United States
Environmental Protection
Agency
Office of
Research and
Development
CERI-90-114
December 1990
oEPA The Federal
Technology Transfer
Act
Opportunities for
Cooperative Biosystems
Research and
Development with the
U.S. EPA
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oth the U.S. Environmental Protection Agency (EPA) and private
industry seek new, cost-effective technologies to prevent and control
pollution. In the past, however, legal and institutional barriers have
prevented government and industry from collaborating in develop-
ing and marketing these technologies. Also, the efforts of many com-
panies to develop new technologies have been held back by a lack of
resources, such as scientific experts in particular fields or highly spe-
cialized equipment. The Federal Technology Transfer Act of 1986
(FTTA) removes some of these barriers to the development of com-
mercial pollution control technologies.
The FTTA makes possible cooperative research and development
agreements (CRDAs) between federal laboratories, industry, and
academic institutions. CRDAs set forth the terms of government/ in-
dustry collaboration to develop and commercialize new tech-
nologies. These agreements will, according to the Act, foster the
technological and industrial innovation that is "central to the
economic, environmental, and social well-being of citizens of the
United States."
What can industry gain from signing a CRDA with EPA?
• Access to high-quality science. EPA's 12 research laboratories
employ over 850 scientists and engineers and operate on a budget
of approximately $450 million per year. Many of these
laboratories combine world-class expertise with state-of-the-art
equipment and fully permitted testing facilities. Certain types of
environmental research, such as pollution prevention and control,
require the collaboration of experts in many different fields. This
type of interaction is readily accomplished at EPA laboratories,
which employ a range of pollution control experts.
« Expanded communication channels between government and
the private sector. CRDAs build working relationships between
the government and the private sector. The different perspectives
that government scientists bring to an applied R&D project can
provide a knowledge base that can significantly reduce the time
spent on problem-solving tasks during technology development.
« Exclusive agreements for developing new technologies. Under
some CRDAs, companies are given exclusive rights to market and
commercialize new technologies that result from the collabora-
tion. Until recently, industry had little incentive to cooperate with
federal laboratories because any technologies developed during
joint research remained in the public domain for all to use. Now,
^!g> Printed on Recycled Paper
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exclusive rights can be negotiated for some projects, though other
arrangements of rights are also possible, depending on the type of
CRDA.
The advantages of collaboration have prompted EPA and in-
dustry to set up CRDAs in areas ranging from reducing air pollution
to cleaning up oil spills. Bioremediation, a technology that uses bac-
teria to break down hazardous chemicals such as spilled oil or Super-
fund hazardous wastes, has proved to be a particularly fertile area
for collaboration (see page 6). For years, other methods of cleaning
up wastes have dominated the market, but recently bioremediation
has seen increasing use. CRDAs have resulted in the field application
of this technology, providing an ever-expanding data base of chemi-
cals known to be biodegraded. This technology is, therefore, an excel-
lent example of the benefits of government and industry working
together to develop new means of pollution prevention and control.
Biosystems use microorganisms, such as bacteria or fungi, to
transform harmful chemicals into less toxic or nontoxic compounds.
To the microorganisms, the pollutants are an energy source; they
break down the pollutants in the course of getting the energy they
need to live and multiply. Different organisms metabolize different
chemicals, so, by using an organism in the biosystem that breaks
down a particular pollutant, scientists can tailor the technology to
the pollutants at specific sites. Wherever possible, development
teams use native microorganisms in biosystems that they know are
already metabolizing the pollutants on the site. In these cases, the
number of microorganisms — and thus the speed at which a pol-
lutant is broken down — can be increased by adding nutrients to the
site. In other cases, non-native organisms known to metabolize the
pollutants are introduced.
Biotransformation is an attractive option because:
• It is "natural," and the residues from the process (such as carbon
dioxide and water) are usually harmless.
« It is less expensive and less disruptive than other options
frequently used to remediate hazardous wastes, such as
excavation and incineration.
• It holds a clear advantage over many technologies relying on
physical or chemical processes: instead of merely transferring
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contaminants from one medium to another, biosystems can
destroy the target chemicals.
Potential markets for commercialized biosystems treatment of
hazardous wastes are huge. Across the United States, thousands of
state, federal, and private hazardous waste sites are currently slated
for cleanup under Superfund and the Resource Conservation and
Recovery Act. In addition, approximately 15 percent of the nation's
four to five million underground storage tanks containing
petroleum, heating oil, and other materials are leaking. Many more
underground tanks will begin to leak in the next 5 to 10 years. Yet
another market results from the 10,000-15,000 oil spills that occur
each year, requiring cleanup of contaminated soils and waters.
