2007
Start date of project: 01.01.2005 Duration: 3 years
Organisation name of lead contractor for this deliverable: Universität Lüneburg
Revision [draft, 1, 2, …]
Project co-funded by the European Commission within the Sixth Framework Programme (2002-2006)
Dissemination Level
PU Public PU
PP Restricted to other programme participants (including the Commission Services)
RE Restricted to a group specified by the consortium (including the Commission Services)
CO Confidential, only for members of the consortium (including the Commission Services)
State-Of-The-Art-Report: Enhanced In Situ Bioremediation Technologies 1
CG 6 Technical Reliability
1 Enhanced In Situ Bioremediation Technologies (EISB)

1.1 Introduction
Bioremediation relies on microorganisms to biologically degrade groundwater contami-
nants through a process called biodegradation. It may be engineered and accomplished
in two general ways: (1) stimulating native microorganisms by adding nutrients, oxygen,
or other electron acceptors (a process called biostimulation); or (2) providing supple-
mentary pregrown microorganisms to the contaminated site to augment naturally occur-
ring microorganisms (a process called bioaugmentation) (AFCEE, 2004; ITRC, 2002;
U.S.EPA, 2000a; U.S.EPA, 2004b). This technology mainly focuses on remediating or-
ganic chemicals such as fuels and chlorinated solvents. One approach, aerobic biore-
mediation, involves the delivery of oxygen (and potentially other nutrients) to the aquifer
to help native microorganisms reproduce and degrade the contaminant (U.S.EPA,
2004b). Another approach, anaerobic bioremediation, circulates electron donor materi-
als—for example, food-grade carbohydrates such as edible oils, molasses or lactic acid
whey—in the absence of oxygen throughout the contaminated zone to stimulate an-
aerobic microorganisms to consume the contaminant (AFCEE, 2004; DoD, 2002). In
some cases, pregrown microbes may be injected into the contaminated area to help
supplement existing microorganisms and enhance the degradation of the contaminant,
a process known as bioaugmentation. A potential advantage of bioremediation is its
ability to treat the contaminated groundwater in place with naturally occurring microor-
ganisms, rather than bringing contaminants to the surface. By using native microorgan-
isms, rather than injecting additional ones, cleanup can be more cost-effective at some
sites. However, heterogeneous subsurfaces can make delivering nutrient/oxygen solu-
tions to the contaminated zone difficult by trapping or affecting movement of both con-
taminants and groundwater. Also, nutrients for stimulating the microorganisms can be
consumed rapidly near the injection well, thereby limiting the microorganisms’ contact
with the contaminants, or stimulating biological growth at the injection site. In summary,
this technology avoids the costs associated with bringing water to the surface for treat-
ment; instead, the main costs associated with bioremediation include: delivery of the
amendments to the subsurface (which varies depending on the depth of contamination),
the cost of the amendments themselves, and monitoring of the treatment.
This report focuses on the approach of enhanced in situ anaerobic bioremediation of
chlorinated solvents, because chlorinated solvents are the most common groundwater
contaminants and therefore numerous laboratory studies as well as pilot and field appli-
cations have been or currently are being conducted. To date, enhanced anaerobic bio-
remediation has been applied at over 600 sites (AFCEE, 2004).
1.2 Enhanced In Situ Anaerobic Bioremediation of chlorinated sol-

vents
Enhanced in situ anaerobic bioremediation can be an effective method of degrading
various chlorinated solvents dissolved in groundwater, including chloroethenes, chloro-
ethanes, and chloromethanes. Collectively, these compounds (some of which are deg-
radation products of chlorinated solvents) are referred to as chlorinated aliphatic hydro-
carbons (CAHs) (AFCEE, 2004; Cope and Hughes, 2001; ITRC, 2004; U.S.EPA,
2000a).
Bioremediation of CAHs can occur through natural mechanism (natural attenuation or
intrinsic bioremediation or by enhancing the natural mechanisms (enhanced bioreme-
diation). The addition of an organic substrate to an aquifer has the potential to further
stimulate microbial growth and development, creating an anaerobic environment in
which rates of anaerobic degradation of CAHs may be enhanced. Therefore, a variety of
organic substrates have been applied to the subsurface to promote anaerobic degrada-
tion of CAHs to innocuous end products. In some cases, microorganisms also may be
added (bioaugmentation), but only if the natural microbial population is incapable of per-
forming the required transformations.
The most common chlorinated solvents released to the environment include tetra-
chloroethene (PCE, or perchloroethene), trichloroethene (TCE), trichloroethane (TCA),
and carbon tetrachloride (CT). These chlorinated solvents are problematic because of
their health hazards and their resistance to natural degradation processes. Because
these compounds exist in an oxidized state, they are generally not susceptible to aero-
bic oxidation processes (with the possible exception of cometabolism). However, oxi-
dized compounds are susceptible to reduction under anaerobic conditions by either bi-
otic (biological) or abiotic (chemical) processes. Enhanced anaerobic bioremediation is
intended to exploit primarily biotic anaerobic processes to degrade CAHs in groundwa-
ter (AFCEE, 2004; Suthersan, 2001).
Other common groundwater contaminants that are subject to reduction reactions are
also susceptible to enhanced anaerobic bioremediation. While not addressed in this
report, constituents that can also potentially be treated with this approach include the
following:
• Chlorobenzenes;
• Chlorinated pesticides (e.g., chlordane), polychlorinated biphenyls (PCBs), and
chlorinated cyclic hydrocarbons (e.g., pentachlorophenol);
• Oxidizers such as perchlorate and chlorate;
• Explosive and ordnance compounds;
• Dissolved metals (e.g., hexavalent chromium); and
• Nitrate and sulphate
Detailed informations for the design and implementation of enhanced anaerobic biore-
mediation systems for theses groundwater contaminants listed above are described e.g.
by (ITRC, 2002; Liles et al., 2004; Mikszewski, 2004; Suthersan, 2001).
1.2.1 Degradation Mechanisms

There are several potential reactions that may degrade CAHs in the subsurface, but not
all CAHs are amenable to degradation by each of these processes (Table 1). For ex-
ample, PCE is not amenable to any process of aerobic degradation, while TCE may
only be degraded by aerobic cometabolism that typically requires addition of a substrate
in the presence of oxygen (McCarty et al., 1998). However, anaerobic biodegradation
processes may potentially degrade not only PCE and TCE, but all of the common
chloroethenes, chloroethanes, and chloromethanes.
Altogether, the degradation pathways and microbiology of aerobic and anaerobic
dechlorination of chlorinated ethenes are better studied than for chloroethanes and
chloromethanes (AFCEE, 2004; U.S.EPA, 2000a). This is primarily because they occur
more frequently as contaminants in groundwater.
Anaerobic reductive dechlorination is the degradation process targeted by enhanced
anaerobic bioremediation. Through addition of organic substrates to the subsurface,
enhanced anaerobic bioremediation converts naturally aerobic or mildly anoxic aquifer
zones to anaerobic and microbiologically diverse reactive zones, making them condu-
cive to anaerobic degradation of CAHs.
Biodegradation of an organic substrate depletes the aquifer of dissolved oxygen (DO)
and other terminal electron acceptors (e.g., nitrate or sulfate), and lowers the oxidation-
reduction potential (ORP) of groundwater, thereby stimulating conditions conducive to
anaerobic degradation processes. After DO is consumed, anaerobic microorganisms
typically use native electron acceptors (as available) in the following order of prefer-
ence: nitrate, manganese and ferric iron oxyhydroxides, sulfate, and finally carbon diox-
ide. Figure 1 illustrates a CAH plume where substrate has been injected into the source
area. An anaerobic treatment area is created with the development of progressively
more anaerobic zones closer to the source of organic carbon as electron acceptors are
depleted. Anaerobic dechlorination has been demonstrated under nitrate, iron, and sul-
fate reducing conditions, but the most rapid biodegradation rates, affecting the widest
range of CAHs, occur under methanogenic conditions (Bouwer, 1994).
Table 1: Potential Degradation Prozesses for CAHs (AFCEE, 2004).

Chloroethenes Chloroethanes Chloromethanes
Degradation Process PCE TCE DCE VC PCA TCA DCA CA CT CF MC CM
Aerobic Oxidation N N P Y N N Y Y N N Y P
Aerobic
N Y Y Y P Y Y Y N Y Y Y
Cometabolism
Anaerobic Oxidation N N P Y N N Y P N N Y P
Direct Anaerobic Reduc-
Y Y Y Y Y Y Y Y Y Y Y Y
tive Dechlorination
Cometabolic Anaerobic
Y Y Y Y P Y Y P Y Y Y P
Reduction
Abiotic Transformation Y Y Y Y Y Y Y Y Y Y Y Y
PCE = tetrachloroethene, TCE = trichloroethene, DCE = dichloroethene, VC = vinyl chloride, PCA = tetra-
chloroethane, TCA = trichloroethane, DCA = dichloroethane, CA = chloroethane, CT = carbon tetrachlo-
ride, CF = chloroform, MC = methylene chloride, CM = chloromethane.
N = Not documented in the literature.
Y = Documented in the literature.
P = Potential for reaction to occur but not well documented in the literature.
Figure 1: Reducing zone established downgradient of substrate Injection (AFCEE, 2004).
Under anaerobic conditions, reductive dechlorination mechanisms can effectively bio-

degrade CAHs. Reductive dechlorination generally involves the sequential replacement
of a chlorine atom on a CAH with a hydrogen atom (that is, converting PCE to TCE to
DCE, and so on) and has been observed to occur both directly and cometabolically. In
anaerobic reductive dechlorination (direct), the mediating bacteria use the CAH directly
as an electron acceptor in energy-producing redox reactions. Anaerobic reductive
dechlorination (cometabolic) occurs when bacteria incidentally dechlorinate a CAH in
the process of using another electron acceptor to generate energy. The three general
reactions that may degrade CAHs by anaerobic reductive dechlorination are Direct An-
aerobic Reductive Dechlorination, Cometabolic Anaerobic Reductive Dechlorination and
Abiotic Reductive Dechlorination. More detailed information about the different path-
ways by which microorganisms transform contaminants is given by (AFCEE, 2004;
DoD, 2002; U.S.EPA, 2000a).
1.3 APPLICATION OF ENHANCED ANAEROBIC BIOREMEDIATION
Application of enhanced anaerobic bioremediation starts with a review of site-specific

conditions and evaluation of remedial objectives to determine if this remedial approach
is appropriate for a site. Once enhanced bioremediation is selected as a remedial alter-
native, design criteria for implementation are developed including selection of a sub-
strate and system configuration (AFCEE, 2004). The following subsections describe
some common technology screening criteria, substrate alternatives, and system con-
figurations used for enhanced anaerobic bioremediation.
1.3.1 Technology Screening
The addition of an organic substrate to the subsurface to stimulate and enhance the
anaerobic dechlorination process in situ has been explored at many sites (AFCEE,
2004). Enhanced anaerobic bioremediation has been applied under a broad range of
site conditions, including the following:
Hydrogeologic Settings. Enhanced anaerobic bioremediation has been applied in a

variety of hydrogeologic settings, from low permeability silts and clays to high perme-
ability alluvial sand and gravel deposits to fractured bedrock. Enhanced bioremediation
has been applied at depths up to 400 feet below ground surface (bgs) and with ground-
water velocities ranging from a few feet per year to several feet per day. However, there
are limits to applying the technology in settings with the extremes of very high and very
low rates of groundwater flow. It may be impractical to maintain reducing conditions in
high flow settings, due to the magnitude of groundwater and native electron acceptor
flux. On the other hand, it may be difficult to inject substrates into tight formations, and
under low flow settings mixing of substrate with groundwater due to advection and dis-
persion may be limited (AFCEE, 2004).
Contaminant Levels and Distribution. The technology has typically been applied to
groundwater plumes with concentrations of CAHs ranging from 0.01 to 100 mg/L. Sites
with indications of residual or sorbed DNAPL (dissolved CAH concentrations in excess

of 100 mg/L) also have been successfully treated. However, it may not be realistic to
expect rapid remediation of source areas with DNAPL pools (AFCEE, 2004).
Geochemical Conditions. During anaerobic dechlorination, CAHs function as electron

acceptors in competition with naturally occurring (inorganic) electron acceptors. For ex-
ample, a high rate of groundwater flow coupled with high concentrations of DO may
create an oxygen electron acceptor demand that cannot practically be overcome with
substrate addition. In some cases, adverse site conditions can be mitigated with proper
system design. For example, recirculation systems may be used to impose a hydraulic
gradient and enhance groundwater flow at sites with very low natural hydraulic gradi-
ents. However, when pumping of significant quantities of groundwater is required, the
technology may not be cost competitive with pump and treat; this becomes a site-
specific issue. Once enhanced bioremediation has been selected as an appropriate
technology, there are several substrate alternatives and system configurations to con-
sider (AFCEE, 2004).
1.3.2 Substrate
There are many organic substrates which can be naturally degraded and fermented in
the subsurface that result in the generation of hydrogen. Examples of easily fermentable
organic substrates include alcohols, low-molecular-weight fatty acids (e.g., lactate), car-
bohydrates (e.g., sugars), vegetable oils, and plant debris (e.g., mulch). The substrates
most commonly added for enhanced anaerobic bioremediation include lactate, molas-
ses, Hydrogen Releasing Compound (HRC®), and vegetable oils. Substrates used less
frequently include ethanol, methanol, benzoate, butyrate, high-fructose corn syrup
(HFCS), whey, bark mulch and compost, chitin, and gaseous hydrogen. Table 2 sum-
marizes the attributes of several substrate types. These substrates are classified here
as soluble substrates, viscous fluids and low viscosity fluids, solid substrates, and ex-
perimental substrates. The physical nature of the substrate dictates the frequency of
addition, the addition technique, and potential system configurations.
Table 2: Substrates used for enhanced anaerobic bioremediation (AFCEE, 2004).

1.3.3 System configuration

Enhanced in situ anaerobic bioremediation can be implemented to provide source area
or dissolved plume treatment or containment, or a combination of source area and dis-
solved plume remediation can be used. Enhanced bioremediation and conventional
source treatment or containment approaches (e.g., chemical oxidation or groundwater
extraction) will be subject to the same difficulties associated with mass transfer limita-
tions of a continuing source and preferential flow paths in heterogeneous formations
(AFCEE, 2004). The single largest difference between conventional remedial technolo-
gies and enhanced bioremediation may be that enhanced bioremediation, if properly

implemented, can maintain effectiveness over a longer period of time at a lower overall
cost. This may make enhanced bioremediation an effective remedial approach due to
the substantial challenges associated with significant CAH source mass removal. Typi-
cal system configurations and associated remedial action objectives that engineered
anaerobic bioremediation may be used to address include the following:
Source Zone Treatment: Remediation of source zones where good sub-

strate/contaminant contact is possible.
•Plume Containment using a Biologically Reactive Barrier: Reduction of mass flux

from a source zone or across a specified boundary.
•Plume-Wide Restoration: Total treatment of an entire dissolved plume.
In some cases, several approaches may be combined. For example, a source area may
be targeted for remediation using a grid configuration, combined with a linear barrier
configuration upgradient from a downgradient point of compliance (Figure 2).
Figure 2: Schematic of Source Area and Biobarrier Injection Configurations (AFCEE,

2004).
The appropriate application of enhanced anaerobic bioremediation will be site-specific

and based on a strategy that takes into account final remedial objectives, feasibility of
the application, and regulatory issues. Ultimately however, there will be an economic
limit to the size of a plume that can be treated with a complete plume-wide application of
enhanced bioremediation. For plume sizes greater than 10 to 20 acres, use of contain-
ment strategies combined with other remedial approaches may be more feasible.
Source Zone Treatment

Enhanced anaerobic bioremediation has been used to address source zones either to
limit mass flux from the source zone or to accelerate source mass removal. Mass flux
reduction is achieved by stimulating biodegradation in the dissolved phase, while reduc-
ing contaminant massis available to migrate downgradient. Source mass removal is
achieved by accelerating DNAPL dissolution and then stimulating biodegradation of the
dissolved contaminants. Anaerobic dechlorination is a process that takes place in the
aqueous phase and does not directly attack DNAPL mass. Therefore, enhanced biore-
mediation may be limited in its ability to rapidly treat DNAPL source zone areas
(AFCEE, 2004; DoD, 2002).
On the other hand, treatment to reduce mass flux and to perhaps increase the rate of
dissolution and treatment as compared to natural attenuation or groundwater extraction
may be more achievable. Enhanced bioremediation of DNAPL sources is being re-
searched and may someday be a proven and feasible long-term remedial alternative
(AFCEE, 2004).
Alternatively, injection of a low solubility, persistent carbon source such as vegetable oil
into a source area may serve to reduce mass flux and to effectively sequester the
source due to partitioning and lowering of hydraulic conductivity. However, while degra-
dation of dissolved constituents may be stimulated, this may not accelerate destruction
of DNAPL or sorbed source mass.
Plume Containment using Biologically Enhanced Barrier Systems

For large plumes having poorly defined, widely distributed, or inaccessible source ar-
eas, enhanced bioremediation systems may be configured as permeable reactive barri-
ers (biobarriers) to intercept and treat a contaminant plume. For example, biobarriers
may be employed at a property boundary or upgradient from a point of regulatory com-
pliance to prevent plume migration to potential receptors. Biobarriers typically consist of
either rows of substrate injection wells or a solid-substrate trench located perpendicular
to the direction of groundwater flow. Passive biobarriers typically use slow-release,
long-lasting substrates (e.g., HRC®, vegetable oils, or mulch) that can be either injected
or otherwise placed in a trench, and that are designed to remain in place for long peri-
ods to maintain the reaction zone. Contaminant mass is delivered to the treatment zone
via natural groundwater flow. Capital and operating costs for a passive biobarrier con-
figuration are typically lower than for plume-wide configurations because of a limited
treatment area. However, life-cycle costs could be significant if the source of the CAHs
upgradient of the biobarrier is not addressed. Semi-passive or active biobarriers are
similar to passive biobarriers except that a soluble substrate is typically injected periodi-
cally (semi-passive) or via a recirculation system (active). Soluble substrates migrate
with groundwater flow, are depleted more rapidly, and require frequent addition. How-
ever, these systems offer the advantage of being able to adjust the rate or type of sub-
strate loading over time, and soluble substrates may be easier to distribute throughout
larger volumes of the contaminant plume. Recirculation can improve substrate distribu-
tion, contaminant/substrate mixing, and retention time for treatment; but the overall
groundwater flux downgradient of the system does not change (AFCEE, 2004).
Plume-Wide Restoration
Enhanced bioremediation systems may be configured to treat dissolved CAHs across
an entire contaminant plume. Creating an anaerobic reaction zone across broad areas
of a plume is an aggressive approach that may reduce the overall timeframe for reme-
diation. Plume-wide delivery systems will typically be configured as a large injection
grid, or a recirculation well field may be employed to increase the effective area of sub-
strate distribution. Higher initial capital and operating costs of recirculation systems may
be offset by shorter remedial timeframes with lower monitoring and total long-term op-
erating costs. However, plume-wide applications where substrate is delivered to the en-
tire plume may be cost prohibitive for very large plumes or cost inefficient for low-level
contaminant plumes.
At sites where larger plumes are present (greater than several acres), or the depth of
the plume makes installing injection wells difficult and expensive, multiple treatment
lines can be established perpendicular to the direction of groundwater flow, typically
separated by 6 to 12 months of groundwater travel time. A recirculation approach may
not be practical or cost effective at a large scale due to the large volumes of groundwa-
ter to be processed and ineffective in situ mixing in heterogeneous environments. There
is some controversy as to the cost effectiveness of using enhanced anaerobic bioreme-
diation for plume-wide restoration. For any kind of recirculating system, groundwater
pumping rates may have to be similar to pump and treat methods; the cost of enhanced
bioremediation must be carefully compared to pump and treat. If substrate addition is
done by some kind of multiple point injection relying on natural groundwater flow for
dispersion, this may require very close spacing of injection points and or it may not re-
sult in good mixing of substrate and CAHs in situ (AFCEE, 2004).
1.3.4 Delivery Options

Common substrate delivery options include direct injection or recirculation of fluid sub-
strates, or emplacement of solid substrates in biowall trenches. Where direct-push
methods can be used, substrate may be injected directly through the probe rods. This is
a common approach for both slow-release and soluble substrates. Otherwise, injection
wells are used. Soluble substrates may be injected in batch mode, or in the case of fre-
quent injections, the use of automatic injection systems may be warranted. Recircula-
tion systems may also be employed for distribution of soluble substrates. Figure 3 is an
example of a horizontal circulation system. Recirculation may be continuous or in a
pulsed mode. Substrates are added to the groundwater as it is reinjected into the treat-
ment zone. Recirculation systems may be effective for difficult hydrogeological condi-
tions. For example, recirculation may be used to effectively mix substrate and contami-
nated groundwater at sites with very low hydraulic gradients and low rates of groundwa-
ter flow (AFCEE, 2004).
Figure 3: Schematic of a horizontal recirculation system (AFCEE, 2004).
1.4 Case Studies
Dry Cleaning Facility, Arlington, Texas

