United States
                    Environmental Protection
                    Agency
Robert S. Kerr Environmental
Research Laboratory
Ada OK 74820
                    Research and Development
EPA/600/S2-87/008 Apr. 1987
SERA         Project Summary
                    Leaking  Underground  Storage
                    Tanks:  Remediation  with
                    Emphasis on In  Situ
                    Biorestoration
                    J. M. Thomas, M. D. Lee, P. B. Bedient, R. C. Borden, L. W. Canter, and
                    C. H. Ward
                      The current literature indicates that
                    in situ biorestoration has great potential
                    for remediation of aquifers contami-
                    nated by leaking underground storage
                    tanks. In situ aquifer restoration involves
                    natural  microbial action and the en-
                    hancement of the indigenous microf lora
                    to more efficiently degrade subsurface
                    pollutants.  Aquifers amenable to en-
                    hanced biorestoration are usually those
                    that are perfusable with solutions that
                    carry the nutrients to the zone of con-
                    tamination. The presence of naturally
                    occurring microorganisms that can
                    degrade subsurface contaminants has
                    been demonstrated. However, a period
                    of adaptation is usually required before
                    the subsurface microflora can degrade
                    pollutants.  Factors that may limit de-
                    gradation,  even  in the presence of
                    adapted organisms, include lack of an
                    essential nutrient, substrate concentra-
                    tion, substrate inaccessibility, pH, tem-
                    perature, and the presence of toxicants.
                    Biorestoration of contaminated aquifers
                    often involves the addition of limiting
                    nutrients such as oxygen, nitrogen, and
                    phosphorus. Enriching for microor-
                    ganisms with special  metabolic  cap-
                    abilities has been demonstrated in the
                    laboratory and may be applicable to
                    some in situ biorestoration schemes.
                    Mathematical models of biorestoration
                    have  been developed to simulate pro-
                    gress of the cleanup and provide in-
                    formation on the kinetics of the process.
                    The full report emphasizes the state-of-
                    the-knowledge of In situ biorestoration
                    techniques available for remediation of
                    contaminated ground water.
  This Project Summary was developed
by EPA's Robert S. Kerr Environmental
Research Laboratory, Ada, OK, to an-
nounce key findings of the  research
project that is fully documented In a
separate report of the same title (see
Project Report ordering Information at
back).

Introduction
  One ground-water pollution source
category which has recently received in-
creased attention is underground storage
tanks, specifically, the inadvertent leakage
of liquid products from such tanks into
the unsaturated and saturated zones. This
document was requested by  the U.S.
Environmental  Protection Agency (EPA)
Office  of Underground  Storage  Tanks
(OUST) to aid in planning a program for
restoring ground water contaminated by
leaking underground storage tanks and is
intended to guide OUST in making judge-
ments regarding the efficacy of alternative
technologies for remedial action The full
report specifically focuses on the use of
innovative  biological technologies to
remove or attenuate contaminants.
  Remedial action technologies for leak-
ing underground  storage tanks can be
considered  in  relation  to preventive
measures and contaminant plume man-
agement Preventive measures include
corrosion prevention, proper installation
practices, material selection, and con-
tainment systems. Plume management
techniques may include one or several of
the following:
  (1) ground water pumping to extract
    water from or inject water into wells

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     to capture a  plume or alter  the
     direction  of ground-water move-
     ment;
  (2) subsurface  drains consisting  of
     permeable barriers designed to in-
     tercept ground-water systems;
  (3) vertical underground barriers made
     of low-permeability materials  to
     divert ground-water flow or mini-
     mize leachate generation and plume
     movement; and
  (4) innovative technologies that bio-
     logically  or chemically remove or
     attenuate contaminants  in the
     subsurface.
  The most recent and innovative tech-
niques for  aquifer  remediation  involve
stimulating the indigenous microflora to
degrade subsurface pollutants. Stimula-
tion of the native  microorganisms can
result in the complete destruction of the
contaminants  whereas chemical  or
physical treatment  may result in incom-
plete destruction or transfer of the con-
taminant  to  another  phase  of the
environment.  Biostimulation  is  often
achieved by adding limiting nutrients such
as oxygen, nitrogen, and phosphorus. The
addition of specialized microbial popula-
tions to degrade subsurface  pollutants
has been incorporated into many remedi-
ation projects; however, the role of these
microbes in biodegradation has not been
distinguished from that of the indigenous
microflora.