Our increasing knowledge of the biology of microorganisms that
degrade wastes and of the engineering technologies needed to apply
and control degradation processes has opened the doors for new
biosystems technologies. Microbial treatment has already been suc-
cessfully used both in the United States and in other countries for in
situ treatment of soils contaminated with organic compounds at haz-
ardous waste sites. Biological systems have also been used to treat
soils and aquifers contaminated by hydrocarbons, phenols, cyanides,
and chlorinated solvents such as trichloroethylene. Another approach
to bioremediation is to isolate the enzymes that catalyze the metabo-
lism of pollutants in the microorganisms. Once separated from the or-
ganism, some of these cell-free enzymes have been modified to break
down pollutants, such as organophosphate pesticides.
These achievements have only begun to tap the potential of
biosystems technologies, but they point to the rich opportunities for
the emerging biosystems industry in waste management. To help
develop these opportunities, EPA has initiated a major research
program in this area: The Biosystems Technology Development
Program.
This program is answering key research questions in the develop-
ment of biosystems for treating hazardous wastes (see Figure 1). For
example, can microorganisms be used to treat chemicals commonly
found at waste sites — such as chlorinated solvents, polychlorinated
biphenyls (PCBs), chlorinated phenols, petroleum hydrocarbons,
creosote constituents, and dioxins? Can bacteria act to remove
chlorine substituents (dechlorination) from synthetic chemicals?
What are the limitations of biological treatment? How effectively do
organisms metabolize chemicals under anaerobic as opposed to
aerobic conditions? What types of organisms work the fastest? EPA
invites industry collaboration to answer these questions and solve a
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1. Develop the Biosystem
• Identify
appropriate
organisms
Study
metabolic
process
• Enhance
activity of
organism
Engineer the
biosystem
• Demonstrate
the system
Bacteria
+0z
(Aerobic)
-Oz
(Anaerobic)
Steps in developing a biosystem.
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2. Implement the System
X Characterize the site
n Pollutants found on site
m Depth of water table
• Size of site
m Annual rainfall
3. Technology Transfer
4. Write Protocol and Guidelines
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Alaska Oil Spill Bioremediation Project
In the wake of the Exxon Valdez tanker accident in March
of 1989, EPA signed a cooperative agreement with Exxon to
initiate a bioremediation study in Prince William Sound, Alas-
ka. This innovative study has highlighted the great promise
bioremediation holds for more timely and effective cleanup of
future oil spills worldwide. Even though enhancing the
natural biodegradation of oil has been extensively studied for
the past 20 years, it had never before been tested on such a
large scale in a marine environment.
The first challenge of the project was the need for quick
response. The spilled oil, which spread over an estimated 900
miles of poorly accessible shoreline in Prince William Sound,
Say in Prince William Sound after the Exxon Valdez spill.
spelled grave danger to the area's large populations of seals,
sea otters, fish, and birds. A field bioremediation test had to
be set up as quickly as possible so that, if the results were
favorable, large-scale application could begin before the short
northern summer ended. EPA met that challenge: in less than
2 months from the time of the spill, EPA developed a research
plan for using bioremediation to degrade the residual oil
washed up on the beaches and set up a command center in
Valdez. Field tests began on June 8,6 days after the formal
signing of the CRDA between Exxon and EPA.
On designated beach sites, a team of EPA scientists ap-
plied fertilizer containing nitrogen- and phosphorus-rich
nutrients to stimulate the native organisms on the beach to
degrade the spilled oil. The nutrients were applied in the
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form of three different types of fertilizer. "Reference" test
plots, where no fertilizer was added, were also set up for com-
parison. The striking visual contrast between the reference
and fertilizer-treated plots focused immediate national atten-
tion on biosystems as treatment options for spilled oil. By July
25, large-scale application of the fertilizers began. Eventually,
the fertilizer was applied to over 70 miles of Alaskan coastline.
