United States
           Environmental Protection Agency
Office of Solid Waste and
Emergency Response (5203P)
                                  EPA 542-F-l 0-006
                                  March 2010
Green Remediation Best Management  Practices:
Bioremediation
Office of Superfund Remediation and Technology Innovation
                                      Quick Reference Fact Sheet
  The U.S. Environmental Protection Agency (EPA) Principles for
  Greener Cleanups outlines the Agency's policy for evaluating
  and minimizing the environmental "footprint" of activities
  undertaken when cleaning up a contaminated site.1 Use of
  the best management practices (BMPs) recommended in
  EPA's series of green remediation fact sheets can help project
  managers and other stakeholders apply the  principles on a
  routine basis, while maintaining the cleanup objectives,
  ensuring protectiveness of a remedy, and improving its
  environmental outcome.2
  Overview
Bioremediation actively enhances  the  effects of naturally
occurring  biological processes that degrade contaminants
in  soil, sediment,  and  groundwater.  In  situ  processes
involve    placement   of   amendments   directly   into
contaminated  media  while  ex situ processes transfer the
media for treatment  at  or  near ground  surface. Green
remediation   BMPs   for  bioremediation   address  the
techniques for:
• Biostimulation:    injection   of   amendments    into
  contaminated    media    to   stimulate    contaminant
  biodegradation  by  indigenous  microbial  populations.
  Amendments  may  include  air  (oxygen)  by  way of
  bioventing,  oxygen-releasing  compounds  to keep  an
  aquifer  aerobic, or  reducing agents such as carbon-rich
  vegetable  oil  or  molasses  to  promote  growth  of
  anaerobic microbial populations
• Bioaugmentation:   injection  of  native   or   non-native
  microbes to a  contaminated  area to aid  contaminant
  biodegradation;  successful   bioaugmentation   may
  involve  prior addition of biostimulation  amendments to
  create the conditions favorable for  microbial activity
• Land-based systems: treatment of  contaminated soil or
  sediment through surface  mixing  with amendments or
  placement of soil/sediment in  surface piles or treatment
  cells,  such as composting  or landfarming, and
• Bioreactors:   treatment   of  contaminated   soil   or
  groundwater in a  controlled  environment to optimize
  degradation, such  as  an  in situ  bioreactor  landfill or
  biological permeable reactive barrier (biobarrier) or an
  ex situ batch- or continuous-feed reactor.
  Designing a Bioremediation System
Early   and  integrated   planning   will   help  design  a
bioremediation project involving activities with  a minimal
environmental  footprint.  Effective  design   will  provide
       flexibility  for modified site or engineering  parameters as
       cleanup  progresses  while  continuing to  accommodate
       current or future use  of a site. Options  for reducing the
       footprint   of   bioremediation   implementation  can   be
       affected   by   local,   state,   and   federal   regulatory
       requirements.  Permits for  underground  injections,  for
       example,  vary  considerably   among   state   regulatory
       programs.3 Option evaluation  also examines  the  short-
       and long-term advantages and disadvantages of in  situ
       versus ex situ  bioremediation techniques in  terms of green
       remediation core elements.
                                           Materials
                                           & Waste
      Core Elements of Green Remediation
Reducing total energy use and increasing renewable
energy use
Reducing air pollutants and
greenhouse gas emissions
Reducing water use and negative
impacts on water resources
Improving materials management
and waste reduction efforts, and
Enhancing land management and
ecosystem protection
Energy
       Successful   bioremediation   relies   on   adequate   site
       characterization and development of a good conceptual
       model to assure thorough delineation of the  contaminant
       source  area(s)  and   plumes.  Effective  modeling  will
       typically lower the  potential for unnecessary activities and
       associated   natural  resource   consumption   or  waste
       generation.4"  Techniques   such  as  three-dimensional
       imaging, for example,  can  help optimize placement of
       injection  boreholes. Representative field data are needed
       during   in  situ  bioremediation  design   to  assure:   (1)
       influential factors  such  as aquifer hydraulic  conductivity,
       groundwater geochemistry,  and soil  heterogeneity and
       adsorptive capacity are well  understood,  (2) the radius of
       influence for any  injected  substrates  reaches  the  entire
       target  area  and  spacing   of multiple  injection points
       provides optimal substrate control, and (3) any excavation
       for techniques such as installation of a trenched biobarrier
       are conducted in a surgical manner.4
       Efficiency in  energy and natural resource consumption can
       be achieved through BMPs that optimize initial design of a
       bioremediation  system.  Early bench-scale treatability tests
       on soil collected from the target treatment area will help:
       •  Determine the onsite mass of contaminant parent and
         daughter  products,   other  metabolic  products,  and
         existing microbial populations

