&EPA
United States
Environmental Protection
Agency
Office of Research and
Development
Washington DC 20460
Center for Environmental
Research Information
Cincinnati OH 45268
Technology Transfer
February 1989
CERI-89-11
Bioremediation of
Hazardous Waste Sites
Workshop
Speaker Slide Copies and
Supporting Information
-------
-------
Table of Contents
Section 1
Basic Requirements for Implementing Biological
Systems to Remediate Hazardous Wastes 1-1
Abstract 1-2
Slides 1-9
Section 2
Initial Data Requirements 2-1
Abstract 2-2
Slides 2-11
Worksheets 2-42
Section 3
Example Site for Bioremediation 3-1
Section 4
Reactor Design 4-1
Abstract 4-2
Slides 4-10
Worksheets 4-49
Section 5
In Situ Design 5-1
Abstract 5-2
Slides 5-14
Worksheets 5-47
-------
BASIC REQUIREMENTS FOR
IMPLEMENTING BIOLOGICAL
SYSTEMS TO REMEDIATE
HAZARDOUS WASTES
SECTION 1
Abstract
Slides
1-2
1-9
1-1
-------
BASIC REQUIREMENTS FOR IMPLEMENTING BIOSYSTEMS
John Glaser A1 Venosa Bill Mahaffey
U.S. EPA U.S. EPA Ecova
Cincinnati, Ohio Cincinnati, Ohio Redmond, Washington
I. Introduction
The key to the assessment of the fate of organic chemicals in the
environment is a realistic evaluation of their susceptibility to
biological conversion. In order to make this evaluation rationally, it
is important that the terminology used in the field is understood. The
discussion presents some terms needed to understand the rest of the
presentation. The following terms are defined: mineralization,
biodegradation, recalcitrant compounds, persistent compounds, biogenic
compounds, xenobiotic compounds, and biosystems.
Biological technology development is based on: (1) an adequate
information base, which is derived from an understanding of microbiology,
biochemistry, and genetics; (2) a basic understanding of the metabolic
processes leading to the detoxification of hazardous wastes; and (3) an
understanding and appreciation of the structure and function of natural
microbial communities.
The key word above is "understanding." Without understanding the
underlying microbiology, developing the technology becomes sheer
guesswork. Thus, basic science research must be a part of any program
concerned with biodegradation technology development.
II. The Carbon Cycle
Carbon plays a key role in the structural make-up of protoplasm and
its essentiality in the energy metabolism of heterotrophs. The
biogeochemistry of carbon is interesting because of the vast array of
organic molecules that are involved and the cyclical nature of the
interaction between these compounds and inorganic carbon, a cycle that
describes the movement of carbon from the inorganic to the organic state
and back to the inorganic again. Movement of organic carbon to the
inorganic state is accomplished either through direct combustion or
through the action of microbial biooxidation.
Biotransformation of organic pollutants is accomplished either
aerobically or anaerobically.
A. Aerobic metabolism
1. Aerobic respiration: energy-yielding metabolism involving
oxidation reactions in which hydrogen (electrons) is
transferred to oxidized pyridine nucleotides (NAD and NADP)
1-2
-------
resulting in reduced forms (NADH and NADPH) that either
provide reducing power for biosynthetic reactions or can
transfer the electrons to electron transport chains wherein
high energy bonds of ATP are formed. The final electron
acceptor is molecular oxygen.
2. Compounds devoid of oxygen atoms (alkanes, saturated ring
structures, and unsubstituted benzenes) can still be acted
upon by certain microorganisms by their unique ability to
catalyze oxidations using molecular oxygen. They do this
through the mediation of two types of enzymes, both of which
activate oxygen from the triplet state to the singlet state.
a. Monooxygenases: R-H + NADH + H+ + 02 = R-OH + NAD+ + H20
Monooxygenases yield hydroxyl groups, and all are
extremely specific for their aromatic substrate.
b. Dioxygenases: R + 02 = R02
Dioxygenases are responsible for the fixation of the oxygen directly
into organic compounds. A common use of dioxygenases is to cleave
the benzene rings by inserting both atoms of the molecular oxygen.
Before this can occur, however, the ring must contain two hydroxyl
groups placed ortho or para to each other. Like the monooxygenases,
the dioxygenases are highly specific for their substrates. Once the
ring is cleaved, the product can enter more common degradative
pathways.
B. Anaerobic metabolism. Many compounds can be mineralized
anaerobically, yielding carbon dioxide and methane. The aromatic
ring is first reduced to a cyclohexanone, then cleaved to an
aliphatic acid. Reduced coenzymes must be available for such
reactions.
1. Anaerobic respiration: energy-yielding reactions in which the
final electron acceptor is a compound other than molecular
oxygen, such as sulfate or nitrate.
2. Fermentation: anaerobic reactions in which the final product is
partially oxidized organic compound such as organic acid.
C. Reactions involving organohalides. Organohalides have been around
for millennia, and microorganisms have had a long time within which
to develop methods for dealing with them.
1. In aerobic environments, metabolism of haloaromatic compounds
that contain only one or two halides generally leave the
carbon-halogen bond intact until the aromatic ring has been
cleaved by the oxygenases. Thereafter, dehalogenation usually
1-3
-------
occurs by elimination of the halogen as the hydrogen halide,
with subsequent double-bond formation in the aliphatic
intermediate.
2. Dehalogenations have also been observed in anaerobic
environments from both alkyl and aromatic halides. In both ^
cases the halide is apparently replaced by hydrogen. Mechanisms
have not been worked out yet but obviously require reducing
power.
Some haloorganics appear to require anaerobic conditions for
dehalogenation to occur whereas others require aerobic. This means that
the environment within which biodegradation is attempted may well be a
critical factor in the outcome.
III. Mechanisms for Attacking Xenobiotics
Bacteria can only do those things for which they have a genetic
capability. If biodegradation requires the presence of enzymes, if
enzymes are synthesized in response to the presence of a recognizable
substrate, and if the genetic capability of a bacterium which allows it
to synthesize those enzymes has evolved over time in response to its
environment, how can biodegradation of xenobiotic compounds be achieved?
The answer to those questions lies in the fact that the stereospecificity
of enzymes is not exact.
A. Gratuitous biodegradation: reactions involving enzymes having high
substrate specificity with respect to their catalytic function but
low specificity with respect to substrate binding. It is not
uncommon for enzymes to bind analogs of the natural substrate which
contain xenobiotic functional groups. The success of gratuitous
metabolism depends on:
1. Ability of xenobiotic to induce requisite enzymes.
2. Nature of product
a. More toxic, either to organism or to other organisms.
b. Less susceptible to further microbial attack, leading to
persistence.
c. More susceptible to bioaccumulation.
d. Coordinate induction of many enzymes. May involve whole
pathways through the combined efforts of many organisms
within a community.
B. Cometabolism. In the situation in which an organism cannot extract
energy and reducing power from metabolic reactions, the only way in
which they can effect continual biodegradation of the xenobiotic
compound is through the use of additional carbon and energy sources
supplied externally or from the action of other organisms in a mixed
1-4
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microblal community. Cometabolism 1s the transformation of a
non-growth substrate in the obligate presence of a growth substrate
or another transformable compound. Two key concepts are involved
here.
1. The non-growth substrate is one that will not support cell
division.
2. There must be a growth substrate present in order for the
transformation to occur.
C. Fate of products resulting from gratuitous metabolism and
cometabolism.
1. If the transformed product is more toxic than the original
compound, it will accumulate. If the transformed product is
less toxic, the process may continue until it has been converted
to a biogenic structure that fits into the normal metabolism of
the cell. If the xenbbiotic compound is cometabolized by a pure
culture, then metabolic products will always accumulate. If it
is cometabolized by an organism in a mixed culture, it may well
not result in accumulation but rather be metabolized by other
species in the consortium. Thus, it is possible that the
compound may be completely degraded, even if there is no single
organism in the community that can totally degrade it itself.
THIS MEANS THAT THE CAPACITY TO SERVE AS THE SOLE CARBON AND
ENERGY SOURCE FOR GROWTH OF A PURE (OR ANY) MICROBIAL CULTURE IS
NOT AN APPROPRIATE CRITERION BY WHICH TO JUDGE THE
BIODEGRADABILITY OF A XENOBIOTIC COMPOUND. BECAUSE OF THE
SIGNIFICANCE OF COMETABOLISM AND MICROBIAL INTERACTIONS,
BIODEGRADABILITY CAN ONLY BE ACCURATELY ASSESSED IN
MIXED-CULTURE, MIXED SUBSTRATE SYSTEMS.
D. Requirements associated with the use of mixed-substrate systems.
1. Control of enzyme synthesis acts to conserve carbon and energy
when the cell could not really benefit from having the enzyme
present.
2. Control of enzyme activity is more rapid because it acts to
influence the rates of enzymes that are already present.
Classical batch studies place small inocula of bacteria into
contact with high concentrations of substrate. Consideration of
the above control mechanisms suggests that the presence of high
concentrations of easily degradable substrates could well
prevent the synthesis of the very enzymes needed to degrade a
compound of interest.
1-5
-------
The concentration of the compound being tested for
biodegradability is another factor of importance. The
concentration must be high enough to induce the enzymes needed
for its transformation, but low enough either not to be toxic
itself or its intermediates not to be toxic.
Importance of microbial communities: consortia. The complete
mineralization of a compound may require the sequential
metabolism of two or more organisms because no single species
within the culture contains complete genetic complement of the
whole culture.
a. Typical interaction within communities. Organisms within
microbial communities involved in the degradation of
xenobiotics have been classified by some as falling into two
groups: the primary utilizers and the secondary organisms.
The primary utilizers are those species capable of
metabolizing the sole or major carbon and energy substrate
provided to the system. The secondary organisms cannot use
the major substrate but, instead, rely on the utilization of
products released by the primary utilizers.
b. Importance of communities in adaptation. Mixed microbial
communities have distinct advantages over pure cultures.
This is because the biodegradative capacity of a community
is much greater, both qualitatively and quantitatively,
particularly where xenobiotic compounds are involved.
Furthermore, the resistance of a community to toxic
substances may be much greater because there is a greater
likelihood that an organism that can detoxify them will be
present. Finally, mineralization of xenobiotic compounds
sometimes requires the concerted activity of multiple
species.
If a compound is degraded by the concerted action of several
organisms, it is likely that the community will develop
stepwise. That is, a product may accumulate until an
organism that can degrade it becomes established. This
suggests that development of the community will be expedited
by continually seeding it rather than placing organisms into
it at one time.
c. The Ubiquity Principle states that "...all types of bacteria
are available at all times everywhere..." Hence, natural
population selection mechanisms will always result in the
right biological culture for treatment of a given waste.
1-6
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IV. Requirements for Successful Biodegradation
A. A capable organism or community must be present. With a single
axenic culture, the chances of finding a capable organism are
remote if substrate is the least bit peculiar. With a single
mixed culture inoculum, chances are somewhat better because of
the diverse genetic potential of the inoculum. Long-term
continuous inoculation with organisms from diverse sources offer
best potential for success.
B. Conditions must be adequate for enzyme induction. This is most
likely to occur under carbon-limited conditions. Thus, batch
shaker studies with multiple carbon sources are inappropriate.
A supply of energy is needed for enzyme synthesis. This is best
accomplished with continuous culture wherein the carbon source
concentration is kept low and energy source is constantly
provided.
Induction may require an intracellular inducer, and entrance of
the inducer may require energy. A steady, continuous supply of
energy under carbon-limited conditions is best.
Gratuitous or cometabolic biodegradation favors a supply of an
auxiliary biogenic carbon source. The best course is to supply
a diverse mix of compounds.
C. The concentration of test compound is important. Too high may
be toxic. Too low may be inadequate for enzyme induction.
D. The proper aerobic or anaerobic environment must be provided for
growth of the requisite organisms.
E. The physical-chemical characteristics of the compound must be
considered, including such properties as volatility,
absorbability, and solubility.
F. Methods to enhance biodegradation include: (1) applying
physiological information (i.e., knowledge of the proper
morphological and physiological state of the organism is
essential to achieve enhanced activity); (2) adjusting
environmental conditions; or (3) applying genetic engineering
techniques. The mechanisms of gene transfer will be discussed
here.
V. Reference Reading
The reader is referred to the following references for detailed
discussions of the above information.
1-7
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Grady, C.P.L. 1985. "Biodegradation: its measurement and
microbiological basis." Blotechnol. Bloeng., XXVII, 660-674.
Rehm, H.J. and G. Reed. 1981. "Biotechnology. Vol. 1, Microbial
Fundamentals." Verlag Chemie, Weinheim, Deerfield Beach, FL.
"Technology screening guide for treatment of CERCLA soils and
sludges." Sept., 1988. EPA/540/2-88-004.
"Groundwater handbook." March, 1987. EPA/625/6-87/016.
"Review of in-place treatment techniques for contaminated surface
soils. Vol. 1: technical evaluation." Sept., 1984.
EPA-540/2-84-003a.
Gibson, D.T. 1984. "Microbial degradation of organic compounds."
Marcel Dekker, New York.
Rochkind, M.L., J.W. Blackburn, and G.S. Saylor. Sept., 1986.
"Microbial decomposition of chlorinated aromatic compounds."
EPA/600/2-86/090.
Callahan, M.A., et al. Dec., 1979. "Water-related environmental fate
of 129 priority pollutants. Vols. 1 and 2." EPA-440/4-79-029a and b
1-8
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Toxic/
Hazardous
Waste
Influent
BIOREMEDIATION
"The Black Box"
Clean
Effluent
NOTES
NOTES
OBJECTIVES
• Introduce concepts and terminology
of Btodegradation/BRxemedfetion
• Discuss factors that influence biodegradation
• Discuss the benefits /imitations
of this technology
• Generaly provide an increased comfort
level with this technology
by deimiting the Back Box Concept
ON-STTE TREATMENT AND
REMEDIATION OF TOXIC
AND HAZARDOUS MATERIAL
NOTES
1-9
-------
SITE SPECFIC SYSTEMS
• Biological
• Chemical
• Physical
• On-site engineering
NOTES
NOTES
BENEFITS OF BIOREMEDIAT1ON
• Terminal destruction
• On site
• EnvironmentaOy sound
• Cost effective
MINERALIZATION
The conversion of organic
chemicals to carbon cfoxide
and/or methane, water, and
various inorganic forms.
Cl
r J—>COZ + NH3* + Cl" +• H20 t BIOMASS
NH2
1-10
NOTES
-------
BIODEGRADATION
The biological transformation of an
organic chemical to another form
withouLregardJLO_extent Biologists,
however, usually use biodegradation as
a synonym for mineralization.
NOTES
NOTES
PERSISTENT COMPOUND
A chemical that fais to undergo
biodegradation under a specified set
of conditions. A chemical may be
inherently biodegradable yet persist
in the environment
PCBs IN HUDSON RIVER
AEROBIC
AROCHLOfi 1254
MINIMAL DEGRADATION
-> EXTENSIVE TRANSFORMATION
RECALCITRANT/REFRACTORY
COMPOUND
A chemical that has an
iiherent resistance to any
degree of biodegradation,
Toxaphene, Dieldrin, Endrii
l-il
NOTES
-------
BIOGENIC COMPOUNDS
Naturally occurring compounds
that have been present for
milions of years. Thus, there
are organisms somewhere in the
biosphere that can Initiate their
biodegradatioti.
NOTES
NOTES
XENOBlOTrC COMPOUNDS
Compounds that are "foreign"
to the biosphere, having been
present for only an instant on
the evolutionary tine scale. May
or may not be biodegradable.
ADAPTATION/ACCLIMATION
An increase in the btodegradation rate
of a chemical after exposure of the
microbial community to the chemical
for some period of time.
1-12
NOTES
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IDEALIZED EXAMPLE OF ADAPTATION/ACCLIMATION
o 160 _
R«iult of idiptatlon
Dtyt
CO production
Mlcroblal blomitt
Ch*mlc«l
conc*ntnUon
NOTES
NOTES
BIOREMECHATrON
The manipulation of living systems
to bring about desired chemical and
physical changes in a confined and
regulated environment
BIOREMEDIATION
Hybrid Of:
* Microbiology
• Ecology
• Biochemistry
• Chemical engineering
• Environmental engineering
• In-situ technology (hydrogeology
and soil science)
• Risk management
NOTES
1-13
-------
MICROORGANISMS
OXYGEN
CHEMICAL
PHYSICAL
BIOLOGICAL
NUTRENTS
NOTES
NOTES
BASIC MICROBIOLOGY
Ecology
Physiology
Genetics
BASIC MICROBIOLOGY
Ecology
Interaction of a microorganism
and its environment
(physical, chemical)
NOTES
1-14
-------
BASIC MICROBIOLOGY
Physiology
Processes by which any organism obtains
food and energy for biosynthesis and
performing other work
(Chemical energy- ->Biological energy)
(proteins, enzymes, cell structural parts)
BASIC MICROBIOLOGY
Genetics
The equivalent of a computer
program. Codes of information
which control or dictate the
physiology of an organism in
response to its environment.
(DNA, genes)
1-15
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ATMOSPHERE
HYDROSPHERE
BACTERIAL
PHOTOSYNTHESIS
PLANT
PHOTOSYNTHESIS
CHEMICAL
FIXATION
ORGANIC
CARBON
ORGANIC
CARBON
ANIMAL
CONSUMPTION
FERMENTATION
PUTREFACTION
DECAY
ORGANIC
CARBON
NONLIVING
ORGANIC
CARBON
COMBUSTION
.FOSSIL
FUELS
RESPIRATION
HAZARDOUS
ORGANICS
CARBON CYCLE
BASIC PREMISES OF
BIODEGRADATION
1. Organic compounds are converted to
simpler structures by the action of
microorganisms as part of the con-
tinual cycling of carbon in nature.
2. Microorganisms generally derive the
nutritional and energy requirements
necessary for growth from the com-
pounds they degrade.
1-16
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BASIC PREMISES OF
BIODEGRADATION
(Continued)
3. BkxJegradation occurs in a wide variety
of environments through the action of
microorganisms using processes deter-
mined by environmental factors.
4, Enzymes evolved throughout time for the
degradation of naturally occurring
organics can be recruited to degrade
man-made waste materials.
NOTES
NOTES
BASICS OF PHYSIOLOGY
Cell composed of macromotecutes
(proteins, polysaccharides, iptds,
nucleic adds)
Basic buidmg blocks are amino
acids, carbohydrates, fatty acids,
nucleic acids
BASICS OF PHYSIOLOGY
(Continued)
> Cells synthesize components from
multitude of nutritional and
energy sources
> Intermediary metaboism - - central
mechanism by which eels process
and harness chemical energy to
produce biomass and energy
1-17
NOTES
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INTERMEDIARY METABOLISM
ORGANIC
PROTEINS CARBOHYDRATES LIPIOS CHEMICALS
PYRUVATE
ACETYL CoA
ELECTRONS:
REDUCING EQUIVALENTS
(XH2)
END PRODUCTS
NH3,C02.H20
INTERMEDIATES
AM I NO ACIDS
NUCLEIC ACIDS
FATTY ACIDS
CARBOHYDRATES
ORGANIC C
MICROBIAL
DEGRADATION
\
INTERME
METABO
N 2 OR S '^-'•y — *• * \
NOj OR S04 /^N—XH2 -^— '
ANAEROBIC RESPIRATION
OMPOUNDS
^
f \ ORGANIC
DIARY \ J ^ ACIDS,
FFRMPNTATlON RtLATcu
COMPOUNDS
, X-«^y-**H20 |
^-^-XH2 — 02 ACETATE,
FORMATE,
AEROBIC RESPIRATION C02,H2
C02
CH,
CENTRAL REACTIONS IN MICROBIAL METABOLISM
1-18
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BIODEGRADATTON
PATHWAYS
AEROBIC
RESPIRATION
EXAMPLE
HEXANE
ANAEROBIC BENZOATE
RESPIRATION
FERMENTATION PHENOL
END PRODUCTS
C02 , H20
ORGANIC ACIDS
NO j
ORGANIC ACIDS
C02. CH4
MICROBE
PSEUDOMONAS
PSEUDOMONAS
METHANOGENIC
NOTES
NOTES
AEROBIC RESPIRATION
Energy-yielding metabolism in
which the terminal electron
acceptor for substrate oxidation
is molecular oxygen.
AEROBIC BIODEGRADATION
Oxygen Involved In Two Ways
1. Acceptor of electrons produced from
oxidation reaction resulting in
reduction to water.
Glucose
NOTES
1-19
-------
AEROBIC BIODEGRADATION
Oxygen Involved In Two Ways
(Continued)
2. Important substrate for oxygenase enzymes,
which incorporate molecular oxygen into
relatively unreactive compounds:
EXAMPLES OF OXYGEN INVOLVEMENT IN
AEROBIC BIODEGRADATION
°2 ,CH(CH)4 COOH
CH3 (CH2)6CH3 -^ ^ CH3 (CH2)6 - CH2 OH t H20 1
^S*S*-
XH2 x CH3 CHO
1-20
-------
EXAMPLES OF OXYGEN INVOLVEMENT IN
AEROBIC BIODEGRADATION
PYRUVATE
^
OH.
XH2
ANAEROBIC RESPIRATION
Energy-yielding metabolism in which
the terminal electron acceptor for
substrate oxidation is an inorganic
compound other than molecular
oxygen, such as sulfate or nitrate.
1-21
-------
FERMENTATION
Energy-yielding metabolism that
involves a sequence of oxidation -
reduction reactions in which both
the substrate (primary electron donor)
and the terminal electron acceptor are
organic compounds.
FERMENTATION OF BENZOATE UNDER
METHANOGENIC CONDITIONS
[4HJ
—^+~\ /-O
:OOH
H,0
BENZOATE
CH3 fafl
21 V
COOH •* N
CH4-t-C02
Y \COOH 1—
~(~ \COOH
CYCLOHEX-I-ENECARBOXYLATE 2-H YDROX YCYCLOHEX ANE-2-OXOCYCLOHEXANE-
CARBOXYLATE CARBOXYLATE
CH2
COOH
BUTYRATE •*-
COOH
ACETATE
&H)
CH2
-------
ANAEROBIC BIODEGRADATION
Anaerobes Require Electron
Acceptors Other Than Oxygen
With Reduction To Characteristic
Products:
CO2 ~^ Methane Methanogens
NOs —*> N2 Denitrifiers
804 —> H2S Sulfate reducers
Glucose —> Lactate Fermenters
Ethanol
LIMITED DEGRADATIVE POTENTIAL BUT SEVERAL NOVEL
REDUCTION REACTIONS (DEHALOGENATION, ETHER CLEAVAGE)
GRATUITOUS METABOLISM
Reactions involving enzymes having
high substrate specificity with
respect to catalytic function but
low specificity with respect to
substrate binding
1-23
-------
RELATIONSHIP BETWEEN ENZYME ACTION
AND GRATUITOUS METABOLISM
on luffoce
t
F.e.tniymc
j/me, OCIiv« lilc
on lurloce
tniyme-lubllrow
Eod ptoducit
COMETABOLISM/COOXIDATION
The transformation of a non- growth
substrate in the obligate presence
of a growth substrate or another
transformable compound.
1-24
-------
NON-GROWTH SUBSTRATE
A substrate that will not
support cell division.
There must be a growth
substrate present in
order for the transformation
to occur.