Results and Discussion

Remediation/Restoration Plume
Management Techniques
  This full report  initially presents  a
cursory review of conventional  contain-
ment and treatment techniques  Physical
containment is reviewed relative to re-
moval and to ground-water flow barriers
such as slurry walls, grout curtains, sheet
piling,  block  displacement and liners.
Surface water  control techniques such
as caps, dikes, terraces, channels, chutes,
downpipes, grading, vegetation,  seepage
basins, and ditches are also discussed  A
simple  review of  passive and active
hydrodynamic controls used to manipulate
the hydraulic gradient of ground-water
systems is also presented  Withdrawal
and treatment techniques are reviewed
in terms  of  conventional  wastewater
treatment processes.

In Situ Biological Treatment
  The major emphasis of the full report
concerns  in  situ  biological  treatment
which  is discussed in detail. Microbial
processes may be used to degrade con-
taminants in situ by stimulating the native
microbial  populations.  Another in situ
biostimulation technique which is not yet
demonstrated is the inoculation of the
subsurface with a microbial population
that has specialized metabolic capabilities.
Even in the presence of an indigenous
population which is acclimated to organic
contaminants, degradation may be limited
at high contaminant concentrations or by
some environmental factor. Addition of
electron acceptors, such as oxygen, and
inorganic  nutrients, typically  nitrogen,
phosphorus, and trace metals,  may pro-
vide the  microflora  with essential
nutrients that are limiting in the presence
of high concentrations  of  pollutants.
Inoculation of a  specialized microbial
population may theoretically reduce the
time required for acclimation of the con-
taminants and/or allow the removal of
recalcitrant contaminants. Related pro-
cesses such as the addition of  bioemul-
sifiers  or  surfactants to  increase  the
availability of subsurface contaminants
to the microflora can also be used. When
applicable, biological processes may offer
the advantage  of partial or complete
destruction of the contaminants rather
than simply transferring the pollution to
another phase of the environment.
  The  full  report goes  into significant
detail  concerning microbial activity in
aquifers. In  summary,  the  subsurface
environment contains  microbes  that
degrade many of the organic compounds
that contaminate ground water. The sub-
surface microorganisms  in uncontami-
nated aquifers are likely to be adapted to
use  low  concentrations of organic
materials. The  majority  of  the  micro-
organisms are associated with soil par-
ticles  Even  in the presence of adapted
populations, environmental factors such
as temperature, pH,  dissolved oxygen
levels,  inorganic nutrient concentrations,
and the availability and concentration of
the organic contaminants may limit bio-
degradation of subsurface pollutants.
  A  number  of advantages  and disad-
vantages exist in using in situ biorestora-
tion.  Compounds ranging from petroleum
hydrocarbons to solvents have  been
treated by in situ biorestoration.  Unlike
many aquifer remediation techniques, in
situ bioreclamation can often treat con-
taminants that are  sorbed to soil  or
trapped in pore  spaces.  In  addition to
treatment of the saturated zone, organics
held in the  unsaturated  and  capillary
zone can be treated  when an infiltration
gallery or soil flushing  is used.  Biode-
gradation  in the subsurface can be en
hanced by increasing the concentratior
of dissolved oxygen, through the use o
hydrogen  peroxide,  ozone, or a  colloida
dispersion of air (colloidal gas aphrons)
Complete  biodegradation (mineralization
of organic compounds usually produce:
carbon dioxide, water, and an increase ir
cell mass.  However, incomplete degrada
tion (biotransformation) of organic mate
rials can sometimes produce by-products
that are  more  toxic than  the pareni
molecule.  In situ biorestoration relies or
a microflora that contains few pathogenic
organisms. The  time required  to  treal
subsurface pollution using in  situ bio-
restoration can often be faster than some
withdrawal and treatment  procedures,
since with current technology not all con-
tamination can be physically withdrawn.
In situ biorestoration can also cost less
than other remedial options. The areal
zone of treatment  using biorestoration
can be larger than other remedial tech-
nologies because the treatment moves
with the plume and  can reach areas that
are otherwise inaccessible.
  Advantages  as well as disadvantages
of biorestoration are described through-
out the full report. Major advantages and
disadvantages discussed include:

