CRC for Contamination Assessment and Remediation of the Environment

National Remediation Framework

Technology guide: Bioremediation

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CRC CARE National Remediation Framework Technology guide: Bioremediation

National Remediation Framework

The following guideline is one component of the National Remediation Framework

(NRF). The NRF was developed by the Cooperative Research Centre for
Contamination Assessment and Remediation of the Environment (CRC CARE) to
enable a nationally consistent approach to the remediation and management of
contaminated sites. The NRF is compatible with the National Environment Protection
(Assessment of Site Contamination) Measure (ASC NEPM).
The NRF has been designed to assist the contaminated land practitioner undertaking a
remediation project, and assumes the reader has a basic understanding of site
contamination assessment and remediation principles. The NRF provides the
underlying context, philosophy and principles for the remediation and management of
contaminated sites in Australia. Importantly it provides general guidance based on best
practice, as well as links to further information to assist with remediation planning,
implementation, review, and long-term management.
This guidance is intended to be utilised by stakeholders within the contaminated sites
industry, including site owners, proponents of works, contaminated land professionals,
local councils, regulators, and the community.
The NRF is intended to be consistent with local jurisdictional requirements, including
State, Territory and Commonwealth legislation and existing guidance. To this end, the
NRF is not prescriptive. It is important that practitioners are familiar with local
legislation and regulations and note that the NRF does not supersede regulatory
requirements.
The NRF has three main components that represent the general stages of a
remediation project, noting that the remediation steps may often require an iterative
approach. The stages are:
• Define;
• Design and implement; and
• Finalise.
The flowchart overleaf provides an indication of how the various NRF guidelines fit
within the stages outlined above, and also indicates that some guidelines are relevant
throughout the remediation and management process.
It is assumed that the reader is familiar with the ASC NEPM and will consult other CRC
CARE guidelines included within the NRF. This guideline is not intended to provide the
sole or primary source of information.

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Executive summary

Bioremediation techniques are designed to break down contaminants via the

stimulation of microorganisms which use the contamination as an energy source for
development and growth. Various bioremediation techniques can be applied in situ or
ex situ and under aerobic or anaerobic conditions. Soil, groundwater and vapour are all
able to be bioremediated.
Successful design and implementation of a bioremediation program for soil, for
example, is dependent on the following key technical considerations:
• Physical properties of the soil to be treated.
• Chemical composition of the soil to be treated.
• Chemistry and concentrations of contaminants.
Moisture content, available nutrients, contaminant mass and distribution and
physiochemical parameters are also important factors to consider in assessing whether
soil bioremediation will be effective. For groundwater-related bioremediation,
hydrogeological factors such as aquifer permeability and water quality will play an
important role.
If there is uncertainty as to whether bioremediation will achieve the required outcome,
treatability studies may need to be conducted to resolve the issues. Treatability studies
can be undertaken in three stages – the first stage is feasibility testing to assess the
ability of bioremediation techniques to meet the remediation objectives. This is typically
conducted as a series of bench tests. The second, more detailed stage is to evaluate
the application of the method under the specific site conditions, usually conducted on
site as a pilot trial. The information obtained in stages 1 and 2 is generally sufficient to
enable formulation of a remediation action plan. However, where additional data are
required to enable the remediation system to be designed a third stage of treatability
testing should be undertaken to determine specific operating requirements and
performance criteria to enable completion of a RAP.
References to case studies are provided in Appendix A.
A number of sources of information were reviewed during the formulation of this
document to compile information on potential technologies. These are listed in
references, and provide an important resource to readers.

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Abbreviations

Cooperative Research Centre for Contamination

CRC CARE
Assessment and Remediation of the Environment
NRF National Remediation Framework
ORC Oxygen Releasing Compounds
PAH Polycyclic Aromatic Hydrocarbons
PCBs Polychlorinated Biphenyl
PCDD/F (several) Tetrachloroethylene
PCE Tetrachloroethylene
PCP Pentachlorophenol
PFOS Perfluorooctanesulfonic Acid
pH Power of Hydrogen
PPE Personal Protective Equipment
RAP Remediation Action Plan
Redox potential Reduction/Oxidation Potential
SVOCs Semi-Volatile Organic Compounds
TCE Trichloroethylene
VOCs Volatile Organic Compounds

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Glossary

Aerobic A process that occurs in the presence of free oxygen.

A substance added to a soil to encourage
Amendment
microbiological activity and degradation
A process that occurs in the absence of free oxygen. In
this case it refers to degradation in the absence of
Anaerobic oxygen
The addition to the environment of microorganisms that
Bioaugmentation can metabolise and grow on specific organic
compounds
The transformation of a substance or chemical by
micro- organisms such as bacteria or fungi, resulting in
Biodegradation
a change in chemical structure mass within the
environment.
A bioremediation technology that utilises the action of
soil microorganisms to remove contamination.
Excavated soils are mixed with soil amendments,
Biopile formed into compost piles and enclosed to allow the
microorganisms to metabolise the contamination.
Destruction of contaminants via the stimulation of
microorganisms which use the contamination as an
Bioremediation energy source for development and growth.
A process that increases activity of microorganisms
biodegrading contaminants. For example, addition of
Biostimulation
nutrients, oxygen, or other electron donors and
acceptors
The process of supplying oxygen in-situ to oxygen
deprived soil microbes by forcing air through
Bioventing
unsaturated contaminated soil at low flow rates. This
stimulates biodegradation and minimises volatilisation.

Concentration The amount of material or agent dissolved or contained

in unit quantity in a given medium or system.
A representation of site-related information including
the environmental setting, geological, hydrogeological
and soil characteristics together with the nature and
Conceptual site model distribution of contaminants. Contamination sources,
exposure pathways and potentially affected receptors
are identified. Presentation is usually graphical or
tabular with accompanying explanatory text.
Any chemical existing in the environment above
Contaminant background levels and representing, or potentially
representing, an adverse health or environment risk.

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A site that is affected by substances that occur at

concentrations above background or local levels and
which are likely to pose an immediate or long-term risk
to human health and/or the environment. It is not
necessary for the boundaries of the contaminated site
Contaminated site to correspond to the legal ownership boundaries.
The presence of a substance at a concentration above
Contamination background or local levels that represents, or potentially
represents, a risk to human health and/or the
environment.
A chemical capable of accepting / donating electrons
Electron acceptor / donor
during oxidation-reduction reactions.
The addition of microorganisms or nutrients to the
subsurface environment to accelerate the natural
Enhanced bioremediation biodegradation of contaminants.
The government agency in each state or territory that
Environment(al) protection has responsibility for the enforcement of various
authority / agency jurisdictional environmental legislation, including some
regulation of contaminated land.
A Latin phrase that translates literally to "off site" or "out
of position". It refers to remediation that is performed on
the contamination following removal, ususally the
Ex-situ excavation of soil.
A group of diverse and widespread unicellular and
Fungi multicellular eukaryotic organisms. Some species are
important in the decomposition of plant litter.
Water stored in the pores and crevices of the material
below the land surface, including soil, rock and fill
Groundwater material.
Indigenous Naturally occurring at that location
A Latin phrase that translates literally to "on site" or "in
position". It refers to remediation that is performed on
the contamination while it is in place, without excavating
In-situ soil.
A remediation method that involves tilling the
contaminated soil, with or without additives, to aerate
the soil and encourage biological activity to remediate
Land farming the contamination.
Destruction of contaminants via the stimulation of fungi
which use the contamination as an energy source for
Mycoremediation development and growth.
A chemical that oxidises other substances, causing
Oxidant them to loose electrons.