1-25
-------
COOXIDATION EXAMPLE
a;
. COOH
H OH Ring fiction _HO
"OH
PYRUVATE
aOOH
H
~
^
PYRUVATE
R- CI,S03,CH3
INDUCIBLE ENZYMES
Enzymes produced by a cell in
response to a specific compound
which is referred to as the
inducer.
1-26
-------
CONSTITUTIVE ENZYMES
Enzyme(s) always produced by a
cell regardless of the nature
of the medium. An inducer
compound is not required
for the enzyme(s)
formation.
NOTES
NOTES
.14.
MINERALIZATION OF C LABELLED
BENZANTHRACENE BY BEIJERINCKIA B1
I «
"-* O
Constitutive
I I I I I I I I I I T
0 2 4 • a 10 12 14 1« 18 20 22 24
Tim. (Hourt)
ENVIRONMENTAL FACTORS
LIMITING BIODEGRADATION
Biological
• Active viable biomass
• Physiological limitations
• Electron acceptors
• Predation
1-27
NOTES
-------
ENVIRONMENTAL FACTORS
LIMITING BIODEGRADATION
Physical
• Temperature
• Availability of chemical
• Surface adhesion
• Access to substrate
• Light
Properties of Some PAH Compounds
Aq. Log Log
COMPOUND SOLUBILITY Kow Koc
9/1
CO
31.7 3.37 3.11
NAPHTHALENE
1.29 4.46 4.36
PHENANTHRENE
0.135 5.32 4.92
r
PYRENE
0.0038 6.04 6.65
BENZO(o)PYRENE
1-28
-------
ENVIRONMENTAL FACTORS
LIMITING BIODEGRADATION
Chemical
• pH
• Salinity
• Organic nutrients (vitamins
cofactors, substrates)
• Redox potential (02,
, C02 )
ENVIRONMENTAL FACTORS
LIMITING BIODEGRADATION
Chemical
(Continued)
* Major inorganic nutrients
(N, P, S, Mg, K, etc.)
• Trace elements (Fe, Zn,
Mn, Mo, Co, Cu, Ca)
• Toxic chemicals
• Chemical mixtures
-------
Contaminated soil
• xcavatlon
SOLID PHASE BIODEGRADATION
Oversized material
to special handling
Perforated
Drain pipe -
SOLID PHASE TREATMENT
Soil layer
Sprinkler system
Source: Ecova Corp
1-30
-------
FACTORS CONTROLLING BIOOEGRADATION
(Liquids and Solids)
Factors
Effect
Data Needs
Variable waste
composition
Water solubility
B1odegradab1l1ty
Temperature
outside 25-70'C
range.
Inconsistent blodegradatlon caused
by variation In biological activity.
Contaminants with low solubility are
harder to blodegrade.
Low blodegradablllty Inhibits
process.
Larger, more diverse mlcroblal
population present In this range.
Haste
composition
Solubility
Chemical
constituents,
presence of
metals/salts,
bench-scale
testing
Temperature
monitoring
Nutrient deficiency Lack, of adequate nutrients for
C/N/S ratio
Oxygen deficiency
Moisture content
pH outside
4.5-7.5 range
Microbial
population
Presence of
elevated levels of:
• Heavy metals
• Highly
chlorinated
organics
biological activity (although nutrient
supplements may be added).
Oxygen depletion slows down the
process.
A moisture content of greater than
79% affects bacterial activity and
availability of oxygen. A moisture
content below 401 severely Inhibits
bacterial activity.
Inhibition of biological activity
If Indigenous microorganisms not
present, cultured strains can be
added.
Can be highly toxic to
microorganisms.
Oxygen
monitoring
Ratio of air
to water 1n
interstices,
porosity of
composting
mass
Sludge pH
testing
Culture test
Analysis for
contaminants
1-31
-------
FACTORS CONTROLLING BIODEGRADATION
(Solids)
Factors
Effect
Data Needs
• Some
pesticides.
herbicides
• Inorganic salts
Water and air
emissions and
discharges
(composting only)
Compaction of
compost
(composting only)
Nonunlform
particle
(composting only)
Unfavorable soil
characteristics
• Low permeability
« Variable soil
conditions
• Low soi1 pH
(< 5.5)
• Low soil organic
content
• Low moisture
content (< 101)
Unfavorable site
hydrology
Potential environmental and/or
health Impacts (control achieved
through air scrubbing, carbon
filtration, forced aeration, cement
liner).
Particles tend to coalesce and
form an amorphous mass that is not
easily maintained in an aerobic
environment (wood chips or
shredded tires may be added as
bulking agents).
Waste mixtures must be of uniform
particle size.
Concentrations
of
contaminants
Oetermlne
integrity,
physical nature
of material
Particle size
distribution
Hinders movement of water and Percolation
nutrients through contaminated testing
area.
Inconsistent biodegradation due Soil mapping
to variation in biological activity.
Inhibition of biological activity Soil pH testing
Lack of organic substrate for Soil humus
biological growth. content
Subsurface biological growth Soil moisture
requires adequate moisture. content
Groundwater flow patterns must Site
permit pumping for extraction hydrogeology
and relnjection. must be well
defined.
1-32
-------
FACTORS CONTROLLING BIODEGRADATION
(Groundwater)
Factors Effect Data Needs
Unfavorable
groundwater
quality parameters
• Low dissolved Oxygen necessary for biological Dissolved
oxygen growth. oxygen In
groundwater,
determine
amount of hy-
drogen per-
oxide needed to
satisfy oxygen
demand.
• Low pH, Inhibition of biological activity. pH and alkalinity
alkalinity of groundwater
1-33
-------
COMPARISON OF AVAILABLE TECHNOLOGIES
FOR SOIL TREATMENT
Technology
Organic
Contaminant
Halogenated vdatfec
Hatogenated senivoJaties
Nonhatogenated votettes
Nonhatogenated
semtvolatfes
PCBs
Pesticides
Organic cyanides
Organic corrosives
D. ill uitjuuilvnixu J
1 del i toman ateu
Rotary In -Situ
Kiln Chemical
Incin. Treat.
D
D
D
D
D
D
D
D
effectiveness;
N
N
N
N
N
N
P
P
In -Situ
Bio. Bio.
p
p
p
p
p
p
p
X
p
p
p
p
p
p
p
X
P» potential effectiveness;
N=no offocHvonossj X= potential advorso
to process or
envroranent
bnpacts
COMPARISON OF AVAILABLE TECHNOLOGIES
FOR SOIL TREATMENT
Technology
Rotary In-Situ
Organic Kiln Chemical In-Situ
Contaminant Incin. Treat. Bio. Bio.
Volatile metals X N X X
Nonvolatile metals N N X X
Asbestos N N N N
Radioactive N N X X
materials
Inorganic N P X X
corrosives
Inorganic cyanides P P X X
P=potential effectiveness; N= no effectiveness;
X=potential adverse impacts to process or environment
1-34
-------
COMPARISON OF AVAILABLE TECHNOLOGIES
FOR SOIL TREATMENT
Technology
Rotary In-Situ
Organic Kiln Chemical In-Situ
Contaminant Incin. Treat. Bio. Bio.
Oxidizers
Reducers
D=demonstrated effectiveness; P=potential effectiveness;
X=potential adverse impacts to process or environment
1-35
-------
EXAMPLES OF CONSTITUENTS WITHIN WASTE GROUPS
HALOGENATED VOLATILES
Bromodlch1oromethane
Broraoform
Bromomethane
Carbon tetrachloride
Ch1orod1b romomethane
Chlorobenzene
Chloroethane
Chloroform
Chioromethane
Chloropropane
01bromomethane
C1s.l ,3-d1chloropropene
1.1-01Chloroethane
1.2-01Chloroethane
1.l-D1chloroethene
1,2-Olchloroethene
1,2-D1chloropropane
Fluorotrlchloromethane
Methylene chloride
1,1,2.2-tetrachloroethane
Tetrachloroethene
1.1,1-TM Chloroethane
1,1,2-TrfChloroethane
1,2-Trans-d1chloroethene
Trans-1,3-d1chloropropene
1. l.2-tHchloro-l.2.2-tr1fluoroethane
Trlchloroethene
Vinyl chloride
Total chlorinated hydrocarbons
Hexachloroethane
Dlchloromethane
HALOGENATED SEMIVOLATILES
2-chlorophenol
2,4-d1chlorophenol
Hexachlorocyclopentadlene
p-chloro-m-cresol
Pentachlorophenol
Tetrachlorophenol
2,4.5-trlchlorophenol
2,4,6-trlchlorophenol
Bf s-(2-chloroethoxy)methane
Bis(2-chloroethyl)ether
B1s(2-chloro1sopropyl)ether
4-bromophenyl phenyl ether
4-chloroan11Ine
2-chloronapthalene
4-chlorophenyl phenylether
HALOGENATED SEHIVOLATILES (cont.)
81s(2-chloroethoxy)phthalate
Bis(2-chloroethoxy)ether
1,2-b1s(2-chloroethoxy)ethane
NONHALOGENATED VOLATILES
Acetone
Acroleln
Acrylonltr1le
Benzene
2-butanone
Carbon dlsulflde
Cyclohexanone
Ethyl acetate
Ethyl ether
Ethyl benzene
2-hexanone
Isobutanol
Methanol
Methyl Isobutyl ketone
4-methyl-2-pentanone
n-butyl alcohol
Styrene
Toluene
Trimethyl benzene
Vinyl acetate
Xylenes
NONHALOGENATED SEMIVOLATILES
Benzole add
Cresols
2,4-dimethyl phenol
2,4-dinltrophenol
2-methylphenol
4-methylphenol
2-nltrophenol
4-n1trophenol
Phenol
Acenaphthene
Acenapthylene
Anthracene
Benzldlne
BenzoCa)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo(gh1)perylene
Benzyl alcohol
B1s(2-ethylhexyl)phtha1ate
1-36
-------
EXAMPLES OF CONSTITUENTS HITHIN HASTE GROUPS (cent)
HALOGENATED SEHIVOLATILES (cont.)
1,2-dlchlorobenzene
1,3-dlchlorobenzene
1,4-dichlorobenzene
3,3-d1chlorobenz1d1ne
Hexachlorobenzene
Hexachlorobutadlene
1,2.4-tr1chlorobenzene
PESTICIDES
Aldrln
Bhc-alpha
Bhc-beta
Bhc-delta
She-gamma
Chlordane
4,4'-DOO
4.4'-DOE
4.4'-OOt
Oleldrln
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrln
Endrln aldehyde
Ethlon
Aluminum
Heptachlor
Heptachlor epoxlde
Ma lathi on
Methylparathlon
Parathlon
Toxaphene
NONHALOGEHATED SEMIVOLATILES (cont)
4,6-d1n1tro-2-methylphenol
2,4-d1n1trotoluene
2,6-dlnltrotoluene
D1-n-octyl phthalate
1,2-d1phenylhydraz1ne
Fluoranthene
Fluorene
Indeno(l,2.3-cd)pyrene
Isophorone
2-methy1napthalene
Napthalene
2-n1troan1l1ne
3-n1troan1l1ne
4-n1troan1l1ne
Nitrobenzene
n-n1trosod!methyl amine
n-nltrosodl-n-propylamine
n-n1trosodlphenylamine
Phenanthrene
Pyrene
Pyrldlne
2-methynaphthalene
B1s phthalate
Phenyl napthalene
Ethyl parathlon
Butyl benzyl phthalate
Chrysene
D1benzo(a,h)anthracene
Dibenzofuran
01 ethyl phthalate
Dimethyl phthalate
Di-n-butyl phthalate
VOLATILE METALS
Arsenic
Bismuth
Lead
Mercury
Tin
Selenium
OTHER CATEGORIES
Asbestos
1-37
-------
EXAMPLES OF CONSTITUENTS WITHIN HASTE GROUPS (cont)
INORGANIC CORROSIVES
Hydrochloric add
Nitric add
Hydrofluoric add
Sulfurtc add
Sodium hydroxide
Calcium hydroxide
Calcium carbonate
Potassium carbonate
PCBs
PCS (Arochlor)-1016
PCB (Arochlor)-1221
PCB (ArochloD-1232
PCB (Arochlor)-1242
PCB (ArochloD-1248
PCB (Arochlor)-1254
PCB (ArochloD-1260
PCB NOS (not otherwise specified)
ORGANIC CORROSIVES
Acetic Add
Acetyl chloride
Aniline
Aeromatic Sulfonlc adds
Cresyllc add
Formic acid
HONMETALLIC TOXIC ELEMENTS
Fluorine
Bismuth
NONVOLATILE METALS
Aluminum
Antimony
Barium
Beryl 11 urn
Bismuth
Cadmi urn
Calcium
Chromium
Copper
Cobalt
Iron
Magnesium
Manganese
Nickel
Potassium
Selenium
Sodium
Vanadium
Zinc
RADIOACTIVES
Radioactive Isotopes of
Iodine, barium, uranium
Radium
Gamma radioactivity
ORGANIC CYANIDES
Organonitrlles
OXIDIZERS
Chlorates
Chromates
REDUCERS
Sulfides
Phosphides
Hydrazine
INORGANIC CYANIDES
Cyanide
Metallic cyanides
(e.g.. ferrlcyanlde.
sodium cyanide)
1-38
-------
RELATIVE DEGRADABILITY
Classes of chemicals that are good candidates
for treatment at hazardous waste sites
• Monochlorinated aromatic compounds (A)
• Benzene, toluene, xyiene (A or AN)
• PhenoEcs (nonhalogenated) and
cresols (A or AN)
• Polynuclear aromatic hydrocarbons
(creosotes) (A)
• Akanes and alkenes (fuel oP) (A)
(A) using aerobic biodegradation processes
(AN) using anaerobic biodegradation processes
NOTES
NOTES
RELATIVE DEGRADABLITY
Classes of chemicals that, with further
research (short term), could be candidates for
biological treatment at hazardous waste sites
• Polychlorinated biphenyls (A and AN)
• Pentachlorophenol (A or AN)
• Nitrogen heterocyclics (A)
* Chlorinated solvents (alkanes and
akenes) (A and AN)
(A) using aerobic biodegradation processes
(AN) using anaerobic biodegradation processes
Phenanthrene Degradation During Pilot -Scale
Bbremediation of Styrene Tar Waste
in Soils from a Refining Site
Initial
Treatment (Dav 0)
Control 27,850
Nutrient ' 19,400
Adjusted
Skigle 73,600
Inoculation
Phenanthrene ( PPB )
Final Half Life
(Day 94) Reduction (Davs)
5,725 79.44% 40.8
2,712 86.02% 33.0
5,750 92.19% 25.7
' Nutrients: inorganic nitrogen & phosphorous
NOTES
1-39
-------
I 1
21 SB
TIME(OAYS)
I
94
Reduction in Phenanthrene Concentration
NOTES
NOTES
Effect of Initial Concentration on Phenanthrene
Degradation During Pilot-Scale Bioremcdiation
of Styrene Tar Waste In Soils at a Refining Site
jnrtial_Concentration, PPB
1,000 4,999
5,000 - 9,999
10,000 - 49,999
50,000 - 100,000
greater than 100,000
Average Reduction. %
27.4
33.4
67.2
94.0
96.7
CONCENTRATIONS OF 2.4-0 IN * SIMULATED SOLID-PHASE
eiOBECLArVUION SYSTEM
(ing/kg)
NOTES
Simple Day 0
Sterile T9.7 (±5.0)
Covered 19.7
Uncovered. 19.7
uninocLilated
Uncovered 19.7
» JMP 134 t TF-6
Uncovered 19. 7
« ME-3 t TF-«
Pay 5 Oiv 10 Pay 20
23 23 16
8. 8 (»2.2) 8.1 (»3.1) 2.2 (tO.2)
7.3 (±0.5) B.8 (±3.6) 2.1 (jO.2)
7.7 (±2.0) 6.0 (±1.1) 1.7 (±0.3)
9. B (±1.5) <.0 (±0.2) 1.9 (±0.1)
NOTE: NO - Not detected tt detection Umit of 5.0 ng/kg
Numbers In p#renth«»ej {nd1c*t« r*r»9# of duplic*tt f*mp1«c
1-40
-------
NOTES
(ONCtNIIOMIONS Of HCPA IN * S1MULAHD iOUO-PHAS(
eiORCCLAWllON SYSKH
._. Swlj .
Sterile
Covered
Uncovered.
uninocul tted
Uncovered
. JMP 134 » TF-6
Uncovered
. HE-3 « U-6
.. .0*1 0.. P»> i _ 0,v 10
117 (.40) 121 115
117 71 (.2?) 46 I < 141
117 119 (±1S) 44 (,17)
117 62 (»44) 57 (<12)
117 86 dU) 40 (.1)
. D« ?S.
18
NO
31 <*<)
16 (±3)
24 (*9)
Numbers in F^r«nthei*$ indicate range of duplicate samples
NOTES
DIAUXIE
The response of microorganisms to the
presence of mixed substrates in which
preferential utilization of the substrates
for carbon and energy is observed
PHENOMENON OF DIAUXIC GROWTH WITH A BACTERIAL
CULTURE GROWING ON MIXED SUBSTRATES
SUBSTRATE
SUBSTRATE 2
TIME
1-41
NOTES
-------
CONCENTRATIONS OF 2,4-D AND MCPA IN
A SOIL SLURRY TREATMENT SYSTEM
< o
a: j:
h- x
2 C»
o
o
Importance of Microbial Communities
CONSORTIA
• Typical interactions within communities
• Importance of communities in adaptation
• Changes in the genetic information or
constitution of microorganisms
• The Ubiquity Principle
1-42
-------
CARBON AND ENERGY
SOURCE: PARATHION
PSEUDOMONAS STUTZERI
\
01 ETHYL
THIOPHOSPHATE
p-NITROPHENOL
UNIDENTIFIED
MOTILE ROD
EXCRETED METABOLITES
AND CELL LYSIS PRODUCTS
PSEUDOMONAS AERUGINOSA
PARATHION MICROBIAL COMMUNITY BASED ON COMETABOLISM: *•, SUBSTRATE
UTILIZATION ASSOCIATED WITH GROWTH; ->-, COMETABOLIC TRANSFORMATION
NOT LINKED TO GROWTH.
(SOURCE: SLATER AND LOVATT)
GENETIC APPROACHES TO
ENHANCE BIODEGRADATION
• Increase enzyme yields
• Overcome cell regulatory controls
* Engineer more efficient proteins
• Construct novel foiodegradation
pathways
1-43
-------
r
INITIAL DATA
REQUIREMENTS
SECTION 2
Abstract 2-2
Slides 2-11
Worksheets 2-42
2-1
-------
INITIAL DATA REQUIREMENTS
John Rogers P- Hap Pritchard Paul Flathman
U.S. EPA U.S. EPA OH Materials
Athens, Georgia Gulf Breeze, Florida Findlay, Ohio
Because of the tight time constraints in effecting the cleanup of
Superfund hazardous waste sites it is imperative to make timely decisions
in selecting the appropriate remediation technology. Such decisions,
however, should be predicated on sound information about the site and
some initial information about the individual remediation processes.
Information on the site can be obtained from the initial site
characterization: Information about the remediation process can be
obtained from published literature as well as from simple laboratory
feasibility studies. The purpose of this portion of the workshop is to
describe what information should be collected during the initial site
characterization to evaluate bioremediation processes and also to
describe some simple feasibility studies that can be used to assist in
the selection process.
At all sites, an initial site investigation is conducted to establish
the identity of chemicals at the site, determine the nature and extent of
the contamination, obtain a description of the environmental
characteristics of the site, and to make an initial appraisal of the
appropriate remediation technologies. This information is used to
determine if the site is hazardous and, if necessary, what action should
be taken to reduce the hazard to a safe level. The amount of information
required to make these decisions is not insignificant. In this
presentation and in these handouts only the information that is required
to evaluate bioremediation has been emphasized.
To facilitate the data review a flow diagram is presented that can be
used to walk through the data analysis. The diagram is divided into six
major areas.
In the first area the problem is defined and the types of
contaminants are identified. The physical and chemical properties of the
compounds that can influence biodegradation are identified and the
literature assessed for information concerning the degradation of the
compounds.
In the second area the distribution of the chemicals within the site
is determined. Examples of specific analytical procedures are presented
in Appendix A. At this point the site is divided into a series of
subsites for further evaluation. Compound concentration becomes
important at this point because concentrations may be toxic and some
pretreatment may be required before bioremediation can be considered.
Pretreatment may consist of dilution of the contaminated area, e.g.,
mixing of wastes.
2-2
-------
In the third area the contaminated environment is characterized.
This characterization extends from gross characteristics such as soil,
sediment, water or subsurface material to more specific characteristics
such as permeability, redox conditions, pH and hydrology. The
characteristic microbiological characteristics of the different
environments are also identified. For example, anaerobic bacteria may
predominate in sediments whereas aerobic organisms would predominate in
unsaturated soils.
In the fourth area any adjustment of the environment that might be
required to permit bioremediation is addressed directly. Such
adjustments could include alteration in pH, preremoval of toxic metals,
and changes in moisture content. In some cases the judgment may be that
bioremediation is not possible because the environment cannot be adjusted
In the fifth area the microbiological needs of the sites are
evaluated. At this point the concern becomes the availability of
nutrients, the potential additions of bacteria with specific degradative
characteristics, and whether the process should be conducted under
anaerobic conditions or aerobic conditions.
In the sixth area a feasibility study is designed to test potential
bioremediation scenarios.
2-3
-------
REFERENCES
Swallow, K. C., N. S. Shifrin, and P- J. Doherty. 1988. Hazardous
organic compound analysis. Environ. Sci. Technol. 22: 136-142.
RCRA Corrective Action Plan: Interim Final, June 1988, Office of Solid
Waste and Emergency Response, U.S. EPA, EPA/530-SW-88-028, Washington,
DC 20460.
RCRA Corrective Action Interim Measurements Guidance: Interim Final,
June 1988, Office of Solid Waste and Emergency Response, U.S. EPA,
EPA-530-SW-88-029, Washington, DC 20460.
Guidelines and Specifications for Preparing Quality Assurance Program
Plans, September 20, 1980, Office of Monitoring Systems and Quality
Assurance, ORD, U.S. EPA, QAMS-004/80, Washington, DC 20460.
Interim Guidelines and Specifications for Preparing Quality Assurance
Program Plans, Dec. 29, 1980, Office of Monitoring Systems and Quality
Assurance, ORD, U.S. EPA, QAMS-005/80, Washington, DC 20460.
Test Methods for Evaluating Solid Waste. Volume 1A: Laboratory Manual
Physical/Chemical Methods, November 1986, Office of Solid Waste and
Emergency Response, U.S. EPA, SW-846 Third Edition, Washington, DC 20460
Test Methods for Evaluating Solid Waste. Volume IB: Laboratory Manual
Physical/Chemical Methods, November 1986, Office of Solid Waste and
Emergency Response, U.S. EPA, SW-846 Third Edition, Washington, D.C.
20460.
Test Methods for Evaluating Solid Waste. Volume 1C: Laboratory Manual
Physical/Chemical Methods, November 1986, Office of Solid Waste and
Emergency Response, U.S. EPA, SW-846 Third Edition, Washington, DC 20460.
Interim Protocol for Determining the Aerobic Degradation of Hazardous
Organic Chemicals in Soil, September 1988, Biosystems Technology
Development Program, U.S. EPA.