This success is striking proof of the potential benefit of
FTTA agreements. EPA demonstrated its credibility as a scien-
tific resource to industry by providing experts in molecular
biology, genetics, ecology, biochemistry, bioprocess engineer-
Aerial photograph of a bioremedial test plot that resembles a clean
rectangle etched upon the surface of the beach. The cobblestone
plot, which was located at Snug Harbor, was treated with oleophilic
fertilizer during the summer of 1989.
ing, risk assessment, microbial ecology, ecological toxicology,
and mathematical modeling. Exxon had few microbiologists
on staff and little experience in field testing of pollution con-
trol technologies. EPA handled the nutrient addition, monitor-
ing, and analyses necessary for carrying out this project.
Under the CRDA, Exxon paid for all the costs of field
operations directly applicable to the bioremediation study. To
ensure the independence of study results, EPA paid and was
responsible for oversight and management of the project as
well as conducting the field work.
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variety of common, real-world pollution problems, such as the 1989
oil spill in Alaska.
Through the Biosystems Technology Development Program, EPA
has established a strong research base in six key areas of biosystems
research — liquid reactors, soil/sediment treatment, combined/se-
quential treatment, ground-water treatment, metabolic processes re-
search, and risk assessment.
Liquid reactors. In liquid reactors, toxic and hazardous pollutants
in liquid form are brought into contact with microorganisms to en-
hance degradation. Landfill leachates are a good example of a type of
liquid waste that could be treated with this type of biosystem. Nearly
130 million tons of solid waste is disposed of each year in landfills
across the nation, and various inorganic and organic chemicals often
leach from the waste into drainage water. One area of EPA research
focuses on using liquid reactors at publicly owned treatment works
(POTWs) to treat this leachate stream.
As POTWs are now operated, many do not handle liquid wastes
well enough: pollutants may still pass through the system un-
changed and be released into salt or fresh water; aeration results in
air stripping of volatile toxicants into the air; and many of the
toxicants associated with the residual sludges are not completely
dechlorinated and destroyed. Through improved engineering and
operational controls, existing facilities may be modified to handle these
wastes. In one current project in this area, EPA researchers are examin-
ing how to treat leachate in a biofilter, where the pollutants are broken
down by microorganisms. Another set of experiments is showing that a
combination of anaerobic and aerobic methods in a bench-scale treat-
ment system can result in 30 to 70 percent removal.
Soil/sediment treatment. The soils at many industrial sites are con-
taminated with complex mixtures of pollutants. Cleaning up these
soils in situ with biosystems is potentially much more effective and
inexpensive than excavating the soils. Three classes of industrial
chemicals are currently the focus of EPA biosystems research on soils
and sediments: pentachlorophenol (PCP), polycyclic aromatic
hydrocarbons (PAHs), and polychlorinated biphenyls (PCBs).
The first of these, PCP, is a common soil contaminant at wood-
preserving facilities. EPA researchers have shown that a white-rot
fungus, Phanerochaete chrysosporium, can reduce the level of PCP in
the soil at these sites. A field study was set up at a former tank farm
where for 12 years aboveground storage tanks were filled with wood
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preservatives and diesel fuel, which then leaked into the surround-
ing soil. Two different strains of the fungus both noticeably depleted
the levels of PCP at the field site. Another EPA project showed that
when anaerobic conditions were induced and maintained, PCP and
other chlorinated compounds were degraded in freshwater sediments
from areas as diverse as Georgia ponds, the East River in New York
Gty, and Lake Borek in the Soviet Union.
High-molecular-weight PAHs, some of which are carcinogens,
are associated with wood-preserving plants, coal gasification sites,
and petroleum refineries. The estimated 700 wood-preserving plants
in this country, for example, use more than 4,960 tons of creosote
per year, some of which leaks from holding tanks and treatment
areas into the soil. Creosote is a complex mixture of over 200 in-
dividual compounds. Research on Pseudomonas paucimobilis under
a CRDA shows that this bacteria can metabolize these chemicals
(see page 10).
Another class of toxic compounds, PCBs, was used heavily for
about 50 years in a number of industrial applications. In the 1960s
and 1970s, researchers discovered that PCBs, which resist biological
degradation, were accumulating in the fatty tissues of animals and
fish. Even though their use has been largely discontinued, an es-
timated half million tons of PCBs remain in landfills. One EPA
project is assessing the rate at which two different bacterial strains
metabolize PCBs and pinpointing the enzymes that are responsible
for initiating the degradation.