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  Demonstrate  specific biodegradation  mechanisms  of
  potential  microbial  cultures,  chemical  substrates,  or
  amendments
  Evaluate  potential  delivery  methods  and  dispersion
  characteristics   under  simulated  aquifer  conditions,
  including   use  of  options  such  as   biodegradable
  surfactants
  Select the  most suitable  reagents  or  amendments and
  optimal concentrations or proportions, and
  Determine any  need for supplemental  technologies  to
  destroy contaminants in hot spots or areas anticipated
  to involve lengthy periods of microbial acclimation.
 Profile: Bioaugmentation at MAG-1 Site,
         FortDix, NJ
 * Began bioaugmentation design through laboratory tests on
   MAG-1 groundwater samples to evaluate efficacy of a
   commercial bacterial culture in degrading targeted
   chlorinated volatile organic compounds (CVOCs) that were
   resistant to degradation by native bacteria
 * Dispersed the microbial inoculant through a groundwater
   recirculation system, which minimized construction of new
   wells and associated resource consumption
 * Optimized the system within six months of the first (of two)
   injections to reduce the initially high volume of buffering
   agents and extensive well fouling, resulting in reduced
   material consumption and equipment maintenance
 * Decreased CVOCs nearly 99% within one year of project
   startup without negative impacts to natural groundwater
   conditions
Natural   resource   efficiencies   also   are   gained  by
conducting an  onsite pilot test that evaluates methods for
delivering  the  selected substrate or  amendment  to a
portion of the  treatment area.  Green  remediation  BMPs
applied  during  a   bioremediation  pilot  test  will   help
optimize full-scale  operations and  may identify adverse
environmental impacts  in the field; for example, improper
addition of nutrients in certain aquatic environments  could
quickly cause algal  blooms.
Use  of innovative  reagents  from non-traditional sources
can  significantly reduce consumption  of virgin  natural
resources while beneficially using various waste products.
For  instance,  enzymes are often   introduced into  the
remedial  process   to  additionally  stimulate  microbial
degradation  of contaminants. These enzymes commonly
exist in agricultural  or  industrial byproducts  that may be
readily available from  local  sources.  One  example is
manure  compost,  which  can  provide   various enzymes
depending  on   the feedstock  and  maturity.  Another
byproduct  gaining   use for bioremediation  purposes is
spent-mushroom compost, which can be supplied at little
or no cost by local producers. Evaluating potential  use of
products  often  considered  to   be  waste  will include
examining  the product's  traditional fate and  demand  in
markets other than site remediation.
Land-based   systems   and   in  situ   bioreactors   can
particularly   benefit  from  use  of  commercial  waste.
"Supermulch"  contains  common  byproducts  such   as
municipal biosolids, wood ash,  and paper sludge that can
be included in recipes for soil amendments or placed in a
permeable   reactive  barrier   to   enhance  activity   of
indigenous microbial populations. This  approach can also
be   integrated  with   phytoremediation  to  encourage
contaminant   degradation   and   volatilization    while
enriching  soil for revegetation  in significantly  disturbed
areas such as mining sites.
Project  designers  can  establish a schedule for periodic
review of the selected bioremediation process  and related
decision points to:
• Determine if any improvements to field operations could
  reduce   natural   resource   consumption   and  waste
  generation while maintaining  bioremediation efficacy
• Identify    any   innovative    materials   that   recently
  demonstrated   success  in   biologically   degrading
  contaminants     while     reducing    the    project's
  environmental footprint
• Identify  unanticipated environmental impacts such  as
  uncontrolled production of secondary byproducts, sub-
  optimal  nutrient  levels,  or  changes in  non-targeted
  indigenous microbial populations, and
• Identify    other    processes    that   could   accelerate
  biodegradation  in certain  areas  without  significantly
  increasing  the  project  footprint;  for example,  some
  injection wells could  be equipped with passive air flow-
  control devices and  renewable energy-powered blowers
  to deliver air to the subsurface after bioaugmentation is
  conducted.
Future optimization  may include introduction  of alternate
amendments  to  remediate  portions  of a  site  showing
marginal biodegradation  progress or alternate methods to
increase efficiency of reagent delivery.
     Integrated planning of bioremediation activities at
     Marine Corps Base Camp Lejeune enabled injections of
     emulsified vegetable oil and sodium lactate in four
     borings to be completed within only one week, which
     reduced field redeployment and associated fuel use.