Pesticide Assessment Guidelines Subdivision N Chemistry: Environmental
Fate, October 1982, Office of Pesticides and Toxic Substances, U.S. EPA,
Washington, DC 20460.
795.54 Anaerobic Microbiological Transformation Rate Data for Chemicals
in the Subsurface Environment, June 1988, Federal Register, Vol. 53,
no. 115, 22320-22323.
Crip, C. R., W. W. Walker, P. H. Pritchard, and A. W. Bourquin. 1987. A
Shake-flask test for Estimation of Biodegradability of Toxic Organic
Substances in the Aquatic Environment. Ecotox. Environ. Safety.
14: 239-251.
2-4
-------
Shelton, D. R. and 0. M. Tiedje. 1984. General Method for Determining
Anaerobic Blodegradation Potential. Appl. Environ. Microbiol.
47: 850-857.
Owen, W.F. et al. 1979. Bioassay for monitoring biochemical methane
potential anaerobic toxicity. Water Res, 13:485-492.
Protocol Development for the Prediction of the Fate of Organic Priority
Pollutants in Biological Wastewater Treatment Systems. (AEROBIC AND
ANAEROBIC MULTI-LEVEL BIODEGRADABILITY TESTING PROTOCOLS)
E.G. Kirsch, C.P.L. Grady Jr. and R.F. Wukasch, Purdue University, West
Lafayette, Indiana 47507 and Henry H. Tabak, U.S. EPA, Water Engineering
Research Laboratory, AWBERC, ORD, Cincinnati, Ohio 45268.
EPA/600/S2-85/141 February 1986
Protocol for Determination of Biodegradation Kinetics Through the Use of
Electrolytic Respirometry
C.P.L. Grady, J.S. Dang, D.M. Harvey, A. Jobbagy and X.-L. Wang, Clemson
University, Clemson, South Carolina 29634, and Henry H. Tabak, U.S. EPA,
Risk Reduction Engineering Laboratory, AWBERC, ORD, Cincinnati, Ohio
45268.
Presented at the 14th Biennal Conference of International Association on
Water Pollution Research and Control, Brighton, England 17-23 July 1988.
To be published in the Water Science and Technology Journal. July. 1989.
Protocol for Evaluation of Biodegradation Kinetics with Respirometric Data
C.P.L. Grady, J.S. Dang, D.M. Harvey, A. Jobbagy, Clemson University,
South Carolina, Clemson, South Carolina 29634, and Henry H. Tabak, U.S.
EPA, Risk Reduction Engineering Laboratory, AWBERC, ORD, Cincinnati, Ohio
45268.
Presented at the 61st Annual Conference of the Water Pollution Control
Federation, October 2-6, 1988, Dallas, Texas, and submitted for
publication October, 1988 to the Journal of Water Pollution Control
Federation.
Protocol for the Determination of Biodegradability and Biodegradation
Kinetics of Toxic Organic Compounds with the use of Electrolytic
Respirometry
Henry H. Tabak, Risk Reduction Engineering Laboratory, U.S. EPA, ORD,
AWBERC, Cincinnati, Ohio 45268, Rakesh Govind and Sanjay Desai,
University of Cincinnati, Cincinnati, Ohio 45221 and C.P.L. Grady,
Clemson University, Clemson, South Carolina 29634.
2-5
-------
Presented at the 61st Annual Conference of Water Pollution Control
Federation, October 2-6, 1988, Dallas, Texas and submitted for
publication in December 1988, to the Journal of Mater Pollution Control
Federation.
"Assessment of Bioaugmentation Technology and Evalution Studies on
Bi©augmentation Products"
Henry H. Tabak, U.S. EPA, Wastewater Research Division, Water Engineering
Research Laboratory, ORD, Cincinnati, Ohio 45268.
Presented at the Tenth United States/Japan/NATO/CCMS Joint Conference on
Sewage Treatment Technology, October 15-18, 1985, Cincinnati, Ohio.
Published in the Proceedings of the Tenth United States/Japan Conference
on Sewage Treatment and NATO/Committee on the Challenges of Modern
Society (NATO/CCMS) Conference on Sewage Treatment Technology. Volume I.
Part B. United States Papers p. 431-499. 1986. EPA/600/9-86/015b.
NTIS PB87-110631.
Screening Protocol for Assessing Toxicity of Organic Chemicals to
Anaerobic Treatment Processes (MULTI-STEP SCREENING ANAEROBIC INHIBITION
PROTOCOL)
James C. Young, University of Arkansas, Civil Engineering Department,
Fayetteville, Arkansas and Henry H. Tabak, U.S. EPA, Risk Reduction
Engineering Laboratory, AWBERC, ORD, Cincinnati, Ohio 45268.
Presented at the AWMA/EPA International Symposium on Hazardous Waste
Treatment: Biosystems for Pollution Control. February 20-23, Cincinnati,
Ohio 45202 and accepted for publication in the Air & Waste Management
Association Journal. 1989.
2-6
-------
APPENDIX A
CHEMICAL ANALYSIS OF TEST CHEMICALS AND/OR WASTE SAMPLES
The selection of a suitable extraction procedure for a given
combination of analyte(s) and soil matrix generally requires some method
development (Coover et al. 1987). For example methods that successfully
recover a compound from one medium may not adequately recover the same
chemical from similar media (Albro 1979). Also, extraction recoveries
from a given set of structurally similar media may vary (Albro 1979).
Where possible it is recommend that the existing and established
analytical methods described in Test Methods for Evaluating Solid Waste
(USEPA SW-846 3rd Edition November 1986) be used.
The recommended SW-846 methodology for selected analytes are:
Gas Phase Volatiles
Method 0010 Modified Method 5 Sampling Train
Method 0020 Source Assessment Sampling System (SSAS)
Method 0030 Volatile Organic Sampling Train (VOST)
Method 5040 Protocol for Analysis of Sorbent Cartridges from
Volatile Organic Sampling Train.
Soil Phase Volatiles
Method 5030 Purge and Trap
Method 8010 Halogenated Volatile Organics
Method 8015 Non-Halogenated Volatile Organics
Method 8020 Aromatic Volatile Organics
Method 8030 Acrolein, Acrylonitrile, Acetonitrile
Selected Non-Volatiles
Method 8040 Phenols
Method 8060 Phythalate Esters
Method 8080 Organic Pesticides and PCB's
Method 8090 Nitroaromatics
Method 8100 Polynuclear Aromatic Hydrocarbons
Method 8120 Chlorinated Hydrocarbons
Method 8140 Organophosphorous Pesticides
Method 8150 Chlorinated Herbicides
Recommended extraction/concentration techniques (soils and sediments)
are:
Method 3540
Method 3550
Soxhlet Extraction
Sonication Extraction
2-7
-------
Other published methods for Soxhlet extraction (Anderson et al. 1985,
Bossert et al. 1984, Coover et al. 1987, Eicemen et al. 1986, Kjolholt
1985, Grimalt et al. 1986), sonication extraction (de Leevw et al. 1986,
Sims 1982) and homogenization and extraction (Coover et al. 1987, Fowlie
and Bui man 1986, Lopez-Avila et al. 1983, Sims 1982, Stott and Tabatabai
1983, and U.S. EPA 1982a, and extraction of materials from treatability
studies (Brunner et al. 1985, Russell and McDuffle 1983) are available
for reference and special applications.
Soil spiking and recovery studies should be conducted to determine
the effects of soil, test substance(s), and soil test substance(s) matrix
on chemical extraction and recovery efficiency. Soil samples should be
sterilized using a method such as mercuric chloride, causing minimal
change in soil physical and chemical properties (Fowlie and Bulman
1986). The sterile soil should be spiked with the test substance(s) to
achieve a range of initial oil concentrations (Coover et al. 1987). The
range of concentration should include the highest concentration and less
than one-half of the lowest initial concentration to be used in
degradation evaluations. Extractions of the soil/test-substance(s)
mixtures using the selected procedure will allow the evaluation of the
effect of test substance(s) soil concentrations on recovery efficiency.
The effect of soil concentration was evaluated and found to be
significant for anthracene and benzo[a]pyrene by Fowlie and Bulman (1986),
Extracts of the soil and complex wastes should be spiked with test
substance(s) of interest to evaluate the effect of these matrices on
chemical identification and quantification. Interferences due to the
extract matrix may be identified. Extraction procedures or
instrumentation used for identification and quantification may then be
changed if necessary.
Standard curves should be prepared using primary standards of the
test substance(s), or chemicals in the test substance, dissolved in a
suitable solvent that does not interfere with chemical identification and
quantification. Standard curves should be generated using at least six
points ranging from the highest concentration anticipated to the
detection limit for the chemical.
2-8
-------
REFERENCES
Albro, P.W. 1979. Problems In analytical methodology: Sampling
handling, extraction, and cleanup. Ann. N.Y. Acad. Sci. 320:19-27.
Anderson, O.W., G.H. Herman, D.R. Theilen, and A.F. Weston. 1985.
Method verification for determination of tetrachlorodibenzodioxin in
soil. Chemosphere 14: 1115-1126.
Bossert, I., N.M. Kachel, and R. Bartha. 1984. Fate of hydrocarbons
during oil sludge disposal in soil. Applied and Environmental Micro.
47:763-767.
Brunner, W., F.H. Sutherland, and D.D. Focht. 1985. Enhanced
biodegradation of polychlorinated biphyenyls in soil by analog enrichment
and bacterial inoculation. 0. Environ. Qual. 14:324-328.
Coover, M.P., R.C. Sims, and W.J. Doucette. 1987. Extraction of
polycyclic aromatic hydrocarbons from spiked soil. J. Assoc. Off. Anal.
Chem. 70(6):1018-1020.
de Leevw, J.W.E., W.B. de Leer, J.S. S. Damste, and P.J.W. Schuyl.
1986. Screening of anthropogenic compounds in polluted sediments and
soils by flash evaporation/pyrolysis gas chromatography-mass
spectrometry. Anal. Chem. 58:1852-1857.
Eiceman, G.A., B. Davani, and J. Ingram. 1986. Depth profiles for
hydrocarbons and polycyclic aromatic hydrocarbonss in soil beneath waste
disposal pits from natural gas production. J. Environ. Sci. Technol.
20:500-514. Federal Register. 1979. 44(53): 167-16280 (Friday,
March 16).
Fowlie, P.O.A., and T.L Bulman. 1986. Extraction of antharacene and
benzo(a)pyrene from soil. Anal. Chem. 58-721-723.
Grimalt, 0., C. Marfil, and J. Albaiges. 1986. Analysis of hydrocarbons
in aquatic sediments. Int. J. Environ. Anal. Chem. 18:183-194.
Kjolholt, J. 1985. Determination of trace amounts of organophorous
pesticides and related compounds in soils and sediments using capillary
gas chromatography and a nitrogen-phosphorus detector. Journal of Chrom.
325:231-238.
Lopez-Avila, V., R. Northcutt, J. Onstot, M. Wickham, and S. Billets.
1983. Determination of 51 priority organic compounds after extraction
from standard reference materials. Anal. Chem. 55:881-889.
Russell, D.O., and B. McDuffie. 1983. Analysis for phthalate esters in
environmental samples: Separation from PCSs and pesticides using dual
column liquid chromatography. Int. J. Environ. Anal. Chem. 15:165-183.
2-9
-------
Sims, R.C. 1982. Land application design criteria for recalcitrant and
toxic organic compounds in fossil fuel wastes. PhD dissertation. North
Carolina State University, Raleigh, NC.
Sims, R.C., D.L. Sorensen, W.J. Doucette, and L. Hastings. 1986.
Waste/soil treatibility studies for hazardous wastes: Methodologies and
results. Vols. 1 and 2. U.S. Environmental Protection Agency, Robert S
Kerr Environmental Research Laboratory, Ada, OK. EPA/6—/6-86/003a and
b. NTIS No. PB87-111738.
Stott, D.E. and M.A. Tabatabai. 1985. Identification of phospholipids
in soils and sewage sludges by high-performance liquid chromatography.
J. Environ. Qual. 14:107-110.
2-10
-------
UNOISIKAM.E
CHA«ACTERIiTIC»T
•ELECTION Of
im ro«
•IONEMEOIATION
(ELECT
ALTERNATIVE
TECHNOLOGY
2-11
-------
ENVIRONMENTAL
ADJUSTMENT
FEASIBILITY
HCAVY
MCTAL
REMOVAL
TOXIC
ORGANIC
REMOVAL
CHANCE IN
ENVIRONMENTAL
CONDITIONS
SELECT
ALTERNATIVE
TECHNOLOGY
SPECIAL
EMOMEERMQ
CONSBERATIONS
L
INITIAL STRATCOY
FOR MOREMEDIATION
INITIATE
SCALE UP
STUDIES
2-12
-------
[
UNMENT
CERN3
GOALS OF
REMEDIATION
"^
^^
SITE
CHARACTERIZATION
I
REGUL/
REQUIRE
• -•"" OROA
--._ PRE8
NOTES
NOTES
MINIMUM REQUIREMENTS FOR QA/QC
* Project description
• Project organization
• QA objectives
• Sample custody
• Internal QC checks
• Performance and system audits
• Preventative maintenance schedule
MINIMUM REQUIREMENTS FOR QA/QC
(Continued)
• Data assessment procedures
• Corrective actions
• QA reports
• Sampling plan
NOTES
2-13
-------
DOCUMENTATION REQUIREMENTS FOR QA
• Accepted sampling techniques
• Field actions contrary to QAPP
• All pre-field activities
• QC for field measurement data
• Field activities
* Post-field activities
• Quality control samples
(generation & use)
NOTES
NOTES
QA FOR ANALYTICAL PROCEDURES
• Duplicate spike
• Reagent blank
• Documentation of fill samples
* Analytical procedures for surrogate
compounds
• Recovery efficiency for columns
• Detection limits and data reduction
QA FOR ANALYTICAL PROCEDURES
(Continued)
• Internal QC checks
• Performance and system audits
• Equipment calibration
• Extraction and sample preparation
procedures
NOTES
2-14
-------
DEFINE
PROBLEMS
I
AINMENT
CERNS
1
,
GOALS OF
REMEDIATION
^rx
MENTf /
\\N SITE \A\
S. CHAR ACTERIZATION \X
]
REGULl
REQUIRE
••"' ORQA
'-.._ PRESI
NOTES
NOTES
SITE CHARACTERIZATION
• Description of facility
• Identification of contaminants
• Extent of contamination
NOTES
2-15
-------
DESCRIPTION OF FACILITY
• Geographic location; property lines.
topography and surface drainage
• Infrastructure present
• Description of hazardous waste treatment.
storage, disposal and spill areas
* Surrounding land uses
• Production and groundwater monitoring wells
NOTES
NOTES
IDENTIFICATION OF CONTAMINANTS
• Organic/inorganic
• Chemical classes (metals,
halogenated volatiles,
pesticides)
• Mixtures
INITIAL MATERIAL CHARACTERIZATION
• Organics: GC or GC/MS. HPLC
• Group analysis: priority pollutants,
fuels analysis, EP-Toxicity
• Metals: AA. ICP
• General chemistry: TOG, COD. BOD.
TPH, Oil & Grease (IR or GC).
TKN, N03, TP. P04, S04
* Optional radioisotope analysis: isotopically
labeled substrate studies,14CO2
NOTES
2-16
-------
GENERAL CHEMISTRY
Analysis Price Per Sample
Total Organic Carbon (TOO 40
Total Kjeldahl Nitrogen (TOO 50
Chromium VI 25
Cyanides 50
Phenol $ 50
Orthophoshates 20
Total Phosphorous 35
Nitrate 20
SulMde 25
Oil and Grease 40
Total Suspended Solids (TSS) 15
Chemical Oxygen Demand (COO) 35
Ion Chromography 65
(Bromide. Chloride. Fluoride. Nitrate.
Nitrite. Phosphate. Sulfate)
MtcrotOK Price on Request
Radio Isotope Analysis (Liquid Scintillation) Price on Request
NOTES
NOTES
ORGANICS
Analvsls
GC/MS
Volatile Organic Analysis
Acid/Base Neutrals
Confirmation by GC/MS
GC
Pestlddes/PCBs
PCBs In 011
Herbicides
Phenols
Pentachlorophenol (PCP)
Polynuclear Aromatic
Hydrocarbons (PNA)
Hydrocarbon Fuels
(gasollne/dlesel)
Creosote
Price
Hater
240
420
100
150
50
200
100
90
115
110
90
For Sample
Solids
280
475
150
200
250
100
90
130
130
90
Method
Hater
624
625
60S
604
604
610
Number
Solids
8240
8270
8080
8150
8040
8100
GROUP ANALYSES
Analysis
Priority Pollutants
Acid/Base Neutrals (37)
Volatile Organic Analysis (31)
Pesticides & PCBs (28)
Metals (13)
Cyanides
Phenols
EP-ToxIclty
Sample Prep and Extraction
Metals
(Ag, As, Ba. Cd, Hg, Pb. Se)
Herbicides and Pesticides
(2.4-D. 2,4.5-TP. Endrln. Llndane.
Methoxy Chlor. Toxaphane)
Fuels Analysis
8TX (Benzene. Toluene. Xylene)
EOS (Ethyl Dlbromlde)
Tetraethyl Lead (total)
Characterization of Fuels by
GC (Gasoline and Diesel)
Price Per Sample
Hater Solids
1195 1295
450
90
100
35
110
450
100
120
35
130
NOTES
2-17
-------
METALS
Price Per Element
Method of Analysis 20
Graphite Furnince '3
AAS 30
Hydride 30
Cold Vapor
Price Per Sample
ICP Multl Element Analysis
(Ag, Al, B, Ba, Be. Ca. Cd
Co. Cr, Cu. Fe. K, Hg. Mn,
Mo. Na, Ml. Pb. Se. SI. Sn
Tl. V. Zn)
1-12 Elements 90
13-24 Elements US
Sample Preparation Price Per Sample
Hater 14
Soil/Hater/Sludge 20
EP-Tox Extraction 95
Price Per Sample
Group Metal Analysis Hater SolIds
Priority Pollutant Metals 160 199
(Ag. As. Ba. Cd. Cr. Co. Hg
HI. Pb. Sb. Se. Tl. Zn)
RCRA Metals Analysis 130 130
(Ag. As. Bs. Cd. Cr. Hg. Fe, Se)
Hazardous Substance Listed Metals (Non CLP) 200 215
(Ag. Al. As. Ba. Be. Ca, Cd, Co, Cr.
Cu. Fe. Hg. K. Hg. Hn. Na, Ml, Pb. Sb.
Se. Tl. V. Zn
REPRESENTATIVE FIELD SAMPLES REQUIRED
FOR BIOTREATABILITY STUDIES
• Evaluation of many samples to
obtain a bioactivity site matrix
• Field composite to define
any site bioactivity
• Field background samples essential
for material characterization
2-18
-------
EXTENT OF CONTAMINATION
• Groundwater
Plume size and movement
Contaminant concentration profiles
• Soil contamination
Distribution and concentration
• Surface water contamination
Horizontal and vertical distribution
• Sediment contamination
Horizontal and vertical distribution
I VV ^ '•••'* lv k \. W N
\^ PROPERTIES Of\VO
> THE CONTAMIMANTS\\
ENVIRONMENTAL
CHARACTERISTICS
SELECT
ALTERNATIVE
TECHNOLOGY
2-19
-------
PROPERTIES OF CONTAMINANTS
Physical/ Chemical Characteristics
• Solid, liquid or gas
• Powder, oily sludge
• Acid, base, valence or
oxidation state
• Molecular weight
• Density
• Boiling poirrt
NOTES
NOTES
PROPERTIES OF CONTAMINANTS
Physical / Chemical Characteristics
(Continued)
• Viscosity
• Solubility in water
• Cohesiveness
• Vapor pressure
• Flash poirit
PROPERTIES OF CONTAMINANTS
Safety Considerations
• Toxicity (human, microorganisms)
• Flammability
• Reactivity
• Corrosiveness
• Oxidizing or reducing
characteristics
NOTES
2-20
-------
PROPERTIES OF CONTAMINANTS
Environmental Fate Characteristics
• Sorption
• Biodegradability
• Photodegradability
• Hydrolysis
• Chemical transformation
NOTES
NOTES
Je.
ENVIRONMENTAL
CHARACTERISTICS
GROUND
WATER
SOILS
WATER-
SEDIMENT
SELECT
ALTERNATIVE
TECHNOLOGY
ENVIRONMENTAL CHARACTERISTICS
OF THE SITE
Groundwater
• Flow characteristics
• Hydrogeological units
• Water level and movement
• Man-made influences
NOTES
2-21
-------
ENVIRONMENTAL CHARACTERISTICS OF THE SITE
Surface Water And Sediments
• Physical characteristics (location,
velocity, depth, surface area, etc.)