TECHNOLOGY: Hydrogen Release Compounds (HRC®)
SCALE: Full
GEOLOGY: Clayey silt underlain by medium to dark grey shale
CONTAMINANTS: PCE, TCE, cDCE, VC
The site is located in Arlington, Texas, covering approximately seven acres. A former
dry cleaner was situated in a small suite and operated at the site between 1982 and
1992. Environmental site assessments were conducted in 1996 and the site was admit-
ted into the TCEQ (Texas Commission on Environmental Quality) Voluntary Cleanup
Program. An unknown amount of chlorinated solvents was released at the site and had
contaminated soil and groundwater. The concentrations of contaminants of concern
(COC) in subsurface soils at a depth of approximately eight feet below the building were
in excess of site target concentrations. Site investigations revealed that the groundwater
with COC concentrations above the site target levels occupied approximately 3,500
square feet directly beneath and downgradient of the source area (U.S.EPA, 2004).
Immediately prior to injection of HRC®, the groundwater concentrations of COCs were
4,500 µg/L for PCE, 1,000 µg/L for TCE, 7,300 µg/L for cis-1,2-DCE, and 870 µg/L for
VC.
The underlying rock at the site is predominantly medium to dark grey shale, which read-
ily weathers to a thick, clayey soil. Site geology consists of a very low permeability light
to dark brown, soft, moist clay from the ground surface to approximately 22 feet bgs.
groundwater is encountered at 7 feet bgs.
Table 3 shows the target cleanup concentrations at the site, established by Conceptual
Environmental Assessment Model (CEAM), a health-based risk assessment for
groundwater.
The cleanup approach included soil excavation from the source area, performed in
1998, followed by in situ bioremediation using HRC® in 2002. In May 2000, HRC® was
injected into 45 borings that are located within the shallow aquifer area to a depth of
approximately 22 feet bgs. Of the 45 injection borings, 29 borings were advanced per-
pendicular to the ground surface, and 16 borings were advanced at angles of 15 to 30
degrees from vertical to extend beneath the building’s foundation. A total of approxi-
mately 7,000 pounds of HRC® were injected. Quarterly sampling of groundwater was
conducted as a part of the response action.
Table 3: Cleanup criteria

CEAM Cleanup Criteria for
Contaminant
Groundwater [µg/L]
PCE 500
TCE 500
cDCE 7,000
VC 200
Approximately 18 months after HRC injection, PCE, TCE, cDCE, and VC in the monitor-
ing well located nearest to the contaminant source area had decreased to 408 µg/L,
87.4 µg/L, 438 µg/L, and 132 µg/L, respectively. Based on these results, the site re-
ceived a Conditional Certificate of Completion from the TCEQ Voluntary Cleanup Pro-
gram on October 26, 2001.
INEEL, Test Area North, Idaho Falls, Idaho

TECHNOLOGY: Sodium lactate
SCALE: Full
GEOLOGY: Deep-fractured basalt aquifer
CONTAMINANTS: PCE, TCE, radionuclides
The historical injection of liquid wastes and concentrated sludges into the Snake River
Plain Aquifer using Well TSF-05 has resulted in a TCE, PCE, and tritium plume in
groundwater at the TAN facility of the Idaho National Engineering and Environmental
Laboratory (INEEL). The depth to water at TAN is approximately 200 ft. The aquifer and
most of the unsaturated zone are comprised primarily of layered basalt flows, interca-
lated with sedimentary interbeds deposited during periods of volcanic quiescence.
Groundwater flow in the aquifer is controlled by the highly transmissive zones that occur
at the contacts between individual basalt flows, and to a lesser extent, by fractured
zones within flow interiors. The scale of the basalt geology dictates that preferential flow
can be very important at spatial scales less than approximately 100 m (330 ft), after
which a transition to continuum behaviour occurs and the aquifer can be thought of es-
sentially as a macro-porous medium (ITRC, 2004).
Figure 4 shows the TCE plume at the TAN facility. A very large, low-concentration fringe
surrounds and emanates from a much smaller, high concentration core. Within the core
is a very small residual source area that continues to contaminate fresh groundwater
flowing through from up-gradient. The transition from the scale of the residual source,
where preferential flow is significant, to the scale of the fringe, where sufficient vertical
communication has been present along the flow path to create a relatively well-mixed,
predictable groundwater plume.
The residual source of contamination in the aquifer near TSF-05 is believed to be com-
posed primarily of the sludge that was injected into the well over 15–20 years. The pore
water of the sludge probably contains large amounts of TCE, with PCE and tritium also
present in significant amounts. Given the organic content of the sludge, sorbed PCE
and TCE are also likely to be present. TCE concentrations as high as 300,000 µg/L
have been measured in groundwater from well TSF-05. Some of the sludge has been
shown to have TCE concentrations as high as 3%. The sludge therefore represents a
long-term source of contamination to the aquifer.
Figure 4: Medial zone of TCE plume at TAN (TCE-concentrations in µg/L).
The 1995 Record of Decision (ROD) identifies the primary Remedial Action Objectives
(RAO) for TAN, which is to restore the contaminated aquifer groundwater by 2095 (100
years from the date of the ROD) by reducing all contaminants of concern to below
MCLs and a 1 × 10-4 total cumulative carcinogenic risk-based level for future residential
groundwater use and for noncarcinogens, until the cumulative hazard index is less than
1. The ROD selected pump and treat as the default remedy for the residual DNAPL
source area, but because this approach was unlikely to restore the site in a meaningful
time frame, it also identified five alternative technologies to be evaluated for their poten-
tial to enhance or replace the default remedy. One of the technologies identified was
enhanced in situ bioremediation (ISB).
Field activities for enhanced ISB began in November 1998 with the shutdown of the in-
terim pump and -treat facility that was operating in the hotspot and the initiation of the
first phase of ISB field operations. The primary performance objective of the field opera-
tion was to determine whether anaerobic reductive dechlorination of TCE in the residual
DNAPL source area could be enhanced through the addition of an electron donor (so-
dium lactate). Initially, baseline sampling and a conservative tracer test were performed,
followed by initiation of weekly lactate injections into the original disposal well and bi-
weekly groundwater monitoring for the parameter sets described above.
Seven months after sodium lactate injections began, significant dechlorination activity
was stimulated to distances of >40 m from the injection well. For example, TCE concen-
trations in well TSF-05 decreased to <10 µg/L, with increases in ethene of up to 2,500
µg/L. Dechlorination was strongly correlated with elevated electron donor concentra-
tions and the establishment of strongly reducing conditions. In particular, dechlorination
of TCE to cis-DCE coincided with sulphate reduction, while dechlorination of cis-DCE to
vinyl chloride and ethene was observed only in the presence of methanogenesis. Elec-
tron donor concentrations [as chemical oxygen demand (COD)] of >4,000 mg/L and
>5,800 mg/L were observed in wells TAN-25 and TAN-26 (refer to Figure 4 for well lo-
cations). By the end of the field evaluation, methane concentrations in these wells were
20,000 µg/L and 17,000 µg/L, while TCE concentrations had dropped to 68 µg/L and
nondetect.
A significant outcome of the field evaluation work was the strong evidence indicating
that the injected sodium lactate solution significantly enhanced the bioavailability of TCE
in the residual DNAPL source area, thereby accelerating degradation of the source.
This effect was evidenced by the near 21-fold increase in TCE concentrations that cor-
responded to arrival of lactate in well TAN-26, followed by complete dechlorination of
this newly bioavailable TCE to ethane.
ISB has been demonstrated to be a much more effective technology for treatment of
residual source areas than previously believed. Figure 5 shows the pre-lactate TCE
concentrations in the residual source area, and the TCE concentrations after 21 months
of lactate injections. Based on these substantial concentration reductions within and
immediately downgradient of the DNAPL residual source area, coupled with the evi-
dence for accelerated degradation of the residual DNAPL source, an amended ROD
was signed by the State of Idaho and EPA Region 10 to replace pump and treat with
bioremediation for restoration of the TCE source area at TAN. Operations conducted
since the conclusion of the field evaluation have been focused on optimization (the cur-
rent phase of operations) and long-term implementation of ISB as defined by the objec-
tives described below.
Figure 5: TCE concentrations before lactate injection (a) and TCE concentrations after 21
months of lactate injection (ITRC, 2004).
Two stages of operations remain for the TAN ISB hotspot remedy-optimization activities
and long-term operations. The primary performance objective for ISB optimization activi-
ties, which is the current phase of ISB operations, is to distribute electron donor
throughout the entire residual source area to eliminate VOC migration beyond the ISB
treatment cell. The performance metric for this objective, as it was for the ISB field
evaluation, is changes in groundwater concentration. Once down- and cross-gradient
migration has been eliminated, then the ISB remedy will move into its final phase, long-
term operations. The primary performance objective of ISB long-term operations is to
maintain the sufficiently large biologically active zone for a period of time such that the
residual source material is degraded to a point where natural attenuation processes will
address the residual levels of groundwater contamination.
The performance objective for ISB optimization activities is to distribute electron donor
throughout the entire residual DNAPL source area to completely eliminate contaminant
migration to down-gradient wells. During the field evaluation, a relatively constant sup-
ply of lactate was provided through weekly injections. During initial optimization activi-
ties, however, the presence of propionate and acetate combined with the absence of
lactate resulted in increased dechlorination efficiency as evidenced by conversion of all
accumulated cis-DCE to ethene throughout the residual source area. Therefore, the
primary operational change during optimization activities was to inject larger pulses of
lactate less frequently, approximately every six to eight weeks, to increase periods
when propionate was the primary electron donor. This has resulted in significant pro-
gress towards meeting the performance objective.
Although substantial progress toward the performance objective for optimization activi-
ties has been made, the objective has not yet been achieved. To achieve this objective,
a second injection well, was incorporated into ISB operations in December 2003. Al-
though the ISB remedy has not yet moved into long-term operations, progress toward
this objective can still be measured by monitoring chloroethene and ethene groundwater
concentrations in ISB wells. During optimization activities, the same pattern of en-
hanced TCE dissolution in response to sodium lactate injections, followed by complete
dechlorination of the newly bioavailable TCE to ethene that was observed in the field
evaluation still persisted. During optimization activities, TCE concentrations in well TSF-
05 have routinely increased from nondetect to 500–1000 µg/L in response to lactate
injections, but then have been subsequently reduced below detection by 4–5 weeks
after injections.
Overall, significant progress towards the performance objective for both optimization
activities and long-term operations has been made at TAN. Additional optimization ac-
tivities are needed to distribute electron donor throughout the entire residual DNAPL
source area to completely eliminate flux to down-gradient wells, evaluate alternative
electron donors that may decrease long-term costs, and study microbial competition to
determine whether the system can be optimized to favour the organisms of interest.
Lessons Learned
Enhanced in situ bioremediation can be very effective even in complex source
areas,
toxicity of chlorinated solvents to bacterial communities, once thought to be a
problem for ISB of DNAPLS, may not be an issue. In fact, dechlorinating bacteria
may have an ecological niche in these high concentration areas,
reductive dechlorination is strongly correlated both to electron donor distribution
and redox conditions, and,
dechlorination efficiency can be controlled in the field by the electron donor addi-
tion strategy and type.
NASA’s Launch Complex 34 (LC34) at Cape Canaveral

TECHNOLOGY: Biostimulation and Bioaugmentation through ethanol and a consortium of natu-
rally occurring microorganism (KB-1TM)
SCALE: Demonstration
GEOLOGY: Surficial fill and silty sands
CONTAMINANTS: TCE, DCE
The purpose of the project was to evaluate the technical and cost performance of the
biostimulation and bioaugmentation technology when applied to dense, nonaque-

ousphase liquid (DNAPL) contaminants in the saturated zone. This demonstration was
conducted at Launch Complex 34, Cape Canaveral Air Force Station, Florida, where
chlorinated volatile organic compounds (CVOCs), mainly trichloroethylene (TCE), are
present in the subsurface as DNAPL. Smaller amounts of cis-1,2-dichloroethylene
(DCE) and vinyl chloride (VC) also are present as a result of the natural degradation of
TCE. The part of the source zone used as a test plot for the demonstration is entirely
underneath the Engineering Support Building (Figure 6).
Figure 6: Location map of Launch Complex 34 site (Battelle, 2004).
The biostimulation and bioaugmentation project was conducted under the National
Aeronautics and Space Administration (NASA) Small Business Technology Transfer
Research (STTR) Program. For this project, the Small Business Concern vendor was
GeoSyntec Consultants (GeoSyntec). This demonstration was independently evaluated
by Battelle under the United States Environmental Protection Agency’s (U.S. EPA’s)
Superfund Innovative Technology Evaluation (SITE) Program (Battelle, 2004).
A sequential process of biostimulation and bioaugmentation is a promising remediation
technology for enhancing the extent and rate of degradation of CVOCs. Biostimulation
involves stimulating indigenous microbial cultures by adding nutrients (i.e., biostimula-
tion), whereas bioaugmentation involves introducing microbial cultures that are particu-
larly adept at degrading these contaminants into the target aquifer. The premise is that
although many aquifers contain native microorganisms that can degrade CVOCs, the
native microorganisms can be supplemented by specific cultures that enhance the deg-
radation of chlorinated solvents. Natural microorganisms, such as Dehalococcoides
ethenogenes, can be separately cultured and introduced into the aquifer to enhance the
degradation rates and extent of degradation that would normally be achievable by natu-
ral attenuation or by biostimulation (addition of nutrients) alone. Bioaugmentation using
specific cultures is claimed to be particularly effective in (1) degrading byproducts of
reductive dehalogenation, such as cis-1,2- DCE and VC, which would otherwise accu-
mulate in the aquifer; and (2) completing dechlorination processes to non-chlorinated
products such as acetylene, ethene, ethane, and methane.
This demonstration involved biostimulation followed by bioaugmentation in the same
test plot. During the biostimulation phase of treatment, an electron donor (ethanol) was
added to provide nutrients for indigenous microorganisms and stimulate CVOC degra-
dation. During the bioaugmentation phase, KB-1™, a consortium of naturally occurring
microorganisms known to completely dechlorinate high concentrations of TCE to
ethene, was added to the test plot. At Launch Complex 34, the DNAPL source zone
was not large enough to conduct a control demonstration using biostimulation alone for
comparison. Therefore, the sequential treatment of biostimulation and bioaugmentation
was evaluated at Launch Complex 34 in the same test plot. Bioaugmentation was cho-
sen as a second treatment phase to determine if complete dechlorination of a TCE-
DNAPL source zone was possible.
Based on pre-demonstration groundwater and soil sampling by Battelle, a test plot was
identified for biostimulation and bioaugmentation that was 20 ft long × 20 ft wide × 20 ft
deep (saturated thickness). The Upper Sand Unit, where the treatment was targeted, is
the shallowest part of the surficial aquifer, and extends down to a depth of 26 ft. The
water table at the site occurs at about 5 to 6 ft below ground surface (bgs), thus provid-
ing about a 20-ft-thick zone of aquifer for treatment. The Upper Sand Unit is underlain
by the Middle Fine-Grained Unit, which is made up of finer sand and silt, and constitutes
somewhat of a hydraulic barrier to the Lower Sand Unit below. These three stratigraphic
units constitute the surficial aquifer. The Lower Clay Unit forms a thin aquitard under the
surficial aquifer.
The bioaugmentation treatment was particularly targeted at depths of 16 to 24 ft bgs in
the Upper Sand Unit, where most of the DNAPL appeared to be present. The pre-
demonstration soil and groundwater characterization was done in January 2002, before
the vendor began installing the treatment system.
Prior to beginning the demonstration, the vendor installed a recirculating groundwater
system to establish a controlled hydraulic flow field. This was done to facilitate the dis-
tribution of electron donor, simplify the placement of monitoring points, and accelerate
the degradation process to a point where it could be monitored in the reasonable time-
frame allotted to this demonstration. The groundwater was recirculated from the extrac-
tion wells to the injection wells for several weeks to establish hydraulic control. During
this testing and modification period (May 23 to September 12, 2002), the recirculated
groundwater was passed through carbon canisters and treated prior to reinjection.
CVOCs were removed from groundwater in the treatment plot during this time. Prior to
beginning the biostimulation phase of the treatment, the carbon canisters were removed
from the recirculating system. The electron donor (ethanol) was injected inside the plot
to begin the biostimulation phase of the demonstration (October 23, 2002). Approxi-
mately 14 weeks later (February 6, 2003), the KB-1™ culture was injected in the aquifer
to begin the bioaugmentation phase. Groundwater sampling was conducted in Decem-
ber 2002 (one month after electron donor injection) and March 2003 (one month after
KB-1™ culture injection).
Figure 7: KB-1™ Dechlorinator Culture Containers (Battelle, 2004).
Post-demonstration soil and groundwater characterization was done in June 2003.

Performance assessment activities for the biostimulation and bioaugmentation demon-
stration included pre-demonstration investigations, installation of wells, operation, moni-
toring, and post-treatment evaluation. Battelle conducted detailed soil and groundwater
characterization activities to establish the DNAPL distribution and mass inside the test
cell. The vendor conducted additional operational measurements. The objectives of the
performance assessment were to:
Determine changes in total TCE (dissolved and free-phase) and DNAPL mass in
the test plot due to the biostimulation and bioaugmentation treatment;
Determine changes in aquifer quality due to the treatment;
Determine the fate of TCE, the primary DNAPL contaminant; and,
Determine operating requirements and cost of the technology.
Changes in Total TCE and DNAPL Mass

Detailed pre-demonstration and post-demonstration soil sampling was the main tool for
estimating changes in total TCE and DNAPL mass in the plot due to the treatment tech-
nology. In general, the eastern portion of the plot had the highest predemonstration TCE
concentrations. TCE concentrations were higher at approximatelyi 26 ft bgs, which is at
the interface between the Upper Sand Unit and Middle Fine- Grained Unit. The rest of
the plot appeared to contain mostly dissolved-phase TCE. The soil sampling results
were evaluated using both linear interpolation and kriging to obtain mass estimates for
the entire treatment zone (i.e., Upper Sand Unit). Linear interpolation indicated that, un-
der pre-demonstration conditions, 25.5 kg of total TCE (dissolved and free phase) was
present in the Upper Sand Unit. Approximately 2.6 kg of the total TCE was estimated to
be DNAPL. Following the demonstration, soil sampling indicated that 0.4 kg of total TCE
remained in the Upper Sand Unit; the post-demonstration mass of TCE-DNAPL was
estimated as 0.0 kg because no post-demonstration TCE concentrations were observed
above the threshold of 300 mg/kg. Therefore, the overall decrease in TCE mass due to
the treatment, as indicated by linear interpolation, was 98.5% for total TCE and >99%
for DNAPL in the Upper Sand Unit.
Kriging of the soil data indicated that the total TCE mass in the target zone before the
biostimulation and bioaugmentation treatment ranged from 17.6 to 46.6 kg, with an av-
erage of 32.1 kg. After treatment, the total TCE mass in the plot ranged from 0.1 to 0.3
kg, with an average of 0.2 kg. The decline in TCE mass due to the biostimulation and
bioaugmentation treatment ranged from 98.6 to 99.7%, with an estimated average de-
cline of 99%. Because few data points were available for DNAPL estimation, only the
total TCE data were subjected to kriging. These estimated TCE mass ranges are based
on an 80% confidence level and incorporate the uncertainty and spatial variability in the
data. The linear interpolation estimates are within the range of the kriging estimates.
These results indicate that the biostimulation and bioaugmentation treatment caused a
significant decrease in total TCE and DNAPL mass in the target treatment zone.
Changes in Aquifer Quality

Dissolved TCE concentrations, as measured in the monitoring wells, declined substan-
tially in the Upper Sand Unit of the demonstration area following the bioaugmentation
treatment. DCE levels increased following biostimulation, and then decreased after bio-
augmentation. Vinyl chloride levels increased immediately after biostimulation and bio-
augmentation, and then decreased during subsequent post-demonstration monitoring.
Ethene concentrations increased substantially toward the end of the demonstration.
These changes indicate sequential degradation of TCE to DCE, and ultimately to vinyl
chloride and ethane during the demonstration. In order to verify that the DNAPL source
had been substantially reduced and that the CVOC reductions observed during the
demonstration could be sustained (without encountering rebound), one further round of
groundwater monitoring was conducted in January 2004, almost one year after injection
of the KB-1™ culture. This long-term monitoring showed further substantial reductions
in TCE (to below detection), cis-1,2- DCE, and vinyl chloride. These results show that
DNAPL mass was substantially removed by the treatment and that the reduced CVOC
levels were sustainable.
Oxidation-reduction potential (ORP) and dissolved oxygen (DO) levels decreased in the
demonstration area after biostimulation began. The decreases continued through the
bioaugmentation phase of the demonstration and post-demonstration sampling. These
data indicate that strongly reducing anaerobic conditions were created in the Upper
Sand Unit during the demonstration. Groundwater pH in the shallow wells remained
relatively steady. Dissolved iron concentrations in the center well of the test plot gener-
ally decreased after the bioaugmentation treatment. The secondary drinking water limit
for iron is 0.3 mg/L, which was exceeded in the majority of wells before, during, and af-
ter the demonstration. Chloride levels in the monitoring wells, which were already high
partly due to saltwater intrusion in the aquifer, showed a slight increase over the course
of the demonstration. Anaerobic reductive dechlorination of TCE, cis- 1,2-DCE, and VC,
which was observed in this demonstration, releases chloride from contaminant mole-
cules and leads to increases in chloride levels in groundwater. Increases in dissolved
methane, as well as decreases in sulfate concentrations, indicate that an increase in
biological activity occurred as a result of the biostimulation and bioaugmentation treat-
ment. Biological oxygen demand (BOD) levels in the groundwater increased, indicating
an increase in the bioavailable organic matter in the aquifer, most likely due to the addi-
tion of a carbon electron donor to the recirculating groundwater. Total organic carbon
(TOC) levels also increased, probably as a result of the carbon electron donor addition.
The hydraulic conductivity of the Upper Sand Unit does not appear to have been af-
fected by the treatment, suggesting that the addition of electron donor and KB-1™ cul-
ture did not noticeably affect the aquifer. There were no substantial changes in perme-
ability in the test plot according to slug tests conducted in the center well before and
after the demonstration.
Fate of TCE/DNAPL in the Aquifer