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  Profile:  Soil Composting at Former JolietArmy
          Ammunition Plant, Will County, IL
  *  Conducted pilot-scale field tests on compost windrows to
    optimize the designed soil amendment recipe, amendment
    timing, loading rate, and turning frequency
  •  Constructed a 20-acre composting facility to treat 280,000
    tons of excavated explosives-contaminated soil with
    amendments such as manure, wood chips, stable bedding,
    and spent biodigestor waste from local producers
  •  Installed a one-million-gallon basin to capture stormwater
    runoff for onsite aquifer infiltration
  •  Began early transfer of uncontaminated acreage to the
    U.S. Forest Service in 1997 to the newly formed Midewin
    National Jallgrass Prairie, with subsequent transfers of
    additional parcels as remediation progressed; by 2002, all
    (19,000) targeted acres were conveyed to the Prairie
  •  Completed soil cleanup in 2008, three years ahead of
    schedule, through implementation of an  integrated cleanup
    and reuse plan for 3,000 acres now under development as
    business parks and an engineer training  center
  Constructing a Bioremediation System
Best management practices initiated during  bioremedi-
ation design can  continue in the construction phase and
during operation  and  maintenance (O&M). A significant
portion of the environmental footprint left by construction
of a bioremediation  system involves the installation and
testing of wells  used  to deliver the selected  reagents and
monitor performance. Recommended practices include:
• Using direct-push technology for constructing temporary
  or permanent wells rather than typical rotary methods,
  wherever feasible,  to eliminate the need for disposal  of
  cuttings  and improve efficiency of substrate delivery into
  discrete vertical  intervals
• Maximizing reuse of existing or new wells and boreholes
  for injections to avoid a range of wasted resources, and
• Using  groundwater   recirculation  processes  allowing
  multiple passes  of groundwater through fewer wells.
Recommended  practices for designing,  constructing, and
operating  wells, such as  those  used for in  situ injection
and  groundwater  recirculation, are provided  in:  Green
Remediation Besf  A/lonogemenf Practices: Pump and Treat
Technologies.4c Additional  practices  for  subsurface  air
delivery   are  provided   in   Green  Remediofion   Besf
A/lonogemenf  Practices:  Soil  Vapor  Extraction  & Air
Sparging.Ad
Project managers  of land-based bioremediation systems
can reduce the project footprint through  BMPs such as:
• Constructing  a  retention   pond  within   a  bermed
  treatment  area  to  store, treat, use, or release diverted
  stormwater
• Reclaiming  clean  or  treated  water  from  other  site
  activities  for  use in  injection  slurries or  as injection
  chase water
• Integrating a  landfarm rain shield  (such  as  a  plastic
  tunnel)  with  rain  barrels  or  a  cistern  to  capture
  precipitation for potential onsite use, and
• Evaluating the need for a  leachate collection  system for
  a landfarm (along with a  leachate treatment  system) to
  fully preserve the  quality  of  downgradient  soil  and
  groundwater.
Land  disturbance   during  bioremediation  construction,
particularly  at sites involving  ex situ techniques, can be
reduced through practices such as:
• Maintaining specific areas for different activities such as
  materials  mixing  or waste sorting, which will also avoid
  cross-contamination
• Covering  ground surfaces of work areas with mulch to
  prevent soil  compaction  caused  by  activities such as
  front-loader application of soil amendments
• Establishing   well-defined  traffic  patterns  for   onsite
  activities,  and
• Employing rumble grates  with a  closed-loop  graywater
  washing system (or an advanced, self-contained wheel-
  washing system)  to  minimize onsite and offsite trackout
  by delivery vehicles.
Emission of  greenhouse gas (GHG) and  particulate matter
from mobile sources can be reduced through BMPs such
as reducing  engine idling, fueling  heavy machinery  with
ultra low-sulfur  diesel fuel, and retrofitting equipment  with
diesel  oxidation   catalysts  or  other   advanced   diesel
technology.   More  practices  are  outlined  in  Green
Remediation Best Management Practices:  Clean  Fuel &
Emission Technologies for Site Cleanup.4"1
Contributors to the Bioremediation Footprint at
Romic East Palo Alto

Energy
Potable water
CO2 equivalent
Sulfur oxides
Particulate matter
Air toxics
Total Estimated
Footprint
23,000 million Btu
6,800,000 gallons
5,000,000 pounds
22,000 pounds
800 pounds
200 pounds
Attributed to
O&M

1 00%




   O&M activities account for much of the environmental
   footprint of bioremediation recently initiated at the Romic
   RCRA site in East Palo Alto, CA. Site investigation, remedy
   construction, and future decommissioning also contribute
   but to a lesser extent. Although onsite contributors are
   relatively small in comparison to offsite factors such as
   "upstream" materials manufacturing, they may hold
   greater importance to the local community.