• Seasonal fluctuations
• Temperature stratification
• Flooding tendencies'
• Drainage patterns
• Evapotranspiration
• End use of water
NOTES
NOTES
ENVIRONMENTAL CHARACTERISTICS
OF THE SITE
Water/Sediment Chemistry
• pH
• Total dissolved solids
• Biological oxygen demand
• Alkalinity
• Conductivity
ENVIRONMENTAL CHARACTERISTICS
OF THE SITE
Water/Sediment Chemistry
(Continued)
• Dissolved oxygen profiles
• Nutrients NHs. NO3/NO2.PO4
• Chemical oxygen demand
• Total organic carbon
-3
NOTES
2-22
-------
ENVIRONMENTAL CHARACTERISTICS
OF THE SITE
Distribution And Soil Structure
• SCS soil classification
• Surface soil distribution
• Soil profile ASTM classification
• Depth to water table
NOTES
NOTES
ENVIRONMENTAL CHARACTERISTICS OF THE SITE
Physical Characteristics Of Soils
• Hydraulic conductivity
• Relative permeability
• Bulk density
• Porosity
• Particle size distribution
• Moisture content
• Infiltration
• Vertical flow
ENVIRONMENTAL CHARACTERISTICS OF THE SITE
Chemical Characteristics Of Soils
• Soil stratigraphy
• Soil sorptive capacity
• Ion exchange capacity
• Soil organic content
• Soil pH
• Mineral content
NOTES
2-23
-------
NOTES
NOTES
ENVIRONMENTAL
ADJUSTMENT
FEASIBILITY
CHANGE IN
ENVIRONMENTAL
CONDITIONS
w
PROPERTIES
ASSESSED
1
v\ TREATABILITY\V
"O PROTOCOL \\
V INFORMATION \N
\\ \ \ i. >, •> s ^\^
^
f \f
PROTOCOL AVAILABLE
COMPONENTS PROTOCOLS
>
J
INITIAL THIATASILITY
STUDY DESION
1
NOTES
2-24
-------
TREATABILITY PROTOCOLS
Properties Assessed
• Biodegradability of contaminants
- aerobic
- anaerobic
• Effectiveness of nutrient amendments
- inorganic supplements (N. P. S)
- electron acceptors
- organic supplements
NOTES
NOTES
TREATABILITY PROTOCOLS
Properties Assessed
(Continued)
• Effectiveness of inocula
- cultures of natural organisms
- specific degraders
• Nondegradative losses
- volatilization
- sorption
- leaching
• Genotoxicity of the waste
PROTOCOL COMPONENTS
Scope and approach
Summary and method
Collection and sampling of site materials
- sample selection
- sample collection
- sample characterization
- sample transportation
- sample preservation
- sample holding times
NOTES
2-25
-------
PROTOCOL COMPONENTS
(Continued)
* Apparatus and materials
- reactor components
- reactor design
* Procedures
- reactor setup
- reactor operation
- analysis of reactor contents
- reactor configurations
minimal treatment
intermediate treatments
complete treatment
NOTES
NOTES
PROTOCOL COMPONENTS
(Continued)
• Data recording and analysis
- data to be reported
- determination of degradation rates
• References
- general
- chemical analysis
- sampling
AVAILABLE TREATABILITY
PROTOCOLS
NOTES
2-26
-------
PROTOCOLS
SOILS
Aerobic
• Interim protocol for determining the
aerobic degradation potential of hazardous
organics in soil. September 1988. Biosystems
Technology Development Program. U. S. EPA
• Uses four reactor configurations
• no tillage
- periodic tillage
- forced aeration
- soil slurry
NOTES
NOTES
PROTOCOLS
SOILS
Aerobic
(Continued)
• Measures loss of target chemicals
• Corrects for volatile losses
• Requires psuedo-mass balance
PROTOCOLS
SOILS
Anaerobic
Pesticide assessment guidelines
subdivision N chemistry: Environmental Fate,
October 1982. Office of Pesticides and Toxic
Substances. U.S. EPA. Washington, D.C. 20460
Uses waterlogged soils (30 days)
One reactor design
Measures loss of product
Strict anaerobic conditions optional
NOTES
2-27
-------
PROTOCOLS
SUBSURFACE
Aerobic
• Not available
NOTES
NOTES
PROTOCOLS
SUBSURFACE
Anaerobic
• 795.54 Anaerobic microbiological
transformation rate data for chemicals in
the subsurface environment. June 1988. Federal
Register, Vol. 53, no. 115. 22320-22323
• Methanogenic
• Sulfate reducing
• Serum bottles for reaction vessels
• Requires strict anaerobic techniques
PROTOCOLS
SUBSURFACE
Anaerobic
(Continued)
* Designed for subsurface materials
• Uses 20% (w/v) slurries
• Could be modified for denitrifying conditions
• Measures loss of hazardous compound
NOTES
2-28
-------
PROTOCOLS
SEDIMENTS
Aerobic
• Under development
NOTES
NOTES
PROTOCOLS
SEDIMENTS
Anaerobic
• 795.54 Anaerobic microbiological
transformation rate data for chemicals in
the subsurface environment. June 1988, Federal
Register. Vol. 53. no. 115, 22320-22323
• Methanogenic
• Sulfate reducing
• Serum bottles for reaction vessels
• Requires strict anaerobic techniques
PROTOCOLS
SEDIMENTS
Anaerobic
(Continued)
• Designed for subsurface materials
• Uses 20% (w/v) slurries
• Could be modified for denitrifying conditions
• Measures loss of target chemicals
NOTES
2-29
-------
PROTOCOLS
WATER
Aerobic
• Under development
NOTES
NOTES
PROTOCOLS
WATER
Anaerobic
• Shelton. D.R. and J.M. Tiedje. 1984. General
method for determining anaerobic biodegradation
potential. Appl. Environ. Microbiol. 47: 850-857
• Methanogenic
• Serum bottles for reaction vessels
• Requires strict anaerobic techniques
PROTOCOLS
WATER
Anaerobic
(Continued)
• Measures gas production
• Sludge dependent
• Could be modified to include loss of
hazardous chemical
NOTES
2-30
-------
NsViiflAL TREAT ABILITY
\\\ STUDY DESIGN v V x\
X\\\xx.\\\ \ \\\\\
INITIAL
TREATABILITY
STUDY
NOTES
NOTES
,\\x X X
\\ INITIAL TREATABILITY ,.
^\\'T"°A\\\^
SELECT
ALTERNATIVE
TECHNOLOGY
SPECIAL
ENGINEERING
CONSIDERATIONS
INITIAL STRATEGY
FOR BIOREMEDIATION
INITIATE
SCALE UP
STUDIES
Control
(No •mondm«ntt)
lnt»rm*dlat«
Chang* pH
1
Maximal
• ChingtpH
• Add nutrient*
• Add mlor«*
• Mix
NOTES
2-31
-------
CO
w
o
o
a
E
o
O
Control
Maximal
Time
NOTES
NOTES
W
CO
o
_l
•o
c
3
O
Q.
E
o
o
Control
Maximal
Time
EXPERIMENTAL DESIGN
Controls: sterile, no treatment.
field background, number?
Replicates: duplicate or triplicate?
all time points? all controls?
Treatments: what are the questions
you want answered?
How are you going to optimize
the degradation process?
NOTES
2-32
-------
EXPERIMENTAL DESIGN
(Continued)
• Treatment time, how long should
the study be performed?
• Types of analysis! bulk measurements?
waste specific?
• Data reduction: raw data?
massaged data? QC/QA?
• Cost considerations: how will it
limit scope of test?
NOTES
NOTES
RELATIVE RATES OF REDUCTION AND OXDATON
2 3
Numb.r of Att«ch«d Chlorlnoi
Inoroailnf Extent of Halogonitlon
for M«th*n««, Ethanoe, Ethonoi
AERBOBIC BIODEQHADATION OF AROCLOR 1284
INCREASING NUMBER OF CHLORINES
in it « tt ttt tut IN in
». »utr«|Mi>i»Hteo
NOTES
2-33
-------
INCREASING NUMBER Of CHLORINES
I* '-
tl >-
s'
.Nlv .Ul P
i.
REDUCTIVE DECHLORffilTION OF POyCHDRINATED BIPHENYLS
BY ANSERC81C MICfiOORGANISMS FROM SEDIMENTS
NOTES
NOTES
RELATIVE BODEGRADATION of POLYCYCUC
AROMATIC HYDROCARBONS (PAH)
3 4
Numb«r ol Rlngl/PAH
Inetxtlng Mol«cul«r Weight
0«cr«»lng Aqu«oui Solubility
MICROBIOLOGICAL DEGRADATION
NOTES
2-34
-------
FATE OF POLYNUCLEAR AROMATIC
CONTAMMATES M CREOSOTE WASTE
DURMG LAND TREATMENT
4 Month Study
PNA Class % Reduction Hatf-Ufa
2 Rng Structure 90
(Naphthalene)
3 Rhg Structure 80
(Phenaphthalene)
4 Ring Structure 25
(Pyrene)
Total PNA 65
33 Days
47 Days
235 Days
100 Days
NOTES
NOTES
PHYSIOLOGICAL BARKERS TO BIODEGRAOATION
A contaminate wl be a poor substrate if:
No active microorganism is present, therefore,
no avaiabte enzymatic machinery
Kticroorganisms present, but.
• Substrate is a poor hducer
• Substrate concentration is too low
• Substrate fate to enter eels
• Cel lacks essential nutrients
' WitoHJon/tOHCity of enzymes
by substrate or products
• Other necessary microbes are absent
ENVIRONMENTAL BARRIERS TO BKXJEGRADATtON
Potentiaty Limiting Environmental Factors
• PH
• SaEnity
• Other synthetic chemicals
• Heavy metals
* Osmotic pressure
• Hydrostatic pressure
• Free water Imitations
• Radtation
NOTES
2-35
-------
GENETIC BARRERS TO BIODEGRADAT1ON
• No genetic coding for
contaminant degradation
• No genetic ccwfng for
transport into eel
• Genetics for biodegradation exist
but not hductote or
dtebursed on genome
• Low level of expression
NOTES
NOTES
BIODEGRADATION
Requires
• Suitable electron acceptor
• Organic substrate
• Nutrients: nitrogen,
phosphorous, others
• Trace metals
PERCENT 40
PCP
REMOVED
FROM SOIL 60
BIODEGRADATION OF 20 ppm
PCP IN SOIL UNDER
LABORATORY CONDITIONS
80 -
IOO
NOTES
2-36
-------
MICROBIAL EVALUATION
Reduction of Contaminants During a 4-Week
Incubation of Nutrient Amended Site Samples
Saturated
Soil
Unsaturated
Soil
Ground
Water
Surface
Soil
Compound
Acenaphthene
Anthracene
Beruo (a) Anthracene
Benzo (a) Pyrene
Chrysene
Oibenzofuran
Fluoranlhene
Fluorene
Indeno (1,2,3,-cd) Pyrene
2-Methylnaphthalene
Naphthalene
Pentachlorophenol
Phenanthene
Pyrene
COST BREAKDOWN CASE # 1
17
2
34
2
68
X $450
$30,600
+ 4,000
Field Samples
Replicates
Sample Times (0, 4 weeks)
Samples for Analysis
GC/MS BNA
Analytical Costs
Materials/Labor for Set up
$34,600 Total Cost (est)*
'Note: No Administrative Charges; Data Evaluation;
Report Preparation; QA/QC
2-37
-------
BIOREMEDIATION OF CREOSOTE/POP
Contaminated Sols (Slurry) Case Study #2
mg/kg IttotaaidB)
01 ma
50
• AdM
Una
0 ConM
NOTES
NOTES
BIOREMEDIATION OF CREOSOTE/PCP
Contaminated Sols (Slurry) Case Study 82
• Ac*ra PNA
X Advo Kf
CASE STUDY # 2
1 Single Soil Sample
3 Repficates
X2 Treatments (Active Amend^/ Control)
6
x4 Sample Times (0,2,6.6 wks)
24 Samples 6
$40 Oa/Grease (TJ.) __ j(3 (0.4.8 wks)
W60 18 Samples
GC/MS(BNA1
$8100
$960 + $6100 = $9060 Analytical Costs
for Experimental Section Initial Material
Characterization: TOC, TKN, 0-PO4, NOs, NH3
NOTES
2-3i
-------
CASE STUDY tt 2
(conthued)
170
x 2 Repicates
$340
$9,400
$4,500
Total Analytical Costs
Labor/Materials
$13,900 Total Cost of Treatabiity*
• Note: No administrative charges; data
evaluation, report preparation, QC/OA.
NOTES
NOTES
EFFECT OF SLURRY TREATMENT ON PAH AND PCP
CONCENTRATIONS* IN CREOSOTE/PCP CONTAMINATED SOILS
Comoound
Acenaphthene
Acenaphthalene
Dlbenzofuran
Fluorene
Fluoranthene
Anthracene
Phenanthrene
Pyrene
Chyrsene
Benzo( A) Anthracene
Benzo(A)Pyrene
Pentachlorophenol
Initial
Concentration
(mg/kg)
60 +/- 12
3.4 +/- 0.1
17 +/- 3
37 +/- 6
167 +/- 38
30 W- 3.5
130 +/- 17
177 +/- 38
40 +/- 3
34 +/- 3
19 +/- 1.3
127 +/- 12
4 weeks
(mo/ka)
3.8H
0.8 +/- 0.1
3.8H
3.8H
3.9 +/- 0.8
2.2 W- 0.6
0.5 +/- 0.1
26 +/- 18
5.9 +/- 1.1
1.7 +/- 0.2
9.8 +/- 1.3
24 +/- 2.0
8 weeks
(nta/ka)
3.8W
2.1J
3.8H
3.8H
3.6 +/- 0.3
6.7 +/- 1.2
0.7 +/- 0.1
10.6 +/- 1.5
3.5J
1.9 +/- 0.2
10.6 +/- 2.1
31.6 +/- 5.0
a Average of triplicate analysis +/- variance.
H Undetected at the noted concentration.
0 Estimated concentration. Sample data was less than the quantHation
Hm1t but greater than zero.
PARAMETERS MOMTORED DURING
THE PILOT TEST OPERATION
Parameter
Sol temperature
Sol pH
Bang?
54 F to 82 F
7.0 to 8.9
Sol moisture content 11% to 14% by weight
NOTES
2-39
-------
TOTAL OIL AMD GREASE CONCENTRATIONS
Sample
Treatment Nunbtr
CONTROL 1
2
3
Average
Standard Deviation
51 LOADING RATE 1
2
3
Average
Standard Deviation
5t LOADING RATE AND
NUTRIENT-ADJUSTED 1
2
3
Average
Standard Deviation
5t LOADING RATE.
NUTRIENT-ADJUSTED 1
AND INOCULATED 2
3
Average
Standard Deviation
101 LOADING RATE 1
2
3
Average
Standard Deviation
IN SOIL MICROCOSMS (og/kg)
Week
510.000
470.000
460.000
480.000
26.458
33.000
33.000
26.000
30.667
4.041
38,000
43,000
22.000
34.333
10.970
22,000
26.000
28,000
25,333
3,055
47,000
66.000
46.000
53.000
11.269
410.000
440.000
450.000
433.333
20.817
34.000
26.000
31.000
30.333
4.041
18.000
19.000
16.000
17.667
1.S28
26.000
26.000
59.000
37.000
19.053
47.000
87.000
56.000
63.333
20.984
510.000
550,000
510.000
523.333
23.094
35.000
28.000
34.000
32.333
3.786
18.000
18.000
22.000
19.333
2.309
37.000
29.000
21.000
29.000
8,000
41.000
43.000
48,000
44.000
3.606
530.000
510.000
460.000
500.000
36.056
30,000
32.000
30,000
30,667
1.155
14.000
16.000
15.000
15,000
1.000
18,000
25.000
18,000
20.333
4,041
42.000
31.000
34,000
35.667
5.686
TOTAL OIL AND GREASE CONCENTRATIONS (ng/kg) IN SOIL
MICROCOSMS SIMULATING SOLID PHASE BIOREMEDIATION
OF SLUDGE MATERIAL
Treatment
Control
51 Loading Rate
+pH Adjust
51 Loading Rate
+ Nutrients + pH Adjust
51 Loading Rate
+ Nutrients + pH Adjust
+ Inoculated
101 Loading Rate
+ Nutrients + pH Adjust
Time (weeks)
480,000
30.667
34.333
25.333
53,000
433.333
30,333
17.667
37,000
63.333
523.333 500.000
32.333 30.667
19,333 15.000
29.000 20.333
44,000 35,667
2-40
-------
NOTES
SUMMARY
Ctearty define the scope of work
Look for wel controlled studtes
Look for statisticaly vafd experimental design
Always look at the raw data and
formulate your own opinion
Beware of the Imitations of standard methodologies
Always seek expert opinion and
independent evaluation
2-41
-------
WORKSHEET FOR HAZARDOUS WASTE SITE CHARACTERIZATION
1. What information is important to the facilities description?
2. What are the most important aspects of the general site description?
3. What can the history of the ownership and operation tell us?
4. What site characteristics should be considered?
5. What chemicals are present at the site?
6. How many different contaminated areas are within the site?
7. Where is the contamination located?
Site 1.
Site 2.
Site 3.
Site 4.
Site 5.
site 6. ^I^ZIZ^ZI^Z^IZIZIZ^ZI^^ZZZ!
Site 7.
2-42
-------
8. What is the extent (e.g. ppm) of the contamination at each site?
Site 1.
Site 2. I^^^ZIIIZZI^IZZZIZIIZZZZ^^I^ZZ
site 3. !^^^^IZZZ^ZZZIZIZI^IZZZZZZZZZZI
site 4. ZZ^^ZZZZHZIZZZIIZZrZIZI^IZIIZIZZ
site 5. ZZZZZZ^Z^ZIZ^IIIZ^^ZZZIZZII^ZZ
site e. ZZIZZIZIZZZZZ^^ZZ^^ZZZIIIIZIIZIZI
Site 7. ~~~~~~^~~~
9. What do we need to know about the site to estimate the extent of
contamination?
10. What are the important hydrogeological aspects?
11. Do you anticipate movement from these locations? If so, how could
that impact treatment?
12. What aspects of chemical identification should we be most concerned
about?
2-43
-------
13. What are the important aspects of quality assurance?
14. What are the principal analytical tools used for the identification
and quantification of hazardous organic chemicals and for which
groups of compounds?
15. Where should you look for extraction and sample preparation
procedures?
16. What do you need to know to ensure the validity of the analytical
procedures?
17. Are the chemicals potentially biodegradable?
Site 1.
Site 2. ~
Site 3. ~
Site 4. ~
Site 5.
Site 6. ~~
Site 7.
2-44
-------
18. Are any of the contaminants potentially toxic to microblal
degradation processes?
Site 1.
Site 2. ^ZI^ZZ^ZIZZZIZ^^^^^^^IIZZZ
site 3. IIIZIZ^^ZZIZIIZZZZZIZ^IIZZIZIZI^ZII
site 4. ^ZZZHHZZ^^^^ZZIZI^ZIZIIIZI^^^;
site 5. ^^^^ZZHZZZZI^^ZZIZZZIII^^ZZZZZ
site 6. ZZZHHZZZ^^I^^ZZIZI^ZZZZIZZIIII
Site 7. ————————-^
(Could you pretreat the waste so It could be degraded biological)
19. Is the environment appropriate for biological treatment or can the
environment conditions be adjusted to make it more appropriate for
biological treatment?
Site 1. Site 5.
Site 2. Site 6.
Site 3. Site 7.
Site 4.
20. Should aerobic or anaerobic biotreatment be considered?
Site 1. Site 5.
Site 2. Site 6.
Site 3. Site 7.
Site 4.
21. How would you design a treatability study(ies) and what protocols
would you use to encompass all of the contaminated areas?
2-45
-------
22. What Information will be obtained from a treatability study and how
will it be used?
23. What type of information should be sought before final technology
selection?
2-46
-------
EXAMPLE SITE
FOR
BIOREMEDIATION
3-1
-------
HAZARDOUS WASTE SITE FOR BIOREMEDIATION
Background
The operations at a 25 acre industrial waste complex located near
factories and various chemical processing plants have contributed to a
seven acre hazardous waste disposal area located on site. Figure 1
represents the general layout of the industrial complex. To the north of
the site a residential area has been developed. Over the past forty
years, organics and inorganics generated from the on-site factory and
other nearby industries have been dumped into the hazardous waste
disposal site. During drought conditions, local water wells have been
found to be contaminated by materials from the hazardous waste site. In
response, a site investigation was completed to determine the
contaminants present in each media, their approximate concentrations, and
where each contaminant zone was located.
Site Description
The hazardous waste disposal area is approximately seven acres and is
located in the southwest corner of the industrial complex as illustrated
in Figure 1. It contains a one acre pit in which contaminated soils and
sludges were deposited and a three acre pond containing miscellaneous
liquid wastes. An underground storage tank containing diesel fuel,
located between the pit and pond was abandoned when dredging of the pond
was discontinued. An additional source of contamination identified was
the tank farm area, where trucks had spilled their contents during
loading and unloading operations.
The site geological setting, as determined from existing surveys of
the area, is as follows. The surface soil layer at the site is a sandy
soil with high permeability and a depth of 3-5 feet. The subsurface has
been characterized as a silty and sandy clay that is moderately permeable
and has a depth of approximately 30 feet.
Based on field investigations, a cross section of the site was
developed as shown in Figure 2. The depth to groundwater from the
surface averages 30 feet across the site, and the depth to bedrock is
approximately 65 feet. The bedrock consists of an impermeable
limestone. Table 1 lists additional information about each contaminated
media.
The climate in this area is very humid and has an average temperature
of 72°F and an annual precipitation of 53.4 inches. The high and low
temperatures in Jaunary are 74°F and 49°F and in August are 92°F and
72°F, respectively.
3-2
-------
Figure 1. Industrial Complex
-*•
O
U)
I
oo
Tank Farm
O
O
O
O
O
O
Truck
Loading
Area
Pipe Line
-, Underground
i Storage Tank
-*-
Factory
Drivewav
-rr
O
N
-------
TABLE 1. ADDITIONAL SITE INFORMATION
System 1 — Contaminated Surface Soil
Estimated Volume — 2000 cubic yards
Estimated Size (50 ft x 200 ft x 5 ft)
System 2 — Pit Containing Contaminated Sludges and Soils
Surface Area Depth Volume
Pit Size (overall) 1 acre 5 feet 8000 cubic yards
Haste Volume 1 acre 4 feet 6400 cubic yards
System 3 — Leaking Underground Storage Tank
Estimated Volume of Contaminated Soil Beneath the Tank — 410 cubic
yards (approximate size 45 ft x 25 ft x 10 ft)
Estimated Volume of Contaminated Groundwater — 0.5 million gallons
(approximate size 45 ft x 100 ft x 15 ft)
System 4 — Pond
Estimated volume of contaminated water in the pond - 20 million
gallons.
Estimated Volume of Contaminated Soil Beneath the Pond — 91700 cubic
yards (approximate size 660 ft x 250 ft x 15 ft)
Estimated Volume of Contaminated Groundwater — 128 million gallons
(approximate plume size 660 ft x 1300 ft x 20 ft)
System 5 — Mixed Groundwaters - Tank and Pond
Estimated Volume -- 10,000 gallons
System 6 — Broken Pipe Leakage
Estimated volume of contaminated soil — 250 cubic yards
Estimated volume of contaminated groundwater — 500,000 gallons
(approximate size 125 ft x 25 ft x 20 ft)
System 7 — Mixed Groundwater Pipe Leakage and Pond
Estimated volume of contaminated groundwater — 75,000 gallon
3-4
-------
Description of Contamination
During the field Investigation, the hazardous waste site was found to
contain a variety of organic contaminants as well as some Inorganic
contamination. The following 1s a general description of the
contaminants found:
• Pit - The pit contains contaminated soils and sludges. The
material is acidic and is contaminated with methyl ethyl
ketone. In addition, an oil sludge was found at the bottom of
the pit.
• Pond - The liquid in the pond contains water contaminated with
coal tar and its by products including some cyanide.
• Underground Storage Tank - An undergound storage tank located
between the pit and pond was found to be leaking diesel fuel.
• Tank Farm Area - The soil in the area of the loading dock is
contaminated with pentachlorophenol, polychlorinated biphenols
and trivalent chromium. A review of plant history indicated
these spills resulted from loading and unloading operations
prior to the construction of the concrete dock. Groundwater
contaminated with trichloroethylene was identified during the
field investigation. The source of this contamination was
traced to a broken transfer line from the tank farm to the
factory. The broken line was discovered and repairs made two
years ago.
The contaminated leachate plumes from the various sources identified
above are shown in Figures 2 and 3. Table 2 represents concentration
levels for each contaminated system and media and other pertinent
information.
Planning Site Response
The cleanup objectives for each contaminated media are also listed in
Table 2. These objectives offer an end point for remediating the site
when biological and other supporting technologies have been applied.
These clean-up objectives are for the purposes of this workshop only.
Table 3 provides chemical and physical properties of the contaminants
discovered at the industrial complex.