Advantages -
  (1) can  be used to treat hydrocarbons
     and  certain organic  compounds,
     especially water-soluble pollutants
     and  low levels of other compounds
     that  would be difficult to remove by
     other methods;
  (2) environmentally sound because  it
     does not usually generate waste
     products and  typically results in
     complete  degradation  of the
     contaminants;
  (3) utilizes  the indigenous microbial
     flora and does not introduce po..~..
     tially harmful organisms;
  (4) fast, safe and generally economical;
  (5) treatment moves with the ground
     water; and
  (6) good for short-term  treatment of
     organic contaminated ground water.


Disadvantages -
  (1) can  be  inhibited by heavy metalj
     and  some organics;
  (2) bacteria can plug the soil and reduce
     circulation;
  (3) introduction  of  nutrients  couk
     adversely  s4'1""* -o^'b-.1  surfac«
     waters;
  (4) residues may  cause taste and odo
     problems;

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  (5) labor  and maintenance  require-
      ments may be high, especially for
      long-term treatment;
  (6) long-term effects are unknown; and
  (7) may not work for aquifers with low
      permeabilities that do not permit
      adequate circulation of nutrients.
  No technique to remediate environ-
mental contamination  is  universally
applicable.  However,  there should  be
many incidents of  contamination where
biorestoration through a forced co-oxida-
tion is the technology  of choice, either
alone  or  in  conjunction with  physical
containment. Successful application  of
the approach will  require an adequate
understanding of the physiology of the
co-oxidation  biotransformation,  and
quantitative  information  on the  nutri-
tional ecology of the active organisms.

Hydrologlc Considerations and
Mathematical Modeling of
Biorestoration
  The full report considers a number of
methods which  have  been  reported  in
the  literature for  containment  of  con-
taminated ground water through hydrau-
lic control or through injection-pumping
networks  of wells. Biorestoration  of  a
contaminant plume may involve the addi-
tion of nutrients such as dissolved oxygen
or hydrogen peroxide or the addition of
microbes capable of degrading a particular
waste. In order for such additions to  be
successful,  it may be necessary to use
hydraulic controls to minimize the migra-
tion of the plume during the in situ
treatment process.  Thus, hydrologic con-
siderations cannot be  neglected in the
biorestoration of aquifers. Hydraulic con-
trols for the containment of ground water
should  be carefully considered  for any
site where  biorestoration  is a viable
treatment alternative. In particular, in-
jection-pumping well networks offer ad-
vantages for the creation of stagnation
(no-flow) zones or  for the control of the
trajectory of a contaminant plume. Once
the plume has  been controlled  hydrau-
lically,  then application of additional
nutrients,  oxygen,  or microbes can be
better controlled and evaluated in terms
of biodegradation efficiency.
  Mathematical  modeling of biorestora-
tion processes  is  useful in simulating
cleanup progress and can provide insights
into the kinetics of the restoration  process.
Modeling of the hydraulics  of  the site
may also aid in  designing the optimum
injection and production  system. Develop-
ment of mathematical models of the bio-
restoration  process  requires:  1)   a
description of the kinetics of biodegrada-
tion/transformation in the subsurface; 2)
a  description  of  the  abiotic  processes
controlling the transport and availability
of the contaminant and other required
nutrients; and 3) an appropriate procedure
for combining the  processes and pre-
dicting the effect of the biorestoration
technique. Most attempts at quantifying
the transport  and removal of contami-
nants in  ground water have relied on  a
solution  of the  classical form  of the
advection/dispersion equation. This ap-
proach is inappropriate for many applica-
tions.  Biorestoration  models and the
kinetics of biodegradation are considered
in some  detail in the full report. Many
references  and  examples  of  various
aspects  of biorestoration  modeling are
given.
  The current technology for simulating
subsurface biorestoration is  still in its
infancy.  Some progress has been made
in developing kinetic descriptions of the
biodegradation process and combining
these  with available  solute transport
models. Unfortunately, little reliable field
data has  been available to rigorously test
these models. Considerable uncertainty
exists over the importance of simulating
transport into  biofilms or microcolonies.
Also, the effects of variations in aquifer
parameters on  the efficiency of bio-
restoration is unknown. At present, the
technology is  not available to quantita-
tively predict the  efficiency of enhanced
biorestoration but significant advances
are being made in our ability to describe
the process.