Combined and sequential treatment. Most hazardous waste sites con-
tain complex mixtures of persistent organic and inorganic con-
taminants that can be cleaned up only by a combination of treatment
techniques. EPA researchers are developing methods to combine
various physical, chemical, and biological treatment technologies,
and comparing the effectiveness of the various combinations. For ex-
ample, a chemical treatment — adding potassium polyethylene
glycol (KPEG) to the soil at a site — may be used to dechlorinate
PCBs. Then, biological treatment, which is more effective after
dechlorination, can be used to complete the soil restoration. In one
project, researchers are combining KPEG treatment with composting
to enhance biodegradation in contaminated soils. These initial
studies will indicate how effectively composting enhances the meta-
bolic activity of the organisms at the site.
Ground Water. Effective bioremediation of ground water is
limited by the geology, hydrology, and geochemistry of the subsur-
face environment. In this area, EPA researchers are examining how
to supply electron acceptors, such as oxygen or hydrogen peroxide,
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Example of EPA Cooperative Agreement
with Small Business: Southern Bio
Products
In 1989, Southern Bio Products (SBP) signed a cooperative
research and development agreement (CRDA) with EPA's Gulf
Breeze Environmental Research Laboratory in Gulf Breeze,
Florida. Under this agreement, SBP provides funding and
expertise as part of an effort to investigate the use of microor-
ganisms to metabolize, or degrade, polycyclic aromatic
hydrocarbons (PAHs). High-molecular-weight PAHs are
carcinogens and thus represent a risk to human health and the
environment.
Creosote waste sites, coal gasification sites, and petroleum
refinery sites all can contain PAH-contaminated soils. In an
Electron micrograph of Pseudomonas paucimobilis.
(Magnification x 10,000.)
effort to make bioremediation a viable alternative for such sites,
the project team isolated a strain of bacteria that uses PAHs
(e.g., fluoranthene) as a sole source of carbon and energy for
growth. The team then examined the possible pathways by
which the bacteria break down the PAHs.
Next, the team developed a three-phase sequential system
to treat creosote- and similarly contaminated soil and water.
The steps in this process entail soil washing, a membrane ex-
traction process that reduces the waste to be treated by 100- to
100,000-fold, and biodegradation of extracted pollutants. In the
past year, EPA submitted three patents for steps in this new
technology, which will be field tested within one year. Under
this CRDA, EPA will grant an exclusive license to SBP to com-
mercialize the patented technology.
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For Southern Bio Products, a relatively new company, the
CRDA provided a way to develop a product quickly. Accord-
ing to a representative of the company, the cooperative efforts
of the Gulf Breeze laboratory under the FIT A agreement
stimulated information flow and cross-fertilization of ideas bet-
ween EPA and SEP. SBP worked primarily with the microbial
Contaminated
Contaminated
Soil Slurry
Soil Washing
Clean
Overburden
Discarded
i
Clean "Washed"
Soil Discarded
Dewatering of Concentration of
Contaminants Contaminants
Schematic diagram ofSBP/EPA tri-phasic treatment process: scv7
washing, membrane extraction/concentration, and biodegradation.
ecology and biotechnology branches at Gulf Breeze, but also
had access to the ecotoxicology and pathobiology branches.
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to promote aerobic processes in contaminated ground water. They
are also working on ways to develop or improve anaerobic biological
processes for degrading contaminants in the subsurface.
In one ground-water study, researchers are assessing whether
they can stimulate indigenous microorganisms in the subsurface to
remove carbon tetrachloride from contaminated aquifers. Pilot-scale
field evaluations suggest this form of bioremediation will work.
Another project deals with creosote that has infiltrated into the soil
around wood-preserving plants and moved into the water table. Re-
searchers have shown that some of the organic compounds in
creosote can be anaerobically degraded as they move down through
the aquifer.
Metabolic processes research. EPA's metabolic processes research
generates a better understanding of the processes by which microor-
ganisms degrade chemicals, expanding the types of organisms that
can be used in biosystems technologies. Based on the insights gained
from this research, scientists can then choose indigenous organisms
or genetically engineered organisms to meet needs in pollution
cleanup and control. In one research project, EPA is currently
developing systems to biodegrade pollutants formed as combustion
products of synthetic fuel generated from fossil fuels. These
heterocyclic compounds are known to be persistent in the environ-
ment, but researchers are examining the metabolic processes by
which bacteria can break them down. Another study is focusing on a
single pollutant: trichloroethylene (TCE). Examination of bacteria al-
ready shown to degrade TCE led to a better understanding of the me-
tabolic process by which TCE was oxidized. As a result, EPA
scientists have identified seven more strains that can degrade this
pollutant.