3-5
-------
Figure 2. Cross-sectional View of Site
n
System 1 System 2
System
Contaminated/ /
Surface Soil Pipe
Elevation
0 Figure 3. Industrial Complex Showing Plume Delineations
3-6
-------
TABLE 2. CONTAMINATED SYSTEMS
Contaminant Concentrations and Clean Up Objectives
System 1 — Contaminated Surface Soil
Soil
Contaminant
PCP
PCB
Cr+a
Concentration
180 mg/kg
300 mg/kg
900 mg/kg
Clean-up Objectives
50 jag/kg
50 mg/kg
170 mg/kg
System 2 — Pit Containing Contaminated Sludges and Soils
Pit
Contaminant
MEK
Oily sludge
pH*
Solids %
Concentration
400 mg/kg
900 mg/kg
2.5
85
Clean-up Objectives
1 mg/kg
45 mg/kg
6-9
System 3 — Leaking Underground Storage Tank and Related Contaminated
Zones
Soil below
tank
(Soil - 3)
Groundwater
(GW-3)
System 4 —
Pond
Contaminant
Diesel fuel
Contaminant
Diesel fuel
Iron
pH*
Pond and Related
Contaminant
Cyanide
Benzene
Toluene
Xylene
Phenol
Cresol
Naphthalene
Ammonia
pH*
TDS
TSS
TOC
COD
Concentration
50 mg/kg
Concentration
150 mg/ft
25 mg/ft
6.5
Contaminated Zones
Concentration
3 mg/ft
400 mg/ft
280 mg/ft
250 mg/ft
325 mg/ft
45 mg/ft
60 mg/ft
39 mg/ft
9.2
500 mg/ft
100 mg/ft
298 mg/ft
950 mg/ft
Clean-up Objectives
15 mg/kg
Clean-up Objectives
10 mg/ft
NA
6-9
Clean-up Objectives
0.15 mg/ft
10 iag/ft
10 ng/ft
10 yg/ft
10 iag/ft
5 |ig/ft
5 yg/ft
2 mg/ft
6-9
—
50 mg/ft
15 mg/ft
50 mg/ft
3-7
-------
TABLE 2. (continued)
Soil below
pond (Soil-4)
Groundwater
(GW-4)
Contaminant
Cyanide
PCP
PCB
Benzene
Toluene
Xylene
Phenol
Ammonia
Cr+3
Contaminant
Cyanide
Benzene
Toluene
Xylene
Phenol
Ammonia
Iron
pH*
Concentration
1 .7 mg/kg
18 mg/kg
50 mg/kg
250 mg/kg
160 mg/kg
110 mg/kg
190 mg/kg
50 mg/kg
200 mg/kg
Concentration
0.4 mg/a
150 mg/a
80 mg/a
70 mg/a
100 mg/a
80 mg/2.
25 mg/a
6.5
Clean-up Objectives
0.09 mg/kg
50 ng/kg
50 n9/kg
10 jag/kg
10 n9/kg
10 ng/kg
10 ng/kg
2 mg/kg
170 mg/kg
Clean-up Objectives
0.02 mg/a
5 iag/a
5 pg/a
5 pg/Jl
5 ng/a
2 mg/ft
NA
6-9
System 5 — Groundwater Contaminated with Mixture of Pollutants from
Tank, and Pond
Groundwater
(GW-5)
Contaminant
Diesel fuel
Cyanide
Benzene
Toluene
Xylene
Phenol
Ammonia
Iron
pH*
Concentration Clean-up Objectives
150 mg/fi,
0.4 mg/fi,
150 mg/2,
80 mg/a
70 mg/a
100 mg/a
80 mg/a
25 mg/a
6.5
10 mg/a
0.02 mg/a
5
5
5
5
2
NA
6-9
mg/a
3-8
-------
TABLE 2. (continued)
System 6 — Leaking Transfer Piping System
Contaminant Concentration
Soil below pipe
(Soil - 6)
Trichloroethylene 2.50 mg/kg
Clean-up Objectives
10 yg/kg
Groundwater
(GW-6)
System 7 —
Contaminant
Trichloroethylene
Iron
pH*
Concentration CJ
10 mg/fi,
25 mg/fi,
6.5
Groundwater Contaminated Hith a Mixture
ean-up Objectives
5 yig/fi.
NA
6-9
of Pollutants
From the Pipe Leakage and Pond
Groundwater
(GW-4)
Contaminant
Cyanide
Benzene
Toluene
Xylene
Phenol
Ammonia
Trichoroethylene
Iron
pH*
Concentration CJ.
0.4 mg/fi.
150 mg/ft.
80 mg/fi.
70 mg/fi.
100 mg/fi.
80 mg/fi.
10 mg/fi.
25 mg/fi.
6.5
ean-up Objectives
0.02 mg/fi.
5 ng/fi.
5 ng/fi.
5 pg/fi.
5 ng/fi.
2 mg/fi.
5 ng/&
NA
6-9
*standard units
3-9
-------
TABLE 3. PROPERTIES OF CONTAMINANTS
Solubility Soluble
Chemical Class
Halogenated Aliphatics
• Trichloroethylene (TCE)
Halogenated Polycyclic Aromatics
• Polychlorinated biphenyls
Monocyclic Aromatics
• Benzene
UJ
,_!_, • Toluene
o
• Xylene
• Phenol
in
Water
1000
mod.
3.1
low
low
515
mod.
0.3
low
84,000
high
in
Solvents
alcohol ,
ether, acetone,
chloroform
alcohol, ether,
acetone
alcohol ,
ether, acetone,
carbon
tetrachloride
alcohol ,
ether, acetone,
benzene
alcohol ,
ether, acetone,
benzene
water, alcohol,
ether, acetone,
benzene,
chloroform
Soil
Adsorp-
tion
mod.
high
high
high
mod.-
high
low-
mod.
Henry's
Constant
(Volatility)
8.9 x 103
high
1.7 x 10~3
high
high
6.6 x 10~3+
high
6.3 x 10-3+
high
7.0 x 107
low
Biodegrad-
ability Toxicity
R toxic by
inhalation
D,R highly toxic
to ecology
D highly toxic
to ecology
D toxic by
ingestion and
skin adsorption
D toxic by
ingestion and
inhalation
D toxic by
ingestion and
inhalation and
skin absorption
Mobility
mod. - high
in soil-
water systems
1 ow - v . 1 ow i n
water systems,
mod . in ai r
mod. - high in
water systems
mod. - high in
water systems
soil-
v . 1 ow-
soil-
soil-
mod. in soil -water
systems, mod. -
in ai r
mod. - high in
water systems
• high
soil-
(continued)
-------
TABLE 3. PROPERTIES OF CONTAMINANTS (continued)
Solubility
in
Chemical Class Water
• Cresol high
• Pentachlorophenol (PCP) 14
low
Polycyclic Aromatics
• Naphthalene 31.7
low
Alkylated Aliphatics
• Methyl ethyl ketone 353,000*
high
I
t— '
i -•-
Metals
• Chromium III NA
Diesel Oil low
Ammonia high
Soluble
in
Solvents
alcohol, glycol
water
alcohol, ether,
benzene
alcohol, ether,
acetone,
benzene
water, alcohol,
benzene, ether,
acetone
->
hydrochloric
acid,
sulfuric acid
benzene,
toluene
water, alcohol,
ether
Soil Henry's
Adsorp- Constant
tion (Volatility)
, low - low
mod.
high 2.8 x 10~6
low - mod.
high 4.8 x !Q-4+
mod. - low
low 4.35 x 10-5
mod.
NA NA
mod . 1 ow
high high
Biodegrad-
ability Toxicity
D toxic by
skin absorption
R.D highly toxic
to ecology
D toxic by
inhalation
D toxic by
inhalation
( ) ]ow toxicity
D envi ronmental
hazard
toxic by
D inhalation
Mobi 1 i ty
mod. - high in
soil -water systems
high - v. high in
soil -water systems
v. low - low in soil-
water systems
v . 1 ow - mod . in air
mod. - high in soil-
water systems
negligible to v. low in
air, v. low - v. high
in soil and aqueous
systems
mod. - high in soil-
water system
high in air, mod. -
high in soil -water
systems
Solubility = mg/fl. at 20°C (*at 10°C)
Henry's Constant = atm • m3/mo1 at 20°C (+ at 25°C)
Biodegradability (D = degradable, R = refractory, N = non-degradable, ( ) = no information available)
NA = not applicable
-------
REACTOR DESIGN
SECTION 4
Abstract
Slides
Worksheets
4-2
4-10
4-49
-------
REACTOR TREATMENT DESIGN
Evan K. Nyer George J. Skladany
Geraghty and Miller, Inc. JTC Environmental Consultants
Tampa, Florida Gaithersburg, Maryland
Biological processes have successfully transformed organic and
inorganic materials on the earth for billions of years. Biological
processes have been used extensively since the turn of the century to
treat municipal and industrial wastewaters. The use of microorganisms to
treat hazardous materials is a logical extention of applied
microbiology. In the past few years, great progress has been made in
isolating, characterizing, and modifying organisms able to metabolize
materials considered to be hazardous. The successful application of
these microorganisms to commercially available treatment systems falls
within the engineering domain.
In many site remediation projects, it is difficult to determine if a
waste stream (liquid or soil) is amenable to biological treatment, and if
it is, what type of bioreactor design to use. Successful biological
treatment of groundwater, leachate, or industrial process water requires
the combined action of basic microbial processes and sound process
engineering designs. Such a treatment system is then able to both
efficiently and cost effectively remediate the contaminants present. The
decision to consider and use bioremediation at hazardous waste sites,
however, rests with site remediation project managers.
This presentation is designed to provide information about several
subject areas critical to the success of any biological treatment
project, including conceptual process design, basic bioengineering
principles, a review of currently available biological unit processes,
important pretreatment and postreatment factors, and case histories.
While not being comprehensive in detail, the written material given below
(coupled with the oral presentation) should provide class attendees with
a base level of understanding of bioreactor selection and operation.
Conceptual Remediation Approach
One of the first steps to take in selecting remediation equipment is
to define the treatment system needed. Specifically, this requires the
project worker to identify all of the inputs and outputs to a treatment
process. In all cases, the composition of the influent waste and
required discharge standards for the waste stream must be considered.
With a biological treatment system, consideration must also be directed
to any anticipated air emissions and to proper biological and/or
inorganic sludge disposal. Once the treatment parameters have been
defined, attention can be given to the proper selection of remedial
process designs.
4-2
-------
"Life-Cycle Design" is a remediation approach that takes into account
changes in site conditions throughout the duration of the project.
Life-Cycle Design has three major facets:
• Time effect on parameters
• Capital equipment costs
• Operating expenses
The "time effect on parameters" considers that any process design
must be flexible enough to overcome changes over time in the volume of
materials to be treated (such as varying water flow rates), the
appearance or disappearance of specific organic or inorganic
contaminants, and changes in individual contaminant concentrations. A
process designed only for present si^e conditions may become cost
prohibitive or catastrophically fail at some point in the future.
Actual capital equipment costs reflect both the total dollar amount
spent as well as the expected duration of equipment use. Nhile most
municipal projects are designed for 20 years or more of operation, many
environmental projects will have a much shorter period of operation.
Thus, the daily cost for equipment will tend to be higher for hazardous
waste projects. To lower this cost, consideration should be given to
using equipment that is portable and reuseable. Depending on the
project, large permanent installations should be avoided if possible.
Lastly, Life-Cycle Design considers the affect of operating expenses
on the remediation effort. Operating expenses consist of maintenance
items, power costs, consumable supplies, and personnel costs. Personnel
costs can be kept low by utilizing equipment that requires a minimal
amount of operator attention or that is self operating. On many
projects, personnel costs are the major operating expense, especially
with complex treatment systems that require round the clock attention.
High initial capital equipment costs can be quickly offset in many cases
by lower annual operating expenses. The design engineer must consider
operating as well as capital equipment costs when evaluating potential
process equipment designs.
Bioprocess Engineering and Treatment Equipment
The design engineer must create an environment favorable for rapid
microbial growth. In terms of overall treatment processes, bioreactors
can be designed to handle either batch or continuous flows. Contaminants
can be treated in:
• Batch mode with discontinuous flow
• Plug flow mode with continuous flow
• Partially mixed mode with continuous flow
Completely mixed mode with continuous flow
4-3
-------
Each of these treatment modes has advantages and disadvantages from
both microbiological and operational perspectives. The microbial growth
rate (and hence the specific compound removal rate) can be controlled by
the design and operation of the specific bioreactor. For example, a
fixed-film design may be superior to a dispersed growth design if the
reactor needs to be populated with slow growing bacteria. The fixed-film
design effectively separates the microbial residence time within the
reactor from the hydraulic retention time of the water passing through
the system.
Any bioreactor design must also ensure that proper pH, temperature,
oxygen concentration, and inorganic nutrient concentrations (primarily
nitrogen and phosphorus) are maintained. On a practical note, the
hydraulic retention time needed for biodegradation to occur controls the
size of the bioreactor. Suitable microbial populations must be
maintained within the system to keep the hydraulic retention time (and
hence the bioreactor size) to a minimum. Very large tanks are capital
Intensive and have greater operating costs due to power requirements in
mixing and oxygen transfer.
Biological treatment equipment can take many forms, but all designs
employ bacteria growing either dispersed 1n the bulk liquid or attached
as films on some sort of inert support surface. Below are brief
descriptions of several commercially available biological processes for
water treatment:
Activated Sludge
• Suspended growth system
• Completely mixed mode
• Biomass captured 1n clarifler and recycled to reactor
• Contact time between waste and biomass controlled by wasting
excess biomass
Aerated Lagoons
• Suspended growth system
• Completely mixed mode
• Contact time limited to hydraulic retention time
• Limited effluent quality
Extended Aeration
• Suspended growth system
• Completely mixed mode
• Biomass captured in clarifier recycled to reactor
• Long contact time created by enlarging aeration basin
Contact Stabilization
• Suspended growth system
• Completely mixed mode
Waste quickly contacted with biomass in first aeration tank
4-4
-------
• Contaminants adsorbed to clarified biomass are then digested in
second aeration tank
• Total hydraulic residence time held to a minimum
Trickling Filter
• Fixed-film system
• Plug flow mode
• Design based on specific surface area
• Aeration provided by induced or forced draft
Rotating Biological Contactors
• Fixed-film system
• Plug flow mode
• Design based on specific surface area
• Aeration provided by rotating disks
Submerged Fixed-Film Reactors
• Fixed-film system
• Completely mixed or plug flow modes
• Design based on volume
• Aeration provided by air released below media
Powdered Activated Carbon Treatment (PACT)
Hybrid suspended growth/fixed-film system
Completely mixed mode
Biomass suspended and fixed to carbon particles
Carbon particles also adsorb organic contaminants
Clarifier still controls bacterial residence time
Fluidized Bed
• Fixed-film system
• Completely mixed or plug flow modes
• Media fluidized in reactor
The nine treatment systems described above are designed for the
aerobic biodegradation of contaminants. However, some chemicals are more
readily biodegraded under anoxic (low oxygen) or strict anaerobic (no
oxygen) conditions. With the proper engineering modifications, many of
the above mentioned systems can be used for anoxic/anaerobic treatment of
hazardous chemicals. Anaerobic digesters have been used for some time in
combination with aerobic activated sludge to treat municipal waste.
Combination anoxic/anaerobic treatment systems are also in use.
Anaerobic fluidized beds, with and without activated carbon, have shown
promise for use in the hazardous waste treatment field. While much is
known from a microbiological standpoint about the anaoxic/anaerobic
biodegradation of compounds, very few large scale applications of this
technology exist today.
4-5
-------
While the biological treatment of liquid wastes is a fairly well
understood and straightforward process, the biological treatment of
contaminated soils is more complex and difficult to put into practice.
The same factors important to rapid microbial growth in above ground
systems (pH, nutrients, oxygen concentration, etc.) are critical when
treating soils. However, soils are typically quite heterogeneous, as
opposed to the more homogeneous water matrix. It is more difficult for
microorganisms (or physical/chemical reagents for that matter) to gain
equal access to each and every soil particle present. In addition, soils
treatment presents more difficult materials handling problems.
Excavation of contaminated soils may reveal the presence of buried
materials such as pipes or bricks, making it more difficult to homogenize
the soils prior to treatment.
In spite of these difficulties, biological treatment of soils remains
a valuable tool for the remediation specialist. In many cases,
indigenous microorganisms possess the metabolic capability to metabolize
the contaminants present. All that is needed is to further optimize
growth conditions. In some cases, it may be necessary to inoculate the
soils with microorganisms containing the desired metabolic activity. Two
major forms of biological soils treatment are described below:
Contained Above Ground Soils Treatment
Batch mode
Contaminants treated in the heterogeneous soil matrix
Nutrients, moisture and oxygen added as needed
Leachate, runoff, and air emissions must be controlled
Soil left on site when clean
Soil Slurry Reactors
Batch or continuous flow mode
Heterogeneous soils treated in a liquid slurry
Nutrients and oxygen added as needed
Water and soil must be separated after treatment
Soil left on site when clean
Pre and Post Treatment Considerations
There are several factors that must be evaluated prior to and after
using biological treatment. Pretreatment factors are concerned with
creating a suitable microbial growth environment. Apart from the factors
discussed earlier (pH, temperature, oxygen and nutrient concentrations),
attention must be directed at the presence of high concentrations of
toxic or inhibitory compounds. These materials may be organic or
inorganic (such as metals) in nature. In many cases, toxic or inhibitory
concentrations of materials can be effectively treated with the proper
reactor design. For example, toxic concentrations of phenol, will cause
process failure under batch treatment conditions, but may be easily
4-6
-------
biodegraded in a continuous flow completely mixed bioreactor. The
process engineer may need to consult with an environmental microbiologist
when dealing with compounds of known microbial toxicity or inhibition.
Another pretreatment factor to consider is the presence of nuisance
chemicals, such as high concentrations of iron. While iron would not
adversely affect the biological processes taking place, it would oxidize
and precipitate out of solution. This could cause fouling and
degeneration of the biofilm or the production of excess metal-containing
sludge.
Post-treatment factors which need to be evaluated include solids
removal (both biological and inorganic precipitates) and pass through
organics (those organics which cannot be biodegraded or remain as a
result of process efficiency). Not every compound present in the waste
stream may be completely metabolized during biological treatment under a
defined set of conditions. Certain compounds (such as trichloroethylene
or carbon tetrachloride) may pass through completely undegraded.
Metabolic byproducts and cell lysis materials are also produced with any
biological treatment process. These materials may have to meet certain
discharge criteria (Total Organic Carbon, for example) before the treated
water is suitable for disposal. Volatile compounds, especially those
resistant to biodegradation, can be air stripped from biological
treatment systems and may have to be controlled.
Once a decision has been made that a waste stream is amenable to
biological treatment, conceptual process designs can be made. Several
different types of biological treatment systems may be under
consideration. At this point it is important to look at the overall
economics of the project. This encompasses all capital, installation,
and operating expenses (including disposal of any end-product
materials). The expected duration of the project will have an obvious
impact on the overall project costs. Changes in waste volume,
contaminants, and concentrations over the life of the project will also
impact the system design and project costs. It is important to have a
realistic project time estimate and life cycle description in order to
compare the costs associated with different biological treatment systems.
Lastly, there may be important benefits in combining the action of
above ground and in situ biological treatment systems. This is
especially true if treated water from the bioreactor (usually rich in
nutrients, oxygen, and suitable bacteria) can be reinjected into the
subsurface. The combined action of such treatment systems may
considerably shorten the time required to complete a remediation as
compared to above ground or in situ remediation used alone. However,
care must be exercised to ensure that the subsurface injection of
materials does not further solubilize and mobilize the contaminants
present.
4-7
-------
SELECTED ADDITIONAL READING
Alexander, Martin. (1985). "Biodegradation of Organic Chemicals.",
Environmental Science and Technology. 18(2): 106-111.
Atlas, Ronald M., Editor. (1984). Petroleum Microbiology. Macmillan
Publishing Company, New York.
Cerniglia, Carl E. (1984). "Microbial Metabolism of Polycyclic Aromatic
Hydrocarbons.", Advances in Applied Microbiology. Volume 30,
Allen I. Laskin, Editor, Academic Press Inc., New York: 31-71.
Dragun, Games. (1988). The Soil Chemistry of Hazardous Materials.
Hazardous Materials Control Research Institute, Silver Springs, Maryland.
Guady, Anthony F., Or., Elizabeth T. Gaudy. (1980). Microbiology for
Environmental Scientists and Engineers. McGraw Hill Book Co., New York.
Grady, C.P. Leslie, Jr. (1985). "Biodegradation: Its Measurement and
Microbial Basis.", Biotechnology and Bioengineering. 27: 660-674.
Grady, C.P. Leslie, Jr. and Henry C. Lim. (1980). Biological Kastewater
Treatment. Marcel Dekker, Inc., New York.
Kobayashi, Hester and Bruce E. Rittmann. (1982). "Microbial Removal of
Hazardous Organic Compounds.", Environmental Science and Technology.
16(3): 170A-183A.
Leisinger, T., R. Hutter, A.M. Cook, and J. Nuesch, Editors. (1981).
Microbial Degradation of Xenobiotics and Recalcitrant Compounds. Academic
Press, New York.
Metcalf & Eddy, Inc. (1979). Nastewater Engineering: Treatment.
Disposal. Reuse. McGraw-Hill Book Company, New York.
Nyer, Evan K, (1985). Groundwater Treatment Technology. Van Nostrand
Reinhold Company Inc., New York.
Patterson, James W. (1985). Industrial Hastewater Treatment Technology.
Second Edition, Butterworth Publishers, Stoneham, Massachusettes.
Rochkind-Dubinsky, Mellissa L., Gary S. Sayler, and James W. Blackburn.
(1987). Microbiological Decomposition of Chlorinated Aromatic Compounds.
Marcel Dekker, Inc., New York.
Tabak, Henry H., Stephen A. Quave, Charles I. Mashni, and Edwin F. Barth.
(1981). "Biodegradability Studies with Organic Priority Pollutant
Compounds.", Journal. Hater Pollution Control Federation. 53(10):
1503-1518.
4-8
-------
Verschueren, Karel. (1983). Handbook of Environmental Data on Organic
Chemicals. Second Edition. Van Nostrand Relnhold Company Inc., New York.
Wood, John M. and Hong-Kang Wang. (1983). "Mlcroblal Resistance to
Heavy Metals.", Environmental Science and Technology. 17(12): 582A-590A.
Wood, John M. (1982). Chlorinated Hydrocarbons: Oxidation in the
Biosphere.", Environmental Science and Technology. 16(5): 291A-297A.
4-9
-------
DEFINE TREATMENT SYSTEM
Influent
Concentration*
Oltch«rg«
Requirement*
I Sludge
I Dltpoivl
NOTES
NOTES
LIFE-CYCLE DESIGN
• Time effect on parameters
• Capital costs
• Operator expenses
TIME EFFECT ON PARAMETERS
(.••chat*
Source: Nyer, QroundwaUr Treatment Technology
4-10
NOTES
-------
CAPITAL EQUIPMENT COSTS
Daily costs.