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The uptake of contaminants by vegetation roots and

transfer of the contaminants to the vegetation shoots
Phyto-accumulation and leaves.
Phyto-degradation The metabolism of contaminants in vegetation tissues
Remediation utilising the natural process of
phytoaccumulation by encouraging growth of vegetation
that will uptake the contaminant of concern. Particularly
Phytoremediation used for remediation of heavy metals.
The production of chemicals by the vegetation that
immobilises contaminants at the interface between the
Phyto-stabilisation roots and the soil.
Those in the private sector professionally engaged in
Practitioner the assessment, remediation or management of site
contamination.
A person who is legally authorised to make decisions
Proponent about a site. The proponent may be a site owner or
occupier or their representative.
This is short for reduction / oxidation, and describes a
chemical reaction in which electrons are transferred
Redox between chemicals.
An action designed to deliberately break the source-
pathway-receptor linkage in order to reduce the risk to
Remediation
human health and/or the environment to an acceptable
level.
The zone of soil around a plant root where the biology
Rhizosphere and chemistry of the soil are influenced by the root.
The probability that in a certain timeframe an adverse
outcome will occur in a person, a group of people,
plants, animals and/or the ecology of a specified area
that is exposed to a particular dose or concentration of
Risk
a specified substance, i.e. it depends on both the level
of toxicity of the substance and the level of exposure.
‘Risk’ differs from ‘hazard’ primarily because risk
considers probability.
A parcel of land (including ground and surface water)
being assessed for contamination, as identified on a
map by parameters including Lot and Plan number(s)
and street address. It is not necessary for the site
boundary to correspond to the Lot and Plan boundary,
Site however it commonly does.
A chemical that is able to dissolve other substances.
Solvent contaminants include a range of hydrocarbons
used in industrial processes due to their dissolving
Solvent properties.

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A substance that chemically bonds to both oils and

water, and transfers oils into the aqueous phase where
Surfactant they are more readily remediated.
A series of tests designed to ascertain the suitability of
the treatment for the contaminants under the site
Treatability studies conditions
A long line of soil or similar material heaped up by a
machine to aerate, dry out or encourage biological
Windrow activity through composting.

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Measurements

Unit or symbol Expansion

o
C Degrees Celsius
m bgl Metre(s) below ground level
mg/kg Milligram per kilogram
mV Millivolts

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Table of contents

National remediation framework Error! Bookmark not defined.

Executive summary iii
Abbreviations iv
Glossary v
Measurements ix
Table of contents x
1. Introduction 1
2. Technology description and application 2
2.1 Types of bioremediation 2
Bioventing 2
Enhanced bioremediation 2
Phytoremediation 3
Mycoremediation 3
Biopiles or windrows 4
Composting 5
Land farming 5
Slurry phase biological treatment 6
2.2 Comparison of technologies 6
3. Feasibility assessment 11
3.1 Data requirements 12
Physical properties 12
Chemical composition 12
Maximum allowable concentrations 13
Regulatory requirements 14
3.2 Treatable contaminants 14
4. Treatability studies 17
4.1 Bench testing 17
4.2 Pilot trial 18
4.3 Finalising design 18
4.4 Anaerobic bioremediation 19
5. Validation 20
5.1 In-situ bioremediation 20
5.2 Ex-situ bioremediation 21

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6. Health and safety 22

Appendix A – Case studies 25
Appendix B – Treatability case studies 27
Appendix C – References 31

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1. Introduction

The purpose of this guideline is to provide information on bioremediation as a treatment

technology for the remediation of contaminated sites to assist with selection of
remediation options. The document contains information to inform remediation planning
and aid compilation of a remediation action plan (RAP).
While soil, groundwater and vapour are all able to be bioremediated, this document
predominantly provides guidance on the application of bioremediation as a remediation
technology to treat contaminated soil. Readers are directed to the NRF Technology
guide: Monitored natural attenuation for more information on the bioremediation of
contaminated groundwater.
This guidance is primarily intended to be utilised by remediation practitioners and those
reviewing practitioner’s work, however it can be utilised by other stakeholders within
the contaminated sites industry, including site owners, proponents of works, and the
community.
Bioremediation is one of many technologies available for contamination remediation,
and other technologies may be more appropriate. It is assumed that the information
presented within will be used in a remediation options assessment to identify and
select the preferred technologies for more detailed evaluation. This guideline provides
information for both initial options screening and more detailed technology evaluation.
This guideline does not provide detailed information on the design of bioremediation
systems as this is a complex undertaking and should be carried out by appropriately
qualified and experienced practitioners. Readers are directed to the NRF Guideline on
performing remediation options assessment for detailed advice on assessing
remediation options. In addition, the remediation objectives, particularly the required
quality of the soil after treatment, are a critical matter and it is assumed that these have
been determined and considered in the remediation options assessment and selection
process. Readers are directed to the NRF Guideline on establishing remediation
objectives for more detailed advice.
References to case studies are provided in Appendix A.

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2. Technology description and application

Bioremediation techniques aim to break down contaminants via bioaugmentation

and/or biostimulation of microorganisms that use the contamination as a food and
energy source for development and growth. This is done via the introduction of
oxidising or reducing agents, and moisture and/or nutrients to increase the favourability
of the environment for microorganisms. Bioremediation technologies can be conducted
ex-situ or in-situ.

2.1 Types of bioremediation

There are several techniques that fall under the broad heading of bioremediation,
including:
• Bioventing;
• Enhanced bioremediation;
• Phytoremediation;
• Mycoremediation;
• Biopiles or windrows;
• Composting;
• Land farming; and
• Slurry phase biological treatment.
Each of these techniques is described in the section below.
Bioventing
Bioventing is an in-situ remediation technology involving the injection (and sometimes
extraction) of air into the subsurface to enhance microbial activity and facilitate
biodegradation of organic contaminants adsorbed to soils in the unsaturated zone.
Bioventing is different from air sparging, which is typically conducted at higher flow
rates to promote volatilisation, rather than biodegradation of volatile organic
compounds.
During bioventing, air is injected at low rates to increase the oxygen content in the sub-
surface and promote oxidation reactions. Other gases, such as methane or propane,
can also be injected (at concentrations below the lower explosive limit) to promote the
degradation of organic contamination under reducing conditions.
Where high concentrations of contaminants are present, it is possible that the soil
pores can become clogged with additional biomass generated during bioventing,
reducing the oxygen levels. Pulsed air injection can be useful to increase the oxygen
levels under these conditions.
Enhanced bioremediation
Enhanced bioremediation is an in-situ remediation technology involving the addition of
a chemical to the subsurface to enhance microbial activity and facilitate biodegradation
of organic contaminants adsorbed to soils in the unsaturated zone.