The performance assessment indicates that biodegradation was a substantial pathway
accounting for the decrease in TCE, cis-1,2-DCE, and vinyl chloride measured in the
test plot. An increasing trend in dissolved ethene and chloride levels is evidence of
dechlorination reactions in the aquifer. The combination of biostimulation and bioaug-
mentation treatments accounted for the enhanced biodegradation seen in the plot. In
addition, some TCE and other VOCs were likely extracted by the recirculation system
and captured by adsorption in the aboveground carbon canisters. However, an analysis
of the amounts of water and TCE potentially extracted from the test plot by the recircula-
tion system showed that biostimulation and bioaugmentation contributed substantially to
the TCE removal observed in the test plot, even after adjusting for any dilution due to
the water recirculation system and carbon.
Operating Requirements and Cost

In general, the treatment system operated smoothly through the recirculation, biostimu-
lation, and bioaugmentation phases. Relatively good hydraulic control appeared to have
been maintained in the test plot, and the electron donor and KB-1™ culture were well-
distributed in the target zone. The vendor reported that biofouling in the injection wells
became apparent after amending the recirculating groundwater with electron donor. To
mitigate the biofouling, the duration of ethanol was decreased to one concentrated dose
administered daily; the injection wells were scrubbed, surged, and purged on a weekly
basis to removed biofilm from the screen; and the reinjected groundwater was amended
with sodium hypochlorite to inhibit microbial growth in the injection wells. It is unclear
what the long-term effect of the change in electron donor dose/timing and the addition of
sodium hypochlorite into the aquifer had on the microorganisms throughout the demon-
stration plot. Future applications of the biostimulation and bioaugmentation technology
may benefit from a study of optimizing electron donor dosing schedules, and establish-
ing procedures to monitor for biofouling and treat occurrences of biofouling.
A present value (PV) analysis was conducted to compare the cost of DNAPL source
treatment with biostimulation and bioaugmentation to the cost of installing and operating
an equivalent pump-and-treat system for a long period of time (30 years). It was as-
sumed that the biostimulation and bioaugmentation treatment would reduce the DNAPL
presence in the aquifer sufficiently for the rest of the contamination to attenuate natu-
rally. This analysis showed that the cost of source treatment with biostimulation and
bioaugmentation was lower than the PV of the costs of long-term treatment with a
pump-and-treat system at this site.
1.5 Status and Applicability
Enhanced in situ anaerobic bioremediation has emerged in recent years as a remedia-

tion strategy for CAHs in groundwater. Advantages include complete mineralization of
the contaminants in situ with little impact on infrastructure and relatively low cost com-
pared to more active engineered remedial systems (e.g., groundwater extraction, per-
meable reactive iron barriers, or chemical oxidation).
There are many considerations to take into account when selecting and designing an
enhanced bioremediation system. Enhanced anaerobic bioremediation as a remediation
technology may not be appropriate at all sites due to the complexity of chlorinated sol-
vent contaminant plumes (e.g., DNAPL source areas) and potential site-specific limita-
tions (e.g., difficult hydrogeologic conditions). At some sites, it may have utility only
when coupled with other remedial technologies.
Enhanced anaerobic bioremediation may be appropriate at sites where:
• Site-specific data indicate that the contaminants present (including any toxic deg-
radation products) can be readily degraded by native microbial populations under
anaerobic conditions.
• Subsurface conditions (e.g., aquifer permeability) are conducive to adequate
emplacement and distribution of a substrate, and creation of an in situ reactive
zone conducive to anaerobic degradation of the targeted contaminants.
• A cost/benefit analysis indicates that the technology is cost-effective relative to
other remedial measures (e.g., monitored natural attenuation [MNA], air
sparging, groundwater extraction, permeable iron reactive barriers, or chemical
oxidation).
Conditions that may preclude the use of enhanced anaerobic bioremediation are:
• Sites with impacted receptors, or with short travel time or distance to potential
discharge and/or exposure points
• Sites with inaccessible DNAPL sources
• Difficult hydrogeologic conditions that may preclude cost-effective delivery of
amendments, such as low permeability or a high degree of aquifer heterogeneity
• Geochemical conditions (e.g., unusually low or high pH) that inhibit the growth
and development of dechlorinating bacteria
Contaminant Distribution
Enhanced anaerobic bioremediation takes advantage of natural processes that may

already be contributing to the degradation of CAHs. The presence of degradation prod-
ucts that indicate that anaerobic dechlorination of CAHs is occurring, or has occurred,
naturally is a favourable indicator. Conversely, the lack of any dechlorination products is
a “red flag” that either enhanced bioremediation may not be a suitable approach or that
further evaluation is required (AFCEE, 2004).
The release of CAHs is often associated with release of other potential electron donors
such as fuels or landfill leachate. A review of historical records may indicate that an-
aerobic dechlorination occurred in the past, but that the system has stalled (e.g., at cis-
DCE) once the initial electron donor supply was depleted. In this case, complete and
rapid degradation can often be restored by substrate addition.
Enhanced anaerobic bioremediation has been successfully applied to a few sites with
residual or sorbed DNAPL. Application to sites with large quantities of free-phase
DNAPL has yet to be proven effective, and in these instances enhanced anaerobic bio-
remediation may be more suitable to reduce source mass or as a polishing step follow-
ing application of more aggressive source removal technologies (Stroo et al., 2003).
Highly elevated concentrations of solvents may act as toxic inhibitors to biodegradation,
especially for sites where the release is relatively recent (e.g., within 1 to 3 years).
However, dechlorinating bacteria (at least for chloroethenes) are known to be tolerant of
concentrations nearing solubility limits (Yang and McCarty, 2000).
Successful site closures to date (involving enhanced anaerobic bioremediation) typically
have involved relatively small- to moderate-size plumes associated with small commer-
cial operations such as dry cleaners (AFCEE, 2004). An area-wide treatment using en-
hanced anaerobic bioremediation may simply not be economical where treatment areas
exceed tens to hundreds of acres.
Microbiology
Enhanced anaerobic bioremediation of CAHs is targeted at stimulating microbially me-
diated anaerobic reductive dechlorination. The success of the technology largely de-
pends on the presence of appropriate dechlorinating bacteria and the ability to stimulate
sufficient growth and activity to degrade contaminants to the extent (and at a rate) that
meets the intended remedial objectives. Incomplete dechlorination (e.g., cis-DCE or VC
stall) due to insufficiently reducing conditions or lack of appropriate dechlorinating popu-
lations are common microbial issues when applying enhanced anaerobic bioremediation
(AFCEE, 2004).
Initially, a site can fall into one of three microbiological categories:
1. Sites where appropriate dechlorinating microorganisms are present, geochemical

conditions are appropriate for their growth, and sequential dechlorination prod-
ucts (e.g., VC and ethene) are observed.
2. Sites where appropriate dechlorinating microorganisms are present, but at insuf-
ficient quantity or level of activity for complete sequential dechlorination to in-
nocuous end products.
3. Sites where appropriate dechlorinating microorganisms are completely absent
(rare).
In the first case listed above, biostimulation alone can be applied with a high degree of
confidence. In the second case listed above, biostimulation alone may or may not be
successful. It may be difficult to distinguish the second case from the third case, be-
cause detection and identification of appropriate microbial species in these systems is
problematic (AFCEE, 2004).
Hydrogeology
The uncertainty in characterizing subsurface hydrogeology complicates all in situ treat-
ment technologies, and must be considered during the site selection and design proc-
ess. Inadequate characterization of the site hydrogeology can lead to remedial system
failure. However, in many cases, the system can be designed to mitigate difficult hydro-
geologic conditions. Difficult hydrogeologic conditions that may preclude cost-effective
delivery of amendments include excessive groundwater flow velocity, low permeability,
high levels of aquifer heterogeneity, or excessive depth to groundwater (i.e., high drilling
costs) (AFCEE, 2004).
Depth to Groundwater. Depth to water and the vertical thickness of the plume primarily
impact the capital cost of drilling and delivering the substrate to the intended treatment
zone.
The capital expense of installing multiple injection wells in deep settings (e.g., greater
than 100 feet bgs), or across thick formations needs to be compared to the costs asso-
ciated with competing technologies. There are practical limits (perhaps 15 to 20 feet) to
the maximum length of well screen across which a substrate can be uniformly injected;
therefore, large saturated thicknesses may require multiple vertical injection points
(AFCEE, 2004).
Hydraulic Conductivity. Hydraulic conductivity is a primary factor in effective distribu-

tion of substrate in the subsurface. In general, hydraulic conductivities greater than 1
foot per day (ft/day), or approximately 3 x 10-4 centimeters per second (cm/sec), are
suitable for injection of dissolved substrates. It is generally infeasible to effectively dis-
tribute substrates in zones having a hydraulic conductivity less than 0.01 ft/day (3 x 10-6
cm/sec). Alternate injection techniques such as hydraulic fracturing may be used in

some cases, but the timeframe for remediation may still be many years as remediation
of the entire aquifer volume will likely be diffusion-limited (AFCEE, 2004).
Groundwater Flow. Groundwater velocity, flow direction, and horizontal and vertical
gradients will impact the effectiveness of substrate addition. Most applications rely to
some extent on advective groundwater flow or recirculation to distribute substrate uni-
formly throughout the intended treatment zone. Excessively high rates of groundwater
flow (greater than 5 to 10 ft/day) may require large amounts of substrate to overcome a
large influx of native electron acceptors migrating into the reactive zone (AFCEE, 2004).
It may be impractical to maintain sufficiently reducing conditions in high-flow aquifers.
Cross-gradient distribution of soluble substrates in high-flow regimes also may be lim-
ited by lower transverse dispersion. Where rates of groundwater flow are very low (less
than 10 to 30 feet per year [ft/yr]), closer injection well spacing will be required and the
timeframe for remediation may be extended due to reduced mixing of substrate and
contaminant mass.
Groundwater Geochemistry
Redox processes in natural systems are rarely in equilibrium, and the predominant elec-
tron acceptor being utilized by microbial populations to derive energy often varies in
zones across the site. Addition of an organic substrate is intended to consume native
electron acceptors and to maintain optimal conditions for high rates of anaerobic
dechlorination. Excessive levels of competing electron acceptors (e.g., DO, bioavailable
iron, and sulfate) may limit the effectiveness of substrate addition. Groundwater geo-
chemical characteristics across the site should be reviewed to identify any undesirable
conditions (AFCEE, 2004).
Dissolved Oxygen and Oxidation-Reduction Potential. Background levels of DO and

values of ORP are an indicator of the pre-injection redox conditions that must be low-
ered to achieve efficient dechlorination. In general, elevated levels of DO and nitrate in
most aquifer systems can be overcome by providing adequate organic substrate. How-
ever, the problem may be compounded by other factors such as high rates of ground-
water flow (AFCEE, 2004).
Bioavailable Iron. High levels of bioavailable ferric iron (as iron oxide or iron hydroxide
minerals) may inhibit microbial anaerobic dechlorination in a manner similar to other
competing electron acceptors. In particular, it has been theorized that the free energy
associated with electron transfer during reduction of bioavailable iron by iron-reducing
bacteria is greater than that associated with the reduction of cis-DCE. Therefore, an-
aerobic dechlorination of cis-DCE to VC may potentially be inhibited in the presence of

relatively high levels of bioavailable iron because iron-reduction is more energetically
favorable (Evans and Koenigsberg, 2001; Wilson et al., 2003). This may be a temporal
phenomenon until the bioavailable iron is depleted; the concentrations or levels of
bioavailable iron that may inhibit anaerobic dechlorination have not been well docu-
mented or defined. Because bioavailable iron cannot be determined from groundwater
sampling alone, this parameter is frequently underestimated.
Sulfate/Sulfides. Existing guidance documents tend to suggest that, while CAH dechlo-
rination under sulfate reducing conditions is feasible, high sulfate levels are problematic
for CAH bioremediation. High sulfate levels may lower the efficiency at which substrate
is used for anaerobic dechlorination (AFCEE, 2004).
However, there is ample evidence in the literature for dechlorination of a variety of
CAHs at sites containing elevated dissolved sulfate levels (ITRC, 1998; Devlin and Mul-
ler, 1999). ARCADIS (Suthersan et al., 2002) reports successful application of en-
hanced anaerobic bioremediation at sites containing up to 500 to 700 mg/L of sulfate.
Complete anaerobic dechlorination has been stimulated at several high-sulfate Air
Force sites including Altus AFB, Oklahoma (sulfate up to 2,600 mg/L) and Travis AFB,
California (sulfate up to 5,400 mg/L). Therefore, the presence of high sulfate concentra-
tions does not necessarily preclude effective application of this technology (AFCEE,
2004).
Excessive levels of sulfides produced by reduction of sulfate may potentially inhibit an-
aerobic dechlorination. Elevated levels of dissolved sulfides or hydrogen sulfide have
been shown to inhibit sulfate reducing bacteria and methanogens, as well as some fer-
mentation reactions that produce hydrogen. The levels of sulphide that may potentially
inhibit dechlorinating microorganisms (and whether these levels are commonly encoun-
tered in the field) are not well documented. In general, dissolved sulphide and hydrogen
sulfide are rapidly co-precipitated with ferrous iron (a byproduct of ferric iron reduction),
but this may not be sufficient to reduce sulfide levels at high sulfate/low iron sites, where
there is insufficient iron to react with the sulfides (AFCEE, 2004).
pH and Alkalinity. A pH close to neutral (i.e., 6 to 8) is the most conductive to the pro-
liferation of healthy, diverse microbial populations. Low pH conditions (<5) are detrimen-
tal to sulfate-reducing, methanogenic, and dechlorinating bacteria. Fermentative organ-
isms favour lower pH conditions, and therefore will out-compete sulfate-reducing and
methanogenic bacteria in more acidic environments; this can result in the formation of
undesirable byproducts of fermentation, such as ketones, alcohols, and aldehydes. In
such cases, pH buffering, typically using common basic salts such as sodium bicarbon-
ate, may be used during implementation to raise and/or neutralize pH against further
decreases (AFCEE, 2004). Sites with pH outside of the 5 to 9 range may require more
thorough biological screening (e.g., using microcosm studies) to evaluate the effect of
pH manipulation on the existing dechlorinating microbial populations. In practice, care
must be taken in evaluating site-specific behaviour. For example, if groundwater pH is
below 5 but complete dechlorination is observed in the field, then it may be clear that
the local microbial population has adapted to low pH conditions. Aquifer systems with
lower buffering capacities are more susceptible to decreases in pH. Alkalinity is a gen-
eral indicator of the buffering capacity of an aquifer system. However, because of the
importance of the aquifer solids in establishing buffering capacity, groundwater alkalinity
may underestimate the true buffering capacity. From a practical standpoint, alkalinities
greater than 300 mg/L are generally sufficient to buffer against adverse pH changes.
Alkalinity less than 100 mg/L is cause for concern, and pH should be monitored care-
fully.
Lowering of pH and problems with adequate buffering are more likely to occur where
organic acids (e.g., lactic acid), organic acid salts (e.g., sodium lactate), or soluble sug-
ars (e.g., HFCS or molasses) are used. Substrate selection, substrate loading rate, and
the addition of buffering reagents should be carefully evaluated at sites with low alkalin-
ity or in response to field observations of excessive drop in pH (AFCEE, 2004).
The Air Force Center for Environmental Excellence and the Naval Facilities Engineering
Service Center (AFCEE, 2004) has reviewed 17 case studies to evaluate common at-
tributes of either successful or unsuccessful applications of enhanced anaerobic biore-
mediation. Site conditions and system design were evaluated to identify factors that
contributed to a successful application of enhanced bioremediation, or that limited the
effectiveness of the application. The case studies reviewed include the application of
different substrate types and bioaugmentation. Overall six field applications of soluble
substrates were reviewed, including applications of lactate and molasses, and a site
with an acetate/fructose combination. Eight applications of HRC® and vegetable oil were
reviewed to evaluate the performance of slow-release (viscous fluid) substrates. The
solid substrate case studies reviewed include two applications of bark mulch biowalls for
remediation of chlorinated ethenes and furthermore the study included two bioaugmen-
tation cases studies. Table 4 (attached) contains a summary of these sites. In the fol-
lowing the main statements of this evaluation are summarized.
Soluble Substrate Applications

The soluble substrates used in the preceding cases studies were readily injected and
distributed in the subsurface, and may be particularly effective in difficult hydrogeologic
settings. The applications at Test Area North and Hanscom AFB used only a single well
to distribute substrate throughout the treatment zone. Recirculation systems have the
potential to impact even larger treatment zones, which may be necessary at sites with
complex hydrogeology or low rates of groundwater flow.
The primary disadvantage of using these substrates is the requirement for frequent in-
jection and resulting higher operations and maintenance (O&M) costs relative to slow
release substrates. The cost of O&M alone is a large percentage of the life-cycle costs
of soluble substrate systems.
Factors that resulted in incomplete dechlorination observed in some cases was either
due to 1) an inadequate supply (low concentration) of substrate, resulting in insufficient
reducing conditions, or 2) a lack of microorganisms capable of complete anaerobic
dechlorination. Limiting the substrate loading rate in order to avoid methanogenesis
may result in large areas of the treatment zone not becoming sufficiently reducing. Al-
though soluble substrates require frequent injection, the substrate concentration and
volume can be modified to increase the substrate loading rate, as needed. However,
this also may result in extra cost and time to monitor and adjust the substrate volume,
concentration, and frequency of injection until an optimal scenario is obtained.
Furthermore, difficult hydrogeologic conditions are the cause of failure at some sites,
including a high flux of aerobic groundwater, unexpected groundwater flow direction,
and inadequate characterization of complex stratigraphy. For example, the demonstra-
tion project conducted at Hanscom AFB required modifications to the injection scenario
over a period of approximately 1 year to fully deplete native electron acceptors and to
induce methanogenic conditions under which degradation of DCE and VC was ob-
served. Temporal fluctuations in the direction of groundwater flow also complicated
substrate distribution at this site. The substrate limitation was overcome by adjustments
to the system, and system performance objectives were achieved when sufficient sub-
strate loading was accomplished.
Advantages of soluble substrate systems include the following:

Soluble substrates are readily mixed and distributed in the subsurface relative to
other substrate types. This may be beneficial for deep treatment zones where di-
rect-push techniques cannot be used. In this case, the high cost of well installa-
tion may be offset by the use of soluble substrates that require fewer injection
wells for distribution of substrate over larger volumes of the aquifer.
The ability to vary the concentration, volume, and frequency of injection allows
for optimizing substrate delivery and manipulating the geochemical conditions in
the reaction zone over time.
Recirculation systems may be used for hydraulic containment and/or for a
greater residence time of contaminants in the reaction zone. Recirculation sys-
tems also may treat larger aquifer volumes with fewer wells.
Disadvantages or limitations of soluble substrate systems include the following:

O&M costs are high relative to long-lasting substrates as a result of the need for
frequent injection.
Biofouling of injection wells may require additional maintenance.
The use of potable water to make up large volumes of substrate mixture may re-
sult in dilution and displacement of the contaminant plume.
Low-molecular-weight substrates (e.g., lactate or butyrate) can be more expen-
sive than bulk food-grade products, while the presence of impurities may require
the use of higher grades of molasses or the use of high fructose corn syrup.
Slow –release viscous fluid substrate systems

HRC® Applications
HRC® has been demonstrated to be an effective substrate for enhanced anaerobic bio-
remediation, with over 474 field applications since 1999 (Willett et al., 2004). Complete
degradation of PCE and TCE parent compounds to VC and ethene was observed at
both the Fisherville Mill Site and the Springdale Cleaners Site. Although the ability to
reach federal MCLs has yet to be demonstrated at these sites, significant contaminant
reductions were achieved. The results obtained for the Springdale Cleaners Site sug-
gests the potential for remediating DNAPL source areas using HRC® products. Con-
versely, application of HRC® at the Atlas 10 Site in Nebraska ((USACE), 2003) did not
stimulate significant reduction of TCE, largely due to a high flux of native electron ac-
ceptors (DO, nitrate, and sulfate). Limited substrate distribution and a consequent inabil-
ity to induce highly reducing conditions was observed at the Naval Air Station Dallas
Site (CH2M Hill Constructors, 2001) due to low aquifer permeability and lack of ground-
water flow. These limitations are not unique to the application of HRC®; rather, they can
be problematic with enhanced anaerobic bioremediation applications in general.
Vegetable Oil Applications