300
250
200
150
100
50
0 1 2 4 6 8 10
Time for write-off, years
Assume: 2100,000 capital equipment costs and 12% interest rate
Source: Nyer, Groundwater Treatment Technology
OPERATIONAL EXPENSES
Assume:
* $100,000 capital cost
• 10 year life of equipment
• 12% interest rate
• 15 hp for power ($0.06/kWh)
• $3/day chemical cost
* $10/hour for operator
4-11
-------
OPERATIONAL EXPENSES
WITH NO OPERATOR ATTENTION
• Chemicals 4Z
EO Power 362
IB Equipment 60Z
Source: Nyer, Groyndwater
Treatment Technology
OPERATIONAL EXPENSES WITH
8 HOURS/DAY OPERATOR ATTENTION
• Chemicals 2%
m Power 18Z
H Equipment 302
0 Operator 50Z
Source: Nyer, Groundwater
Treatment Technology
4-12
-------
OPERATIONAL EXPENSES WITH
24 HOURS/DAY OPERATOR ATTENTION
Chemicals 1%
Power 9%
Equipment 152
Operator 752
Source: Nyer, Groundwater
Treatment Technology
OPERATIONAL EXPENSES SUMMARY
Daily costs Si/day
•100
300
100
10 20
Man-hours/day
• Source: Nycr. Gjiqund_waJer.Tjreatme_nLTep.h.l1
-------
OPERATIONAL EXPENSES WITH
$500,000 CAPITAL EQUIPMENT AND
24 HOURS/DAY OPERATOR ATTENTION
9 Chemicals 2.32
m Power 22.7%
S Equipment 37.52
E3 Operator 37.52
Source: Nyer, Groundwater
Treatment Technology
DIFFERENT DESIGN CONFIGURATIONS
Based On Practical Solution
To Two Issues
* Microorganisms residence time and
the relative effect on effluent
concentration
• Oxygen transfer
4-14
-------
BIOREACTOR DESIGN
Flow Considerations
• Batch
• Plug flow
• Continuous flow completely mixed
• Continuous flow partially mixed
NOTES
NOTES
PLUG FLOW
9 Ideally no mixing
• Equal treatment for all materials present
• Subject to shock load upsets
6 Influent concentrations decrease with passage
CONTINUOUSLY STIRRED TANK REACTOR (CSTR)
* Evennea* of treatment dependent upon reaction
time within reactor
* Influent concentration* Instantaneously diluted
Into bulk liquid
* Effluent concentration equala bulk liquid concentration
• Good with ehock lotdt and with toxic/Inhibitory
concentrations of chemlcata
4-15
NOTES
-------
ARBITRARY FLOW
Somewhere between plug flow and CSTR
Usually more representative of what
actually happens
NOTES
NOTES
BIOREACTOR DESIGN
Environmental Conditions
• Temperature
• pH
• Oxygen
• Inorganic nutrients
• Toxics
REACTOR DESIGN
Practical Considerations
• Hydraulic residence time
• Bacterial residence time
• Mixing
• Oxygen transfer
• Bacteria/organics contact
NOTES
4-16
-------
6OO
I 2 4 « 6 IO 20 40 60 100 200 400 800
Spoce Tim«. T-tirs
Effect of Space Time on the Performance of a Single CSTR.
Source: Grady and Lun, Biological Wastowater Treatment
500
"2 4 7 10 20 40 70 IOO 200 4OO
Meon Cell Residence Time -$c-hr«
Effect of Mean Cell Residence Time on Oxygen Requirement and
Excess Microorganism Production Rate in a Single CSTR with Cell Recycle.
Source: Grady and Lim, Biological Wastewater Treatment
4-17
-------
AERATED LAGOON
• Biomass kept suspended
in liquid
• Contact time limited to
hydraulic residence time
• Limited effluent quality
NOTES
NOTES
AERATED LAGOON
Mixing and
Oxygen Transfer
ACTIVATED SLUDGE
• Biomass kept suspended in liquid
• Biomass captured in clarifier
recycled to reactor
• Contact time between waste
and biomass controlled by
wasting excess biomass
4-18
NOTES
-------
COMPLETELY MIXED ACTIVATED SLUDGE
Excesc Biofnas*
NOTES
NOTES
EXTENDED AERATION
• Biomass kept suspended in liquid
• Biomass captured in clarifier
recycled to reactor
• Long contact time created by
enlarging aeration basin
EXTENDED AERATION
w»u
A 7
^
,
dr
Blom«i* R«cyel«
\
njy ciiriri
Ijf E(flu«
*~
Exc**t Blomits
NOTES
4-19
-------
CONTACT STABILIZATION
• Biomass kept suspended in liquid
• Waste quickly contacted with
biomass in first aeration tank
• Clarified biomass/waste is then
stabilized in second aeration tank
• Total hydraulic residence time
held to a minimum
NOTES
NOTES
CONTACT STABILIZATION
Mixing and
Oxygen Tranif«r
Mixing and
/^N. Oxyg«n Tr«ntf«r
Stabilization
Exe««a Biomai*
SUSPENDED GROWTH REACTORS
Advantages
• Intimate contact between biomass
and waste
* Several methods available for
adjusting performance
• Very low concentrations of
specific organics in effluent
• Large scale system relatively
inexpensive
4-20
NOTES
-------
SUSPENDED GROWTH REACTORS
Disadvantages
• Relies on clarifier for
performance
* Relative high operator attention
NOTES
NOTES
TRICKLING FILTER
Biomass retained in reactor on inert
support
Design based on specific surface area
Plug flow
Aeration provided by induced or
forced draft
EFFECTIVE
SURFACE
AEROBIC
MICROBES
MEDIA SURFACE
-SUBSURFACE
ANAEROBIC
MICROBES
NOTES
4-21
-------
TRICKLING FILTER
Flow Dl*(rlbutor
Exc*t« Blomasi
NOTES
NOTES
ROTATING BIOLOGICAL
CONTACTORS
• Fixed film keeps biomass
in system
• Design based on specific
surface area
* Aeration provided by rotating
disks
* Plug flow
ROTATING BIOLOGICAL CONTACTOR (RBC)
Routing DUki
\/
A 7
Clirlll.d
Effluent
Exc««t Blomast
4-22
NOTES
-------
FIXED FILM REACTORS
Advantages
Low operator attention
Retention of slow growing bacterial
population
Low cost oxygen transfer
NOTES
NOTES
FIXED FILM REACTORS
Disadvantages
Plug flow
Limited operation at high influent
concentration
Hard to adjust operation
SUBMERGED FIXED FILM
• Biomass retained in reactor
on inert support
• Design based on volume
• Completely mixed
• Aeration provided by air
released below media
4-23
NOTES
-------
SUBMERGED FIXED FILM
ln»rt Support
NOTES
NOTES
SUBMERGED FIXED FILM REACTORS
Advantages
* Combines advantages of suspended
growth and fixed film systems
* Portable design possible
* Can be run in low-concentration
mode
SUBMERGED FIXED FILM REACTORS
Disadvantages
* Does not scale well - expensive
for large scale system
* Relatively expensive for oxygen
transfer
4-24
NOTES
-------
SUBMERGED FIXED FILM
Case Study:
Industrial Landfill Leachate
Source: DETOX, Inc. (Dayton, OH)
M—C
I
H
. I
.C—C—H
I
H
META-TOLUIC ACID
SOLUBILITY'- 340 ppm
H
I
O
M-C'
M—C
H—C.
PARA-TOLUIC ACID
SOLUBILITY: 850 ppm
I
ORTHO-TOLUIC ACID
SOLUBILITY: 1180 ppm
TOLUIC ACID STRUCTURES
4-25
-------
TREATMENT OPTIONS
• Off-site disposal
$0.20/gallon
• On-site activated carbon
$0.08/gallon
• On-site biological treatment
<$0.01/gallon
(Based on toluic acid concentrations of
300-400 ppm and flow rates of up to 5 gpm.)
NOTES
NOTES
LABORATORY TREATABIL1TY
STUDIES
Microbial Toxicity/
Growth Inhibition
pH 6.6 and 8.7
LABORATORY TREATABILITY
STUDIES
Aerobic Biodegradation
Study
• 37 days
• 60 ppm toluic acids to <1.5 ppm
• Toluic acid plate counts
4-26
NOTES
-------
LABORATORY TREATABILITY
STUDIES
Anoxic Biodegradation
Study
• 37 days
• pH from 7 to >9.5
• 60 ppm toluic acids to
approximately 55.5 ppm
EXISTING CARBON COLUMNS AND TANKS
HpH- HIGH pH
HHpH' HIGH HIGH pH
FC< FLOW CONTROL
LFA« LOW FLOW ALARM
LPA' LOW PRESSURE ALARM
LTA= LOW TEMPERATURE ALARM
EQUIPMENT PROCESS DESIGN
4-27
-------
£ 400 _
COD OPERATING DATA
I I
100 ISO
Day of Project
I
200
I
260
300
NOTES
NOTES
Contaminant
Influent
o-Tolu1c
Effluent
o-Tolu1c
Influent
m & p-Tolulc
Effluent
n & p-Tolulc
TOLUIC ACID CONCENTRATIONS
8/20/87
43
<0.5
-------
LOW CONCENTRATION (<25 PPM)
SUBMERGED FIXED-FILM BIORE ACTOR
Case Study:
Source: DETOX, Inc. (Dayton, OH)
<_>
8 «H
UJ
£ M
Id
o
-smli
2 4 6 8 10 12
LENGTH ALONG COLUMN (cm)
2 4 e e 10 iz
LENGTH ALONG COLUMN (cm)
Examples of acetate removal under both steady-state and
unsteady-state biofilm conditions.
4-29
-------
0 10 2O 30 40 50 6O 7O 80 9O 100 HO
DAYS OF OPERATION
BENZENE REMOVAL DURING LABORATORY BIOREACTOR DEVELOPMENT.
KEY: INFLUENT BENZENE <•); EFFLUENT BENZENE (•).
NOTES
NOTES
GASOLINE STATION
5 gpm
25 ppm total hydrocarbons
BIOLOGICAL TREATMENT OF
BENZENE FRACTION
E
a.
Q.
O
O
O
LU
LU
N
LU
m
20 40 60 80 100
DAYS OF OPERATION
NOTES
4-30
-------
BIOLOGICAL TREATMENT OF PETROLEUM HYDROCARBONS
300
40 60
Day of Operation
60
100
NOTES
NOTES
Compound
Benzene
Toluene
Xylenes
Average
Removal
> 93%
> 96%
> 91%
SERVICE STATION
• Flow: up to 6 gpm
• Influent BTX: 15-30 ppm
4-31
NOTES
-------
I
Bioreactor COD and BTX Data Summary
300
-o Influent COD
-*- Effluent COO
-•- Influent BTX
Effluent BTX
75 100
Days of Operation
150
POWDERED ACTIVATED
CARBON TREATMENT (PACT)
• Biomass suspended and fixed
to carbon particles
• Carbon particles also adsorb
organic material
• Clarifier still controls bacterial
residence time
• Completely mixed
4-32
-------
POWDERED ACTIVATED
CARBON TREATMENT (PACT)
Powd«r*d
Activated Carbon
Wait*
Mixing and
Oxyg*nTranif«r
Exc*i* Blomata
•nd C*rtaon
NOTES
NOTES
POWDERED ACTIVATED CARBON
TREATMENT(PACT)
Case Study:
Bofors-Nobel, Inc.
Muskegon, Ml
Source: Zimpro Passavant (Rothschild. Wl)
SITE BACKGROUND INFORMATION
• Herbicides and organic chemicals
produced
• 1.2 mgd of groundwater from
abandoned landfill
• 0.6 mgd of production process
waters
* Wasted biomass and spent carbon
treated onsite by wet air
oxidation (WAO)
4-33
NOTES
-------
TREATMENT OPTIONS
• Biological treatment
• Liquid phase activated carbon
• Biological treatment followed by
activated carbon
• Chemical oxidation
• Sorption onto bentonite/clay
NOTES
NOTES
IDENTIFIED GROUNDWATER
CONTAMINANTS
Compound
ortho-Chloroanaline (OCA)
Benzene
Dichlorobenzene isomer
Toluene
1.2-Dichloroethane
Concentration (ppb)
13.000
4,900
2.500
1,500
420
IDENTIFIED GROUNDWATER
CONTAMINANTS
(Continued)
Compounds
Ethyl benzene
Chlorobenzene
Bis (ethyl hexyl) phthalate
3.3 -Dichlorobenzidine (DCB)'
3-Chloroanaline
Concentration (ppb)
220
150
100
86
68
4-34
NOTES
-------
IDENTIFIED GROUNDWATER
CONTAMINANTS
(Continued)
Compound
Benzidine isomer
Phenol
Cresol
Tetrachloroethylene
ortho-Chlorophenol
Concentration (ppb)
65
6
5
5
4
NOTES
NOTES
TREATABILITY STUDY RESULTS
(All concentrations are in ppm)
Parameter
BOO
COO
TOC
Influent Biological Carbon Combined
Cone. Treat. Treat. Treat.
30 to 40
0 to 5
N« Data
20 to 3<5
Suspended SolWs 25
N« DaU
9 to 10 N« Data
0 lo 5
70 to 80 5 to 10 No Data S to 10
TREAT ABILITY STUDY RESULTS
(AH concentrations are in ppb}
Influent Biological Carbon Comb.
—Treat. Treat
100 75 <5 <5
Dichlorobenzidine
ortho-Chioroanaline 30
Benzidine 90
Ethytenedichloride 24
Toluene
130 12
ND' 300 ND
ND 15 ND
7 80 3
30
4-35
12
NOTES
-------
TREATMENT SUMMARY
• Over 135 chemicals treated
• Over 780 million gallons of
combined wastes treated to
date(March 1983 to March 1987)
• COD reductions >98%
(6,000 ppm to <100 ppm)
• Ortho-chloroanaline concentrations
from 6.500-53.000 ppb to <100 ppb
• Dichlorobenzidine concentrations from
400-12,000' ppb to <2 ppb
'Solubl* DCS only-(ydem «l»o racelves DCS In solid form
NOTES
NOTES
PACT SYSTEM OPERATION
• PAC concentration 4.000 to
12.000 mg/l
• Mixed liquor composition:
-PAC: 50%
-Biomass: 40%
-Ash: 10%
SYSTEM OPERATING COSTS
• 1986 total operating costs (solids
disposal, neutralization, ground
water pumping, and county wastewater
charges)were approximately $1,000.000
• $2.00 per 1,000 gallons treated
• <$0.10 per pound of COD treated
• Onsite carbon regeneration/solids
disposal budgeted for $300,000 per
year
• Offsite carbon disposal costs estimated
to be over $1.000.000 - and liability
would still exist
4-36
NOTES
-------
TREATMENT PROCESS DIAGRAM
HIGH STRENGTH
TOXIC WASTES""}
•ASH TO LANDFILL
DILUTE
PROCESS WASTE,
LANDFILL
LEACHATE,
CONTAMINATED
GROUNDWATER
VIRGIN
CARBON
MAKEUP
MUSKEGON
COUNTY
WWTP
BIOLOGICAL SEQUENCING
BATCH REACTOR (SBR)
Case Study:
Source: Occidental Chemical Corp.
(Grand Island, NY)
4-37
-------
HYDE PARK LANDFILL
« Used from 1953 to 1975
* Contains 73.000 metric tons
of chemical wastes
* Clay liner installed in 1978
• Tile leachate collection
system installed in 1979
• Leachate trucked to Niagara
plant and mixed with plant
wastewaters
NOTES
NOTES
ORIGINAL TREATMENT PROCESS
• pH adjustment
• Suspended solids settling
• Filtration through 50 micron bag
• Two-stage activated carbon
RAW LEACHATE CHARACTERISTICS
pH 4.3
TOC 3.500
COD 10.040
BOD 7.500
SS 900
VSS 300
IDS 25.700
(Major organics include phenol, benzoic acid,
and isomeric chlorobenzoic acids)
4-38
NOTES
-------
\
SBR TREATMENT STAGES
Draw
S.ttl.
NOTES
NOTES
RESULTS OF 500 LITER PILOT SBRs
TOC(mg/ll COOImg/ll TOXImg/l)
Influent Feed
Effluent A
(5 CkyHTTi
6000 mg/lkLSS)
2.000
140(83%)
EffluentB 120(94X1
10.000 mg/l MSS1
Effluent C 536(73X1
9.300
329
510(90X1 110(66X1
400(92X1 105(661)
1,700(66X1 235(26X1
YEARLY TREATMENT
< Based On 1984 Dollars
Activated Carbon Alone.-
($1.65/kg)
SBR Operation:
(At 173 kg/day)
Activated Carbon:
EXPENSES SUMMARY
And 10 Years Operation)
$715.111
$116.900
$71,511
Total: $188.411
Net Savings Per Yean
$526.700
NOTES
4-39
-------
FLUIDIZED BED
• Bacteria attached to support media
• Media fluidized in reactor
• Plug flow
NOTES
NOTES
FLUIDIZED BED
Oxygon Trlncfor
Excoii Blomm
CONTAINED ABOVE GROUND SOILS TREATMENT
• Contaminants treated in the soil
matrix
* Nutrients, moisture, and oxygen
added as needed
* Leachate, runoff and air emissions
must be controlled
• Soil left on site when clean
4-40
NOTES
-------
CONTAINED ABOVE GROUND SOILS TREATMENT
Air Emissions
Control System
Air Emissions
Contamlnated-Soll
Mixing and
Oxygen Transfer
Leachate
Leachate
Control
System
SOIL SLURRIES
• Contaminants treated in a soil slurry
• Nutrients and oxygen added as needed
• Water and soil must be separated
after treatment
• Soil left on site when clean
4-41
-------
Water
SOIL SLURRIES
Water for Reuse
or Disposal
Mixing and
Oxygen Transfer
Soil/Water
Separator
Clean
Soli
Contaminated
Soil
FIELD PILOT SOIL WASHING
Case Study:
NPL Wood Treating Facility
Minnesota
(Oct.-Nov. 1987)
Source: BioTrol (Chaska, MN)
4-42
-------
CONTAMINANTS
* Oil
• Creosote
• Pentachlorophenol
• Polynuclear aromatics
SITE SOIL CHARACTERISTICS
• Silty, fine to medium grained sands
with intermediate and laterally
discontinuous silt and sand lenses
4-43
-------
BIOTROL SOIL TREATMENT SYSTEM
(BSTS)
CONTAMINATED SOIL
CONTAMINATED WATER
/WATER TREATMENT
/BIOLOGICAL
/PHYSICAL, CHEMICAL
RECYCLE
CONCENTRATED ORGANIC ^
CONTAMINATION
INORGANIC FINES
INORGANICS (ROCKS,METALS)
OPTIONS
SOIL
CLASSIFICATION
*— x
s*\
OVERSIZE \)
„ __\ \
0 \
AsiZE REDUCTIONV
V a )
PHYSICAL TREATMENT
(SOIL WASHING)
REUSE
INCINERATION
GRAVITY
SEPARATOR
ORGANICS
OPTIONS
SCRUBBING
RESIDUALS
MANAGEMENT
INCINERATION-
CLEAN SOIL
PROCESS DIAGRAM FOR SOIL
WASHING SYSTEM
4-44
-------
PILOT SOIL WASHING EQUIPMENT
• 42' semi-trailer
* Soil feed rate up to 500 pounds
per hour (dry weight)
* Soils initially screened and
classified
* Countercurrent soil washing
using water
NOTES
NOTES
PILOT SOIL WASHING EQUIPMENT
(Continued)
• Contaminated water treated with
aerobic biological treatment system
• Decontaminated water recycled to
unit
* Sands and clays separated and
treated
• Large debris treated separately
PENTACHLOROPHENOL SOIL
WASHING RESULTS
(All concentrations are in ppm)
« of Dry Feed Influent Treated Percent
Soi Tests (bs/hr) Cone. Cone. Reduction
81
82
«3
4 282 1,498 80 >94
(+X-77) (+/-55S) (93
443 215 24 >88
(+/-51) (+/-1D (+/-4)
NOTES
4-45
-------
ESTIMATED TREATMENT COSTS
• $100 per cubic yard
• Final cost depends upon:
-volume of soil to be treated
-specific contaminants present
-composition of soils
-required effluent concentrations
NOTES
NOTES
PRETREATMENT FACTORS
Nonaqueous phase neat material removal
- specific gravity <1
- specific gravity >1
pH
Nutrients
• Toxicity
- organic
- inorganic
• Nuisance substances
- iron
- suspended solids
POST TREATMENT FACTORS
• Solids removal and disposal
• Effluent organics
- persistent compounds
- metabolic by-products
• Air emissions
4-46
NOTES
-------
ECONOMICS
• Capital equipment
• Design/engineering
• Installation expenses
• Operational expenses
NOTES
NOTES
OPERATIONAL EXPENSES
• Supplies/reagents
• Energy
• Operating personnel
• Disposal of end-products
PROCESSES FOR SELECTING
BIOREACTOR DESIGNS
• Applicability
• Technical/regulatory
• Cost effectiveness
4-47
NOTES
-------
NOTES
COMBINED ABOVE GROUND
AND
IN-SITU BIOLOGICAL SYSTEMS
4-48
-------
REACTOR TREATMENT DESIGN WORKSHEET
The following worksheet should be used to develop the Information
necessary for evaluating the suitability and design of biological
treatment systems.
I. Waste Characterization
1. List the contaminants and the concentrations present.
2. List the required effluent concentration for each contaminant
3. Which contaminants are biodegradable (aerobic or anaerobic),
inhibitory or toxic, or non-biodegradable?
4. What are the physical and chemical properties of the
contaminants (density, solubility, etc)?
5. Are the observed contaminant concentrations and locations
consistent with the properties of the chemicals?
II. Life-Cycle Design Considerations
6. Define the treatment system needed (include all inputs and
outputs)-
4-49
-------
7. Will site conditions change during the life of the project? If
so, how will these changes affect any proposed treatment system?
8. What is the expected duration of the project?
III. Conceptualized Process Design and Bioreactor Selection
9. Will the material be treated in place or moved to another
location?
10. What method of collecting and conveying the wastes should be
used?
11. What volumetric treatment rate will be required to process the
wastes?
12. Will the waste stream be treated with a single unit process or
several?
13. Do we need pretreatment to allow biological treatment to occur
(adjust pH, remove toxics, addition of nutrients, etc.)?
4-50
-------
14. What specific aerobic or anoxic/anaerobic biological processes
are best for this situation?
15. What is the development status of the processes selected
(demonstrated on similar site and situation, demonstrated in
other applications, developmental, or conceptual)?
16. What organic/inorganic residues will be produced from the
system? Are they hazardous? What equipment is required to
remove the residues? What is the final disposal of these
materials?
17- Draw process diagrams for the proposed treatment systems.
18. Will the proposed treatment systems meet or exceed all required
effluent discharge requirements?
19. What are the overall advantages of the proposed treatment
systems?
20. What are the overall disadvantages of the proposed treatment
systems?
4-51
-------
21. How do the biological unit processes interact with any needed
non-biological unit processes?
IV. Project Economics
22. List the site conditions needed for the proposed treatment
systems (space requirements, power requirements, etc.).
23. Will laboratory and/or field pilot treatability work be
required? How much should be budgeted?
24. What operating expenses will be incurred during treatment
(consumables, maintenance, byproduct disposal costs, and
operating personnel)?
25. Is it possible to reduce the manpower requirements for the
proposed treatment systems?
4-52
-------
IN-SITU DESIGN
SECTION 5
Abstract
Slides
Worksheets
5-2
5-14
5-47
5-1
-------
TREATMENT DESIGN - SURFACE AND SUBSURFACE
John T. Wilson Ronald C. Sims
U.S. EPA Utah State University
Ada, Oklahoma Logan, Utah
Surface Soil Treatment
Bioremediation of surface soils involves the use of naturally
occurring microorganisms to treat specific chemicals associated with the
soil environment at a site. The subject of bioremediation of
contaminated soils, including applications and limitations of the
technology, has been addressed at several recent scientific meetings and
conferences identified in the references section. Three aspects that are
important for consideration in order to accomplish in situ bioremediation
include: (1) site-soil-waste characterization, (2) microbial activity,
and (3) treatment system design and monitoring to evaluate treatment
effectiveness. Information concerning mechanisms involved in vadose zone
(soil) treatment and laboratory and field scale demonstration results
provide a significant information base concerning the applications of
this treatment approach. References are included to assist the reader in
obtaining additional information. The goals of on-site bioremediation of
contaminated soils are presented in Figure 1.