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Aerobic enhancement comprises the addition of oxygen (an electron acceptor) to the
subsurface to increase the population of microbial organisms to assist with the
biodegradation of contaminants in the soil or groundwater.
Although the introduction of oxygen releasing compounds (ORC) is more commonly
used to enhance aerobic bioremediation of groundwater, ORC can also be applied to
the unsaturated zone. The ORC can be a proprietary oxidant, or substances such as
hydrogen peroxide or ozone.
Anaerobic enhancement comprises the addition of an electron donor (such as
hydrogen or hydrocarbons) to the subsurface to increase the population of microbial
organisms to assist with reductive dechlorination processes (anaerobic degradation) in
groundwater. The direct addition of hydrogen is rare, as during anaerobic
biodegradation hydrogen is normally indirectly generated via fermenting organic matter.
Other nutrients such as nitrate and sulphate can be added to groundwater to enhance
anaerobic biodegradation of petroleum hydrocarbons.
Phytoremediation
Phytoremediation is an in-situ remediation technology that involves the use of plants to
remove or stabilise contaminants in soil and, to a lesser extent, groundwater. An
example of phytoremediation in the wastewater industry is the use of reed beds for on-
site biological treatment of sewage effluent.
The following mechanisms are used in the process of phytoremediation:
• Enhanced rhizosphere biodegradation: the release of natural substances from
plant roots to supply nutrients to microorganisms which increases biological
activity;
• Phyto-accumulation: the uptake of contaminants by plant roots and transfer of
the contaminants to the plants shoots and leaves;
• Phyto-degradation: the metabolism of contaminants in plant tissues; and
• Phyto-stabilisation: the production of chemical by the plant that immobilises
contaminants at the interface between the roots and soil.
Hardy species, such as eucalyptus, fern, rye and fescue grasses, are often selected for
phytoremediation in Australia due to their fast growing and robust nature and ability to
survive in saline and water-logged soils.
Mycoremediation
Mycoremediation is a form of in situ bioremediation that uses fungal material
(mycelium) to accumulate and degrade contaminants to remediate contaminated soils
and groundwater. Mycelium is the dense network of branching white hyphae that make
up the fungi. The mycelia deliver the enzymes required to break down the
contamination; as such, the reaction is extra-cellular (outside rather than within the
fungi). Fungi can be effective in breaking down petroleum hydrocarbons and some
chlorinated compounds and are able to stimulate native microbes and enzymes in-situ.
Heavy metals can also bioaccumulate in fungi and the contamination can be removed
during harvesting.
The type of fungi used in mycoremediation is affected by the temperature, soil pH and
the availability (or lack) of oxygen. Typically, a mycelium-treated substrate such as

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wood chips and straw is spread over contaminated soils which produce enzymes
capable of decomposing contaminants over time.
Some of the common fungi used in mycoremediation and the contaminants they can
treat are presented in Table 1 below.
Table 1: Common fungi used in mycoremediation

Type of fungi Target contaminants

Shaggy Mane • Arsenic,
• Cadmium, and
• Mercury
Elm Oyster • Dioxins,
• Wood preservatives
Phoenix Oyster • Cadmium,
• Mercury,
• Copper
Pearl Oyster • Polychlorinated biphenyls (PCBs)
• Polycyclic aromatic hydrocarbons (PAHs)
• Cadmium,
• Mercury,
• Dioxins
Shitake • PAHs,
• PCBs,
• Pentachlorophenol
Turkey Tail • PAHs,
• Organophosphates,
• Mercury
Button Mushrooms • Cadmium
King Stropharia • E-coli and other biological contaminants

Biopiles or windrows
Biopiles or windrows are an ex situ application of bioremediation where petroleum
hydrocarbon impacted soils are excavated and placed in a treatment area where
agents are usually mixed into the contaminated soils to enhance the degradation
process. The soil can be placed in stockpiles (biopiles) or rows (windrows). The
excavated soil needs to be aerated and moisture, temperature, oxygen and pH can be
adjusted to make the process more effective. A leachate barrier and collection system
is required to avoid contamination leaching into the soil and groundwater below the
treatment area.

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Biopiles can also be engineered and contain ventilation piping and blower, irrigation
piping and/or sump and pump systems to facilitate aeration and drainage to maximise
degradation rates.
Composting
Composting is an ex-situ remediation technology that involves the biological
decomposition of wastes under controlled conditions to a state in which it can be
handled, stored and / or applied to land without adversely affecting the environment.
Contaminated soils are added to the compost process, and the contaminants are
degraded together with the degradable waste material into humus and inert by-
products (such as carbon dioxide, water and salts).
Composting is a special type of decomposition for which the conditions are established
to allow for optimal microbial activity. Conditions that are important include the correct
proportions of carbon and minerals in the compost mix (e.g. carbon to nitrogen ratio),
good aeration and adequate moisture content. When the conditions are right, microbial
(bacteria, including actinomycetes, and fungi) activity is very rapid and a large amount
of heat is produced and the temperature rises.
It is generally accepted that if the whole of the composting mass has been held at 55oC
or more for three consecutive days, the compost can be termed a pasteurised product
with significantly reduced numbers of plant and animal pathogens and plant
propagules. The composting process can be extended to produce a mature product
with a lower level of phytotoxicity and a higher degree of biological stability than
pasteurised compost.
When the material is heavily contaminated or odorous, different systems will be
required, such as enclosed trenches or rotating drums where odours can be captured
during the composting process and treated. All systems require air to be drawn through
the contaminated medium to provide suitable conditions for the microorganisms to
survive.
In the initial stage, microbial activity will be at its peak and the temperatures will be
highest, and the most degradable contaminants will be consumed. Following this initial
stage, the temperatures will drop until heat is no longer generated and the material is
now a compost product. The material has a high microbial diversity (higher than
healthy fertile soils), and this expedites degradation of the contaminants.
Land farming
Land farming is an ex-situ remediation technology that involves spreading impacted
soils in thin layers across a prepared surface and regularly turning the material to
enable air flow through the soil matrix (introducing oxygen to facilitate degradation).
The soil material is placed on a lined surface, with drainage control and bunding, to
minimise the potential for leaching and run-off of contaminants. The soil conditions are
controlled to maximise the degradation rate, including moisture content (via
irrigation/spraying), aeration (by tilling) and pH (buffered to neutral by adding acid or
alkali).
Where land farming is carried out in the open and volatile contaminants (such as
petrol) are involved, volatilisation can be a significant contributor to loss of
contaminants. Where volatile emissions and odours are possible, the requirements for
emission management must be addressed as part of such remediation works. If the

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process involves only volatilisation without degradation, some regulatory agencies will
not accept land farming as an acceptable treatment option.
Land farming can also be conducted in-situ to treat soils up to approximately 1 m bgl.
Soils are mechanically agitated to introduce oxygen to the subsurface and facilitate the
addition of nutrients and lime to reduce the soil acidity.
Slurry phase biological treatment
Slurry phase biological treatment is an ex-situ remediation technology that is performed
in a reactor to remediate a mixture of water and excavated soil. The soil is mixed with
water to a slurry that is determined by the proportions of the contaminants in soils, the
rate of biodegradation, and the physical nature of the soils. If the soil is prewashed, the
contaminated fines and wash water are treated in the reactor. Readers are directed to
the NRF Technical guide: Soil washing for more information on soil washing.
The slurry contains between 5% and 40% solids by weight depending on the nature of
the biological reactor. The soil is suspended in a reactor vessel and mixed with
nutrients and oxygen. Microorganisms, acid or alkali may be added depending on
treatment requirements. When biodegradation is complete, the soil slurry is dewatered
and the liquids filtered and clarified.

2.2 Comparison of technologies

The advantages and disadvantages of various bioremediation technologies, along with
the soil types and conditions for which each technology may be suitable, are listed in
Table 2.