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Operating and Monitoring a System
Energy  consumption  and  associated  emissions  during
bioremediation O&M can be reduced by:
• Introducing    biostimulation     or   bioaugmentation
  amendments to  the  subsurface  via  gravity feed  in
  existing   wells,   when    high-pressure    injection   is
  unnecessary  to  assure  proper  distribution   in  certain
  geologic units
• Evaluating   feasibility  of  using   pulsed  rather  than
  continuous injections when  delivering air, to increase
  energy efficiency
• Employing  portable  units  or  trailers   equipped   with
  photovoltaic  panels  to  generate  electricity  or  direct
  power for equipment such as air blowers, and
• Investigating delivery of industrial byproducts  needed  in
  high volumes by way of rail rather than trucks.
Environmentally  preferable  purchasing  in  the  context  of
bioremediation includes products such as:
• Tarps  with recycled  or  biobased  contents  instead  of
  virgin  petroleum-based  contents,  for  protection   of
  ground surfaces in  staging areas and coverage of soil
  undergoing ex situ treatment
• Soil  nutrients and  other treatment-related  materials
  available in bulk quantities  and packed  in  recyclable
  containers and drums, to reduce packaging waste
• Treatment liquids in concentrated form  if  a  product  is
  locally unavailable  (and the concentration  process does
  not involve additional energy consumption),  to  reduce
  long-distance shipping volumes  and frequencies,  and
• Biodegradable cleaning products effective  in cold water
  applications,  to   conserve   energy  while   avoiding
  introduction of toxic chemicals in environmental media.
     Composting of mining waste-contaminated soil and
     sediment with municipal hiosolids and lime along the
     Upper Arkansas River in Colorado resulted in 100%
     vegetative cover in most previously denuded areas
     within ten years, due to increased microbial functions
     combined with phytoremediation and reduced leachate.
Green  remediation  relies  on  continually  improving  a
project's  natural  resource  efficiencies  and  scouting  for
novel approaches. At the Distler Brickyard Superfund site
in  Kentucky,  for  example,  chitin  (a  natural biopolymer
derived from shrimp  and crab shells) was injected into an
aquifer as a source of volatile fatty acids to promote VOC
degradation. Another example is  provided at the Naval
Amphibious  Base   Little  Creek   in   Virginia,  where
bioremediation  involved  injection of diluted cyclodextrin (a
simple  sugar)  that  could  be  recycled.  Information  on
reagent options  and evaluation  of related  factors  is
provided in various demonstration  reports compiled by the
Environmental Security Technology Certification Program
(ESTCP).5
Opportunities  to reduce the  environmental  footprint  of
long-term   actions  can   be  further  reduced   through
optimization  of  the   monitoring   program.   Periodic
reevaluation  can  help   identify   potential   monitoring
changes  such  as  reduced  sampling  frequency,  fewer
sampling  locations, or routine sampling  of a  smaller well
network as a contaminant plume collapses over time.6
  Green Remediation:  A Sampling of Success Measures
               for a Bioremediation System
  "  Reduced fuel consumption due to transport of high-bulk
    reagents via rail rather than trucks
  *  Reduced GHG emissions as a result of using gravity-fed
    injection systems rather than fuel-fed pumping
  *  Protection of nearby and downstream surface water
    through construction of bermed retention ponds that
    capture and treat contaminated stormwater runoff
  *  Beneficial use of industrial waste or surplus byproducts as
    bioremediation reagents
  •  Reduced soil compaction during system construction as a
    result of using well-defined work areas
References [Web accessed: 2010, February 28]
1 U.S. EPA; Principles for Greener C/eonups; August 27, 2009;
 http://www.epa.gov/oswer/greencleanups
2 U.S. EPA; Green Remediation: Incorporating Sustainable Environmental
 Practices into Remediation of Contaminated Sites; EPA 542-R-08-002,
 April 2008
3 Interstate Technology and Regulatory Council; In Situ Bioremediation of
 Chlorinated Ethene: DNAPL Source Zones; June 2008
4 U.S. EPA; Green Remediation Best Management Practices:
 "Site Investigation; EPA 542-F-09-004, December 2009
 b Excavation and Surface Restoration; EPA 542-F-08-012, December
  2008
 c Pump and Treat Technologies; EPA 542-F-09-005, December 2009
 dSoi/ Vapor Extraction & Air Sparging; EPA 542-F-l 0-007, March 2010
 "Clean Fuel & Emission Technologies for Site Cleanup; EPA 542-F-l 0-
  008, April 2010
5  ESTCP Environmental Restoration Projects and Related Efforts;
  http://www.estcp.org/Technology/ER-Chlorinated-Solvents.cfm
6  U.S. EPA and U.S. Army Corps of Engineers; Roadmap to Long-Jerm
  Monitoring Optimization; May 2005, EPA 542-R-05-003
             Visit Green Remediation Focus online:
               http://cluin.org/gresnremediation
                 For more information, contact:
      Carlos Pachon, OSWER/OSRTI (pachon.carlos@epa.gov)
              U.S. Environmental Protection Agency

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