Vegetable oil is currently being developed as a low-cost alternative substrate, designed
to induce or enhance anaerobic dechlorination of CAHs for several years with a single
injection. Vegetable oil applications to date have mostly been performed to achieve
source reduction using grid configurations, or to construct biobarriers using rows of in-
jection points. Applications at Altus AFB, Travis AFB, and Cape Canaveral Air Force
Station have demonstrated that concentrations of chlorinated ethenes can be reduced
by several orders of magnitude to below federal drinking water MCLs. Applications of
vegetable oil at Travis AFB and Cape Canaveral Air Station indicate anaerobic dechlo-
rination can be stimulated for periods of at least 3 to 4 years with a single application.
Some difficulty in achieving effective substrate distribution has been encountered, and
uniform distribution of the substrate may be complicated by low permeability soils or by

a high degree of aquifer heterogeneity. Distribution problems may be expected with in-
jection of neat vegetable oil, and generally this is no longer recommended. Low-
viscosity microemulsions are currently the state-of-the-art for distribution of vegetable
oils, which allows more uniform distribution. Stable emulsions with very fine droplet
sizes may be difficult to prepare in the field. Alternately, more expensive commercial
emulsion products are available.
Advantages of using slow-release viscous fluids include the following:

Application of HRC® or vegetable oil emulsions can be very cost-effective for
treating shallow groundwater plumes using inexpensive direct-push techniques,
where close injection point spacing (5- to 15-foot centers) can be utilized.
The use of fast-acting HRC®-primer and long-lasting HRC-XTM products pro-
vides design alternatives for varied site conditions.
Vegetable oil emulsions are easily modified to fit site-specific conditions. Oil-in-
water emulsions can be modified to include fast-acting soluble substrates or to
modify the effective oil saturation.
The effective lifespan of a single application of these substrates may last from 1
to 5 years.
Slow-release substrates also may be used in trenches and excavations to sup-
plement solid substrates (e.g., spraying vegetable oil onto sand or mulch, or
backfilling with a layer of HRC®).
Disadvantages of using slow-release viscous fluids include the following:

Slow-release substrates typically require more injection points and may not be as
cost-effective as soluble substrate applications under very deep or difficult hy-
drogeologic conditions where injection costs are high.
High rates of groundwater flow or high rates of native electron acceptor flux may
require higher loading rates and additional injections. In some cases (e.g., the At-
las 10 site), the native electron acceptor flux may be too high to overcome.
HRC® products are relatively expensive compared to other substrate types.
However, a cost-benefit analysis should be conducted that considers other cost
factors and the overall life-cycle costs.
Creating stable emulsions in the field with an appropriate droplet size may be dif-
ficult in practice. Use of pre-mixed commercial microemulsion products will in-
crease cost.
Bark Mulch Solid Substrate Systems

Two applications of bark mulch in a permeable biowall configuration were reviewed to

evaluate the performance of solid substrate systems. The Offutt AFB and Altus AFB
biowalls were both capable of effectively reducing concentrations of TCE. Reduction of
TCE concentrations to below federal drinking water MCLs occurs in the immediate vicin-
ity of the Altus AFB biowall. Both sites exhibited substantial degradation of cis-DCE
without an accumulation of VC or ethene. Degradation processes other than biotic an-
aerobic reductive dechlorination of TCE, cis-DCE, and VC are likely occurring, including
abiotic degradation by reactive iron-monosulfides. Humic acids in the mulch and com-
post mixtures may also serve as electron acceptors in energy yielding reactions that
result in the oxidation of cis-DCE and VC under anaerobic conditions (Bradley et al.,
1998). Although exhibiting decreasing trends, cis-DCE has persisted at concentrations
above initial conditions and above its MCL at both biowall sites. Nonetheless, overall
mass destruction rates for total CAHs are impressive, ranging from 64 percent (Offutt
AFB pilot-scale biowall) to 86 percent (Altus AFB biowall). Both of the biowalls appear to
be effective at treating shallow groundwater plumes in highly heterogeneous formations
having a low to moderate permeability. It is yet to be determined whether retention time
and substrate loading using mulch biowalls is sufficient for degrading concentrations of
CAHs in excess of 10 to 100 mg/L, or for flow rates greater than 1.0 ft/day. The use of
wider trenches (greater than 2 feet in width) or multiple parallel trenches may be neces-
sary to treat higher CAH concentrations at sites with high rates of groundwater flow or
high rates of native electron acceptor flux.
In summary, the advantages and disadvantages of solid substrate mulch and compost
biowalls are listed below.
Advantages related to the use of mulch and compost biowalls include the follow-
ing:
Effective for shallow groundwater plumes in low to moderate permeability or
highly heterogeneous formations. The continuity of the trench eliminates the po-
tential for groundwater bypass due to preferential flow paths, or non-uniform dis-
tribution of substrate that may occur with delivery of liquid substrate via injection
wells.
Mulch, compost, and sand are relatively inexpensive when purchased in bulk
quantities. Tree mulch can often be obtained for the cost of shipping and han-
dling alone.
Mulch biowalls require no O&M other than periodic performance monitoring.
However, it has yet to be determined how many years biowall systems will be
able to sustain anaerobic reductive dechlorination of CAHs.
Trenches can be modified to include wells or perforated pipe for addition of liquid
substrates to supplement carbon loading, if necessary. In addition, the relatively
small treatment volume of the trench (relative to other substrate configurations)

makes biowall systems ideal candidates for relatively low-cost inoculation with
bioaugmentation cultures.
Disadvantages or limitations of mulch biowalls include the following:

The depth that can be trenched in a practical and cost-effective manner is limited
to approximately 35 feet bgs. Excavation of a bench for the trenching equipment
may provide for an additional 5 to 10 feet of depth.
Trenching may interfere with site infrastructure and utilities.
The contaminant retention time in the trench and substrate loading capacity (i.e.,
rate at which organic carbon is added to the groundwater passing through the
trench) may be insufficient to treat concentrations of CAHs in excess of 10 to 100
mg/L. Use of wider trenches or multiple parallel trenches may be necessary to
treat higher CAH or to deplete high concentrations of native electron acceptors.
The effective life-span of mulch biowalls has yet to be determined.
Bioaugmentation Systems
The bioaugmentation applications at the Aerojet Facility in California and the Bachman
Road site in Michigan demonstrate that bioaugmentation can be highly effective with
small-scale recirculation systems. It has yet to be shown whether bioaugmentation can
be used successfully with large-scale recirculation or passive enhanced bioremediation
systems. The ability to increase the scale of bioaugmentation systems will be depend-
ent to a large extent on the ability to uniformly distribute the culture over large volumes
of the treatment zone. Most bioaugmentation demonstrations performed to date have
used low-molecular-weight soluble substrates (e.g., lactate or ethanol) in recirculation
configurations to carefully control aquifer redox conditions. The fermentation reactions
for low-molecular weight substrates are generally better understood than for more com-
plex substrates (e.g., molasses or vegetable oils). However, there is no reason to be-
lieve that bioaugmentation cannot be effective with all substrate types. Commercially
available bioaugmentation cultures consist predominantly of the microbial strain
Dehalococciodes ethenogenes. Dehalococciodes ethenogenes is the only microbial
strain that has been shown to be capable of complete sequential dechlorination of PCE
and TCE to cis-DCE, VC, and ethene. The ability of these bioaugmentation cultures to
completely dechlorinate chloroethanes and chloromethanes is less well understood.
The advantages and disadvantages of bioaugmentation are listed below.
Advantages of bioaugmentation include the following:

Bioaugmentation can be used to reduce acclimation periods and/or to provide

populations of known dechlorinating microorganisms where dechlorination is in-
complete.
Bioaugmentation may be used to increase the confidence of using an enhanced
anaerobic bioremediation approach. In some cases, bioaugmentation may be
cost-effective in that the overall time for remediation can be decreased, thereby
reducing costs for O&M and performance monitoring.
Bioaugmentation in a carefully controlled reaction zone may result in complete
anaerobic dechlorination without inducing strongly reducing (i.e., methanogenic)
conditions. Avoidance of strongly reducing conditions may not be necessary in
most cases, but this capability can be used to advantage where strict adherence
to drinking water standards is being enforced.
Limitations of bioaugmentation include the following:

Enriched cultures may quickly attach to the aquifer matrix and do not migrate as
readily as soluble substrates. Therefore, it may be difficult to distribute the cul-
tures throughout large volumes of an aquifer.
Although the cost of bioaugmentation cultures is decreasing as more commercial
vendors enter the market, the cost of the recirculation systems commonly used to
carefully control geochemical conditions is high. The ability to inoculate passive
treatment systems in a barrier configuration may represent a more cost-effective
use of the technology.
The following subsections describe some common limitations and reasons for failure of
enhanced anaerobic bioremediation applications that have been encountered in this
survey of case studies.
Hydrogeology and Substrate Delivery

High rates of groundwater flow, low aquifer permeability, and low hydraulic gradient
have all resulted in ineffective applications of enhanced anaerobic bioremediation. In
general, a high flow rate results in a high native electron acceptor flux and difficulty in
establishing a highly reducing environment. For example, a high flux of DO, nitrate, and
sulfate at the Atlas 10 Missile site in Nebraska is thought to have resulted in native elec-
tron acceptor demand that could not be overcome by two applications of HRC®. High
groundwater flow rates may also limit the effective residence time of CAHs in the reac-
tive zone. This may be a critical design factor for systems such as mulch biowalls,
where the reaction zone and residence time may be relatively short.
Low permeability limited the effectiveness of an application of HRC® at the Naval Air
Station Dallas site. HRC® and vegetable oil have been applied successfully in low per-
meability settings at other sites (e.g., the Travis AFB), but it is typical for very long lag
times to occur and for distribution of the soluble constituents of these substrates to be
diffusion limited (a slow process). A low hydraulic gradient and slow rates of groundwa-
ter flow (less than 10 to 30 ft/yr) may similarly inhibit effective distribution of substrate in
passive systems. While it is possible that slow migration of soluble substrate via diffu-
sion may occur over a period of several years, for most applications this may not be an
acceptable timeframe. The use of recirculation systems that enhance hydraulic gradi-
ents and the rate of groundwater flow should be considered for treatment of dissolved
plumes in low gradient or low permeability groundwater systems.
Geochemistry, Redox Conditions, and Substrate Loading

Insufficient substrate loading and high native electron acceptor flux can result in redox
conditions that are not sufficiently reducing to stimulate effective rates of anaerobic
dechlorination. Although anaerobic dechlorination has been demonstrated to occur un-
der sulfate-reducing conditions without methanogenesis (e.g., methane less than 1
mg/L), the number of sites that exhibit complete anaerobic dechlorination under
methanogenic conditions would suggest that an attempt to limit substrate loading, in
order to prevent methanogenesis and increase substrate utilization, may be counter
productive. The cost of additional substrate necessitated by diminished utilization of
CAHs for anaerobic dechlorination is small compared to the cost incurred for longer
O&M and monitoring due to slower rates of anaerobic dechlorination. Due to aquifer and
microbial heterogeneity, controlling subsurface redox conditions with precision is diffi-
cult. Such control typically requires frequent substrate injection and/or recirculation, and
possibly also temporal variation in loading rates, both of which are labour and cost in-
tensive.
Furthermore, the soluble (aqueous) substrate applications at several sites show that,
anaerobic dechlorination of cis-DCE or VC to ethene did not occur until methanogenic
conditions were observed. Methanogenic conditions are often an indication that
bioavailable iron and sulfate have been adequately depleted, and that high rates of fer-
mentation are occurring. These conditions favour high rates of anaerobic dechlorination,
and should not be considered detrimental. At Naval Support Activity Mid-South, produc-
tion of VC and ethene were only observed after a high concentration slug of acetate
was injected at the end of a recirculation test that had utilized a lower substrate loading
rate.
Furthermore, it has been suggested that rapid stimulation of strongly reducing (i.e.,
methanogenic) conditions is one way to expedite an evaluation of microbial sufficiency.
If a complete dechlorination pathway is not observed after inducing strongly methano-
genic conditions for several months, then it is likely that the requisite microbial species
are not present.
In contrast, the Aerojet Facility application illustrated in Appendix E.9 is an example of
where bioaugmentation in a carefully controlled reaction zone was successful in stimu-
lating complete anaerobic dechlorination without inducing methanogenic conditions or
creating undesirable levels of dissolved metals or other fermentation products. The bio-
augmentation culture was capable of complete dechlorination of TCE to ethene without
the need to cultivate methanogenic conditions.
Microbial Sufficiency
Microbial sufficiency is likely the most difficult condition to assess during the site selec-
tion process. Multiple conditions may cause an accumulation of intermediate dechlorina-
tion products (e.g., cis-DCE, VC, or 1,2-dichloroethane [DCA]). Examples of incomplete
or slow dechlorination of cis-DCE or VC include the Point Mugu IRP Site 24and Seal
Beach IRP Site 40.
No singular analysis, including determining the presence or absence of Dehalococ-
coides, can be used to confirm the lack of a sufficient microbial population. For exam-
ple, Dehalococcoides was detected at the Atlas 10 site in a well that had elevated levels
of TOC and metabolic acids. Yet the presence of this species, combined with the pres-
ence of reducing conditions and elevated organic carbon levels, did not result in signifi-
cant degradation of TCE or evidence of complete dechlorination. Quantitative, real-time
polymerase chain-reaction (PCR) analysis may provide estimates of the quantity or
concentration of Dehalococcoides in a sample. However, it is not well known what con-
centrations are required to effect efficient and complete anaerobic dechlorination. Fur-
thermore, molecular screening methods cannot currently identify the strains of Dehalo-
coccoides that may be present. Isolation of different strains in the laboratory indicates
they have different dechlorinating capacities. In addition to the absence of appropriate
dechlorinating microorganisms, insufficient substrate loading, failure to achieve suffi-
cient reducing conditions, inhibition due to high levels of native electron acceptors (e.g.,
sulfate), inhibition due to preferential degradation of more highly-chlorinated compounds
in the contaminant mixture, and kinetic disparity all may contribute to accumulation of
intermediate dechlorination products. At many sites, a relatively high percentage of the
CAH mass may be present in the form of DNAPL or sorbed to the aquifer matrix. Con-
stant dissolution of CAH mass into the groundwater reaction zone may occur. In these
cases, cis-DCE and VC may be constantly produced by dechlorination of the parent
compounds (PCE and TCE). However, unless the rate of dechlorination of cis-DCE and
VC is greater than the rate of dechlorination of PCE and TCE, cis-DCE and VC will ac-
cumulate or persist until the mass of PCE and TCE is depleted. Because cis-DCE and
VC are typically dechlorinated more slowly than PCE and TCE, this rate disparity should
be anticipated.
Given the current state-of-the-practice, it is important to evaluate all potential causes of

the accumulation of intermediate dechlorination products and of incomplete degradation
pathways before considering the need for bioaugmentation. While bioaugmentation
should not be a default remedy for poorly designed or poorly implemented biostimula-
tion applications, there are sites that will benefit from its application. Bioaugmentation
has been successfully applied at several sites, and is a potential option for sites with an
absence, or insufficient quantities or activity, of appropriate dechlorinating microorgan-
isms.
Application of enhanced anaerobic bioremediation using bioaugmentation at the Aerojet
Facility in California and the Bachman Road site in Michigan benefited from bioaugmen-
tation. It should be noted that these projects were specifically designed to demonstrate
bioaugmentation using small-scale recirculation systems. Application of a biostimula-
tion-only test typically uses a much different design and allows for a much longer accli-
mation period. Nonetheless, bioaugmentation was required to meet the project-specific
goals of achieving complete dechlorination within a specified time period at these sites.
1.6 References
AFCEE, N., 2004. Principles and Practices of Enhanced Anaerobic Bioremediation of chlorinated sol-
vents. 022/738863/28.doc, Air Force Center for Environmental Excellence, Brooks City-Base,
Texas
Naval Facilities Engineering Service Center, Port Hueneme, California
Environmental Security Technology Certification Program, Arlington, Virginia.
(AFRL), A.F.R.L., 2001. Reductive Anaerobic Biological In-Situ Treatment Technology (RABITT), Battelle
Memorial Institute, Cornell University, USEPA, NFESC.
ARCADIS Geraghty and Miller, I.A., 2003. Final Report: In-situ Substrate Addition to Create Reactive
Zones for Treatment of Chlorinated Aliphatic Hydrocarbons, Hanscom Air Force Base., Prepared
for the Air Force Center for Environmental Excellence and the Environmental Security Technol-
ogy Certification
Program.
Battelle, 2004. Demonstration of Biodegradation of Dense, Nonaqueous-Phase Liquids (DNAPL) through
Biostimulation and Bioaugmentation at Launch Complex 34 in Cape Canaveral Air Force Station,
Florida, Battelle Press, Columbus, OH.
Bouwer, E.J., 1994. Bioremediation of Chlorinated Solvents Using Alternate Electron Acceptors.
Publishers. In: R.D. Norris, R.E. Hinchee, R. Brown, P.L McCarty, L. Semprini, J.T. Wilson, D.H. Kamp-
bell, M. Reinhard, E.J. Bouwer, R.C. Borden, T.M. Vogel, J.M. Thomas, and C.H.Ward (Editor),
Handbook of Bioremediation. Lewis Publishers, pp. 149-175.
Bradley, P.M., Chapelle, F.H. and Lovely, D.R., 1998. Humic Acids as Electron Acceptors for Anaerobic
Microbial Oxidation of Vinyl Chloride and Dichloroethene. Applied Environmental Microbiology,
64: 3102-3105.
CH2M Hill Constructors, I.C.M.H., 2001. Technical Memorandum, HRC Pilot Study Evaluation at SWMU
138 - Boat Ramp Naval Air Station Dallas, TX.
Cope, N. and Hughes, J.B., 2001. Biologically-enhanced removal of PCE from NAPL source zones. Envi-
ron. Sci. Technol., 35: 2014-2021.
Dean, S., Wiseman, L., Accashian, J., Edgar, M. and Callender, J., 2001. Design of an Enhanced Biore-
mediation System for a Low Permeability Aquifer, Proceedings of the Sixth In-Situ and On-Site
Bioremediation Symposium, San Diego, California.
DoD, 2002. Evaluation of performance and costs associated with anareobic dechlorination techniques-
Revision 02 (Phase 1 site survey), Prepared for: Environmental security technology certification
programm, Arlington, Virginia.
DoE, 1999. In Situ Bioremediation for the Hanford Carbon Tetrachloride Plume.
French, J. et al., 2003. Phased In situ Biostimulation/Bioaugmentation Pilot Testing in a Coastal Aquifer,
Proceedings of the Seventh International Symposium of In Situ and On-Site Bioremediation. Bat-
telle Press, Columbus, Ohio, Orlando, Florida, June 2003.
GeoSyntec, 2002. Pilot Test for In Situ Bioremediation of Perchlorate & Trichloroethene in Groundwater
Using an Active Biobarrier.
Granade, S., D.P. Leigh, and C.D. Johnson., 2003. Chlorinated Solvent Bioremediation: 3 Case Studies,
Proceedings of the Seventh International Symposium of In Situ and On Site Bioremediation. Bat-
telle Press, Columbus, Ohio, Orlando, Florida, June 2003, pp. Paper A-13.
Haas, P., Gonzales, J., Cork, P. and Henry, B., 2003. Remedial Performance of Organic Mulch Biowalls
at Two Geochemically Distinct Sites, Proceedings of the 2003 AFCEE Technology Transfer
Workshop, San Antonio, Texas, Air Force Center for Environmental Excellence, Brooks City-
Base, Texas. February.
Henry, B.M. et al., 2003. Permeable Mulch Biowall for Bioremediation of Chlorinated Solvents, Proceed-
ings of the Seventh International Symposium of In Situ and On-Site Bioremediation. Battelle
Press, Columbus, Ohio, Orlando, Florida, June 2003, pp. Paper K-03.
Horst, J.F., Beil, K.A., Burdick, J.S. and Suthersan, S.S., 2000. Comparison of Natural and Enhanced
Rates Through Substrate Amendments, Proceedings of the Second International Conference on
Remediation of Chlorinated and Recalcitrant Compounds, Monterey, California.
ITRC, 2002. Technical/Regulatory Guidelines: A Systematic Approach to In Situ Bioremediation in
Groundwater Including Decision Trees on In Situ Bioremediation for Nitrates, Carbon Tetrachlo-
ride, and Perchlorate, Interstate Technology and Regulatory Council In Situ Bioremediation
Team.
ITRC, 2004. Strategies for Monitoring the Performance of DNAPL Source Zone Remediation.
Koch, S.A., Rice, J.M., 2001. In Situ Enhanced Reductive Dechlorination of PCE, Proceedings of the
Sixth International In-Situ and On Site Bioremediation Symposium, San Diego, California, pp.
181.
Lee, M.D. and Lieberman, M.T., 2002. Low Cost Emplacement of Insoluble Organic Substrate for En-
hanced In Situ Reductive Dechlorination, Altus Air Force Base.
Lee, M.D. et al., 2003. Pilots to Enhance Trichloroethene Reductive Dechlorination and Ferrous Sulfide
Abiotic Transformation, Proceedings of the Seventh International Symposium of In Situ and On-
Site Bioremediation. Battelle Press, Columbus, Ohio, Orlando, Florida, June 2003, pp. Paper K-
14.
Leigh, D.P. et al., 2000. Enhanced Anaerobic In Situ Bioremediation of Chloroethenes at NAS Point
Mugu, Proceedings of the Second International Conference on Remediation of Chlorinated and
Recalcitrant Compounds, Monterey, California, pp. 229-235.
Lendvay, J.M. et al., 2001. Plume control using bioaugmentation with halorespiring microorganisms,
Groundwater Quality 3rd International Conference, Sheffield, UK.
Liles, D.S., Lut, C.C., Page, G.B. and Suthersan, S.S., 2004. In Situ Chemical Stabilization of Metals and
Radionuclides Through Enhanced Anaerobic Reductive Precipitation: Application of a Commer-
cial Technology to DOE Needs. http://www.arcadis-
us.com/resources/insituchemicalstabilization.pdf.
Maierle, M.S. and Cota, J.L., 2001. Complete PCE Degradation and Site Closure Using Enhanced Re-
ductive Dechlorination, Proceedings of the Sixth International In-Situ and On-Site Bioremediation
Symposium, pp. 149-156.
Martin, J.P., Sorenson, K.S. and Peterson., L.N., 2001. Favoring Efficient In Situ Dechlorination through
Amendment Injection Strategy, Proceedings of the Sixth International In-Situ and On Site Biore-
mediation Symposium, San Diego, California, pp. 265-272.
McCarty, P.L. et al., 1998. Full-Scale Evaluation of In Situ Cometabolic Degradation of Trichloroethylene
in Groundwater Through Toluene Injection. Environ. Sci. Technol., 32(1): 88-100.
Mikszewski, A., 2004. Emerging Technologies for the In Situ Remediation of PCB-Contaminated Soils
and Sediments: Bioremediation and Nanoscale Zero-Valent Iron, National Network for Environ-
mental Management Studies Fellow for U.S. Environmental Protection AgencyOffice of Solid
Waste and Emergency Response Office of Superfund Remediation and Technology Innovation
Technology Innovation Program Washington, DC www.clu-in.org.
Murray, W., Dooley, M. and Koenigsberg, S., 2001. Enhanced Bioremediation of Chlorinated Solvents,
Proceedings of the Sixth International In-Situ and On-Site Bioremediation Symposium, San
Diego, California, pp. 197-204.
Murt, V., Huscher, T., Easley, D., 2000. A Reductive Dechlorination Treatability Study of a Shallow Allu-
vial Aquifer, Proceedings of the Second International Conference on Remediation of Chlorinated
and Recalcitrant Compounds, Monterey, California.
Parsons, 2002. Final Phase II Field Feasibility Test for In Situ Bioremediation of Chlorinated Solvents Via
Vegetable Oil Injection at Hanger K Area, Cape Canaveral Air Force Station, Florida.
Parsons, 2004. Final Project Completion Report for a Field Feasibility Test for In Situ Bioremediation of
Chlorinated Solvents Via Vegetable Oil Injection at Site SS015, Travis Air Force Base, California.
Peeples, J.A., Warburton, J.M., Al-Fayyomi, I. and Haff, J., 2000. Enhanced Reductive Dechlorination of
Ethenes Large-Scale Pilot Testing, Proceedings of the Third International Conference on Reme-
diation of Chlorinated and Recalcitrant Compounds, Monterey, California.
Sandefur, C.A., Parrett, K. and Lapus, K.A., 2002. Bioremediation of a PCE Plume at a Dry Cleaner Site.
In: A.R.G.a.A.S.C. Chen (Editor), Proceedings of the Third International Conference on Remedia-
tion of Chlorinated and Recalcitrant Compounds. Battelle Press, Columbus, Ohio, Monterey, Cali-
fornia, May 2002, pp. Paper 2B-52.
Sorenson, K.S., 2000. Biodegradation of TCE Improved with Lactate Injection in Deep, Fractured Rock.
Groundwater Currents, 38.
Sorenson, K.S., 2003. Enhanced Bioremediation for Treatment of Chlorinated Solvent Residual Source
Areas. In: S.M.H.a.S.D. Warner (Editor), Chlorinated Solvent and DNAPL Remediation: Innova-
tive Strategies for Cleanup. ACS Symposium Series. Vol. 837, pp. 119-131.
Stroo, H.F. et al., 2003. Remediating Chlorinated Solvent Source Zones. Environ. Sci. Technol., 37(11):
224A-230A.
Suthersan, S.S., 2001. Natural and Enhanced Remediation Systems. ARCADIS. Lewis Publishers, Boca
Raton, Florida.
U.S.EPA, 2000a. Engineered Approaches to In Situ Bioremediation of Chlorinated Solvents: Fundamen-
tals and Field Applications. EPA 542-R-00-008
http://www.epa.gov/clu-in.org.
U.S.EPA, 2000b. Cost and Performance Report: Bioaugmentation (Accelerated Anaerobic Bioremedia-
tion) at Area 6 of the Dover Air Force Base, Dover, Delaware.
U.S.EPA, 2004. DNAPL Remediation: Selected projects approaching regulatory closure. EPA 542-R-04-
016.
U.S.EPA, 2004b. How to Evaluate Alternative Cleanup Technologies for Underground Storage Tank
Sites: A Guide for Corrective Action Plan Reviewers. EPA 510-R-04-002.
(USACE), U.S.A.C.o.E., 2003. Project Management Plan for Former Lincoln AFB, Atlas Site 10, York,
NE.
Willett, A., Tseng, J., Gillespie, R. and Koenigsberg, S., 2004. Hydrogen Release Compound (HRC®): A
Review of Published Papers and Case Histories 1999-2003.
Yang, Y. and McCarty, P.L., 2000. Biologically enhanced dissolution of tetrachloroethene DNAPL. Envi-
ron. Sci. Technol., 34(14): 2979-2984.
Table 4: Summary of case studies results (AFCEE, 2004)