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Table 2: Comparison of bioremediation technologies

Bioremediation Advantages Disadvantages Treatable medium/applicable

technology conditions
Bioventing • Fast degradation rates (in • Contaminants may volatilise during • Permeable soils
comparison to other treatment posing a potential vapour
• Unsaturated soils
bioremediation methods) exposure risk and increased
greenhouse gas emissions • Can be applied under aerobic and
• No excavation required
anaerobic conditions
Enhanced bio- • Can support rapid degradation • Requires correct oxygen/hydrogen and • Can be applied to soils with high and
remediation rates nutrient dosing and may need several low permeability
trial stages
• May be used concurrently to • Can be applied under aerobic or
address groundwater anaerobic conditions (groundwater)
contamination
• May require addition of
• Low cost dehalococcoides bacteria for reductive
dechlorination
• Minimal exposure
• Can be applied in situ or ex situ
• No excavation required

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Bioremediation Advantages Disadvantages Treatable medium/applicable

technology conditions
Phytoremediation • Generally low operation & • Where the process involves uptake and • Low permeability soils
maintenance cost concentration of contaminants in the
• Saturated/high moisture content and
plant, the resulting plant matter
• Capable of treating large soil water retaining soils
(following remediation) is another
and wastewater volumes
waste stream that requires treatment
• Environmentally sustainable –
• May not be suitable for high
can improve air quality
contaminant concentrations, or if
• Potentially more agreeable to contaminants toxic to plants are
stakeholders or adjacent land present
users (no equipment, low
• Slow process
maintenance and high
aesthetic outcome) • Weather/season dependent (less
effective during winter months)
• Will not be able to treat contamination
at significant depth as plant roots are
generally contained to shallower soils
Mycoremediation • Low cost and minimal Technology still in development – • Can be applied to near surface soils
maintenance required comparatively small number of with high and low permeability
successful case studies available
• Environmentally sustainable • Aerobic application
• Minimal disturbance
(agreeable to stakeholders)
• Delivers fast results (odours
often immediately mitigated)
and is generally completed in
weeks/months

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Bioremediation Advantages Disadvantages Treatable medium/applicable

technology conditions
Biopiles/windrows • Generally low operation & • Potential exposure risks during • Permeable soils
maintenance cost excavation. Potential odour and air
• Aerobic application
emissions may require management
• Leachate may be an issue and base
liner and/or bunding may be required to
prevent migration of contaminants to
the water table
Composting • Low cost • Bulking agents necessary • Permeable soils
• Generates heat (naturally) • Potential exposure risks during • Aerobic application
excavation
• Residual contamination will require
treatment
• Leachate may be an issue and base
liner and/or bunding may be required to
prevent contamination migration to the
water table
• Treated material may not be suitable
for reuse or building over if retained on-
site (dependent on physical properties
at completion)

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Bioremediation Advantages Disadvantages Treatable medium/applicable

technology conditions
Land farming • Low cost. • May not be suitable for high • Permeable soils.
contaminant concentrations.
• Simple design and set up. • Aerobic application.
• Potential dust, odour and vapour
exposure during spreading of the soil
and aeration.
• Needs a large treatment area (reducing
treatable volume).
• Runoff collection facilities must be
constructed and monitored.
• Leachate may be an issue and base
liner may be required to prevent
contamination migration to the water
table.
Slurry phase • Operational parameters can be • High cost • Can be applied to soils with high and
biological adjusted easily. low permeability.
• Treatable volume (and rate) limited by
treatment
• Fast degradation rates. size of equipment used. • Can be applied under aerobic or
anaerobic conditions.
• Potential exposure risks during
excavation. • Surface contamination.

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3. Feasibility assessment

Key site-specific considerations that will often determine the feasibility of

bioremediation include:
• Whether the contaminants are sufficiently biodegradable and there is
confidence that the remediation targets will be met within an acceptable
timeframe.
• Whether the bioremediated material will be suitable for future use or disposal,
taking into account the amendment material added, other contaminants
present, and the byproducts and residuals of the treatment.
• Whether the extent and distribution of contamination is sufficiently well known.
• Whether the physiochemical composition and heterogeneity of the soil will
allow sufficient uniformity of treatment to meet the remediation targets.
• Whether biodegrading organisms are naturally present or need to be added.
Appropriate remediation data must be collected to evaluate the applicability of a
bioremediation technology. If there is reasonable confidence that the selected
bioremediation method will achieve the required treatment outcome, then other issues
will need to be considered to determine if it is likely to be an appropriate technology for
the site. These include:
• Are there sufficient microorganisms present and is the contaminant
bioavailability sufficient to enable degradation?
• Will the relevant regulatory agencies accept the bioremediation technology as
a viable means of remediation?
• Will it be able to be confirmed that the contaminants have degraded, and have
not been simply diluted by the material added or mixing operations, or
volatilised and lost in ambient air, if these loss mechanisms are not acceptable
to the regulatory agency?
• Are there planning or regulatory approvals required to use these
technologies?
• Will the treated material be of a form and with contaminant concentrations that
will allow the material to be reused as backfill on the site or as clean fill
elsewhere, or will subsequent treatment (e.g. stabilisation) or landfill disposal
be required? Could there be remnant biodegradable material present that
would give rise to methane or carbon dioxide concerns, or a geotechnical
concern (physical stability).
• What are the break-down products of the parent compound/s? Are they more
toxic than the parent compound/s and does this risk require additional
assessment? Does the breakdown product require a different treatment
method (such as the production of vinyl chloride during reductive
dechlorination of PCE)?
• Is there any risk of contamination migrating to other environmental segments
through the use of this technology (e.g. incorrect controls during land farming
resulting in transfer of contaminants from soil to the atmosphere)?

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• Is it likely that other stakeholders (such as local government or the public) will
accept the use of the technology, particularly those stakeholders that can
have a significant bearing on whether the technology is applied at the site?
• Are there sensitive sites nearby that would not be compatible with the
proposed operation?
• Is there a time constraint, and can the bioremediation application meet this
constraint?
• Is the expected order of cost of treatment acceptable?

3.1 Data requirements

Successful implementation and design of a bioremediation system, whichever
approach is used, is likely to be dependent on the following key technical
considerations:
• The physical properties of the soil;
• The chemical composition of the soil; and
• The chemistry, concentrations and distribution of contaminants within the soil
materials.
Physical properties
The physical composition of the material to be treated needs to be well characterised.
Important factors include:
• Soil type and heterogeneity: differing grain sizes and the presence of coarse
fragments of material (such as concrete or bricks) will impact upon air, water
and contaminant migration pathways and can prevent the distribution of
oxygen or nutrients through the contaminated soils;
• Organic matter: Where there is a significant quantity of organic matter present,
this can result in depleted oxygen for microorganisms, slowing the
biodegradation (in aerobic applications); and
• The permeability and plasticity of the material: low permeability soils will
hinder the migration of oxygen and nutrients, both vertically and horizontally.
In situ applications are unlikely to be appropriate for low permeability soils.
Chemical composition
The composition of the material to be treated needs to be well characterised. Important
factors include:
• The distribution, concentrations and mass of contaminants in the soil at the
site, as the requirement will be to locate and treat contamination that exceeds
certain concentrations;
• Range of contaminants, their concentrations and physical form and their ability
to degrade, volatilise or inhibit the rate of microbial degradation. Volatility is
important for slurry phase biological treatment where the contaminants could
volatilise in the reactors before degradation. Certain contaminants (such as
heavy metals) can have a toxic effect on microorganisms, and inhibit
degradation. The form of the contaminant can be important as to whether
biodegradation will occur (e.g. non-aqueous phase liquid will not sustain