Institutional Limitations on
Ground-Water Pollution Control
  The  development and application of
technologies  for  the  prevention and
regulation of leaking underground storage
tanks is a complex and interdisciplinary
science, as is the abatement and cleanup
of the ground-water contamination such
leaks produce.' This science  is  further
complicated, however, by the institutional
limitations that control the rate at which
technology can advance  and  its use in
the development  of regulations. Institu-
tional limitations may be the factor in
determining how the technology will be
applied to leaking underground storage
tank regulations and remediation. Institu-
tional limitations addressed in  the full
report  include: (1) the scientific under-
standing  of the nature of leaking under-
ground  storage  tanks  and released
products; (2) public opinion; (3) business
community attitudes; (4) environmental
 interest group concerns; and (5) govern-
 mental acceptance, use, and implementa-
 tion of the remedial technology.


 Conclusions
  Of  the available  biological aquifer
 remediation techniques, the most effec-
 tive methods  are  enhancement of the
 native population  and withdrawal  and
 treatment by various wastewater treat-
 ment processes.  Before any aquifer
 remediation technique  can  be  imple-
 mented, a thorough understanding of the
 hydrogeology  and  contamination prob-
 lems  of the site must be obtained and
 used  to  design the  treatment system.
 Costs for treatment may range from tens
 of  thousands  up  to  tens of millions,
 depending upon the extent and nature of
 the contaminants, the nature of the site,
 and the desired cleanup levels.
  Addition  of  oxygen, nitrogen,  phos-
 phorus, and trace minerals  stimulates
 the  acclimated indigenous   microbial
 population  to  aerobically degrade the
 contaminants. In situ biorestoration has
 been  chiefly used to treat gasoline con-
 taminated aquifers, but also has been
 employed with ethylene glycol and sol-
 vents including acetone, tetrahydrofuran,
 methylene chloride, n-butanol, dimethyl
 aniline, and isopropanol. Biorestoration
 effectiveness will  be  affected by  toxic
 levels of organics and heavy metals. In
 general, in situ bioreclamation has been
 effective in reducing the quantity of the
 contaminants,  but not  in  completely
 eliminating them. The treatment moves
 with the plume allowing treatment of
 trapped or sorbed contaminants or, by
 using soil flushing or an infiltration gal-
 lery, microbial treatment can reach areas
 that are  not accessible by other tech-
 niques. Biorestoration has been used in a
 number of aquifers, but may be of limited
 usefulness in those with low permeabili-
 ties. Undesirable metabolic and inorganic
 nutrients may escape from the treatment
 zone and affect ground water or surface
 water quality. Alternative oxygen sources
 such as ozone, hydrogen peroxide, pure
 oxygen, and air flooding or soil venting
 may speed the removal of the organic
 contaminants, but their impact  on the
 microbial population and the geochemis-
 try  of the site  is not  fully understood.
 Innovative processes such as treatment
 beds or land treatment can be used in
 some situations. In the presence of an
acclimated  microbial population,  many
aquifers will be anaerobic because the
 microorganisms will have depleted the
dissolved oxygen. It will be possible to