Site Name and Observed Effectiveness of Site Status Criteria for Success Factors Contributing to References
Location Dechlorinaton Substrate Success or Failure
Pathway Distribution References
Soluble Substrates – Lactate
Test Area North, TCE to cis-DCE to Substrate distribution System is being Demonstrate that enhanced MCLs were achieved for TCE and DCE (AFCEE, 2004; Martin
Idaho National VC to ethene. was effectively expanded with larger bioremediation of a DNAPL at the base of the aquifer. Concentra- et al., 2001; Soren-
Energy and attained with complete volume injections and source area can reduce tions of VC are generally less than 15 son, 2003)
Environmental dechlorination a second injection CAHs to below MCLs. Dem- µg/L (October 2003) and VC has not
Laboratory observed up to 150 feet well. onstrate that enhanced bio- accumulated. The
downgradient of the remediation can replace pilot-scale application resulted in a
Idaho injection well. groundwater extraction as a ROD amendment (approved
final remedy for the source September 2001) to switch from
area. groundwater pump and treat to en-
hanced in situ bioremediation.
IRP Site 24, Naval TCE to cis-DCE Substrate distribution Pilot test monitored Pilot test to demonstrate that Dechlorination of VC was deemed too (AFCEE, 2004;
Base Ventura to VC to ethene was effective within the for 3 years, then lactate addition can remedi- slow to meet remedial objectives, and Granade, 2003; Leigh
County, Point (degradation of recirculation cell. converted to aerobic ate TCE and DCE to ethene. the system converted to aerobic come- et al., 2000)
Mugu VC to ethane rela- Substrate was cometabolism. Sequential tabolism. VC was effectively oxidized by
tively slow). allowed to be Full-scale sequential dechlorination of TCE to DCE cometabolic processes. It is unknown
California depleted after last anaerobic/aerobic to VC and ethene were ob- whether injection of additional substrate
injection at day 57. treatment is planned. served over after 57 days would have stimulated
approximately 36 months. faster dechlorination of VC.
IRP Site 40, Naval PCE and TCE to Substrate was added by Biostimulation pilot Pilot test to demonstrate that Strongly reducing conditions were (AFCEE, 2004;
Weapons Station cis-DCE. After 8 direct injection into a test conducted for 8 lactate addition can stimulate induced (sulfate reduction and French et al., 2003)
Seal Beach, months of single well, and sub- months. A bioaug- complete dechlorination of methanogenesis), but after 8 months of
biostimulation, strate distribution was mentation pilot test is PCE and TCE to ethene. lactate injection, dechlorination of PCE
California transformation of effective within the planned. and TCE stalled at cis- DCE. Dehalo-
DCE to VC and treatment zone. coccoides species were not detected by
ethene was not molecular screening. A bioaugmenta-
observed. tion pilot test is planned.
Soluble Substrates – Molasses
Washington PCE to TCE to cis- Substrate was effec- Conditional closure Achieve closure under Wis- Closure with MNA was granted by (AFCEE, 2004;
Square Mall DCE to VC to tively distributed using with MNA. consin Department of Natural WDNR 30 months after the initial appli- Maierle and Cota,
ethene/ethane. an initial injection into Resources (WDNR) flexible cation. Closure was based on complete 2001)
Wisconsin 182 direct-push points closure rules (negotiated dechlorination of PCE to ethene and
and during follow-on preventative action limits). ethane. Residual levels of DCE and VC
injections using a limited were sufficiently low to allow for closure
array of 12 injection with MNA.
points.
Hanscom AFB TCE to cis-DCE to Substrate distribution Pilot test was com- Performance objectives Concentrations of total CAHs above (ARCADIS Geraghty
VC to ethene. was limited initially; pleted in September included 1) for concentrations 200 µg/L were reduced by over 80% at and Miller, 2003)
Massachusetts frequent injections were 2002. Future remedial of total CAHs >200 µg/L – most locations. Concentrations of total
required to extend the actions are under reduce concentration by 80% CAHs between 50 and 200
zone of influence. Loca- evaluation. within 1 year.; 2) for concen- µg/L were reduced by at least 75% at
tions without adequate trations of total CAHs from only a few locations. Complete dechlo-
substrate showed only a 50-200 µg/L – reduce con- rination to ethene was observed at
relatively small decrease centration by 75% within 1 ocations that received a continuous and
in TCE concentration, year; 3) for concentrations of adequate supply of substrate.
and incomplete dechlo- total CAHs <50 ppb – reduce
rination. concentration by 50% within
1 year. Show that TCE can
be completely dechlorinated
to ethane.
Other Soluble Substrates (Fructose/Acetate)
Anaerobic/Aerobic TCE to DCE during The system was effec- System is inactive Demonstrate complete Concentrations of PCE and TCE were (AFCEE, 2004)
Pilot Study, Naval pilot test. tive in recirculating pending regulatory dechlorination of PCE and reduced by only 40 to 60% during
Support Activity Further dechlorina- groundwater, although approval of a final TCE to DCE and VC in an active recirculation, with accumulation
Mid-South tion of DCE to VC distribution of substrate remedy for the site. anaerobic reactive zone, with of cis-DCE. The rate and efficiency of
observed 6 months appeared to be limited MNA with hotspot complete oxidation of residual substrate loading may not have been
Tennessee after a final pulsed as levels of TOC aver- reduction using direct DCE and VC in a natural adequate to completely degrade TCE,
injection of acetate. aged less than 20 mg/L injection of sodium downgradient aerobic treat- or for dechlorination of DCE to VC and
throughout the treatment acetate is proposed. ment zone. ethane.
zone.
Slow-Release Substrates – HRC®
Fisherville Mill TCE to cis-DCE to Substrate was effec- Results of the pilot Demonstrate that application Concentrations of TCE declined by 88 (AFCEE, 2004;
Site VC to ethene. tively distributed test are being of HRC® can remediate PCE, to 98%, coupled with dechlorination of Murray et al., 2001)
throughout the treatment evaluated for TCE, and dechlorination cis-DCE and VC to ethene (completed
Massachusetts zone. Levels of TOC consideration of a products to ethene. Demon- dechlorination pathway). Elevated
and metabolic acids full-scale strate that application of levels of TOC and metabolic acids
remained elevated for a application. HRC® can control the migra- persisted for at least 27 months. How-
period of at least 27 tion of CAHs to potential ever, concentrations of TCE, cis-DCE,
months. downgradient receptors. and VC within and downgradient of the
treatment zone remain above MCLs.
Springdale TCE to cis-DCE to Substrate was effec- Pilot testing complete. Demonstrate that application Sequential increases and decreases (AFCEE, 2004;
Cleaners, Portland VC to ethene. tively distributed Full-scale application of HRC® can remediate PCE, of TCE, cis-DCE, VC, and ethane were Sandefur et al., 2002)
throughout the treatment has been approved TCE, and daughter products observed over approximately 18
Oregon zone. Levels of TOC and is pending fund- to ethene. Obtained regula- months, indicating that PCE was co
and metabolic acids ing for the project. tory approval for full-scale pletely dechlorinated to ethene. Co
remained elevated for a application. centrations of dechlorination products
period of approximately were observed to rebound after ap-
18 months (HRC®) to 40 proximately 18 months, indicating that
months (HRC-XTM). the HRC® substrate had been de-
pleted. Regulatory approval for a full-
scale application was granted based
upon results of the pilot scale applica-
tion.
Atlas 10 Site Limited dechlorina- Elevated levels of TOC Site conditions were Remediate chloroethenes in Significant reductions in TCE concen- ((USACE), 2003)
tion of TCE to cis- and organic acids were determined to be groundwater to federal drink- tration did not occur. Only limited pro-
Nebraska DCE only. only observed in the unsuitable for en- ing water MCLs. duction of cis-DCE was observed in one
immediate vicinity of the hanced in situ biore- location towards the end of the pilot
injection array during the mediation. Groundwa- test. Development of a stable and
11-month test. ter extraction and ex highly reducing reaction zone was not
situ treatment using achieved. It is believed that high levels
aeration are being of DO (> 6 mg/L), nitrate (> 10 mg/L),
considered. and sulfate (>20 mg/L), combined with
a high rate of groundwater flow, inhib-
ited formation of adequate reducing
conditions.
SWMU 138, Limited dechlorina- Elevated levels of sub- Pilot test completed, Reduce concentrations of A lack of groundwater flow and mixing (CH2M Hill Construc-
Naval Air Station tion of PCE and strate were not observed full-scale expansion PCE and TCE to below MCLs is thought to have resulted in insuffi- tors, 2001)
TCE to cis-DCE. in any monitoring loca- was not recom- within the pilot test treatment cient distribution of substrate. Hydro-
Dallas, Texas No significant tions during the pilot mended. zone. geologic conditions at this site do not
concentration test. Distribution ap- appear to be suitable for use of slow-
trends could be pears to have been release substrates. The contractor
discerned in 9 limited by low permeabil- determined that HRC® did not offer an
months of monitor- ity soils and a lack of effective means to provide short term
ing. advective groundwater (less than 1 year) degradation of chlo-
flow. roethenes.
Slow-Release Substrates – Vegetable Oil
Site SS-17, Altus TCE to cis-DCE Injection of the vegeta- Pilot test results Determine if a vegetable oil Complete dechlorination of TCE to VC (AFCEE, 2004; Lee
AFB to VC to ethene. ble oil emulsion was continue to be evalu- inwater emulsion can be and ethene was observed. Concentra- and Lieberman, 2002;
readily accomplished, ated. effectively distributed and a tions of TCE were reduced to below Lee et al., 2003)
Oklahoma but uniform distribution reaction zone maintained in a detection along the centerline of the
was not observed at all barrier configuration. Deter- treatment zone. Although a complete
locations downgradient mine whether complete dechlorination pathway has been ob-
of the treatment zone. dechlorination of TCE to served, concentrations of cis-DCE and
Distribution appears to ethene can be stimulated. VC remain elevated above MCLs after
be controlled by secon- 13 months of monitoring. Substrate
dary permeability. distribution appears to be controlled by
aquifer heterogeneity and secondary
permeability.
SS-015, Travis TCE to cis-DCE to Elevated levels of TOC The site is approved Demonstration project to Complete dechlorination of PCE and (Parsons, 2004)
AFB, VC to ethene. were observed through for MNA and is being determine if vegetable oil is TCE to ethene was observed, with over
out the entire treatment redeveloped by the capable of complete dechlo- 80% reduction in total CAH mass site
California area, indicating that Air Force. Monitoring rination of TCE to ethene in a wide. Although MCLs for PCE, TCE,
substrate was distrib- will continue after high sulfate, low permeability cis-DCE and VC were attained at multi-
uted effectively. construction. environment, and to evaluate ple locations, this site may require
different vegetable oil injec- several years to attain uniform reduc-
tion scenarios. tion in CAHs to MCLs due to low per-
meability and slow desorption of CAH
mass.
Hangar K, Cape PCE to TCE to cis- Based on monitoring Field test complete. Demonstration project to Sequential increases and decreases of (Parsons, 2002)
Canaveral Air DCE to VC to data for TOC and meta- Continue to monitor determine if vegetable oil is TCE, cis-DCE, VC, and ethane were
Force Station ethene. bolic acids, substrate source area for long- capable of complete dechlo- observed over approximately 18
was effectively distrib- term depletion of rination of PCE and TCE to months. Approximately 40 months after
Florida uted throughout the substrate. ethene in a source area injection, MCLs were attained for PCE,
source zone area. application. Results were TCE, and cis-DCE at all locations within
evaluated to determine if the treatment zone. MCLs for VC were
federal drinking water MCLs also attained at multiple monitoring
could be achieved. locations.
Solid Substrates – Mulch Biowalls
Building 301, TCE to cis-DCE, Bark mulch was uni- Full-scale biowall Determine effectiveness of a TCE was dechlorinated to cis-DCE, with (AFCEE, 2004; Haas
Offutt AFB with limited evi- formly distributed within system was installed mulch biowall to completely limited evidence of dechlorination to VC et al., 2003)
dence of cis- DCE the biowall using a in 2001, with contin- dechlorinate TCE to ethene. to ethene. VC did not accumulate. A
Nebraska to VC and ethe- continuous one-pass ued long-term moni- Develop operations and cost full-scale biowall was installed in 2001
ne/ethane. trencher. toring. data to support a full-scale based on results of the pilot test. The
application. extent of dechlorination may be limited
by the residence time of CAHs in the
reactive zone.
OU1, Altus AFB Primarily TCE to Bark mulch was uni- Long-term process Intercept and contain a shal- Concentrations of TCE within and (Haas et al., 2003;
cis-DCE. Low formly distributed within Monitoring continues. low TCE and DCE groundwa- immediately downgradient of the biowall Henry et al., 2003)
Oklahoma levels (< 10 ug/L) the biowall using a Fullscale biowalls are ter plume in order to prevent have been reduced to below the MCL
of VC were ob- continuous trencher. currently being in- surface water discharge and for TCE. TCE was dechlorinated to cis-
served. Abiotic Elevated levels of TOC stalled for other CAH off-base migration. Evaluate DCE, with no accumulation of VC. The
degradation of TCE (greater than 20 mg/L) plumes at Altus AFB. the technology for application average reduction in concentrations of
and cis-DCE were observed up to 30 at other Air Force sites. total CAHs is 86 percent, although
is also occurring. feet downgradient of the elevated concentrations of cis-DCE
biowall. persist.
Bioaugmentation
Aerojet Facility PCE to TCE to cis- The recirculation system Additional pilot testing Determine if TCE and per- Substrate addition alone was incapable (AFCEE, 2004; Geo-
DCE to VC to was effective at distribut- was recommended. chlorate can be completely of completely dechlorinating TCE to Syntec, 2002)
California ethene. Also per- ing substrate within the System is being degraded through substrate ethene within the 90-day test period.
chlorate to chlorate pilot test area. Distribu- evaluated for full- addition. Because microcosm PCE was successfully dechlorinated to
to chlorite to chlo- tion of the bioaugmenta- scale application. studies indicated that a suit- ethane with the addition of the bioaug-
ride. tion culture was limited able microbial population may mentation culture. The pilot test was
to the immediate area of not be present, demonstrate also successful in degrading perchlo-
injection. the effectiveness of bioaug- rate to innocuous end products.
mentation to increase dechlo-
rination rates. Assess feasi-
bility of full-scale application.
Bachman Road PCE to TCE to Recirculation systems Full-scale application Determine relative effective- A 92% conversion of chloroethenes to (AFCEE, 2004)
Residential Wells cis-DCE to VC to were effective at distrib- under consideration. ness of bioaugmentation ethene was observed in the bioaug-
Site ethene. uting substrate and (lactate plus bioaugmentation mentation test plot within 43 days of
bioaugmentation culture. culture) versus biostimulation inoculation. Dechlorination of PCE and
Michigan alone (lactate only) to in- TCE to ethene was observed in the
crease dechlorination rates of biostimulation test plot after appr. 3
PCE and TCE to ethene. months from the start of lactate injec-
tion. The lag time for dechlorination of
cis-DCE to VC and ethene, and overall
dechlorination rates were enhanced by
addition of the bioaugmentation culture.
Field Applications of In Situ Enhanced Bioremediation