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biodegradation, whereas free phase contamination that is distributed as an

adsorbed phase may sustain biodegradation);
• Ion exchange and filtration mechanisms of the soil to be treated to assess
whether it will be necessary to improve the transport of water, electron
acceptors (such as oxygen), nutrients and microorganisms to assist the
biodegradation process (via injection wells or pumping etc); and
• Physicochemical parameters – pH, electron acceptors, nutrients, temperature
and toxicity to assess what effect these will have on microorganisms and
which strain will be most effective at treating the contaminants present, such
as:
- pH can affect contaminant solubility and bioavailability of nutrients for
microorganisms in the degradation process. Bioremediation processes
generally perform optimally in the pH range 6 to 8
- Redox potential and oxygen content indicates whether conditions are
oxidising or reducing
- Availability of nutrients that are required for microbial growth (and cell
division) – these can be added where needed
- Contaminant bioavailability depends on the contaminants ability to sorb to
solids and be diffused in soil macropores (bioavailability for microbial
reactions is lower for contaminants that are strongly sorbed to soils, or are
within macropores and are less bioaccessible)
- Temperature affects the rate of microbial metabolism and degradation.
The biodegradation rate generally increases with temperature.
Maximum allowable concentrations
The maximum allowable concentration and variation in concentration of the
contaminants and by-products of treatment in the treated soil needs to be determined.
If very stringent clean up criteria are applicable, then bioremediation technologies may
not be sufficient to meet the criteria and additional ‘polishing’ stages of treatment may
be required. For example, criteria for substances such as chlorinated organics may be
very stringent (e.g. < 1 mg/kg of total chlorinated organics) and may not be readily
achievable by bioremediation. In such circumstances, additional cycles of treatment
may be required resulting in prohibitive cost, or the requirement for subsequent
treatment by another technique that, by itself, could achieve the criteria more cost
effectively than biodegradation. Specific concentration considerations include the
following:
• The maximum allowable concentrations of contaminants (such as heavy
metals or asbestos) that will remain after treatment, and which could preclude
the intended use or disposal of the treated soil or inhibit the biodegradation
processes;
• The maximum allowable concentrations of volatile components, so that
volatiles will not cause an environmental or human health risk during
excavation or treatment;
• The maximum allowable quantities and concentrations of other material such
as contaminated concrete or plastic clay that may restrict the distribution of
microorganisms, or the movement of gases or nutrients in the subsurface
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which are needed to facilitate biodegradation, and hence affect the uniformity
(and completeness) of treatment;
• The maximum allowable concentrations of reagents and biodegradable
material that are added to the soil to facilitate biodegradation, and which will
remain after treatment (e.g. will the residual concentrations or gases that
result (such as methane or carbon dioxide) affect the intended use or disposal
of the treated material);
• Whether there is a concern regarding the microorganisms that will be present
during or after treatment and could pose a risk to persons or the environment,
and the maximum allowable concentrations that could apply; and
• If phytoremediation is under consideration, the contaminant concentrations
and their depth will be a key factor – high concentrations of certain
contaminants may inhibit plant growth, limiting the viability of phytoremediation
to achieve the remediation objectives. If contamination is deep and extends
beyond the root zone, then remediation of this material may not occur.
Regulatory requirements
The regulatory agencies (particularly the agencies responsible for protection of the
environment, town planning, and licensing treatment facilities) should be consulted to
determine the specific requirements that relate to obtaining the necessary approvals
and licences, and controls that can be expected. This is particularly relevant where
emissions to the environment are planned or possible.

3.2 Treatable contaminants

Bioremediation technologies are potentially able to treat a wide range of:
• Volatile organic compounds;
• Semi-volatile organic compounds; and
• Petroleum hydrocarbons.
Bioremediation of PAHs is possible in engineered biopiles or windrows with the
addition of compost, nutrients, and surfactants to release contaminants to the aqueous
phase. High molecular weight PAHs and aged petroleum products are of low
bioavailability and are therefore not generally suitable for treatment by bioremediation,
particularly phytoremediation, unless only minor reductions in concentration are
required.
Some higher-boiling-point halogenated compounds such as PCBs, dioxins and furans,
and fluorinated compounds such as PFOS can be very difficult to degrade and are
unlikely to be suitable for treatment by bioremediation.
Information on the biodegradability of contaminant groups and the preferred conditions
(anaerobic or anaerobic) is summarised in Table 3.

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Table 3 Contaminant Biodegradability and the Preferred Conditions

Contaminant Microbial Degradability Preferred conditions

High Low No
Short-chain mineral oil hydrocarbons X Aerobic
Mineral oil
Long-chain/branched mineral oil hydrocarbons X Aerobic
hydrocarbons
Cycloalkanes X Aerobic
(Mono)aromatic hydrocarbons X Aerobic

Monoaromatic Phenols X Aerobic

hydrocarbons Cresols X Aerobic
Catechols X Aerobic
Polycyclic 2 to 3 ring PAHs (e.g. naphthalene) X Aerobic
Aromatic
4 to 6 membered ring PAHs (e.g. benzo(a)pyrene) X Aerobic or Anaerobic/Aerobic
Hydrocarbons
Tetrachloroethene, tricloroethane X Anaerobic
Chlorinated
Aliphatic Trichloroethene X Anaerobic
Hydrocarbons
Dichloroethane, dichloroethene, vinyl chloride X Anaerobic/Aerobic
Chlorophenols (superchlorinated) X Anaerobic
Chlorophenols (low chlorinated) X Anaerobic/Aerobic
Chlorobenzenes (super chlorinated) X Anaerobic
Chlorinated
Aromatic Chlorobenzenes (low chlorinated) X Anaerobic/Aerobic
Hydrocarbons
Chloronaphthalene X Anaerobic/Aerobic
Polychlorinated biphenyls (PCBs) (super chlorinated)* X Anaerobic
Polychlorinated biphenyls (low chlorinated) X Anaerobic/Aerobic

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Contaminant Microbial Degradability Preferred conditions

High Low No
Mono- and dinitroaromatics X Anaerobic/Aerobic
Nitroaromatic
Trinitrotoluene (TNT) X Anaerobic/Aerobic
compounds
Trinitrophenol (picric acid) X Anaerobic/Aerobic
Nitroaliphatic Glycerol trinitrate Aerobic
X
Compounds
g-hexachlorocyclohexane (lindane) X Anaerobic/Aerobic
Pesticides b-hexachlorocyclohexane** X Anaerobic/Aerobic
Triazines X Aerobic
PCDD/PCDF X Anaerobic/Aerobic
Dioxins
2,3,7,8-PCDD/PCDF X Anaerobic/Aerobic
Free cyanides X Aerobic
Complex cyanides X Aerobic
Inorganic
Ammonium X Aerobic/Anaerobic
compounds
Nitrate X Anaerobic
Sulphate*** X Anaerobic
* The degradation process and susceptibility to degradation are different for planar and non-planar highly chlorinated PCB congeners
** Microbially transformable but not degradable
*** Activity of sulphate-reducing bacteria results in precipitation of metal sulphides or production of hydrogen sulphide gas

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4. Treatability studies

If it is uncertain whether bioremediation will achieve the remediation objective or be

applicable for the specific site conditions, it may be necessary to conduct a treatability
study. Treatability studies also allow estimates of remedial costs and technology
efficiency to be refined.
Designing the treatability study may require input from a number of technical specialists
including environmental scientists/engineers, chemical engineers, mechanical
engineers and air quality specialists to ensure that the study is designed to obtain the
data required to enable the most appropriate implementation strategy to be developed.
The requirements for additional information should be determined through a review of
the specific bioremediation technology being proposed. If the technology has been
widely applied and the results are readily available, it may be possible to extrapolate
information from previous case studies to avoid duplication and reduce project costs.
If it is determined that treatability testing is required to adequately assess the
application of bioremediation or to assist in cost appraisal, consideration needs to be
given to the staging and level of detail of the testing. Typical stages of testing can be
directed to remediation screening, remediation selection and remediation design,
irrespective of whether aerobic or anaerobic bioremediation is involved.
The process involved in each of these stages is outlined below, for the case of aerobic
bioremediation, and this process is generally applicable to anaerobic degradation.
The objectives of the treatability testing should be determined at the outset and,
although these will be site specific, treatability testing should determine:
• Will the proposed bioremediation technology meet the remediation objectives
in a timely and cost effective manner?
• Are there sufficient contaminant degrading microorganisms present?
• What are the optimum moisture, nutrients and pH conditions to facilitate
contaminant degradation?
• That any reported decreases in contaminant concentrations are the effect of
the bioremediation treatment and not a result of volatilisation, leaching of
contaminants from the soil or adsorption.
References to treatability case studies are presented in Appendix B.