In situ treatment involves the controlled management and manipulation
of soil microbial processes and of soil physical and chemical processes
that affect natural soil microbial processes to achieve degradation and
detoxification of waste chemicals. Successful application of in situ
treatment requires information and understanding of site, soil and waste
characteristics identified above. Specific waste, site, and soil
characteristics that are important for determining the potential success
for in situ treatment are summarized in Tables 1 and 2, and discussed in
detail in the reference "Contaminated Surface Soils In-Place Treatment
Techniques".
Table 3 identifies contaminated sites that are currently using
bioremediation as the only remediation process or as one process in a
"treatment train" to obtain the goals of on-site bioremediation
identified in Figure 1. Management techniques that are currently being
used for in Situ bioremediation of surface soils at the sites, identified
in Table 3, involve the manipulation of factors influencing biological
activity including: oxygen, nutrients, moisture, and pH, and addition of
carbon and energy sources. Addition of amendments to surficial soils
generally have fewer restrictions with regard to mass transfer than
amendments applied to deeper soils, including microorganism inoculations.
With respect to microbial activity enhancement, when considering the
potential application of on-site bioremediation of contaminated soils,
there are several issues that should be considered as part of a
5-2
-------
PROTECTION OF PUBLIC HEALTH AND ENVIRONMENT
TREATMENT OF WASTE CONSTITUENTS TO AN ACCEPTABLE LEVEL
t
GROUNDWATER
SURFACE WATER
ATMOSPHERE
\
I SOIL SYSTEM I
DEGRADATION
TRANSFORMATION
IMMOBILIZATION
Figure 1. The Goals of Onsite Bioremediation of Contaminated Soils.
5-3
-------
TABLE 1. SITE AND SOIL CHARACTERISTICS IDENTIFIED AS IMPORTANT IN
IN SITU TREATMENT (Reference 5)
Site location/topography and slope
Soil type and extent
Soil profile properties
boundary characteristics
depth
texture*
amount and type of coarse fragments
structure*
color
degree of mottling
bulk density*
clay content
type of clay
cation exchange capacity*
organic matter content*
pH*
Eh*
aeration status*
Hydraulic properties and conditions
soil water characteristic curve
field capacity/permanent wilting point
water holding capacity*
permeability* (under saturated and a range of unsaturated
conditions)
infiltration rates*
depth to groundwater*. including seasonal variations
flooding frequency
runoff potential*
Geological and hydrogeological factors
subsurface geological features
groundwater flow patterns and characteristics
Meteorological and climatological data
wind velocity and direction
temperature
precipitation
water budget
*Factors that may be managed to enhance soil treatment
5-4
-------
TABLE 2. SOIL-BASED WASTE CHARACTERIZATION (Reference 5)
Chemical class
add
base
polar neutral
nonpolar neutral
Inorganic
Soil sorptlon parameters
Freundlich sorptlon constants (K, N)
sorptlon based on organic carbon content (Koc)
octanol/water partition coefficient (Kow)
Soil degradation parameters
half-life (t!/2)
rate-constant (first order)
relative biodegradability
Chemical properties
molecular weight
melting point
specific gravity
structure
water solubility
Volatilization parameters
air/water partition coefficient (Kw)
vapor pressure
Henry's law constant (1/KW)
sorptlon based on organic carbon content (Koc)
water solubility
Chemical reactivity
oxidation
reduction
hydrolysis
precipitation
polymerization
Soil contamination parameters
concentration in soil
depth of contamination
5-5
-------
TABLE 3. PROPOSED/ACTIVE BIOREMEDIATION SITES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Site Name
L.A. Clark & Sons
American Creosote
Brown Wood Preserving
Crosby
Wilmington
Burlington Northern
North Cavalcade Street
Old Inger
Brio Refining
Joplin
Baxter/Union Pacific
Burlington Northern
Libby
ARCO
Koppers Company
J.H. Baxter
Region
3
4
4
4
4
5
6
6
6
7
8
8
8
8
9
9
Contaminant
1*
1
1
1
1
1
1
2**
2
1*
1
1
1
3***
1
1
* Wood Preserving
*** Coal Gasification
5-6
-------
preliminary evaluation. Bioremediation is often limited by factors that
include: (1) distribution of the waste which may limit microorganism
access to the waste, (2) supply of nutrients required for metabolism,
(3) toxicity of the waste due to concentration and/or type of
constituents present, (4) formation and accumulation of toxic byproducts,
(5) inadequate population(s) of requisite microorganisms,
(6) non-competitiveness of non-survivability of inoculated cultures, and
(7) inadequate management of the system. Prior to the application of on
site bioremediation, the factors identified above should be addressed.
The importance of conducting treatability experiments with
appropriate controls and conducting a site characterization to identify
environmental, soil, and ecological factors that will affect the process
under field conditions cannot be overemphasized. Evaluation of
commercial claims should involve side-by-side comparisons in time using
appropriate and statistically rigorous control experiments that
faithfully duplicate the commercial process but without inclusion of the
commercial product.
Monitoring of treatment effectiveness in the vadose zone involves the
evaluation of chemical and toxicity changes with time. Both soil core
and soil-pore liquid samples are recommended, and in some cases, air
monitoring is recommended. Monitoring strategies can be based upon
information obtained in the characterization and treatability phases of
the bioremediation of a site.
Subsurface Treatment
In general, biodegradation of hazardous organic chemicals in
groundwater is not limited by the metabolic capability of
microorganisms. However, the prospects for biodegradation is severely
limited by the stoichiometry of microbial metabolism, and by mass
transport limitations of the rate of supply of essential nutrients.
These limitations determine the cost to remediate a site, the time
required, and the level of remediation that can be attained. Practical
application of biotechnology in the subsurface depends on an accurate
three-dimensional understanding of the position and concentration of the
contaminants, of the hydrology of the contaminated material, and an
estimate of quantity of oxygen or other electron-acceptor required to
remediate the site. This challenge is well illustrated in a
demonstration project supported by the U.S. EPA and the U.S. Coast Guard
on the in situ bioremediation of a fuel spill. Aviation gasoline was
spilled from an underground storage tank at the Coast Guard Air Station
at Traverse City, Michigan. The gasoline drained through unconsolidated
sands until it reached the water table, then it spread laterally.
Groundwater flows through the material contaminated with gasoline, and
carries a plume of alkylbenzenes and other fuel hydrocarbons away from
the original spill area. The Coast Guard and EPA plan to remediate the
spill by perfusing it with oxygen and hydrogen peroxide. The
alkylbenzenes are the object of the regulatory concern, and the
5-7
-------
bioremediation will be finished when their concentration is brought to a
level specified by the Michigan Department of Natural Resources.
The spill was cored to identify the depth interval that was
contaminated, and the highest concentration of fuel hydrocarbons. The
cores were extracted with methylene chloride, then analyzed by gas
chromatography. The gasoline was confined to a narrow interval between
15 and 17 feet below the land surface. This interval corresponds closely
with the seasonal high and low water table at the site. The
concentration of fuel hydrocarbons in the most contaminated interval
averages 7,500 mg/kg aquifer material. The porosity of the contaminated
sand is 0.4, and its bulk density is 0.2 g/cm3. Therefore, the water
content of the aquifer is 0.2 liter/kg, and each liter of pore water is
in contact with 37,500 mg of fuel hydrocarbons. The oxygen demand for
microbial respiration of total fuel hydrocarbons was estimated assuming
the following stoichiometry:
CH2 + 1.5 02 —^ C02 + H20
The oxygen demand of the alkylbenzene fraction alone was estimated from:
CH + 1.25 02 —*- C02 + 0.5 H20
Monitoring wells were installed 31 and 50 feet down gradient from the
injection wells. Of the 31 feet between the injection wells and the
first monitoring well, 15 feet was considered to be contaminated. Of the
50 feet to the next monitoring well, 35 feet was consider to be
contaminated. The concentrations of hydrocarbons, the length of the
contaminated portion of the flow path, and the assumed stoichiometry for
microbial respiration were used to estimate the total oxygen required to
remediate the flow paths to the two monitoring wells (Table 4). The
spill was cored in August, 1987 to provide information to design the
demonstration, then cored again in March, 1988, just before the
demonstration began, to define the initial conditions. The concentration
of alkylbenzenes in the spill declined dramatically over the time
interval (Table 5). This was probably due to anaerobic microbial
degradation.
For the first 140 days of the demonstration, the injected water
contained 40 mg/liter oxygen. Then the oxygen was replaced with
80 mg/liter hydrogen peroxide for 20 days. Then the concentration of
hydrogen peroxide was stepped up to 160 mg/liter for 50 days, and finally
to 360 mg/liter for 80 days. Concentrations of alkylbenzenes and oxygen
or hydrogen peroxide was monitored in the wells. The interval between
the injected wells and the monitoring well at 31 feet was remediated
after 220 days, and the interval to the monitoring well at 50 feet after
270 days.
5-8
-------
TABLE 4. STOICHIOMETRY OF AEROBIC BIOREMEDIATION OF A FUEL SPILL
Estimated demand based on:
Total Fuel Hydrocarbons
Alkylbenzene content only,
when sampled in 8/87
Alkylbenzene content only,
when sampled in 3/88
just before the start of
the demonstration
Actually required
Oxygen and Hydrogen Peroxide Demand along
Flowpath to Monitoring wells at:
31 feet 50 feet
—(mg oxygen/liter pore water)—
62,212
8,710
2,364
2,989
90,000
12,000
3,420
2,952
5-9
-------
TABLE 5. QUANTITIES OF ALKYLBENZENES AND TOTAL FUEL HYDROCARBONS
REMAINING IN AN AQUIFER AFTER BIOREMEDIATION USING OXYGEN
AND HYDROGEN PEROXIDE.
Parameter
Total fuel
hydrocarbons
Toluene
m+fi-Xylene
o-Xylene
Benzene
Before
Remediation
8/87
6,500
544
58
42
0.3
Just Before
Remediation
3/88
( mn /kn anin f pr m
\ my / isy OLU u i i c i in
1,200*
37
"
8.4
0.6
After
Remediation
10/88
la-Hprial ^
tl LCI 1 Ol 1 /
8,400
<0.3
<0.3
<0.3
<0.3
*A composited sample containing clean as well as contaminated material.
It is not surprising that the non-aromatic fraction of the spill
remained in the aquifer. A very minor fraction of their oxygen demand
had been supplied when the aquifer was cleansed of alkylbenzenes.
5-10
-------
A tracer test was done with chloride to determine the seepage
velocity in the flow path from the injection wells to the monitoring
wells. The velocity was multiplied by the concentration of oxygen or
hydrogen peroxide along the flow path. The flux was multiplied by the
time required for remediation to determine the actual oxygen demand for
remediation (Table 4).
Aviation gasoline is composed primarily of branched chain alkanes.
The material spilled at Traverse City was 38 percent 2,2,4-trimethyl-
hexane, 7 percent 2,3-dimethylhexane, and 5 percent 2,4-dimethylpentane.
Only 10 percent of the original spill was alkylbenzenes.
The aquifer was purged of alkylbenzenes very quickly. The quantity
of oxygen and hydrogen peroxide required to remove alkylbenzenes from the
wells agree closely with the projected oxygen demand of the alkylbenzenes
alone. This selective removal of alkylbenzenes may result from their
relatively high water solubility. Projected from Raoult's Law, the
expected concentration of toluene in water in equilibrium with the fuel
was 15 mg/liter. The expected concentration of 2,2,4-trimethylpentane is
only 0.2 mg/liter.
Shortly after remediation, the area near the monitoring well at
31 feet was cored and analyzed for alkylbenzenes and total fuel
hydrocarbons. Results were compared to earlier cores to determine
whether the contaminants were removed from the aquifer material itself
(Table 5).
5-11
-------
REFERENCES
1. Omenn, G.S. 1987. Environmental Biotechnology - Reducing Risks from
Environmental Chemicals through Biotechnology. Proceedings of
Conference held July 19-23 at the University of Washington, Seattle,
Washington. Plenum Press, New York. ISBN 0-306-42984-5. 505pp.
2. Engineering Foundation. 1988. Biotechnology Applied to Hazardous
Wastes. Conference held in Longboat Key. Florida, October 31 -
November 4.
3. Hazardous Materials Control Research Institute (HMCRI). 1988. Use
of Genetically Altered or Adapted Organisms in the Treatment of
Hazardous Wastes. Conference held in Washington, D.C., November 30 -
December 2.
4. U.S. EPA. 1986. Waste-Soil Treatability Studies for Four Complex
Industrial Wastes. Methodologies and Results. Volumes 1 and 2.
EPA-600/6-86-003 a,b. October. EPA, Robert S. Kerr Environmental
Research Laboratory, Ada, OK.
5. Sims, R.C., J.L. Sims, O.K. Sorensen, J.E. McLean, R.J. Mahmood, and
J.J. Jurinak. 1986. Contaminated Surface Soils In-Place Treatment
Techniques. Noyes Publications, Park Ridge, New Jersey. 536pp.
6. Woodward, R.E. 1988. Bioremediation Feasibility Studies for
Hazardous Waste. Pollution Engineering 20(7): 102-103.
7. U.S. EPA. 1986. Permit Guidance Manual for Hazardous Waste Land
Treatment Demonstrations. Office of Solid Waste, Washington, D.C.
EPA-530/SW-86-032. February.
8. Martin, J.P., R.C. Sims, and J.E. Matthews. 1986. Review and
Evaluation of Current Design and Management Practices for Land
Treatment Units Receiving Petroleum Wastes. Hazardous Wastes and
Hazardous Materials, 3(3):261-280.
9. U.S. EPA. 1981. A Survey of Existing Hazardous Waste Land Treatment
Facilities in the United States. U.S. EPA, Contract No. 68-03-2943.
10. Sims, R.C. 1986. Loading Rates and Frequencies for Land Treatment
Systems. In: Land Treatment: A Hazardous Waste Management
Alternative (R.C. Loehr and J.F. Malina, Eds. Water Resources
Symposium Number Thirteen, Center for Research in Water Resources,
College of Engineering, The University of Texas at Austin.
11. Loehr, R.C., J.H. Martin, and E.F. Neuhauser. 1983. Disposal of
Oily Wastes by Land Treatment. Report to 38th Annual Purdue
Industrial Waste Conference, Purdue University, West Lafayette,
Indiana, May.
5-12
-------
12. S1ms, R.C, and LM.R. Overcash. 1983. Fate of Polynuclear Aromatic
Compounds (PNAs) in Soil-Plant Systems. Residue Reviews. 88:1-68.
13. K.W. Brown and I.E. Duel. 1982. An Evaluation of Subsurface
Conditions at Refinery Landfarm Sites. Prepared for the American
Petroleum Institute and the U.S. EPA, Grant No. CR-807868.
14. U.S. EPA. 1988. Treatment Potential for 56 EPA Listed Hazardous
Chemicals in Soil. Robert S. Kerr Environmental Research Laboratory,
Ada, OK. EPA/600/6-88-001.
15. Mahmood, R.J., and R.C. Sims. 1986. Mobility of Organics 1n Land
Treatment Systems. Journal of Environmental Engineering
112(2):236-245.
16. Overcash, M.R., K.W. Brown, and G.B. Evans. 1987. Hazardous Waste
Land Treatment: A Technology and Regulatory Assessment. Prepared
for the U.S. Department of Energy by Argonne National Laboratory,
September 22.
17. U.S. EPA. 1983. Hazardous Waste Land Treatment. Revised Edition.
SW-874. Office of Solid Waste and Emergency Response, U.S. EPA,
Washington, D.C.
18. Zitrides, T. 1983. Biodecontamination of Spill Sites. Pollution
Engineering. 15(ll):25-27.
19. Lee, M.D., Thomas, J.M., Borden, R.C., Bedient, P.B., Wilson, J.T.,
and Ward, C.H. 1988. Biorestoration of Aquifers Contaminated with
Organic Compounds. CRC Critical Reviews in Environmental Control.
18(l):29-89.
20. Goldstein, R.M., L.M. Mallory, and M. Alexander. 1985. Reasons for
Possible Failure of Inoculation to Enhance Biodegradation. Applied
and Environmental Microbiology. 50:977.
21. Nyer, E.K. 1985. Groundwater Treatment Technology. Van Nostrand
Reinhold Company, Inc. ISBN: 0-442-26706-1. 188pp.
22. Wilson, J.T. and D.H. Kampbell. 1989. Challenges to the Practical
Application of Biotechnology for the Biodegradation of Chemicals in
Ground Water. Preprint Extended Abstract, American Chemical Society,
Division of Environmental Chemistry, April 9-14, Dallas, Texas.
23. Wilson, J.T., L.E. Leach, M. Henson, and J.N. Jones. 1986. In Situ
Biorestoration as a Ground Water Remediation Technique. Ground Water
Monitoring Review, pp. 56-64. Fall.
5-13
-------
DISTINCTION BETWEEN
SURFACE AND
SUBSURFACE REMEDIATION
* surface treatment: dominant electron acceptor is
oxygen supplied directly (rom the atmosphere
* subsurface treatment: electron acceptor is supplied
by perfusing the contaminated material with water or
air
NOTES
NOTES
IN SITU TREATMENT OF
CONTAMINATED SOIL
Typical Volumetric Composition Of Soil
Water
15-35%
Inorganic
38-45%
Adapted From Overcash & Pal, 1979
NOTES
5-14
-------
FATE
OF HAZARDOUS CONTAMINANTS IN SOIL
[VOLATILIZATION ) | HAZARDOUS CONTAMINANT |
A
WlM-KALf/ATlON •
BIOMASS |
n
INTERMEDIATE 1
PRODUCTS 1
,
N 1
SOIL INTI RAO'IONS 1
P IASI-S SOLID LIQUID GAS 1
J
-EACJ1ING 1 ^ '
1
NOTES
NOTES
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Site Name
PROPOSED/AC!
j GOALS OF IN SITU TREATMENT ]
I i
PROTECTION OF PUBLIC HEALTH AND ENVIRONMENT
TREATMENT OF WASTE CONSTITUENTS TO AN ACCEPTABLE LEVEL
t
( I
1 SOIL SYSTEM 1
DEGRADATION
DETOXIFICATION
IMMOBILIZATION
rIVE BIOREMEDIATION SITES
NOTES
Reaion Contaminant
L.A. Clark & Sons
American Creosote
Brown Wood Preserving
Crosby
Wilmington
Burlington Northern
North Cavalcade Street
Old Inger
Brio Refining
Jopl in
Baxter/Union Pacific
Burlington Northern
Libby
ARCO
Koppers Company
J.H. Baxter
3 1"
4 1
4 1
4 1
4 1
5 1
6 1
6 2**
6 2
7 1*
8 1
8 1
8 1
8 3"**
9 1
9 1
Wood Preserving
*** Coal Gasification
5-15
-------
CHARACTERIZATION
NOTES
NOTES
SOIL-BASED WASTE CHARACTERIZATION
Chemical
Class
Acid
Base
Polar Neutral
Nonpolar Neutral
Inorganic
Soil Sorption
Parameters
Freundlich Sorption
Constants (K.N)
Sorption based on
Organic Content (KJ
Octanol water partition
Coefficient (K J
Soil Degradation
Parameters
Half-life (tj
Rate Constant
Relative bio-
cleg radability
Chemical
Properties
MolecularWeight
Melting point
Specific Gravity
Structure
Water Solubility
SOIL-BASED
WASTE CHARACTERIZATION
Volatilization
Parameters
Air:water partition
coefficient (KJ
Vapor pressure
Henry's law constant
(1/KJ
Sorption based on
organic carbon
content (Koc)
Water solubility
Chemical
Reactivity
Oxidation
Reduction
Hydrolysis
Precipitation
Polymerization
Soil Contamination
Parameters
Concentration in soil
Depth ot Contamination
NOTES
5-16
-------
BIOLOGICAL DEGRADATION
Half-life of a PAH Compound:
t _ 0.693
1/2 ~ k
Where
t M - half-life of PAH compound in soil (time)
k - first-order rate constant (time'1) for
microbial degradation
NOTES
NOTES
IMMOBILIZATION
R = 1 + -flSL
e
soil bulk density
partition coefficient
volumetric moisture content
INTERPHASE TRANSFER
POTENTIAL
NOTES
5-17
-------
Influent
Purg« Gat
Efflu«nt Purg« Gej-v
i
SoU/Wo»ta
Mixture
Sorbent
Tubas
Capillary Flow
Control
Constant
Flow
Samplt
Pump
Effluent Purge Cas
Laboratory flask apparatus used for mass balanct measur«ments.
Trenjoort end oorliuonlna rejotlonthlos wUhtn soil control volumes used in modified R'TZ model
of Conslllutnt(s).
I Oicfiy
2. Leeched wh«n fn »^ter moving
p«sl Ooltom of Treoimenl. Zone
3. Lost lo olmo«phere when
vopor enlert lower olmoson«re
Action within Control Volume.
I. Decoy or Constituent In «ll
Phtiej
2.Tronsferof Constituent smong
Phcsei until Equlllbnum reached
now ZONE •
lOUIERTHEHTMENTZONE
~l
Action between Control Volumes:
I. Downward movement of
Constituent with Water
2. Upward end Downward
movement of Constituent In pore
• pace driven Dy concentration
gradient end properties of
Constituent
oil
soil
5-18
-------
DETERMINATION OF
CONTAINMENT REQUIREMENTS
PROBLEM FOR ASSESSMENT
If the rate of transport (leaching) is significant
compared with the rate of biodegration,
both factors must be considered (degradation
and leaching)
The constituent(s) may reach a "critical depth"
in the soil before being degraded
5-19
-------
FIELD STUDY SITE PROFILE
5-20
-------
ENHANCEMENT OF
MICROBIAL ACTIVITY
NOTES
NOTES
REMEDIATION BASED ON ASSESSMENT
Increasing the degradation factor allows
faster reduction in mass flow of the parent
compound(s) and degradation products
through the soil system toward ground water
and surface water receiver systems.