Site Application Contaminants Description of Remedial Objective Removal Effi- Substrate Comments Contacts/
Name/Location Date/Project Concentrations Geology ciency/Post Cost References
Status level Treatment System Design treatment con-
centrations
Soluble Substrates – Lactate
Bachman Road Sept. 2000 TCE, DCE, VC Relatively Groundwater remediation TCE: ND NA Contaminant concentration John Lenvay, Univ of
Residential Well 140 days TCE: 0.13 mg/L uniform glacial DCE: ND reduction, as measured in San
Site injection DCE: 0.19 mg/L outwash sands Pilot-scale VC: ND one monitoring well, was Francisco
2
VC: 0.25 mg/L and gravels. Surface area: 270 ft , depth: Ethene: 0.36 nearly 100% for TCE, Erik Petrovskis, HSI
Oscoda, MI 8 ft mg/L DCE, and VC. Geotrans
Two injection wells; one
extraction well; network of
monitoring points around the (DoD, 2002; Lendvay
plot. Lactate was injected as et al., 2001)
a 6% solution in water.
Bioaugmentation after 20
days
Amendments: nitrogen,
phosphorous
Area 6, Dover April 1998 to TCE, DCE Sand with Technology demonstration; PCE: ND NA As of March 1998, 98.5% David Ellis, DuPont
AFB June 1999 varying groundwater remediation TCE: 75 µg/L of TCE and DCE in 302-892-7445
PCE: 0.046 mg/L amounts of DCE: 45 µg/L groundwater were con-
Dover, DE TCE: 7.5 mg/L clay, silt and Pilot-scale (Sodium lactate: VC: 20 µg/L verted to ethene and 75% (U.S.EPA, 2000b)
DCE: 2 mg/L gravel. 200 mg/L) to 80% of the TCE and
2
VC: 0.034 mg/L Surface area: 2,400ft , depth: DCE mass had been
5 ft recovered as ethene.
3 injection wells, screen 38-
48' bgs. Closed loop recircu- Biofouling had been re-
lation cell w/ 3 recovery wells. ported as a problem, af-
Cyclic injection of nutrients fecting the consistency of
and unamended GW. injections, the injection
Bioaugmentation with a concentration, and the
microbial culture from the consistency of injection
DOE Pinellas Site rates.
Amendments: ammonia, and
phosphate
Former Dry- Aug.1997 to PCE 20-feet of Source reduction by 50% PCE: 220 µg/L NA PCE concentrations were Stacey Koch, RMT
cleaning Facility April 2000 glacially depos- TCE: 3 µg/L reduced by approximately Inc.
PCE: 520 µg/L ited sand and Full-scale DCE: 28 µg/L 58% while TCE and DCE
WI TCE: ND gravel overly- concentrations increased (Koch, 2001)
DCE: ND ing sandstone Lactate: 200 mg/L, amended form below method detec-
bedrock with 2 mg/L yeast extract, tion limits to concentrations
sodium sulfite greater than the Federal
Injection wells MCL for each contaminant.
centrations
The four injections after the
first were made under pres-
sure to reduce biofouling.
Test Area North Pilot Nov. TCE Fractured Groundwater remediation TCE: <5 µg/L $54,000 Within 6 weeks from the Kent Sorenson, Nor-
(TAN), INEEL 1998 basalt bedrock. total start of lactate injection thwind
TCE: 3.2 mg/L Full-scale nearly 100% of TCE con- Environmental
2
ID Surface area: 700 ft , centrations in groundwater
depth:200 ft had been dechlorinated to (ITRC, 2004; Soren-
300 gallons of sodium Lac- DCE and complete dechlo- son, 2000)
tate was injected weekly for 8 rination was occurring
months. within 4 months.
Ogallala Super- April 1998 PCE, TCE Approximately Technology assessment PCE: 60 µg/L Na Contaminant reduction Vicki Murt, Nebraska
fund Site 26 to 27 feet of TCE: 1 µg/L calculations could not be Department of Envi-
PCE: 30 µg/L fine to medium Pilot-scale DCE: 140 µg/L performed due to the ronmental
2
Ogallala, NE TCE: ND grained sand. Surface area: 420 ft , VC: NA limited availability of ana- Quality
DCE: ND depth:10 ft lytical data.
VC: ND Sodium lactate injected at (DoD, 2002; Murt,
about 60% lactate in water in 2000)
3 to 4 week intervals over 10
months.
One extraction well, two
injection wells, 6 monitoring
wells installed. Substrate
injection was performed
concurrently with groundwa-
ter extraction from the down-
gradient extraction well to
form a closed recirculation
system.
NAS Fallon 1998 PCE, TCE, DCE Approximately Compare enhanced and PCE: <25 µg/L NA Contaminant concentration Victor Magar, Battelle
500 days 4-feet of sandy intrinsic biodegradation rates TCE: 100 µg/L reduction in groundwater
Fallon, NV PCE: 680 µg/L soil underlain DCE: 250 µg/L ranged from a minimum (DoD, 2002)
TCE: 915 µg/L by approxi- Pilot-scale VC: 1.5 µg/L 65% in the case of VC to a
DCE: 866 µg/L mately maximum of 99.7% in the
VC: 4.3 µg/L 2-feet of clay- Recirculation with nutrient case of PCE.
rich silts and addition
sands. Bottom (Yeast extract, vitamins)
is sandy silt,
and clay
layer greater
20 feet thick.
Seal Beach Site PCE: 415 µg/L Surficial soils , PCE: ND NA Strongly reducing condi- Kent Sorenson,
40 TCE: ND sands and silty Pilot-scale TCE: ND tions were induced (sulfate Northwind
centrations
2
DCE: ND sands interca- Area: 1,300 ft , depth: 15 ft DCE: 485 µg/L reduction and methano-
CA VC: ND lated with low Sodium lactate: 3 g/L VC: ND genesis), but after 8
permeable One injection well, with 6 months of lactate injection,
intervals of monitoring wells interspersed dechlorination of PCE and
clay, silt and within a 20 ft radius. Weekly TCE stalled at cis- DCE.
silty clay. injections for 2 months, then Dehalococcoides species
none for 2 months, lastly were not detected by
every 3 weeks. molecular screening. A
bioaugmentation pilot test
is planned.
Tennessee Air 2002 Groundwater remediation to Analytical data was not Kent Sorenson,
national Guard reduce total VOC to < 500 provided for this site. Northwind
ppb
TN
Pilot-scale
Sodium lactate
Multiple manifold wells, bi-
monthly injections
Black’s Drycleaner 2001 PCE, TCE Technology demonstration Analytical data was not Kent Sorenson,
provided for this site. Northwind
OR Pilot-scale
Sodium lactate
Single well injection, one time
for demo.
Pinellas Northeast Analytical data was not
Site Pilot-scale provided for this site. (DoE, 1999)
Sodium Benzoate, sodium
Largo, FL lactate and methanol
Former PEC 2001 PCE, TCE, DCE, The surficial Groundwater remediation NA The well installation and (Dean et al., 2001)
Industries Site VC aquifer consists baseline monitoring
of three sand Pilot-scale phases have been com-
Orlando, FL PCE: 43 µg/L units separated Sodium lactate pleted as of May 2001. The
TCE: 17 µg/L by sandy clay Amendments: Ammonia and injection phase was
DCE: 8.1 mg/L and clayey ortho-phosphate scheduled for 2001 while
2
VC: 112 µg/L sand aquitards. Area: 12,000 ft , depth: 10 ft monitoring was to be
Each treatment cell consists extended through 2002.
of 4-5 extraction wells on cell
perimeter around central
injection well.
Cape Canaveral March 1999 TCE, DCE, VC Poorly sorted Treatability study TCE: ND NA Rapid dechlorination of Jeff Morse, Battelle
Facility 1381 coarse to fine DCE: 970 µg/L TCE and cDCE to VC,
149 days TCE: 1.97 mg/L sands and shell Pilot-scale VC: 625 µg/L followed by slower subse-
2
FL DCE: 11.1 mg/L material to 35 Area: 340 ft , depth: 10 ft quent dechlorination of VC ((AFRL), 2001)
VC: 1.25 mg/L feet. to ethene under sulfate
centrations
Lactic Acid and methanogenic condi-
tions.
Two communicating (recircu- RABITT protocol slightly
lation) wells, 13 tri-level modified because Florida's
groundwater monitoring UIC regulations do not
probes, and upgradient and allow for reinjection of
downgradient monitoring contaminated groundwater.
wells. Reduction of TCE, DCE,
and VC by 88.7%, 90.6%,
and 66.3%, respectively
Watertown Indus- July 1997 TCE; DCE Industrial site Feasibility study PCE: 100 µg/L NA Contaminant concentration Willard Murray, Hard-
trial Area adjacent to TCE: 1 mg/L reduction ranged from 7% ing
PCE: 1.5 mg/L river, 20 ft of Pilot-scale DCE: 3 mg/L in the case of PCE to 92% Lawson Associates
Watertown, MA TCE: 12 mg/L silty sand on VC: 1 mg/L in the case of TCE.
DCE: 3.5 mg/L top of thin Lactic Acid Contaminant mass reduc- (DoD, 2002)
VC: 100 µg/L till and tions were measured in a
bedrock. Amended with ammonia single well (IN-2).
chloride, tripolyphosphate, After 8 months of anaero-
yeast extract, and sodium bic operation, the system
hydroxide. was converted to an aero-
bic circulating cell for 3
System of injection and recir- months.- ORC,methane
culation cells addition => 70% reduction
of total ethenes in 11
months.
Aerojet Superfund Sept. 2000 TCE Technology demonstration TCE: < 5 µg/L TCE concentrations were Evan Cox or David
Site DCE: NA reduced by nearly 100% Major,
TCE: NA Pilot-scale VC: NA 125 days of injection. Geosyntec Consul-
CA DCE: NA Lactic acid tants
VC: NA
Bioaugmentation with a
commercially produced mi- (DoD, 2002)
crobial augment (KB-1)
Naval Air Station Dec. 1998 TCE, DCE, VC Upper uncon- Technology demonstration TCE: < 5 µg/L Within approximately 180 Daniel Leigh, IT
Point Mugu IRP fined aquifer: DCE: <5 µg/L days TCE and DCE con-
Site 24 1 year TCE: 1.7 mg/L sand and Pilot-scale VC: 15 µg/L centrations had been (Leigh et al., 2000)
DCE: 750 µg/L gravel overly- Lactic acid reduced by nearly 100% at
CA VC: 1 µg/L ing approxi- one well. However VC
mately 4 feet of One extraction well, one concentrations increased
clay. injection well and 5 monitor- by a factor of 15 within the
Lower confined ing wells. Groundwater was same time period.
aquifer: mas- circulated in a Pilot test monitored for 3
sive sand unit. closed loop between the years, then converted to
centrations
extraction well and the injec- aerobic cometabolism for
tion well. faster VC degradation
Full-scale sequential an-
aerobic/aerobic treatment
is planned.
Weyerhauser Pilot test PCE, TCE Sandy silts and Feasibility study PCE: 200 µg/L The PCE, TCE, and DCE Sanjay Vanchees-
Sycan Mainte- complete silty sands TCE: 160 µg/L concentrations were re- waran, CH2M
nance Shop PCE: 1.7 mg/L Pilot-scale DCE: 1400 µg/L duced by approximately Hill
TCE: 450 µg/L Lactic acid VC: 90 µg/L 88%, 64%, and 61% re-
Beatty, OR DCE: 3.6 mg/L spectively while the VC (DoD, 2002)
VC: 90 µg/L Injection and monitoring concentration stayed about
system was placed to inject the same.
into and monitor changes
within the upper aquitard.
One well was used for
groundwater extraction,
substrate injection, and proc-
ess monitoring.
Lauderick Creek Pilot test PCA, TCA Treatability study PCA: 3.5 mg/L Within 14 weeks the PCA Andrew Jackson,
Area of Concern, complete TCE: 1.8 mg/L concentration at one well Texas Tech
Main Plume, PCA: 20 mg/L Pilot-scale DCE: 3.9 mg/L was reduced by 83% while University
Aberdeen Proving TCE: 1 mg/L VC: ND the TCE and DCE concen-
Ground DCE: ND Lactic acid trations increased. (DoD, 2002)
VC: ND
Amendments: Vitamin B12,
MD yeast extract
Soluble Substrates – Molasses
Avco Lycoming Pilot: Novem- TCE Glacial sandy Groundwater remediation TCE: 1.4 µg/L Daniel Jacobs, AR-
Superfund Site ber 1995 TCE: 48 µg/L silt overburden DCE: 2 µg/L CADIS
Full: January DCE: 10 µg/L overlying frac- Pilot, Full VC: <1 µg/L 215-752-6840
Williamsport, PA 1997 to July VC: <1 µg/L tured limestone
2
1998 bedrock Area: 5,000 ft , depth: 11 (Horst et al., 2000)
20 injection wells
Cedarburgh Dry- PCE, TCE, DCE Poorly drained Soil and groundwater reme- NA Jim Drought, ARCA-
cleaners silty clay topsoil diation DIS
PCE: 6.8 mg/L with little sand 414-276-7742
Cedarburgh, WI TCE: 810 µg/L and gravel (1- Full-scale
DCE: 1 mg/L 4'). Silty clay (DoD, 2002)
2
and clayey silts Area: 11,500 ft , depth 25-28’
(4-16'). Silty
clay with trace Geoprobe, infiltration gallery
sand and
gravel (15-39')
centrations
overlying silty
find sands.
Industrial Site Phase I: DCE, VC Sand and Contaminant mass reduction DCE: 25 µg/L Phase I was designed to Jim Peeples, formerly
November gravel in a downgradient groundwa- VC: 4 µg/L confirm the capability of Parsons
Central Ohio 1996 DCE: 265 µg/L ter plume enhancing reductive
Phase II: VC: 15 µg/L dechlorination in-situ. (Peeples et al., 2000)
September Pilot-scale Phase II was designed to
1998 Area: evaluate amendment
2
Phase III: Phase 1: 500 ft distribution strategies.
2
June 1999 Phase II: 4,500 ft Phase II was designed to
2
Phase III: 32,000 ft the maximum volume of
Depth: 25 ft aquifer that could be im-
pacted by a single well
Phase I consisted of 1 injec- injection. Contaminant
tion well, 1 injection well, and concentration data were
1 monitoring well. Phase II collected during Phase I
consisted of 2 injection wells only.
and 9 monitoring wells.
Phase III used the same
injection wells installed for
Phase II but added 20 more
monitoring wells.
Washington August '98; 6 PCE Approximately Source reduction with resid- PCE: ND Site closure from WDNR, Michael Maierle,
Square Mall months - 14 feet of clay ual natural attenuation TCE: ND less than 2.5 yrs after ARCADIS
Pilot PCE: 1.5 mg/L and silt overly- DCE: 400 µg/L initiating the in situ reaction 414-276-7742
Germantown, WI March '99; 6 ing a 2 to 5 foot Full-scale VC: 100 µg/L zone.
2
months - thick saturated Area: 30,000 ft , depth: 5 ft (Maierle and Cota,
Full silt and sand 2001)
bed, clay till Molasses (47% carbohydrate
aquitard below during pilot; 66% during full)
the
saturated Geoprobe, infiltration gallery.
silt/sand bed
Manufacturing Pilot - March CT Silty sand and Site closure CT: 5 µg/L $30,000/yr.
Facility 1998 sandy silt
Full - October CT: 0.19-1.2 overlying a Pilot-scale, Full-scale
2
Greenville, SC 2001 mg/L highly weath- Area: 170000 ft , depth: 25-
2 years ered transition 73 ft
zone.
Fully automated
Crestwood Site 1998 PCE: 44 mg/L Approximately Mass reduction, natural NA
4 to 12 feet of attenuation for low-level
Glendale, WI fill material residuals
overlying 4 to 8'
centrations
of sand; clay Full-scale
till aquitard
underlies the Geoprobe, infiltration gallery
sand bed.
Emeryville Manu- Pilot - August TCE, DCE Interbedded Reduce average concentra- TCE: <5 µg/L $3 per Closure, all contaminant
facturing Facility 1995 sand and clay tion of total chromium and DCE: <5 µg/L gallon concentrations were below
Full - April TCE: 6.5 mg/L to a depth of TCE in groundwater by more VC: 21 µg/L MCLs.
Emeryville, CA 1997 DCE: 590 µg/L 24 feet under- than 90%
VC: 10 µg/L lain by imper-
meable clay Pilot-scale, Full-scale
91 injection points with Geo-

probe
Manufacturing Pilot - October PCE 10-20' thick Full-scale PCE: 1 µg/L $5,000/year Zero valent iron proposed
2
Facility 1998 overburden of Area: 450000 ft , depth: 50- TCE: 1 µg/L for source area.
Full - April PCE: 80 mg/L glacial sands, 190 ft DCE: 1 µg/L Full-scale containment in
Northeastern USA 2000 TCE: 1 mg/L silts and clays; VC: 2 µg/L place in mid-plume 2000 to
DCE: 1 mg/L water table in Molasses, 200 gal; 2002. Full scale source
2 to 5 years of VC: ND underlying 10%carbohydrates in each area injection program
injection bedrock. well began in October 2001.
Batch delivery under pres-

sure to 11 injection wells.
Nuclear Fuel August 2000 PCE, TCE 0-30' Alluvial Reduce VOC concentrations, PCE: 850 µg/L Within 6 months following
Services Site 8 months of silts, clays, contain plume migration TCE : 320 µg/L substrate injection PCE,
injection PCE: 12.4 mg/L clayey sand, DCE: 67.8 mg/L and DCE contaminant
Erwin, TN TCE: 1.14 mg/L sand w/ gravel Pilot-scale VC: 13 mg/L mass had been reduced by
DCE: 660 µg/L and cobbles 93% and 82% respectively.
VC: 130 µg/L underlain by Within the same time
weathered, period DCE, VC, and
fractured, ethene concentrations
steeply. increased by approxi-
mately 2 orders of magni-
tude. The pilot scale appli-
cation was considered a
success and full
scale applications are
scheduled for "designated
source areas" in the 2nd
quarter of 2002.
TRW Automotive 4 months of PCE 0-30' Alluvial Reduce VOC concentrations PCE: 2 µg/L
injection silts, clays, TCE: <2 µg/L
Rogersville, TN PCE: 22 mg/L clayey sand, Pilot-scale DCE: 8.6 µg/L
centrations
TCE: 5 µg/L sand w/ gravel VC: 22.6 µg/L
DCE: 42 µg/L and cobbles
VC: 1 µg/L underlain by
weathered,
fractured bed-
rock.
Hanscom AFB October 2000 TCE, DCE, VC Upper and Technology demonstration TCE: 1.9 mg/L Post-treatment concentra-
lower aquifers DCE: 5.3 mg/L tions are those from exist-
Bedford, MA TCE: 1.9 mg/L consisting of Pilot-scale VC. 1.3 mg/L ing monitoring well after 19
2
DCE: 5.3 mg/L glacial till Area: 5000 ft , depth: 50 ft months of demonstration
VC. 1.3 mg/L overburden testing.
overlying One injection well. Manual,
bedrock and pressurized injections. Post-
separated by a injection water pushes.
stiff.
Vandenberg AFB February TCE Alluvial marine Technology demonstration TCE: 19 µg/L Concentrations from exist-
2001 sands (with ing monitoring well show-
Lompoc, CA TCE: 4 mg/L gravel, silt, and Pilot-scale ing good treatment.
2
clay) over Area: 1000 ft , depth: 30 ft
siltstone Molasses: variable frequency
and amount based on injec-
tion well process monitoring
data.
3 injection wells. Manual,
pressurized injections.
Ohio Industrial 1999 PCE, TCE Porous, high Plume restoration by reactive PCE: < 1 µg/L Within approximately 440
Site carbonate barrier TCE: < 1 µg/L days following substrate
PCE: 500 µg/L aquifer VC: < 1 µg/L injection PCE, TCE, DCE,
Southwestern TCE: 700 µg/L Full-scale VC concentrations in
Ohio VC: 3 µg/L groundwater had been
5 to 10% molasses solution reduced by nearly 100%.
biweekly for 6 months, inter- At day 180 DCE concen-
mittent thereafter trations peaked and began
to decline while ethane
Reactive barrier configuration concentrations peaked at
day 240.
Manufacturing 1999 TCE Sandy gravel Plume restoration, partially TCE: 14 µg/L Since substrate injection in
Facility alluvium overly- underneath the facility DCE: 20.8 mg/L 1999 the average TCE
TCE: 22 mg/L ing a thick VC: 4.5 mg/L concentration has de-
Southeast Eng- DCE: 12 mg/L regional aqui- Full-scale creased by nearly 100%
land VC: 45 µg/L tard known as while VC and ethane
the London A total of 53 injection wells concentrations have in-
Clay were installed under the creased by 1 and three
future site of a new building. orders of magnitude re-
centrations
Each injection well was piped spectively and the average
separately to the injection DCE concentration has
system installed in a separate increased by approxi-
building so that substrate mately 73%..
injection could be controlled
by well. 7 monitoring wells
were installed up and down
gradient from the injection
area.
Former Manufac- 2001 PCE, TCE Silty clay Technology Demonstration NA PCE and TCE concentra-
turing Facility tion decreases of 89% and
1 year PCE: 11 mg/L Full-scale 94% respectively, were
North Carolina TCE: 180 µg/L reported although analyti-
cal data was not provided.
Soluble Substrates – Butyrate