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  use anaerobic degradation to remove
  contaminants, although the technology
  for  this treatment  has not yet been
  developed. Reducing the interfacial ten-
  sion between the hydrocarbon and ground
  water with surfactants, dispersants, or
  emulsifiers will mobilize the contaminants
  and may make them available for micro-
  bial degradation. Combinations of in situ
  biorestoration treatment with  other
  chemical, physical, or biological treatment
  processes have been successfully utilized
  in aquifer remediation.
    Treatment by biological wastewater
  processes  is a proven technology. The
  biological  processes  include  activated
  sludge, lagoons, waste stabilization ponds,
  fluidized bed reactors, trickling filters,
  rotating biological discs, and sequencing
  batch reactors. All of these processes are
  dependent upon extraction of the con-
  taminated ground water from the subsur-
  face.  Combinations  of  conventional
  wastewater  treatment processes and
  other  water treatment processes  have
  also been successful.
    Techniques for simulating the subsur-
  face biorestoration  process are  under
  development, but little reliable field data
  has been generated that can be applied
  to these models. Some of the major con-
  siderations in simulating  transport and
  biodegradation  of organic contaminants
  in the subsurface are poorly understood;
  these include the importance of transport
  of organics to the bacteria and the vari-
  ability in aquifer parameters.

  Recommendations
    As  biorestoration  develops as a  re-
  mediation technology, many technical
  research needs can be identified. Exam-
  ples of these needs include:
    - Can microbial degradation processes
      in  the vadose  zone be optimized
          through the controlled addition  of
          nutrients, enzymes, and  bacterial
          seed organisms?
          What are the optimum nutrient con-
          centrations  for achieving in situ
          biorestoration of different classes of
          organic compounds?
          What are the optimum environmental
          conditions (pH,  oxidation-reduction
          potential, micronutrients, etc.) neces-
          sary to achieve in situ biorestoration
          of  different classes  of organic
          compounds?
          Can laboratory  development of ac-
          climated microorganisms enhance in
          situ biorestoration, and what  labor-
          atory tests/procedures are necessary
          to achieve this acclimation?
-  What design approaches andA
  laboratory tests can be used to opt
  mize microbial population densitit
  for in situ biorestoration?
-  What are the  best methods fc
  achieving in situ mixing of the bai
  terial populations,  nutrients an
  micronutrients, and organics?
-  What are the optimum (most cos
  effective) combinations  of in sit
  biorestoration and interdiction wel
  and  surface treatment to achiev
  ground-water remediation?
-  Can we predict clean up efficiencie
  of  biorestoration  processes  usin
  mathematical models that incorporai
  rate coefficients for target pollutan
  that are determined in site specif
  aquifer materials?
         J. M. Thomas, M. D.  Lee. P. B. Bedient, R. C.  Borden.  and C.  H.  Ward are
           with National Center for Ground Water Research, Rice University, Houston,
           TX 77251; and L.  W.  Canter is  with National Center for Ground Water
           Research, University of Oklahoma, Norman. OK 73019.
         Jerry N. Jones and Marion R. Scalf are the EPA Project Officers (see below).
         The complete report, entitled "Leaking Underground Storage Tanks: Remediation
           with Emphasis on In Situ  Biorestoration," (Order No.  PB 87-168 084/AS;
           Cost: $18.95, subject to change) will be available only from:
                 National Technical Information Service
                 5285 Port Royal Road
                 Springfield, VA 22161
                 Telephone: 703-487-4650
         The EPA Project Officers can be contacted at:
                 Robert S. Kerr Environmental Research Laboratory
                 U.S. Environmental Protection Agency
                 Ada. OK 74820
United States
Environmental Protection
Agency
Center for Environmental Research
Information
Cincinnati OH 45268
                                                                                ;    "  /o-f;   j .^-c  j _   ^     ^   1
Official Business
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