SOIL7SITE ASSIMILATIVE CAPACITY (SSAC)
Techniques
(1) Soil incorporation or mixing
(2) Aeration of the soil
(3) Addition of nutrients
(4) Addition of microbial carbon and
energy sources
(5) Water addition (irrigation)
(6) Drainage
(7) Runon and Runoff Controls
(8) pH adjustment
5-21
NOTES
-------
WAYS TO MAXIMIZE
AVAILABLE SOIL OXYGEN
• Prevent Water Saturation
• Presence of Sand, Loam (Not Hvy Clay)
• Moderate Tilling
• Avoid Compaction
• Controlled Waste Loading
NOTES
NOTES
EFFECT OF MANURE AND pH AMENDMENTS ON PAH DEGRADATION
IN A COMPLEX WASTEINCORPORATED INTO SOIL
PAH Compound
Half-Life In Waste:Soil Mixture (Days)
Without Amendments With Amendments
Acenaphthylene 78
Anthracene 28
Phenanthrene 69
Fluoranthene 104
Benz(a)antrhaeene 123
Benz(a)pyrene 91
Dibenz(a,h)anthracene 179
14
17
23
29
52
69
70
EFFECT OF SOIL MOISTURE ON
PAH DEGRADATION
Moisture Half-Life (Days)
(Field Capacity) Anthracene Phenanthrene Fluoranthene
20-40
60-80
43
37
61
54
559
231
5-22
NOTES
-------
TEMPERATURE EFFECT ON DEGRADATION RATE
Half-Life (days)'
Compound 10 C 20 C 30 C
Fluorene 60
(50-71)
Phenanthrene 200
(160-240)
Anthracene 460
(320-770)
Pyrene 1
Benzo(a)pyrene 530
(300-2230)
47
(42-53)
<60
260
(190-420)
1900
(1100-8100)
290
(170-860)
32
(29-37)
<60
200
(170-290)
210
(150-370)
220
(160-380)
• Hall-life (95% conlidence interval)
1 Least squares slopes • zero with 95% confidence
NOTES
11 \J JL 1J O
ACCLIMATION OF SOIL TO COMPLEX
FOSSIL FUEL WASTE
PNA Unacclimated Soil Acclimated Soil
Constituent Reduction in Reduction in
40 days (%) 22 days (5)
Naphthalene 90 100
Phenanthrene 70 83
Anthracene 58 99
Fluoranthene 51 82
Pyrene 47 86
Benz(a)anthracene 42 70
Chrysene 25 61
Benz(a)pyrene 40 50
EVALUATION OF
TREATMENT
NOTES
5-23
-------
PERFORMANCE EVALUATION- MONITORING
• Soil Cores
• Soil-Pore Liquid
• Ground Water
• Runoff Water
• Air
NOTES
NOTES
DEGRADATION IN CLAY
2 % Oil and Grease
C Ta R1
Compound 1*9/0 days
Fluorantnene 351 15 0.966
Pyrene 283 32 0.884
B«nzo(a)anthracone 86 139 0.397
Benzo(g.h.i.)perylene 8 1661 0.006
Indenopyrene 5 69 0.559
C, - Initial Concentration
T - HalMile (first ordor kinetics)
LOAM SOIL
95% Confidence Interval (T,,)
(davsl
Lower Upper
13 18
26 41
87 347
139 ND
43 139
(HC) 7,12-DIMETHYLBENZ(a)ANTHRACENE AND
TRANSFORMATION PRODUCTS IN
A SANDY LOAM SOIL
"C in each Iraction (%)
TKTW Soil Extract
(days)
Parent Translormalion
Compound Products
0 62 (69) 4 (6)
14 26 43
28 20 (60) 53 (11)
Residue CO, Total
12 (13) 0 (0) 78 (88)
16 0 85
17 (16) 0 (0) 90 (87)
Poisoned (control) d,ila in paronthoses
NOTES
5-24
-------
FIELD RESULTS FOR SOIL SAMPLES
C0 019/9)
Compound
AVG SO CV (%}
Naphthalene 186 68 37
Acenaphthena 729 276 38
Phenanthrene 78 28 36
Benz(a)
anthracene 86 42 49
Dibenz(a,h)
anthracene 52 36 69
91 days (ng/g)
AVG SO CV(%)
3 1,8 61
1 1.8 157
2.6 0.6 23
2 0.8 38
ND
C - Inffial So* Concaotrrtion
NOTES
NOTES
REMEDY
SELECTION
FACTORS
SITE CONSTRAINTS
NOTES
5-25
-------
COSTS
Scope Current Dollars
• Laboratory Treatability Study -- 50,000-100,000
• Pilot Scale Study --150,000-200,000
• Full Scale Study -- 400,000 +
NOTES
NOTES
FIELD IMPLEMENTATION COSTS
• Land Area Requirements
• Site Preparation
• Amendments
• Equipment
• Maintenance
• Monitoring
5-26
-------
DISTINCTION BETWEEN
SURFACE AND
SUBSURFACE REMEDIATION
For the purpose of this discussion, treatment will be
considered surface treatment if Ihe dominant electron
acceptor is oxygen supplied directly from the atmosphere,
and subsurface treatment if the electron acceptor is supplied
by perfusing the contaminated material with water or air.
NOTES
NOTES
PRIMARY EMPHASIS IN
SUBSURFACE REMEDIATION
Hazardous wastes that occur as a discrete oily-phase act as
source areas for plumes of contamination in ground water. They
also contaminate the soil air with hazardous fumes. The primary
emphasis in subsurface bioremediation has been the source
areas. Subsurface bioremediation of the plumes is often
technically feasible, but it is usually easier to pump them out and
treat them on the surface.
Leaking Underground:;'.
Storage Tank
NOTES
5-27
-------
NOTES
NOTES
/
PLUME
Hfc
WATER TABLE
GROUND WATER FLOW
^H$
WATER TABLE
N
NOTES
5-28
-------
Groundwale* — •
(cm)
80
NOTES
NOTES
IDENTIFY THE MOST
CONTAMINATED FLOW PATH
Some regions of the source area will clean up faster than others.
One (low path will be the last to clean up. If this flow path can
be identified, then its properties can be used to determine how
much effort is required to remediate the entire source area, and
how long it will take.
INJECTION
WELL
EXTRACTION
WELL
NOTES
5-29
-------
NOTES
NOTES
EXTRACTION
WELL
EXTRACTION
WELL
NOTES
5-30
-------
INJECTION
WELL
EXTRACTION
WELL
NOTES
NOTES
If the supply of mineral nutrients is adequate, the rate of
bioremediation is the rate of supply of electron acceptor. As a
result, the rate of remediation is directly proportional to the
concentration of electron acceptor in the injected water, and
directly proportional to the flow velocity of water through the
source area.
CHARACTERIZATION OF THE
MOST CONTAMINATED
INTERVAL
Time required to
clean most
contaminated
flow path
a
Length of path
through source
area
Concentration of
contaminant along
flow path
Seepage velocity along the
most contaminated flow path
5-31
NOTES
-------
CONTROL OF
HYDROLOGY ON THE
RATE OF REMEDIATION
Seepage Vefocity a Hydraulic Permeability x Hydraulic Gradient
Hydraulic permeability is an intrinsic properly of the
subsurface. It is difficult or impossible to improve it, but
it is easily degraded.
The hydraulic gradient is controlled by the amount of
water available for pumping, and by the difference in
elevation between the source area and the land surface.
NOTES
NOTES
20
FSW417-65 Flowmeter
0.001 0.01 0.1
Hydraulic Conductivity In
Centimeters Per Second
HOW TO PLUG UP AN INJECTION WELL
Add oxygen or hydrogen peroxide to water with
Fe*2
-> get Fe (OH)3
Add oxygen or hydrogen peroxide to water with
Mg/l of organics
-> get biofouling
Add phosphate to aquifer with Ca (Mg) CO., matrix
-> Ca (Mg) PO4
5-32
NOTES
-------
PROBLEMS WITH WELLS
AS MONITORING TOOLS
Treatment can occur In the well Itself. The waler in the well
may not be representative of the water in the aquifer.
A conventional monitoring well produces a composited water
sample. Water from the most contaminated flow path is diluted
by water from many other flow paths that are less contaminated.
A water sample from a well tells nothing about the amount of
hazardous material that is absorbed to aquifer solids or is
trapped as an oily phase.
NOTES
NOTES
BLDG.
CURE 2
ORIGINAL SOURCE
/ \ BUILDING
•FAILED FLANGE
5IY
SCALE
50m
Column with
cont«mlnat*d
• qulf *r • •mpI •
60 ml tyring*
V • I v •
Supply tl»«k
S y r I n g • pump
LEACHING COLUMN CONFIGURATION
5-33
NOTES
-------
I CONCENTRATION
mg/liter
TOLUENE ELUTION FROM A
CONTAMINATED TRAVERSE
CITY CORE
PORE VOLUMES
NOTES
NOTES
]
2
3
4
Teflon wiper Disc
Brass Bushings
Neoprene Seals
Swivel
MODIFIED WIRELINE PISTON DESIGN
Water Table
-20
-JO
11
Gasoline
Saluration
Typical
• Split-Spoon
Sampler
5-34
NOTES
-------
CO-DISTRIBUTION OF CONTAMINATION
AND HYDRAULIC PERMEABILITY IN AN
AQUIFER CONTAMINATED BY
Depth Interval
(tee! below surface)
Interval Cored or
Saeened Interval
15.1 - 15.5
15.5-15.8
15.8- 16.2
16.2- 16.5
16.5-17.2
17.2- 17.5
18.0- 18.3
19.4- 19.6
20.9-21.4
Fuel Hydrocarbons
(mg/kg aquifer)
< 11
39
2370
8400
624
< 13
< 13
A FUEL SPILL
Seepage Velocity
(feel per day)
7.2
9.0
15.6
19.7
NOTES
NOTES
In the most contaminated interval at Traverse City
The concentration of luel hydrocarbons averages
7,500 mg/kg aquifer material, the porosity is 0.4,
and the bulk density is 2.0 kg/dm3.
Each kilogram of aquifer contains 0.2 liter of water, and
each liter of pore water is exposed to 37,500 mg of fuel
hydrocarbons.
The oxygen demand of the hydrocarbons is 128,000 mg
O, per liter pore water.
HYDRAULIC CONTAINMENT
The migration of a plume away from its source area can often be
prevented by capturing the plume with a purge well. The well
must pump hard enough to overcome regional flow in the aquifer.
The flow from purge wells that is necessary to capture a plume
depends on the hydraulic permeability of the aquifer, the regional
hydraulic gradient, and the size of the source area.
NOTES
5-35
-------
NOTES
NOTES
HYDRAULIC CONTAINMENT OF
SUBSURFACE REMEDIATION
Hydraulic containment of a source area can be achieved if more
water is extracted than Injected. II water is recirculated through
the source area, a portion of the extracted water can be discharged
to a sewer of surface drainage, resulting in a net extraction of
water across the entire system.
HYDRAU)Jp CONTQUffS,
812 1012 1112
NOTES
5-36
-------
MODFLOW HYDRAULIC SURFACE
NOTES
NOTES
AQUIFERS AND
NATURAL CONFINING LAYERS
Frequently, geological structures that readily yield water are
layered above or between geological materials that do not readily
transmit water. These non-transmlssive layers can act as
natural containment for subsurface bioremediation. Don't
assume the bed rock is a confining layer; it is often fractured.
Hangar
1
Water
NOTES
5-37
-------
BACKHOE KEYS TRENCH
INTO BEDROCK
BACKFILL
SLOUGHS
FORWARD
NOTES
NOTES
ELEVATION IN INJECTION
ET ABOVE MSL WELI.S\
1
0--
5
0-
i- -
-
V 00 '7 BD-3 1 BD-50BD-62 BD-83 BO-K
— i— _____,, ? i
i, ZONE OF i< H L H 1
6 CONTAMINATION i5 WATER TABLES !
2-24-ea
3-4-88
3-11-88
0 5 10
- HORIZONTAL SCALE III METERS
NOTES
5-38
-------
FEEt
6 1 S
6 10
60S
600-
595
590-
5B5-
ABOVE MSL WELLSv
\
OD 7 BD-31 BD-50 BO-62 OD-B3 BO-1O6
„ ^=^-^ 1 T , 1 i 1 t
Ij ZONE Of 04 I4 04 i4 L
l« CONTAMINATION is WATEH TABLES il
2-24-BB
3-<-B8
3-11-88
0510
HORIZONTAL SCALE IN METERS
NOTES
NOTES
FORMULATION OF
NUTRIENT MIX
* Usually determined empirically
» Not related to C:N:P:S ratios
» Use high concentrations to project significant
concentrations into the aquifer
* Should formulations be related to 0:N:P:S
ratios?
PROPERTIES OF
MOLECULAR OXYGEN
ADVANTAGES
* Low toxicity to acclimated organisms
* Supports removal of many organic compounds
* Inexpensive
DISADVANTAGES
* Low solubility in water
* Will precipitate iron hydroxide
5-39
NOTES
-------
PROPERTIES OF
HYDROGEN PEROXIDE
ADVANTAGES
* Miscible in water
* Supports bioremediation of many organic compounds
* Chemically oxidizes many organic and inorganic
contaminants
* Removes biofouling
DISADVANTAGES
* Toxic at concentrations much above 500 mg/liter
* Will precipitate iron hydroxide
* Relatively expensive
NOTES
NOTES
PROPERTIES OF NITRATE
AS AN ELECTRON ACCEPTOR
ADVANTAGES
* Very soluble in water
* Low toxicity to microorganisms
* Does not cause precipitation of iron hydroxide
* Only aromatic compounds are removed
* Inexpensive
DISADVANTAGES
•* A regulated substance
* Potential for accumulation of nitrite
* Only aromatic compounds are removed
NOTES
COST COMPARISON
OF ELECTRON ACCEPTORS
Electron Acceptors
Sodium Nitrate
Liquid Oxygen
Hydrogen Peroxide
Bulk
Cost
(per kg)
$0.66
$1.46
$1.54
Electrons
Accepted
(moles / kg)
58.8
125.0
58.8
Real Cost
(per moles of
electrons
accepted)
$1.12
$1.17
$2.62
5-40
-------
ADVANTAGES OF
PULSING AMENDMENTS
II more than one amendment is required to promote subsurface-
bioremedialion, they can be injected in alternating pulses. This
prevents undue production of blomass near the injection
system, which would otherwise plug the system.
High concentrations of hydrogen peroxide (>100,000 mg/liter)
can remove bloloullng and restore the efficiency in injection
wells or injection galleries.
Pulses of hydrogen peroxide at high concentration can sterilize
the aquifer and destroy catalase activity, preventing premature
decomposition of the peroxide.
NOTES
NOTES
MONITOR THE OPERATION
OF THE SYSTEM AS WELL
AS ITS PERFORMANCE
* Delivery of mineral nutrients
* Delivery of electron acceptor
* Position in the water table
* Effectiveness of containment
ELEVATION IN INJECTION
FEET ABOVE MSL WELLS\
\^ nD 7 00-31 DO-SO BD-8J 00-03 00-10E
«15-
910
805
800
5»5
590
58S
'.
'•
^='ir=^- 1 , , , ,,,
^--v-^r;r^pg^^|^^^/^^4;
U-JT if< % \4 i>4 04 {4
Ofl 05 WATERTAOLES °5
2-24-88
3-4-88
3-1 1-88
0510
1 , , , , 1 , , , , 1
HORIZONTAL SCALE IN METERS
NOTES
5-41
-------
O)
E
0)
O)
O
o
to
Julian Date
NOTES
NOTES
Pilot Scale Biodegradation Project
Dissolved Oxygen Levels Vs. Time
Well *BO-50B-4
O O
o o
160 210 260
Julian Date Of Sample
Pilot Scale Biodegradation Project
Dissolved Oxygen Levels Vs. Time
Well #BD-50B-3
? -' i Pi P°ciP. °. °. ° P. .0. .OP ° no
u I i n n iOM i i i i i i i i M |" i 'VH i i i i vi iX i i i i i | i i i i i i i i i i i i i i i i i
160 210 260
Julian Date Ol Sample
5-42
NOTES
-------
00-
Pilot Scale Biodegradation Project
Dissolved Oxygon Levels Vs. Time
Wtll *BD-50B-2
OOP O
110 160 J 10 260
Julian Date 01 Satnpl*
NOTES
NOTES
Well # BD-31-2
Jukan Dale
PERFORMANCE OF BIORESTORATION NEAR BD 31
Parameter
(mg/Kg aquifer)
Total Fuel Hydrocarbon
Toluene
£Q *• 0 Xylene
2 - Xylene
Benzene
Before Just Before
8/87 8/88
6.500 1,220'
544 37
58 <1
42 8.4
0.3 0.6
Alter
10/88
8,400
<0.3
<0.3
<0.3
<0,3
• Sample diluted with uncontaminated material.
NOTES
5-43
-------
STOICHIOMETRY OF AEROBIC BIORESTORATION
Oxygen required
BD31-2 BD 500-2
-mg O2 / liler pore water--
Estimated based on:
Total Fuel Hydrocarbons
BTX only (8/87)
BTX only (3/88)
Actually required
62,212
8.710
2,364
2,989
90,000
12,000
3,420
2,952
NOTES
NOTES
HOW OFTEN SHOULD A
MONITORING WELL BE SAMPLED?
The frequency of sampling should be related to the time expected
for significant changes to occur along the most contaminated (low
path.
IMPORTANT CONSIDERATIONS
* Time required for water to move from injection wells to the
monitoring wells
•» Seasonal variations in water-table elevation or hydraulic
gradient.
* Changes in the concentration of electron acceptor.
* Cost of monitoring compared to day-to-day cost of
operation.
FACTORS CONTROLLING THE
RATE AND EXTENT OF
BIOREMEDIATION AT FIELD SCALE
4 Rale of supply of essential nutrients, usually the
electron acceptor
* Spatial variability in flow velocity
» Seclusion ol the waste from the microorganisms
NOTES
5-44
-------
INTERPRETATION OF
TREATABILITY STUDIES FOR
SUBSURFACE REMEDIATION
A good UeatabiMy study determines whether
bioremediation is possible, and whether there are any
biological barriers to attaining the goal for clean-up. It
can also provide an estimate on the rate of remediation
that can be attained if the organisms are not limited by
the rate of supply of some essential nutrient.
NOTES
NOTES
RATES OF OXYGEN CONSUMPTION
IN THE MOST CONTAMINATED
FLOW PATH AT TRAVERSE CITY
Hydrogen Peroxide Injected
7 feet from injection wells
Oxygen Injected
7 feet from injection wells
31 feet from injection wells
50 feet from injection wells
Mg O2 / Liter Day
60
2.20
;> 8.1
;> 7.3
Rates and extent of treatment at field scale should be
estimated with a comprehensive mathematical model
that incorporates
» biological reaction rates
» stoichiometry of waste transformation
* mass-transport considerations
* spatial variability in treatment efficiency
5-45
NOTES
-------
COSTS ASSOCIATED WITH
SUBSURFACE REMEDIATION
SITE CHARACTERIZATION
Wells. Soil Gas Survey, Coring and Core Analysis,
Geological Section, Aquifer Tests, Tracer Tests
REMEDIAL DESIGN
Treatabilily Tests, Mathematical Modeling
SYSTEM DESIGN
Permits, Negotiating trade-offs between cost and time
required
NOTES
NOTES
MORE COSTS
ASSOCIATED WITH
SUBSURFACE REMEDIATION
SYSTEM INSTALLATION
Wells, infiltration galleries, pumps, pipelines, tanks,
conlrol devices, treatment systems
MATERIALS AND OPERATING EXPENSES
Water, electron acceptor, fertilizer, inoculant,
maintenance, power, sewer charges
MONITORING
Monitoring wells and pumps, cores and their analysis
SITE SECURITY AND OPERATIONAL OVERSIGHT
5-46
-------
IN SITU TREATMENT DESIGN - SURFACE AND SUBSURFACE WORKSHEET
Site characterization
A. Surface
1. Hhat are the important characteristics of the following
elements for a "soil-based" characterization?
a) Soil factors
b) Engineering factors
c) Microbiology factors
2. What interphase transfer processes need characterization?
3. How do you use the information on interphase transfer
processes for treatment and monitoring aspects in the
vadose zone?
4. How can you characterize the following?
a) Potential for migration of chemicals at the site
5-47
-------
b) Previous migration of chemicals at the site
B. Subsurface
1. What factors influence three dimensional distribution of
oily phase material?
2. What factors influence three dimensional distribution of
plume in solution?
3. What is the direction of groundwater flow?
4. What is the seasonal variation in direction of flow?
5. What is the seasonal variation in water table elevation?
6. What is the hydraulic conductivity in the most
contaminated interval?
7. What is the frequency distribution of hydraulic
conductivity across the contaminated interval?
5-48
-------
8. What Is the water filled porosity?
9. What is the concentration of oily phase contaminate along
most contaminated flow line?
10. What is the relative concentration of regulated substances
in the oily phase material?
II. Containment Requirements
A. Surface
1. Identify approaches for volatile chemicals
2. Identify approaches for Teachable chemicals
3. How does one assess containment requirements?
B. Subsurface
1. Identify important boundaries in the flow field - rivers,
pumping wells, impermeable layers
5-49
-------
2. Determine if bed rock is fractured, or if it is a good
confining layer
3. Can the system accept sheet piling?
4. Can the system accept a grout curtain?
5. Can the system accept a slurry wall?
6. Can the flow field be modelled as a steady state system?
7. Is there acceptable disposal for extracted water?
III. Appropriateness of in-situ treatment vs in-reactor treatment
A. Surface
1. Pros for in-situ treatment
2. Cons for in-situ treatment
5-50
-------
3. Pros for in-reactor treatment
4. Cons for 1n-reactor treatment
B. Subsurface - Soils
1. Pros for in-situ treatment
2. Cons for in-situ treatment
3. Pros for in-reactor treatment
4. Cons for in-reactor treatment
C. Groundwater
1. Pros for in-situ treatment
2. Cons for in-situ treatment
5-51
-------
3. Pros for in-reactor treatment
4. Cons for in-reactor treatment
IV. Enhancement of microbial activity
A. Surface
1. What factors affect the following biological processes?
a) Metabolism
b) Growth or reproduction
c) Activity
2. Identify important environmental factors
3. Identify important chemical factors
5-52
-------
4. What factors affect the following processes?
a) Rate and extent of "degradation" of a chemical
b) Rate and extent of toxicity reduction
5. Identify approaches to evaluating the enhancement of
microbial activity
B. Subsurface
1. How much electron acceptor is required to reclaim the most
contaminated flow path?
2. What concentration of electron acceptor will the aquifer
accept?
3. How soon must the site be reclaimed? How long can the
interval be between injection and extraction well?
4. Is the nutrient mix compatible with the geochemistry of
the groundwater and the aquifer matrix?
(Can this marriage be saved?)
5-53
-------
5. How much water is available for injection? What is its
quality?
6. Is inoculation required?
V. Evaluation of treatment
A. Surface
1. What types of information can treatability studies provide?
2. What types of information can be obtained from field
monitoring?
3. How do you approach the following elements for evaluation
of treatment?
a) Media to monitor
b) "Things" to monitor
c) When to monitor
5-54
-------
4. Identify "target level" goals at a site
5. Identify factors affecting monitoring data variability
B. Subsurface
1. Does the nutrient mix adequately perfuse the source area?
2. Can the most contaminated interval be cored to evaluate
performance?
3. Is sampling frequency related to flow velocity of water?
To the expected rate of clean-up? To the distance from
the injection wells?
4. Has reclamation left behind organic materials foreign to
the aquifer?
VI. Remedy selection factors
A. Surface
1. How does the "pollutant pathways analysis" assist in
identifying remedy selection factors?
5-55
-------
2. How can time constraints affect remedy selection factors?
3. How can "site size" factors affect remedy selection
factors?
4. Identify specific factors for remedy selection factors
based on the following elements.
a) Characterization of site
b) Treatment evaluation (treatability studies)
c) Constraints on filed implementation
B. Subsurface
1. Will the nutrient mix reduce hydraulic conductivity?
2. Is the treatability study an accurate description of the
proposed technology?
5-56
-------
3. What liability will be generated if containment fails?
4. Will variability in hydraulic permeability preclude
reaching the target clean-up goals?
VII. Economics
A. Surface
1. What is the cost per unit volume of soil treated?
2. What is the cost comparison for treatment with other
technologies?
3. What are the equipment needs at the site?
4. What are the monitoring costs?
5. Identify capital and operation and monitoring (O&M) costs
5-57
-------
6. Identify "alternative" cost for different approaches based
on in situ bioremediation
B. Subsurface
1. What is the most inexpensive electron acceptor?
2. What is the cost to identify and characterize the most
contaminated flow path? How deep? What sort of material?
5-58
•£ U.S. GOVERNMENT PRINTING OFFICE: 1990 — 7 48-159 00469
------- |