Naval Air Station June 1999 TCE, DCE, VC Treatability study TCE: 700 µg/L Water injected with amend-
Alameda DCE: 2.4 mg/L ments contained average
Building 360 (Site 194 days TCE: 24 mg/L Pilot-scale VC: 600 µg/L TCE, cDCE, and VC con-
2
#4) DCE: 8.6 mg/L Area: 45 ft , depth: 3 ft centrations of 81.7 mM, 7.0
VC: 2.2 µg/L Butyric acid (3 mM) at 236 mM, and 3.4 mM, respec-
Alameda, CA gal/day (0.62 L/min) tively. Average TCE con-
Amendments: Yeast extract, centrations were reduced
20 mg/L by 94% by the end of the
demonstration.
Single injection well, single On average, 87% of in-
extraction well, and nine jected chloroethenes could
monitoring points at depths of be accounted for, with
24 to 27 feet bgs ethane levels accounting
for approximately 50% of
the total
chloroethene concentra-
tion.
Fort Lewis East August 2000 TCE, DCE Treatability study TCE: 69 µg/L Injected TCE concentra-
Gate Disposal TCE: 1.5-6.3 DCE: NA tions spiked as high as 169
Yard 179 days mg/L Pilot-scale VC: 217 µg/L mg/L, but this did not prove
2
Area: 120 ft , depth: 3 toxic to the microorgan-
Fort Lewis, WA Butyric acid, 1.5 L/min isms.
Amendments: Yeast extract,
20 mg/L; Sodium bicarbon- Calculated TCE half lives
ate, 279 mg/L of 4.4 to 4.7 hours. cDCE
Three injection wells spaced accumulated with VC
2 ft apart; system of monitor- comprising a small per-
ing wells; existing well in centage of overall mass.
centrations
zone of contamination used Ethene and ethane re-
to supply injection water for mained at or below detec-
test plot. tion.
Marine Corps June 2001 PCE, TCE Sand and clay Treatability study Average instead
Camp Lejeune sediments to of max:
Site 88 180 days NA 75 ft bgs. A Pilot-scale
2
series of sand Area: 120 ft , depth: 3 PCE: 5.5 mg7l
Camp Lejeune, and limestone Butyric acid, 0.65 L/min TCE: 400 µg/L
NC beds occur Amendments: Yeast extract, DCE: 80 µg/L
between 50 20 mg/L VC:<6 µg/L
and 300 ft bgs. Three injection wells, nine
monitoring wells at depths of
45 to 48 feet bgs.
Soluble Substrates – Acetate
Hanford 200 Area January 1995 CT, Nitrate Unsaturated Technology demonstration NA Approximately 2 kg of CT
Site to March zone extends was reported to have been
1996 NA to a depth of Pilot-scale destroyed during the 14
2
Richland, WA 250 feet. Two Area: 400 ft , depth: 10 month pilot test. Within the
permeable Amendments: Nitrate same time period approxi-
zones are mately 30 kilograms (dry
located at 250 One injection well and one weight) of bacterial bio-
to 256 feet bgs extraction well were installed mass was produced.
and 285 to 302 and run continuously as a Acetate and nitrate were
feet bgs with a recirculation system; two injected and recirculated
competant monitoring wells were in- through 2 aquifer zones to
aquitard bet- stalled between the injection treat carbon tetrachloride
ween. and extraction wells for proc- and nitrate contamination.
ess monitoring.
Gillette Company TCE Technology demonstration NA This site had high sulfate
Site (>1000 mg/L), high salinity,
Pilot-scale and tidal impact due to
Eastern US close proximity of the
One injection well and one coast. The target zone was
extraction well were installed. also deep
Acetate was injected to re- >100 ft. TCE degradation
duce sulfate mass and to to DCE was noted early on
push geochemistry to more but DCE degradation and
reducing conditions. 3 VC/ethene accumulation
months after acetate injec- was not observed until 4
tion, bioaugmentation with months after bioaugmenta-
methanol and KB-1 was tion with the
performed. KB-1 commercial
bioaugmentation product.
Manufacturing 1998 PCE, TCE Thin saturated Technology demonstration NA Acetate injection into a
centrations
Facility soil over frac- surficial overburden
tured basaltic Pilot-scale aquifer/fractured bedrock
NJ bedrock. aquifer.
Soluble Substrates –Methanol/Acetate

Building 360, Kelly December, PCE, TCE, DCE 20 to 40 feet of Technology demonstration/ PCE : 810 µg/L Reported a 90% reduction
Air Force 1999 alluvial gravel, source TCE : 19 µg/L in PCE. Cis- 1,2-DCE
Base PCE: 8.1 mg/L sand, and silt reduction DCE : 315 µg/L degradation was observed
320 days TCE: 190 µg/L overlying im- VC : NA only after the addition of
South Central TX DCE: 2.1 mg/L permeable Bench, Pilot-scale the KB-1 microbial consor-
2
VC: ND clay. Area: 90 ft , depth: 25 ft tium amendment.
KB-1 ("A naturally occurring
microbial consortia isolated
by
GeoSyntec and Univ. of
Toronto")
Closed loop circulation sys-

tem
consisting of 3 extraction
wells, 1
injection well and 5 monitor-
ing wells.
Rademarkt Site 35 weeks PCE, TCE, DCE, Technology Demonstration PCE : 600 µg/L Contaminant concentration
VC TCE : 10 µg/L reductions represent site
Groningen, Pilot-scale DCE: 290 µg/L average concentrations
2
Netherlands PCE : 1.28 mg/L Area: 5000 ft , depth 10 ft VC: 2.26 mg/L before injection and 25
TCE : 1.03 0.35% methanol, 1.44% weeks after injection.
mg/LPCE compost This application used
DCE : 2.6 mg/L leachate, 0.9 g/L ammonium methanol and compost
VC : 940 µg/L chloride leachate as a substrate to
10 Injection wells were in- augment reductive dechlo-
stalled rination already
upgradient of 5 extraction occuring at the site.
wells.
Groundwater was extracted,
amended, and reinjected.
Soluble Substrates –Lactose
Former Municipal 1997-2000 - PCE, TCE, DCE Upper fine Groundwater remediation to PCE: <1 µg/L
Waste Water pilot sand and silt potable water standards TCE: <1 µg/L
Treatment Facility 2000-2003 - PCE: 20 µg/L unit (5-20'); DCE: 2.7 mg/L
(Lactose) full-scale TCE: 300 µg/L Lower sandy Full-scale VC: 300 µg/L
2003-pursue DCE: 8.4 mg/L and silt unit
NH closure with VC: 1.5 mg/L (30-50' bgs) Lactose (approx. 70%) and
centrations
MNA Yeast
Extract (approx. 30%) (pro-
prietary
blend)
Injection of blend pumped
into 3
locations: 5,050 lbs biostimu-
lant per 65,950 gallons in-
jected during pilot; 22,100 lbs
per 375 gallons injected
during full-scale
Soluble Substrates –Fructose
Naval Support March 2000 TCE Fluvial deposits Feasibility test for NA
Activity Mid- aquifer groundwater restoration
South (Fructose,
Acetate) Pilot-scale
Amendment: Acetate
Millington, TN Recirculation system with one
downgradient extraction well.
Soluble Substrates –Sodium Benzoate

New England January 1998 DCE, Technology demonstration for NA Degradation rates were
Super Fund Site Tetrahydrofuran, enhanced bioremediation as calculated for cis- 1,2-DCE
Sages Drycleaner 2 years Fuel Hydrocar- a cost effective alternative to and VC based on produc-
bons pump and treat tion rates of ethene and
New England ethane for the first two
Pilot-scale years of operation. The
Sodium Benzoate, injected as average half life calculated
a4 for cis-1,2-DCE was 255
mg/L solution at a rate of 5 days. The calculated aver-
liters per point per day. age half life for
30 electron donor injection VC was 48 days.
points
installed in sets of three
(three injection intervals) in a
barrier configuration. 1 up-
gradient and 3 downgradient
rows of monitoring wells are
used for process
monitoring.
Soluble Substrates –Ethanol
June 1998 PCE DNAPL Technology Demonstration to PCE: 44 mg/L Approximately 43 liters of
remove PCE DNAPL PCE were recovered from
Jacksonville, FL 1 year PCE: 150 mg/L the extracted water/ etha-
centrations
Pilot-scale nol waste stream. This
Ethanol 8,980 gallons in- PCE mass represents
jected as a approximately 63% of the
95% ethanol in water mixture. total PCE mass present in
Amendments: methanol, n the flushed
hexanol area indicating that the
3 injection wells surrounded extraction efficiency is
by 6 recovery wells. Ethanol 63%.
was injected at a total of 15
liters per minute for three
days. Groundwater was
extracted at a rate of 30.4
liters per minute to ensure
containment of injected fluids.
Contemporary PCE, TCE, DCE, Fine grained Groundwater remediation via PCE: 820 µg/L $40,800 1st Within 152 days following
Cleaners 1 year VC quartz sand accelerated anaerobic bio- TCE: 1.3 mg/L application injection PCE, TCE, DCE,
(25-30'), clay 1- degradation DCE: 4.0 mg/L and VC concentrations
Orlando, FL PCE: 19.2 mg/L 12'; fine VC: 1.0 mg/L $40,860 were reduced by 96%,
TCE: 2.5 mg/l grained sand Full-scale 2nd 48%, 38%, and 58%,
2
DCE: 2.4 mg/L and sandy clay Area: 14600 ft , depth: 15 ft application respectively, over 5 wells
20-25 screened in the shallow
HRC; 6,800-lbs was injected aquifer.
into the shallow aquifer Two HRC applications
18 months later 6,810 pounds were performed on this
were injected into the deep site. The first application
aquifer 144 injection points was performed in the
installed within the upper shallow aquifer. The sec-
aquifer on a 10 foot grid ond application was per-
spacing. Second injection formed 18 months later in
consisted of 145 injection the deep aquifer because
points on a 10 foot grid. the first application did not
appear to
be affecting contaminant
mass present in the deep
aquifer.
Decorah Shopping 1999 PCE Sandy silt and Contaminant source removal, NA
Center Dryclean- silty sand with plume stabilization, ground-
ers PCE: 18 µg/L varying a- water
TCE: 4 µg/L mounts of clay restoration
Decorah, WI DCE: <1 µg/L (5'), med
grained sand Full-scale
(5'-11'bgs) silty
sand and
centrations
sandy silt with
silty clay (11-
24'bgs), silty
clay
(24-28'bgs)
Dixie Cleaners June 2000 PCE, TCE, DCE, Silty , fine- Source zone remediation PCE: 42 µg/L Average PCE, TCE, and
VC grained sand TCE: 1.3 mg/L DCE concentrations were
Jacksonville, 1 year (0-18' bgs); Pilot-scale DCE: <5 µg/L reduced by 99%, 90%, and
2
FL PCE: 5.2 mg/L clayey fine- Area: 18400 ft , depth: 10 ft VC: NA 99% 7 months after injec-
TCE: 12.7 mg/L grained sand tion.
DCE: 7.5 mg/L (18-30' bgs); 175 injection points installed
VC: 1.1 mg/L limestone (30- on a 10 foot grid spacing. 30
32' bgs) monitoring wells, 10 installed
within each aquifer zone.
Dover Park Plaza PCE, TCE Shallow sand Source zone and plume PCE: <5 µg/L After approximately 400
Drycleaning Facil- (15-22'), dense remediation TCE: <5 µg/L days following the full scale
ity PCE: 1.4 mg/L silty clay DCE: 220 µg/L application PCE and TCE
TCE: 20 µg/L (30') Pilot-, Full-scale VC: <5 µg/L concentrations were re-
2
Yardville, NJ DCE: 100 µg/L Full: 3,900 ft , depth: 5 ft duced by approximately
VC:<5 µg/L 99% and 75% respectively.
Pilot - 8 direct injection wells
installed in a semicircle 2.5
feet from one monitoring well.
A second monitoring well was
installed 3 feet downgradient
from the outside edge of the
injection point circle.
Full - 159 injection points
installed in 3 grids and 1
barrier.
Former Industrial Aug. 1998 TCE Relatively low Polish source area after TCE: 4.7 mg/L Contaminant concentration
Filter Manufac- permeability removal of 2-Phase extrac- DCE: 18 mg/L data used to calculate
turer TCE: 26 mg/L clayey silts tion system VC: 190 µg/L mass reduction represents
DCE: 340 µg/L maximum values detected
Rochester, NY VC: ND Full-scale in 5 monitoring wells.
2
Area: 900 ft , depth: 15 ft Within 15 months of
injection TCE concentr
21 injection points at 5-ft tions were reduced by
centers using a Geoprobe rig approximately 64% while
DCE and VC concentra-
tions increased by ap-
proximately 288% and
3700% respectively.
Former Landfill Aug. 2000 TCE Determine potential for TCE: ND DCE and VC peaked
centrations
Site 7, Duluth groundwater remediation via DCE: 700 µg/L before starting to decline at
International TCE: 354 µg/L anaerobic biodegradation VC: 20 µg/L end of pilot test.
Airport DCE: 50 µg/L
VC: 10 µg/L Pilot-scale
Duluth, MN
9 injection points using a
Geoprobe rig
Watertown Indus- Feb. 1998 PCE, TCE Appr. 13 feet of Feasibility study for PCE: 8.5 µg/L Average PCE, TCE, and
trial Area sand and enhanced anaerobic TCE: 95 µg/L DCE concentrations were
PCE: 1 mg/L gravel overly- biodegradation DCE: 163 µg/L reduced by 99%, 99%, and
Watertown, MA TCE: 13 mg/L ing approxi- VC: < 200 µg/L 95% at 206 days after
DCE: 3 mg/L mately 7 feet of Pilot-scale injection.
VC: 200 µg/L silty sand. The
surficial mate- HRC-containing canisters
rial is suspended in injection well
underlain by an HRC canisters in wells used
aquitard con- with a recirculation system
sisting of gla-
cial till.
Former Manufac- Jan. 1999 DCE, VC Silty sand Feasibility study for enhanced DCE: 5.6 mg/L Groundwater results
turing Site anaerobic biodegradation VC: 3.2 mg/L showed large amounts of
DCE: 9.9 mg/L lactic acid fermentation.
Walled Lake, VC: 1.9 mg/L Pilot-scale
MI Geoprobe injection
Unocal Wichita Sept. 1999 PCE Silt and clay Feasibility study for enhanced PCE: 40 µg/L PCE was degraded from 6
anaerobic biodegradation TCE: 540 µg/L mg/L to 0.2 mg/L within 30
Wichita, KS PCE: 7 mg/L DCE: 15 mg/L days.
TCE: 840 µg/L Pilot-scale VC: 1 mg/L
DCE: 560 µg/L
VC: ND 15 injection points
FMC Corporation June 1999 to TCE Silty clays (45- Accelerate groundwater TCE: 3.15 mg/L Contaminant concentra-
Site /May 2000 50'); gravelly restoration DCE: 550 µg/L tions used to calculate
TCE: 3.62 mg/L sand (30- VC: 570 µg/L mass reductions represent
San Jose, CA Full-scale DCE: 210 µg/L 35') Pilot-, Full-scale averages of 6 site wells
2
planned VC: 20 µg/L Area: 150 ft , depth: 20 ft used to monitor this pilot
test. Within 6 months of
6 direct-push points and 5 injection the average TCE
monitoring wells. concentration had been
reduced by 13%. DCE, VC,
and ethene concentrations
centrations
increased by 161%,
2750%, and 733% respec-
tively during the same time
period.
Hayden Island May 1999 PCE 20-40 feet of Accelerate groundwater PCE: 200 µg/L PCE concentrations were
Cleaners silty sand restoration TCE: 35 µg/L reduced by approximately
PCE: 800 µg/L DCE: 70 µg/L 75% within 20 months of
Portland, OR TCE: 1 µg/L Full-scale injection, while TCE and
2
DCE: 3.4 µg/L Area: 200 ft , depth: 15 ft DCE concentrations in-
creased within the same
34 injection points in 2 rows time period.
Site closed
Naval Air Station July 2000 – PCE, TCE Fine-grained Groundwater remediation PCE: 79 µg/L
Dallas May 2001 fill, alluvial TCE: 51 µg/L
PCE: 99 µg/L sediments, Pilot-scale DCE: 23 µg/L
2
Dallas, TX TCE: 44 µg/L weathered Area: 1500 ft , depth: 10 ft
DCE: 11 µg/L shale
Naval Air Warfare TCE, DCE Groundwater remediation TCE: 540 µg/L
Center DCE: ND
TCE: 1 mg/L Full-scale VC: 0.5 µg/L
Indianapolis, IN DCE: 202 µg/L
VC: 0.3 µg/L Injection per Regenesis
design
Hurlburt Field Jan. 1999 TCE, DCE The intermedi- Accelerate natural attenua- TCE: 1 mg/L Within 1 year of injection
ate aquifer tion of groundwater to less DCE: 8.5 mg/L TCE concentrations had
Tallahasse, FL TCE: 9.5 mg/L consists of than 5 years VC: 1.5 mg/L decreased by approxi-
DCE: 2 mg/L sand, silty mately 89% while DCE,
VC: <100 µg/L sand, and Pilot-scale VC, and ethene concentra-
2
discontinuous Area: 2500 ft , depth: 30 ft tions increased by ap-
lenses of clay proximately 325%,
and is located 24-25 direct injection points 1,400%, and 400% respec-
from 40 to installed in a grid with 5 foot tively. These concentration
50 feet below spacing. A total of 7 monitor- changes are representative
ground surface. ing wells were installed for of one well installed ap-
process monitoring proximately 5 feet down-
gradient of the injection
area. The highest concen-
trations detected during the
baseline event were de-
tected at this well.
Industrial Site PCE PCE: 775 µg/L
Pilot-scale TCE: 866 µg/L
NJ PCE: 5.28 mg/L DCE: 3.35 mg/L
TCE: 227 µg/L 23 injection points using VC: ND
centrations
DCE: 10 µg/L Geoprobe rig
Closed Industrial Sept. 1999 PCE, TCE 20-25' Uncon- Feasibility study PCE: 500 µg/L Within first month of HRC
Facility solidated silt TCE: 680 µg/L injection, PCE concentra-
1 year PCE: 9 mg/L and clay with Pilot-scale DCE: 17 mg/L tions decreased by up to
CO TCE: 1.4 mg/L occasional 97% in the source zone.
DCE: 560 µg/L lenses of fine Polymer injected at nine
sand and locations using Strataprobe
sandy clay; rig
severely
weathered
claystone
Site 1, Santa Dec. PCE, TCE Interbedded Accelerate groundwater PCE: <20 µg/L
Clara County 1999/Sept. layers of clay, restoration TCE: 71 µg/L
2001 PCE: 27 µg/L silty sand, DCE: 5.23 mg/L
Santa Clara TCE: 11 mg/L and sand (clay Pilot-, Full-scale VC: 14 µg/L
County, CA DCE: 595 µg/L is predominant
VC: <50 µg/L soil type above Injection by Geoprobe rig
water-bearing
zone)
Site 2, Santa Oct. PCE, TCE, DCE Interbedded Accelerate groundwater PCE: 23 µg/L Additional injections to
Clara County 2000/June layers of clay, restoration in source area TCE: 940 µg/L source area after 9
2001 PCE: 220 µg/L silty sand, DCE: 1.5 mg/L months. Both fast and slow
Santa Clara TCE: 820 µg/L and sand (clay Pilot-, Full-scale VC: 1.4 mg/L release HRC
County, CA DCE: 3 mg/L is predominant products were shown to
VC: 300 µg/L soil type above Injection using Geoprobe rig. stimulate anaerobic
water-bearing First, biodegradation of CAHs
zone) HRC-primer (fast-release)
applied, and then the stan-
dard polymer (slowrelease) is
injected few inches away.
Tosco Manufactu- Aug. 2000 PCE, TCE, DCE 30 feet of Accelerate groundwater PCE: 3.1 mg/L Within 159 days following
ring Facility dense clay restoration at hotspots TCE: 2.5 mg/L injection the PCE concen-
PCE: 11.4 mg/L overlying tration in one well was
Burien, WA TCE: 1.7 mg/L approximately Full-scale reduced by 73% while the
2
20 feet of sand Area: 4100 ft , depth: 15 ft TCE concentration in the
same well increased by
33 direct injection points. 44%. DCE and VC data for
HRC was injected at a rate of this site were not provided.
5 pounds per foot from 5 to
20 feet below ground surface.
Strip Mall Dry Dec. 1998 PCE, TCE Groundwater remediation PCE: 34 µg/L PCE and TCE concentra-
Cleaner TCE: 17 µg/L tions had been reduced by
PCE: 67.4 mg/L Full-scale DCE: 20 µg/L nearly 100% within 14
2
Kent, WA TCE: 11.7 mg/L Area: 2000 ft VC: 70 µg/L months of injection at one
centrations
DCE: 360 µg/L source area well. DCE and
VC: 60 µg/L 55 direct injection points and VC concentrations had
three monitoring wells. been reduced by 44% and
88% respectively in one
downgradient well within 2
years of injection.
Dayco Manufactu- Feb. 1998 TCE Feasibility study for HRC in a TCE: 560 µg/L Within 35 weeks of injec-
ring Facility barrier configuration DCE: 1.71 mg/L tion TCE, DCE, and VC
35 weeks TCE: 2.23 mg/L VC: <1 µg/L concentrations were re-
Eldora, IA DCE: 5.03 mg/L Pilot-scale duced by approximately
2
VC: 134 µg/L Area: 250 ft , 15 ft 75%, 66%, and nearly
100% respectively. These
HRC was emplaced using 4 reductions were observed
foot long perforated poly in one well approximately 5
carbonate canisters. A total of feet downgradient from the
4 canisters were installed injection well. Contaminant
successively in four treatment concentration data from
event spanning 1 week. A wells further downgradient
total of 8 monitoring wells from the injection area
were used for process moni- were not provided.
toring activities.
Moen Manufactu- July 1999 DCE, VC Feasibility study to compare DCE: 20 µg/L Original COC was TCE but
ring Facility anaerobic and aerobic deg- VC: 10 µg/L nearly all TCE mass has
180 days pilot DCE: 590 µg/L radation of been reduced naturally to
Elyria, OH tests with VC: 210 µg/L DCE and VC DCE and VC. TCE has not
design of full been detected onsite at
scale applica- Pilot-scale concentrations exceeding
2
tion to follow Area: 450 ft , depth 30 ft the MCL for several years.
Within 180 days of injec-
ORC and HRC tested in two tion DCE and VC concen-
identical side by side field trations in one well 25 feet
applications downgradient from the
injection area were re-
Two pilot test arrays consist- duced by approximately
ing of 4 injection wells and 3 97% and 94% respectively.
monitoring wells per array
Springdale Dryc- Dec. 1999 PCE, TCE DNAPL and source area PCE: <250 µg/L Within 18 months after the
leaners remediation TCE: 300 µg/L pilot scale injection PCE
2-3 years PCE: 98 mg/L DCE: 43.9 mg/L and TCE concentrations
Portland, OR TCE: 35.9 mg/L Pilot-scale VC: 9.5 mg/L had decreased by 99%
2
DCE: <5 µg/L Area: 45000 ft , depth: 3 ft while DCE and VC concen-
VC: <5 µg/L trations had increased
A residual source area was significantly in one well.
treated with 5 injection points
centrations
(80 lb HRC per point) and the
downgradient dissolved
phase plume area was
treated with 22 injection
points (120 lb HRC per point).
Reichhold Chemi- Chlorinated Approximately Groundwater remediation PCP: 540 µg/L Within 280 days after
cals Phenols 400 feet of fine TetraCP: 100 injection PCP and
grained marine Pilot-scale µg/L 2,3,4,6-TCP concentra-
Tacoma, WA PCP: 1.6 mg/L sediments TriCP: 50 µg/L tions had been reduced by
2
TetraCP: 200 Area: 4300 ft , depth: 5 ft DiCP: 50 µg/L approximately 66% and
µg/L 50% respectively at one
TriCP: 50 µg/L A single line of 43 injection monitoring well installed
DiCP: 50 µg/L points were installed at 10 approximately 10 down-
foot intervals. Two monitoring gradient from the line of
wells (10 feet upgradient and injection wells.
10 feet downgradient) were
used to monitor performance.
Acton Mickelson Dec. 2000 Perchlorate, Fine to medium Groundwater remediation of Perchlorate: 1 Within 80 days after injec-
Ordinance Hexavalent silty sand perchlorate mg/L tion perchlorate, hexava-
Facility Chromium, CrVI: <10 µg/L lent chromium, and freon-
Freon 113 Full-scale Freon 113: 20 1113 concentrations had
2
Hollister, CA Area: 1200 ft . depth: 4 ft µg/L been reduced by approxi-
Perchlorate: 7.5 mately 87%, nearly 100%,
mg/L 25 injection points installed in and 92% respectively at
CrVI: 160 µg/L a grid on 5 foot centers. 6 one well installed within the
Freon 113: 250 monitoring wells installed for injection grid. No analytical
µg/L process monitoring. data was provided for wells
installed downgradient
from the injection area.
Pueblo Chemical Feb. 2000 2,4,6-TNT Sand and Groundwater remediation, 2,4-DNT - Within approximately 105
Depot - 1,3,5-TNB clayey sand plume containment 0.0003 mg/l days following injection
SWMU-14 2,4-DNT overlain by a RDX - < 0.0001 2,4-DNT, RDX, and 1,3,5-
RDX clay to silty Pilot-scale mg/L TNB concentrations had
2
Pueblo, CO sand confining Area: 350 ft , depth 1,3,5-TNB - decreased by approxi-
2,4-DNT - or semiconfin- 0.005 mg/L mately 89%, 99%, and
0.0037 mg/L ing unit. HRC was injected into 30 Hydrazine – NA 98% respectively. These
RDX - 0.0093 locations installed in a line. A reduction calculations
mg/L total of 15 represent average concen-
1,3,5-TNB - monitoring wells were in- trations calculated from 4
0.231 mg/L stalled for process monitoring downgradient monitoring
Hydrazine - sampling. wells. Concentration data
0.001 mg/L was provided for only the 4
wells closest to the injec-
tion area (approximately 30
centrations
feet downgradient) out of a
total of 15 monitoring wells
used for process monitor-
ing.
Pueblo Chemical Feb. 2001 TCE Alluvial aquifer Groundwater restoration NA Contaminanat data was
Depot - minimal during first 3
SWMU-14 NA Pilot-scale months of pilot test. How-
ever, and order of magni-
Pueblo, CO Barrier configuration tude reduction in TCE
concentration was ob-
served 50 feet downgradi-
ent of treatment zone after
11 months.
Manufacturing Dec. 1999 TCE Fractured Feasibility study for a bedrock TCE: <5 µg/L Within 13 months of injec-
Facility crystalline site DCE: 8 µg/L tion TCE and DCE concen-
TCE: 225 µg/L bedrock VC: 50 µg/L trations had decreased by
Crozet, VA DCE: 125 µg/L Pilot-scale approximately 98% and
2
VC: <5 µg/L Area: 4900 ft , depth: 15 ft 94% respectively while VC
and ethene concentrations
3 injection wells spaced had increased by approxi-
approximately 5 feet apart mately 900% in one moni-
with one monitoring well toring well installed ap-
installed approximately 10 proximately 10 feet down-
feet downgradient from the gradient from the injection
injection wells. wells.
Manufacturing TCE Feasibility study NA Contaminant concentration
Facility NA data was not provided.
Pilot-scale
2
NJ Area: 4900 ft , depth: 15 ft
15 direct push points installed