4.1 Bench testing

Initial screening or bench scale treatability studies comprise the first stage of treatability
testing, to assess whether biodegradation is a viable option to treat the specific
contaminated material.
The screening tests are generally low cost and relatively fast to complete
(approximately a few weeks or a few months to obtain the results). The initial screening
tests are carried out using simple equipment in a laboratory, such as shake flasks, soil
pans or slurry reactors, and are undertaken using saturated and unsaturated soils,
slurries and various aqueous solutions. The microorganisms can be indigenous to the
specific site, cultured, a commercially available mixture, or a combination of all of
these.

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Various parameters can be adjusted during the initial screening, including pH,
contaminant loading rates, and oxygen and nutrient availability, to improve the potential
success rate.
Setting the data quality objectives for the screening treatability testing at the outset is
vital to obtain the desired results. Usually the main goal of this preliminary treatability
testing is to establish whether biodegradation will occur in the specific contaminated
material. The objectives of the initial screening tests do not normally include assessing
whether the remediation clean up criteria can be met. The testing is normally
concluded after a few weeks when it is evident that the contaminant concentrations
have decreased by a significant percentage, though not necessarily to the level
required to meet the specific clean up criteria.

4.2 Pilot trial

The next stage of treatability testing is to evaluate the application of biodegradation for
the specific site conditions and to establish whether the remediation criteria and clean
up goals are likely to be met. The information obtained in the second stage of testing is
usually sufficient to enable development of the remediation action plan (RAP).
The key objectives for the second stage of treatability testing are:
• Assess contaminant concentrations achieved following treatment (to
determine whether the nominated remediation criteria can be met);
• Determine microorganism populations, oxygen input method and nutrient load
etc. required for efficient treatment; and
• Characterise the composition and physical nature of the final material to
confirm that it can be expected to be suitable for the intended reuse or
disposal.
This stage requires more effort (and cost) than the initial screening testing and
generally takes several months to plan and implement. These tests have the objective
of more closely replicating the physical and chemical parameters of the site under
investigation and the specific bioremediation technology being considered. The tests
should therefore be undertaken using the particular soils to be treated in the full scale
remediation program. However, it should be noted that given the small amount of
material used in the tests (in comparison to the actual volume to be treated), full scale
treatment conditions are likely to differ, particularly where in situ application is being
considered.
At complex sites where in-situ application is being considered, this stage of treatability
testing may be undertaken on-site, ideally in a small section of the area to be
remediated using methods and equipment similar to those proposed for the full scale
application to enable an accurate estimate of whether biodegradation will work and can
meet the remediation criteria.

4.3 Finalising design

If additional data is required to enable the remediation system to be designed, a third
stage of testing may be necessary to establish specific requirements and performance
criteria and provide sufficient information to enable completion of a RAP.
The key objectives of the third stage of treatability testing may be, for example:

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• Obtain all data and information required to enable the remedial program to be
designed;
• Refine the remediation cost estimate;
• Confirm the rate of biodegradation and the concentrations of the treated
medium (to ensure they meet the remediation criteria); or
• Confirm that the composition and physical nature of the treated material will
allow its reuse or disposal.
These studies are usually costly and may take many months to complete, so the
benefit of obtaining more specific operating design parameters and cost estimates
should be weighed against the cost of the overall remedial program.
The tests are usually conducted using a mobile treatment unit brought onto site. The
equipment used should be designed to ensure that the data obtained can be
extrapolated for the full scale unit.

4.4 Anaerobic bioremediation

The process of treatability testing for anaerobic bioremediation can involve a similar
staged approach to that outlined above. In the case of anaerobic treatment, the
available oxygen must be removed (or reduced). This can be achieved by supplying
excess electron donors (reducing agents) to the microorganisms which will consume
oxygen that may be present.
Different electron acceptors (nitrate, carbonate or sulphate) are required to enable
bacteria to facilitate anaerobic, de-nitrifying, sulphate reducing, and methanogenic
conditions, and different by-products and metabolic intermediates will be produced.
Anaerobic tests depend largely on the type of microorganisms that will be used to
achieve biodegradation.
The redox potential of the medium to be treated needs to be reduced to approximately
-150 mV to -350 mV at pH 7 to facilitate the growth of most anaerobic organisms. The
exact redox potential required and the concentration of dosing agent to achieve such a
concentration is dependent upon the specific anaerobic organism.
The redox potential will determine which microorganisms are present and active in the
medium and, as such, the specific anaerobic organisms to be stimulated in the detailed
treatability testing should be determined in the initial screening tests.

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5. Validation

The following information describes the specific validation appropriate for

bioremediation, to assist validation planning within the RAP. Readers are directed to
the NRF Guideline on validation and closure, which among other things, provides
further information on each of the lines of evidence.
Recommended lines of evidence for the validation of bioremediation in soils or
groundwater include:
• The measured decrease in contaminant concentrations;
• An assessment of the microbiota present to break down the contamination;
and
• The geochemistry and biochemistry of the environment
The latter two lines of evidence help demonstrate whether contaminant mass loss is a
result of biodegradation processes or whether it is attributable to non-destructive
processes such as sorption, dilution, or dispersion.
In-situ and ex-situ bioremediation typically require different lines of evidence to
validate:

5.1 In-situ bioremediation

In addition to the COC, amendments such as nutrients as well as carrier fluids injected
into the subsurface should be monitored.
Depending on the nature of the facility, surface and groundwater monitoring may be
required for the duration of the bioremediation operation and for a period post-
completion to verify that contaminants have not migrated during the remediation
process (e.g. displaced by the injection of carrier fluids).
The validation of in-situ groundwater remediation should comprise groundwater
monitoring:
• In the plume,
• Up- and down-gradient of the plume;
• Sufficiently long term to account for seasonal variations.
Depending on the remedial method used, validation monitoring of bioremediation in
groundwater may additionally comprise an assessment of the air distribution and
pressure in an aquifer, oxygenation of the groundwater and vadose zone, changes in
fluid pressures and changes in groundwater chemistry. It is recommended that cross-
sectional contour plots oriented along the path of groundwater flow are prepared as
part of the validation process to understand the distribution and reduction of
contaminants.
It is essential to analyse not only concentrations of the COC, but also biodegradation
products, to establish that the primary contaminants are degrading rather than simply
moving into areas of lower hydraulic conductivity. Testing of biochemical parameters
such as volatile fatty acids and dehydrogenase activity may be useful to determine
whether biodegradation has occurred.