Risk assessment. Biodegradation can result in incomplete
mineralization with the formation of intermediate breakdown
products. Before environmental engineers can adopt a biologically
based remediation technology, researchers must assess the potential
ecological and human health effects associated with each waste
management option. In the area of human health, EPA has estab-
lished a research program to improve risk assessments.
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These projects and others will result in many advances in
bioremediation within the next few years. Enhancing the activities of
microorganisms to hasten the degradation of chemicals is expected to be-
come commonplace. Exploiting cell-free bioproducts, such as enzymes,
to treat a wide range of pollutants will also become an established
technique. Under the FIT A, industry can both participate in and
catalyze this rapid growth of biosystems technologies. By seeking a
synergistic relationship with EPA scientists, industry can achieve
what one researcher called a "perfect union" of expertise, oppor-
tunity, and market savvy.
The procedure for setting up a cooperative R&D or licensing
agreement under the FTTA is designed to encourage collaboration
between industry and EPA laboratories. For industry, the key ad-
vantage of the process is the speed and ease with which the agree-
ments can be negotiated and signed. CRDAs are not subject to
federal contracting or grant regulations. In addition, each laboratory
director has the authority to establish CRDAs for that particular lab,
and this decentralization of the decision-making process reduces the
administrative procedures involved.
Another important advantage is that CRDAs are flexible enough
to fit the goals of many different sizes and types of companies. For ex-
ample, under the FTTA a company can support applied research at
an EPA laboratory while reserving first rights to involvement in any
technology that results. Or, if the scientific mechanism by which a
company's product works is unclear, the company can cooperate
with an EPA laboratory to identify this mechanism. A company can
also share space and equipment with EPA (as well as with a third
party, such as a university or another company) in a combined effort
to develop a new technology. If a CRDA results in a patentable tech-
nology, that invention can be patented by either the federal
laboratory and/or the company (depending on the agreement).
Under these CRDAs, EPA may provide technical expertise,
facilities, equipment, staff, or services. EPA may not provide direct
funding to the outside cooperator, although the cooperator may
provide funds to EPA.
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The FIT A, therefore, provides a potent mechanism for EPA
laboratories to work with the private sector in developing new pollu-
tion control technologies and bringing them to the marketplace. The
achievements of CRDAs in bioremediation have aided EPA in its mis-
sion to remove or minimize the effects of pollutants in the environ-
ment, while at the same time catalyzing the development of the
emerging biosystems industry. The CRDAs already in place suggest
that cooperative agreements between industry and government will
prove highly successful. How could a CRDA help your company
develop a pollution prevention or control technology? Write to the
individuals listed on the next page for further information.
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m Environmental Research Laboratory, Athens, GA (ERL-Athens)
EPA Contact: Dr. John Rogers, U.S. EPA, College Station Rd.,
Athens, GA 30613 (404) 546-3128
« Environmental Research Laboratory, Gulf Breeze, FL (GBERL)
EPA Contact: Dr. Hap Pritchard, U.S. EPA, Sabine Island, Gulf
Breeze, FL 32561 (904) 934-9260
« Risk Reduction Engineering Laboratory (RREL)
EPA Contact: Dr. Al Venosa and Dr. John Glaser, U.S. EPA, 26 W.
Martin Luther King Dr., Cincinnati, OH 45268 (513) 569-7668 and
(513) 569-7568
• Robert S. Kerr Environmental Research Laboratory, Ada, OK
(RSKERL)
EPA Contact: Mr. Dick Scalf, U.S. EPA, P.O. Box 1198, Ada, OK
74820 (405) 332-8800
m Health Effects Research Laboratory (HERD
EPA Contact: Dr. Larry Claxton, U.S. EPA, Research Triangle
Park, NC 27711 (919) 541-2329
m Center for Environmental Research Information (CERI)
EPA Contact: Dr. Fran Kremer, U.S. EPA, 26 W. Martin Luther
King Dr., Cincinnati, OH 45268 (513) 569-7346
For information on the Federal Technology Transfer Act
(FTTA), licensing agreements, or CRDAs, contact:
Mr. Larry Fradkin
FTTA Coordinator
Office of Research and Development
U.S. Environmental Protection Agency
26 W. Martin Luther King Dr.
Cincinnati, OH 45268
(513)569-7960
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