in a barrier configuration
approximately 5 feet apart.
Flemington Manu- March 2001 PCE, TCE, DCE Very dense till Feasibility study PCE: 4 µg/L Within 80 days of injection
facturing consisting of TCE: 9 µg/L PCE, TCE, and DCE
Facility PCE: 110 µg/L clayey silts and Pilot-scale DCE: 22 µg/L concentrations had been
TCE: 100 µg/L some sand VC: NA reduced by approximately
Flemington, NJ DCE: 50 µg/L Three 3.5-inch by 4-foot long 64%, 10%, and 56% re-
VC: NA PVC canisters filled with spectively in one monitor-
gelled HRC were installed in ing well installed an un-
a single 4-inch well. An unknown distance downgra-
known number of monitoring dient from the emplace-
wells were installed for proc- ment area.
ess monitoring. This application involved
centrations
installing canisters of
gelled HRC in a well as a
slowly soluble and there-
fore longer term HRC
source. This site is
very similar to the Central
US Manufacturing Facility
application.
Japanese Electri- 2000 PCE, TCE, DCE Groundwater remediation NA HRC accelerated the
cal Plant natural attenuation proc-
1 year NA Pilot-scale ess.
2
Japan Area: 500 ft , depth: 7 ft
7 injection points in two rows

spaced approximately 13 feet
apart. Row spacing was also
approximately 13 feet.
Two monitoring wells were
installed, 1 upgradient and 1
downgradient.
Coopervision July 2001 1,1,1-TCA Groundwater remediation NA Efforts to inject using direct
Manufacturing push were unsuccessful.
Facility NA Pilot-scale Re-injection using mud
rotary is planned
Scottsville, NY
Invensys Control Pilot - Dec. PCE Groundwater restoration to PCE: 10 µg/L Within 9 months after the
1999 facilitate property transfer pilot scale injection PCE
Old Saybrook Full – Sept. PCE: 13.9 mg/L concentrations had de-
CT 2000 Pilot-, Full-scale creased by 99% in one
well installed an unknown
Full - 55 injection points in an distance from the injection
unknown configuration, un- area. No other data has
known number of monitoring been provided for this site.
points. Electron donor injection
was a viable remedial
approach for chlorinated
VOCs in this low perme-
ability application. 80% of
PCE mass was removed.
Manufacturing TCE Groundwater Restoration NA Conditional Closure has
Facility been granted.
NA Pilot-scale
2
Brighton, NY Area: 560 ft
centrations
24 injection points installed in

a grid with 5 foot spacing. 4
monitoring wells installed for
process monitoring.
Arlington Cleaners May 2000 PCE, TCE, DCE, Groundwater remediation PCE: 410 µg/L Within 18 months after the
VC under voluntary cleanup TCE: 90 µg/L pilot scale injection PCE,
Arlington, TX program DCE: 440 µg/L TCE, DCE, and VC con-
PCE: 4.5 mg/L VC: 130 µg/L centrations had decreased
TCE: 1 mg/L Full-scale by 98%, 91%, 94%, and
2
DCE: 7.3 mg/L Area; 3000 ft , depth: 15 ft 85% respectively at one
VC: 870 µg/L well installed within or
45 direct injection points. 16 downgradient from the
of the points were installed at injection area.
a 15 to 30 degree angle to During this application 1/3
inject beneath a preexisting of the injection wells were
structure. An unknown num- installed at a 15 to 30
ber of monitoring wells were degree angle to inject HRC
installed for process monitor- under an existing building.
ing. Closure under the TNRCC
VCP Program.
Slow-Release Substrates – Vegetable Oil
Former Zipper April 2000 PCE, TCE, DCE Risk-based closure PCE: 34 mg/L
Manufacturing TCE: 11 mg/L
Facility PCE: 120 mg/L Pilot-scale DCE: 27 mg//L
TCE: 8 mg/L VC: 5.3 mg/L
Newport News, DCE: 14 mg/L
VA VC: 1.3 mg/L
Hangar K, Cape Pilot – June PCE, TCE, DCE Sand and silty Technology demonstration PCE: 48 µg/L
Canaveral AFS 1999 sand with TCE: 310 mg/L
Full – July PCE: 1.8 mg/L Pilot-, Full-scale DCE: 190 mg/L
Cape Canaveral, 2000 TCE: 260 mg/L VC: 13 mg/L
FL DCE: 78 mg/L
VC: 950 µg/L
Site N-6, Former Aug. 2000 TCE, DCE, CT Fluvial deposits Field feasibility study TCE: 1.2 mg/L
NSA Mid-South from 40 to 90 DCE: 250 µg/L
TCE: 1.8 mg/L feet bgs con- Pilot-scale VC: ND
Millington, TN DCE: 34 µg/L sisting of sand,
VC: ND silty sand, and
clay stringers
Site SS-015, Pilot – April PCE, TCE Silty clay with Technology demonstration PCE: 430 µg/L
Travis AFB 1999 thin layers of TCE: 2.2 mg/L
Full – Dec. PCE: 1 mg/L silt and silty Pilot-, Full-scale DCE: 4 mg/L
centrations
Travis AFB, CA 2000 TCE: 4.2 mg/L sand VC: 550 µg/L
DCE: 13 mg/L
VC: 17 mg/L
Site FF-87, For- Sept. 2001 PCE Alluvial me- Accelerate groundwater PCE: 740 µg/L
mer Newark AFB dium to coarse restoration TCE: 8 µh/L
PCE: 340 µg/L sand underlain DCE: 36 µg/L
Newark, OH TCE: 13 µg/L by Pleistocene Full-scale VC: 20 µg/L
DCE: 46 µg/L glacial silty
VC: ND clay
Naval Industrial Nov. 2001 PCE, TCE, VC Glacial-fluvial Field feasibility study TCE: 6.2 mg/L At approximately 5 months
Reserve Ord- medium to DCE: 1.8 mg/L after injection
nance Plant, TCE: 6.7 mg/L coarse Pilot-scale VC: 2 µg/L TCE concentrations were
Fridley DCE: 220 µg/L grained sand approximately the same as
VC: ND baseline conditions.
Fridley, MN However, cis-1,2-DCE and
ethane concentrations
detected at 5 months post
injection were substantially
higher than the baseline
event.
Former Radiator Sept. 2000 TCE Clayey sand Field feasibility study TCE:< 1mg/L
Facility DCE: 41 mg/L
TCE: 1.3 mg/L Pilot-scale VC: 8.3 mg/L
IL DCE: 7.4 mg/L
VC: 220 µg/L
Site SS-17 Nov. 2001 TCE, DCE Reddish brown, Field feasibility study TCE: 650 µg/L Within approximately 5
moderately DCE: 1.07 mg/L months the TCE maximum
Altus AFB, OK TCE: 1.66 mg/L plastic, Bench-, Pilot-scale VC: 790 µg/L concentration had been
DCE: 900 µg/L sandy clay to reduced by approximately
VC: 440 µg/L 15 feet below 61%. DCE,
ground VC, and ethene maximum
surface under- concentrations increased
lain by clayey by approximately 19%,
shale with 80%, and 1,470% respec-
occasional tively. TCE concentration
gypsum layers. reductions were observed
in all of the injection wells
and monitoring wells with
per well reductions
ranging from 13 to 99%.
OU-1 Landfill 3 Nov. 2001 TCE, DCE Reddish brown, Field feasibility study TCE: 6.44 mg/L Within approximately 5
moderately DCE: 800 µg/L months the TCE maximum
Altus AFB, OK TCE: 10.4 mg/L plastic, sandy Pilot-scale VC: 240 µg/L concentration had been
DCE: 500 µg/L clay to 15 feet reduced by approximately
centrations
VC:< 120 µg/L below ground 37%. DCE, VC, and
surface under- ethene maximum concen-
lain by clayey trations increased by
shale with approximately 60%, 80%
occasional and an order of magnitude,
gypsum layers. respectively.
TCE concentration reduc-
tions were observed in all
wells except injection well
6, with per well reductions
ranging from 23 to 92%.
Whiteman Air July 2002 TCE, DCE, VC Approximately Field feasibility study NA The first process monitor-
Force Base LF-08 24 feet of ing round is scheduled for
2 years TCE: 2.1 mg/L dense clay Pilot-scale October 2002.
Whiteman AFB, DCE: 33 µg/L overlying a
MO VC: ND clayey gravel
zone which
extends to the
top of weath-
ered
claystone
bedrock at 30
feet below
ground surface.
Solid Substrates – Bark Mulch
Building 301, Pilot – Jan. TCE Stiff to very Field feasibility study to ac- TCE: 1.22 mg/L 61 percent reduction in
Offutt AFB 1999 stiff, reddish celerate groundwater restora- DCE: 98 µ/L mean TCE concentration
Full – July TCE: 1.9 mg/L brown, low tion VC: 4 µg/L downgradient of biowall
Omaha, NE 2001 DCE: 270 µg/L plasticity, silty
VC: 2 µg/L clay Pilot-, Full-scale
OU1 Landfill 3 July 2000 TCE, DCE Reddish brown Full-scale application to Biowall well MP- Concentration data after 4
Upgradient well silty clay prevent surface water dis- 01 weeks of installation.
Altus AFB, OK TCE: 6.2 mg/L charge and prevent potential TCE: 48 µg/L Limited evidence of degra-
DCE: 850 µg/L off-base migration DCE: 640 µg/L dation of TCE to DCE.
VC: ND VC: ND
Full-scale
Naval Weapons 2000 Perchlorate Groundwater restoration for Full-scale System Installed
Industrial Reserve offsite migration via a perme-
Plant able reactive
biobarrier
McGregor, Texas
Bench-, Pilot-, Full-scale
Solid Substrates – Chitin
Distler Brickyard Oct. 2001 TCE, cDCE, VC Relatively low Groundwater remediation to TCE: ND Intermittent wetting of
centrations
Superfund Site permeability potable water standards DCE: 58 µ/L contaminant zone by
TCE: 7 µg/L silts and clays VC: 9 µ/L infiltration
Louisville, KY DCE: 90 µ/L Pilot-scale
VC: 16 µg/L
Gaseous Hydrogen Injection
Launch Complex Feb. 1999 PCE, TCE Silica sand with Technology demonstration TCE: 66 mg/L 50% ( at 15 feet from
15 some shell DCE: 270 mg/L injection point ) to 90%
18 months PCE: 87 mg/L fragments Pilot-scale VC: 67 mg/L (at 3 to 6 feet from injection
Cape Canaveral, DCE: 370 mg/L containing clay point) reduction in total
FL VC: 52 mg/L and organic chlorinated ethenes in
matter to 70 treatment zone over 18
feet bgs months.
Offutt AFB Nov. 1998 DCE Sand Technology demonstration DCE: < 5 µg/L Small scale pilot test,
complete cis-1,2-DCE
Omaha, NE DCE: 430 µg/l Pilot-scale (over 99%) removal.
Twin Cities Army Aug. 2000 TCE Technology demonstration for NA A decline in concentrations
Ammunition use of hollow fiber mem- of TCE and DCE was
Plant 20 months NA branes to deliver hydrogen in observed, with an increase
situ in VC and ethene after a
New Brighton, MN lag period.
Bench-, Pilot-scale

D6-2 Bio 03-Hirfan10

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

D6-2 Bio 03-Hirfan10

Uploaded by

Copyright:

Available Formats

Project no.

Instrument: Coordination Action

Thematic Priority: Global change and ecosystems

D6-2 Status report on technological reliability for demonstrated soil and

Due date of deliverable: November 2005

Start date of project: 01.01.2005 Duration: 3 years

Organisation name of lead contractor for this deliverable: Universität Lüneburg

1 Enhanced In Situ Bioremediation Technologies (EISB)

1.2 Enhanced In Situ Anaerobic Bioremediation of chlorinated sol-

• Nitrate and sulphate

1.2.1 Degradation Mechanisms

Table 1: Potential Degradation Prozesses for CAHs (AFCEE, 2004).

Figure 1: Reducing zone established downgradient of substrate Injection (AFCEE, 2004).

Under anaerobic conditions, reductive dechlorination mechanisms can effectively bio-

1.3 APPLICATION OF ENHANCED ANAEROBIC BIOREMEDIATION

Application of enhanced anaerobic bioremediation starts with a review of site-specific

1.3.1 Technology Screening

Hydrogeologic Settings. Enhanced anaerobic bioremediation has been applied in a

with indications of residual or sorbed DNAPL (dissolved CAH concentrations in excess

Geochemical Conditions. During anaerobic dechlorination, CAHs function as electron

Table 2: Substrates used for enhanced anaerobic bioremediation (AFCEE, 2004).

1.3.3 System configuration

gies and enhanced bioremediation may be that enhanced bioremediation, if properly

Source Zone Treatment: Remediation of source zones where good sub-

•Plume Containment using a Biologically Reactive Barrier: Reduction of mass flux

•Plume-Wide Restoration: Total treatment of an entire dissolved plume.

Figure 2: Schematic of Source Area and Biobarrier Injection Configurations (AFCEE,

The appropriate application of enhanced anaerobic bioremediation will be site-specific

Source Zone Treatment

Plume Containment using Biologically Enhanced Barrier Systems

1.3.4 Delivery Options

Figure 3: Schematic of a horizontal recirculation system (AFCEE, 2004).

1.4 Case Studies

Dry Cleaning Facility, Arlington, Texas

Table 3: Cleanup criteria

INEEL, Test Area North, Idaho Falls, Idaho

Figure 4: Medial zone of TCE plume at TAN (TCE-concentrations in µg/L).

NASA’s Launch Complex 34 (LC34) at Cape Canaveral

biostimulation and bioaugmentation technology when applied to dense, nonaque-

Figure 6: Location map of Launch Complex 34 site (Battelle, 2004).

Figure 7: KB-1™ Dechlorinator Culture Containers (Battelle, 2004).

Post-demonstration soil and groundwater characterization was done in June 2003.

Changes in Total TCE and DNAPL Mass

Changes in Aquifer Quality

Fate of TCE/DNAPL in the Aquifer

Operating Requirements and Cost

1.5 Status and Applicability

Enhanced in situ anaerobic bioremediation has emerged in recent years as a remedia-

Enhanced anaerobic bioremediation takes advantage of natural processes that may

1. Sites where appropriate dechlorinating microorganisms are present, geochemical

Hydraulic Conductivity. Hydraulic conductivity is a primary factor in effective distribu-

cm/sec). Alternate injection techniques such as hydraulic fracturing may be used in

Dissolved Oxygen and Oxidation-Reduction Potential. Background levels of DO and

aerobic dechlorination of cis-DCE to VC may potentially be inhibited in the presence of

Soluble Substrate Applications

Advantages of soluble substrate systems include the following:

Disadvantages or limitations of soluble substrate systems include the following:

Slow –release viscous fluid substrate systems

Vegetable Oil Applications

uniform distribution of the substrate may be complicated by low permeability soils or by

Advantages of using slow-release viscous fluids include the following:

Disadvantages of using slow-release viscous fluids include the following:

Bioaugmentation can be used to reduce acclimation periods and/or to provide