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5.2 Ex-situ bioremediation

The number of samples collected and analysed for validation purposes should be
adequate to provide statistically reliable results, taking into account the intended end
use of the soils .
Where soil remediation involves a static or turning pile (e.g. biopile), sampling should
avoid shallow locations as they may not be representative of the pile. Whilst
heterogeneity is likely to be significant, this will be reduced if the pile is regularly turned.
Following the removal of the soils from the treatment location, the underlying area
should be validated to confirm that contamination has not migrated vertically through
the underlying liner. Where ex-situ treated soils are reinstated on the site, they must
first be validated to ensure that they meet land use and/or validation criteria relevant to
the site and its setting. Where treatment occurs off site, the material must be validated
prior to import back onto the site.
It is noted that monitoring, appropriate cover and potential treatment of vented gases
and leachate should be undertaken. Also excess nutrients where these have been
added.

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6. Health and safety

Bioremediation projects can expose site workers to safety and health hazards via
exposure to the biological agents involved, exposure to vapours, noise, and slip/trip
hazards. The specific remediation technology and design will determine the specific
risks which should be assessed as part of the RAP.
Some of the hazards associated with bioremediation and control mechanisms are
outlined in Table 4. The list is intended to provide an indication of the hazards
potentially associated with soil washing application. They will vary significantly from site
to site and the list is not intended as a substitute for a detailed hazard assessment of
the operation, which should be provided in the RAP.
Readers are directed to the NRF Guideline on health and safety for further information
on health and safety on remediation sites, including risk assessment, the hierarchy of
controls and suggested documentation.

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Table 4 Common bioremediation hazards and controls

Hazard Sources of exposure Suggested controls

Process Chemicals • Splashing or leaking chemicals used to facilitate • Use appropriate storage containers
the biodegradation process
• inspect containers for leaks and damage
• Responding to an emergency release of process
• Install eye wash and emergency shower
treatment chemicals or fuel (for excavators etc)
• Prepare and train for spill containment
• Appropriate bunding to assist with containing any spills
• Ensure use of personal protective equipment (PPE)
Site Contaminants • Off-gassing or releasing contaminants during • Work ‘up-wind’ of disturbed soil, when possible
excavation and spreading/storage and handling of
• Segregate treated feedstock until tested
soil to be remediated
• Routinely monitor work areas; some contaminants require
an initial assessment of exposure (e.g. lead)
• Ensure use of PPE
Dust • Releasing dust while excavating soils and placing • Spray water or use dust suppressants on storage piles
in treatment area/zone, and in the addition of
• Do not operate earth moving equipment during high winds
reagents
• Ensure use of PPE
• Release of pathogens during or after treatment
Ergonomic Risks • Lifting or performing any other movement with too • Provide conveniently located equipment for the job, like
much force and/or in an awkward position, or correctly sized tools
repeating the lift/movement too often
• Train workers on ergonomic risks and prevention

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Hazard Sources of exposure Suggested controls

Slips, Trips and • Storing construction materials or other • Keep walking and working areas free of debris, tools, etc
Falls unnecessary items on walkways and in work areas
• Keep walking and working areas as clean and dry as
• Creating and/or using wet, muddy, sloping, or possible
otherwise irregular walkways and work surfaces
• Perform a Job Hazard Analysis
• Constructing and/or using improper walkways,
• Ensure use of PPE, including fall arrest systems
stairs, or landings or damaging these surfaces
• Train workers on fall hazards and use of ladders
• Creating and/or using uneven terrain in and around
work areas • Use an observer (spotter or signal person) when visibility is
limited
• Working from elevated work surfaces and ladders
• Using damaged steps into vehicles
Moving Vehicles • Moving and stockpiling untreated and treated soils • Train equipment and vehicle operators on limitations of
using earth moving equipment equipment and drivers
• Train equipment and vehicle operators in safe operation
• Set acceptable speed limits and traffic patterns. Ensure that
equipment has, and workers use, back-up alarms, mirrors,
and seat-belts
• Set parking brake and if on incline, chock wheels
• Ensure equipment has required roll-over equipment
• Establish vehicle inspection schedules and procedures
• Do routine maintenance

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Appendix A – Case studies

The following case studies illustrate implantation of bioremedation:

• Transforming Ebbw Vale, Wales, UK:
- http://www.wrap.org.uk/sites/files/wrap/case%20study%20-
%20Ebbw%20Vale.pdf
• Windrowing on Former Industrial Land, Wales, UK:
- http://www.churngold.com/case-studies/remediation/in-situ-and-ex-situ-
bioremediation-of-hydrocarbons-and-japanese-knotweed-treatment.html
• Biopiling at a Former Dairy, England, UK:
- http://www.churngold.com/case-studies/remediation/pcb-mechanical-
segregation-and-ex-situ-bioremediation-of-hydrocarbons-heating-oil.html
• Penny’s Bay Dioxin Site, Hong Kong:
- http://www.thiess.com.au/projects/pennys-bay-dioxin-site/detail
Information on the following case studies can be found in US EPA (1997):
• Land Treatment at the Burlington Northern Superfund Site, Brainerd/Baxter,
Minnesota;
• Composting at the Dubose Oil Products Co Superfund site, Cantonment,
Florida;
• Slurry Phase Bioremediation at the Southeastern Wood Preserving Superfund
site, Canton, Mississippi;
• Cost report: Windrow Composting to Treat Explosives-Contaminated Soils at
Umatilla Army Depot Activity (UMDA);
• In situ Bioremediation Using Horizontal Wells, US Department of Energy, M
Area, Savannah River Site, Aiken, South Carolina; and
• Lasagna Soil Remediation at hat the US Department of Energy Cylinder Drop
Test Area, Paducah, Kennedy.
Information on all the following (and more) US based case studies can be found in the
Federal Remediation Technologies Roundtable case studies database, available at
http://costperformance.org/search.cfm:
• In situ Bioremediation and Soil Vapour Extraction at the Former Beaches
Laundry & Cleaners (2010);
• In situ Bioremediation of Perchlorate and Nitrate in Vadose Zone Soil using
Gaseous Electron Donor Injection Technology in Sediment at Hunters Point
Shipyard Parcel F, San Francisco Bay, California (2010);
• In situ Bioremediation of Chlorinated Solvents Source Areas with Enhanced
Mass Transfer at the Fort Lewis, Pierce County, Washington (2009);
• Enhanced In situ Anaerobic Bioremediation of Chlorinated Solvents at FF-87,
Former Newark Air Force Base, Ohio (2007);

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• Enhanced In situ Anaerobic Bioremediation of Chlorinated Solvents at the

Hangar K Site, Cape Canaveral Air Force Station, Florida (2007);
• Pump and Treat and In situ Bioventing at Onalaska Municipal Landfill
Superfund Site, Onalaska, Wisconsin (2006);
• In situ Bioremediation Using Hydrogen Release Compound (HRC®) at Four
Dry Cleaner Sites, various locations (2005);
• In situ Bioremediation Using HRC® at a Former Industrial Property, San Jose,
California (2004);
• Ex Situ Bioremediation at Two Dry Cleaner Sites, various locations (2001);
• In situ Bioremediation Using Oxygen Release Compounds (ORC®) at an
Active Service Station, Lake Geneva, Wisconsin (2001); and
• In situ Bioremediation at the Texas Gulf Coast Site, Houston, Texas (2000).

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Appendix B – Treatability case studies

Table 5, presented overleaf, includes a summary of documents that may be useful in

conducting treatability studies. References are available in Appendix C.

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Table 5: Summary of treatability case studies

Technology References Comments

Bioventing • US EPA (1995a) • Field studies may include an in-situ respiration test and a soil gas permeability test:
• US EPA (1995b) • In situ respiration test: determines whether microbial activity is occurring and whether oxygen
is limited.
• USAF CEE (2004)
• Soil gas permeability test: assists with determining the permeability and radius of influence and
provides additional in situ respiration data
• If the results of such field tests are positive the installation of an air injection and/or extraction
system may be considered.
Enhanced • USAF CEE (2004) • These documents discuss site selection, design, and performance criteria for enhanced
Bioremediation bioremediation as a remedial alternative, select specific enhanced bioremediation approaches
• US ACE (2010a)
that are suitable for achieving remedial goals, and track the cost and performance of enhanced
• US ACE (2010b) bioremediation applications.
• The addendum involved the evaluation of 15 case studies for system design, operation, and
performance and outlines advances made in the field of enhanced in situ bioremediation of
chlorinated solvents, and provides resources and references that can be used to identify and
mitigate the limiting factors and challenges that practitioners face when applying the
technology.

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Technology References Comments

Phytoremediation • US EPA (2000) • Results of studies pertaining to a wide range of site conditions, plants and contaminants have
been assessed under laboratory and field testing conditions. Information is provided on
undertaking site-specific laboratory tests and small scale field trials, including the growing of
plants in contaminated soil and water collected from the site, rather than from soils that have
been spiked with a known concentration of a contaminant. Pilot studies can include a number
of plant species, soil pH and several chelates, which assists with determining the best
combination of variables.
• In general, treatability studies should provide information regarding the following experimental
factors: reduction in contaminant; phytotoxicity of contaminants at the site; growth of plant
species under site specific conditions and the rate and level of clean up.
Mycoremediation • Singh (2006) This book provides information on techniques to identify, select, and apply fungi for the
remediation of contaminated sites, including degradation of specific waste streams and classes of
contaminants such as: industrial wastewaters; distillery and brewery wastes; dyes; pulp and paper
mill effluents; petroleum hydrocarbons; polychlorinated biphenyls and dioxins; phenols,
chlorophenols, and pentachlorophenol; polycyclic aromatic hydrocarbons; and pesticides.
Information is also provided on fungal biosorption of heavy metals and mycorrhizal fungi in
rhizosphere remediation.
Biopiles / • NAVFAC (1996) Information is provided on the assessment, application and design and operation of biopile
windrows systems. Information is included on determining the feasibility of biopile remediation, including the
data required and a selection process.
Composting • Vallini et al (2002) An overview is provided on the potential use of composting in organic waste recycling and
decomposition of contaminants in contaminated soils and sediments. In the latter case it is
important that the degradation proceeds to the fullest extent with formation of innocuous end
products, and information is provided on evaluating the conditions that can hinder the process in
different situations.

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Technology References Comments

Landfarming • NSW EPA (2014) • The NSW document outlines the requirements for landfarming, including requirements for a
treatability study.
• MDA (2005)
• The Bioremediation Treatability Study Fact Sheet includes information on the requirements for
• NEZ Perce Tribe
treatability studies for bioremediation including landfarming.
(2009)
• The NEZ Perce Tribe Soil Landfarming guidance includes information on treatability studies.
Slurry Phase • US EPA (1993) Information is provided on the planning and carrying out biodegradation treatability studies,
Biological including slurry phase biological treatment.
• US EPA (1995)
Treatment

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Appendix C – References

ICSCS, 2006, Manual for biological remediation techniques, International centre for soil
and contaminated sites, Dessau, Germany.
MDA, 2005, Bioremediation treatability fact sheet, Guidance document no 17,
Minnesota Department of Agriculture, Pesticide and Fertilizer Management
Division, Minnesota.
NAVFAC, 1996, Biopile design and construction manual, Technical Memorandum no
TM-2189-ENV, United States Naval Facilities Engineering Service Centre, Port
Hueneme, CA.
NAVFAC, 2015, Design considerations for enhanced reductive dechlorination,
Technical Report no TM-NAVFAC-EXWC-EV-1501, United States Naval
Facilities Engineering Service Centre, Port Hueneme, CA.
NEZ PERCE TRIBE, Nez Perce Tribe soil landfarming guidance, Nez Perce Tribe
Contaminated Site Cleanup Group,
NSW EPA, 2014, Best practice note: Landfarming, New South Wales Environment
Protection Authority, Sydney.
OSWER, 2010, Green remediation best management practices: Bioremediation, EPA-
542-F-10-006, United States Environmental Protection Agency, Office of Solid
Waste and Emergency Response and Office of Research and Development,
Washington, DC.
SINGH, 2006, Mycoremediation: Fungal biogremediation, John Wiley & Sons, New
York.
US ACE, 2010, Loading rates and impacts of substrate delivery for enhanced
anaerobic bioremediation, ESTCP project ER-0672, United States Army Corps
of Engineers, Washington, DC.
US ACE, 2010, Loading rates and impacts of substrate delivery for enhanced
anaerobic bioremediation: Addendum to the principles and practices manual,
ESTCP project ER-200627, United States Army Corps of Engineers,
Washington, DC.
US EPA, 1993, Guide for conducting treatability studies under CERCLA:
Biodegradation remedy selection: quick reference fact sheet, EPA/540/R-
93/519b, United States Environmental Protection Agency, Cincinnati, OH.
US EPA, 1993, Guide for conduting treatability studies under CERCLA: Biodegradation
remedy selection (interim guidance), EPA/540/R-93/519a, United States
Environmental Protection Agency, Washington, D.C.
US EPA, 1995, Manual: Bioventing principles and practice: Bioventing design,
EPA/540/R-95/534b, United States Environmental Protection Agency,
Washington, D.C.
US EPA, 1995, Manual: Bioventing principles and practice: Bioventing principles,
EPA/540/R-95/534a, United States Environmental Protection Agency,
Washington, D.C.
US EPA, 1997, Innovative uses of compost: bioremediation and pollution prevention,
EPA530-F-97-042, United States Environmental Protection Agency,
Washington, D.C.
US EPA, 1998, An analysis of composting as an environmental remediation
technology, EPA530-R-98-008, United States Environmental Protection
Agency, Washington, D.C.

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US EPA, 2000, Introduction to phytoremediation, EPA/600/R-99/107, United States

Environmental Protection Agency, Washington, D.C.
US EPA, 2010, Fact sheet - Phytotechnologies for site cleanup, EPA 542-F-10-009, US
EPA Office of Superfund Remediation and Technology Innovation, Washington,
D.C.
USAF CEE, 2004, Principles and practices of enhanced anaerobic bioremediation of
chlorinated solvents, United States Air Force, Centre for Environmental
Excellence, Naval Facilities Engineering Service Centre and Environment
Services Technology Certification Program,
USAF CEE, 2004, Procedures for conducting bioventing pilot tests and long-term
monitoring of bioventing systems, United States Air Force, Centre for
Environmental Excellence,
VALLINI, DI GREGORIO, PERA & CUNHA QUEDA, 2002, Exploitation of composting
management for either reclamation organic waste or solid-phase treatment of
contaminated environmental matrices, Environmental Reviews, Vol. 10(4), pp
195-207.
WILLIAMS & KEEHAN, 1992, Bioremediation using composting, Proceedings of 1992
National Waste Processing Conference : fifteenth biennial conference : solid
waste processing into the 21st century Solid Waste Processing Division,
American Society of Mechanical Engineers, New York, N.Y.

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