United States             Office of Research      EPA/600/8-89/073
Environmental Protection   and Development      Revised
Agency                   Washington, DC 20460  July 1990

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           The Alaskan Oil Spill
           Bioremediation Study

                 1990 Update

It has been over a year since the Exxon Valdez spilled
approximately 11  million gallons of crude oil into the
pristine waters off Alaska's Prince William Sound. The
spill, the worst in U.S. history, prompted a monumental
clean-up  effort and  launched significant scientific
research efforts. In the summer after the spill, the U.S.
Environmental Protection Agency's (EPA's) Office of
Research and Development initiated a bioremediation
field demonstration to determine the feasibility of using
nutrients to enhance the microbial degradation of oil
on the Alaskan shorelines. The study, which is the
largest of its kind ever conducted, has already provided
a wealth of datathatwill have far-reaching implications
for mitigating the effects of future oil spills worldwide.
But there are still questions that remain, questions that
EPA hopes to answer through an intensive research
program  of field and laboratory studies this summer.
These studies, some of which are already underway,
complement  the  clean-up efforts planned for the
summer  of 1990, which include  bioremediation by
nutrient application to hundreds of widely scattered
segments of shoreline in Prince William Sound. This
brochuredescribesthe Alaskan Oil Spill Bioremediation
Study of 1989, the research initiated during the winter
following the spill, and the field and laboratory activities
planned for the summer of 1990.

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Prince William Sound, Alaska
                             Back Cover:
                             From the air, a bioremediated test plot resembled a clean rectangle
                             etched upon the surface of the beach. The cobblestone plot, which
                             was located in Snug Harbor, was treated with oleophilic fertilizer
                             during the summer of 1989.

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                 Alaska and Prince William Sound
                     Tanker Aground
                      March 24, 1$89
                                       Prince William    —
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                             Shipping
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ALASKA - The name comes from an Aleutian word meaning "great land." If laid on the
48 lower states, Alaska would cover nearly one-fifth of them. The state is great in
resources as well as land mass. In 1968, enormous quantities of oil were discovered on
Alaska's North Slope in Prudhoe Bay. In 1974, construction began on the Trans-Alaska
Pipeline under the direction of the Alyeska Pipeline Consortium Co., which was formed
by the seven firms that pump crude oil from the North Slope. The pipeline extends
nearly 800 miles with itsterminus in Valdez, Alaska, where a shipping complex and other
facilities are located. Since the pipeline was built, approximately 9,000 shipments of oil
have been transported through Prince William Sound.

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The Aftermath of the Spill

    In March of 1989, the supertanker
Exxon Valdez ran aground on Bligh Reef
in Prince William  Sound, Alaska,
flooding one of the nation's most pris-
tine and sensitive environments in less
than 5 hours with approximately 11
million gallons of crude oil. The spilled
oil affected  up to an estimated 900
miles of shoreline in the Sound. These
islands and their waters are home to a
wide range of wildlife, including deer,
black and brown  bears, seals,  otters,
and whales, as well as an extensive
array of birds and fish. Commercial
salmon fish hatcheries are also located
in the protected bays ringing the Sound.
    In the short term, the oil spill has
taken a toll on the area's diverse wild-
life, and directly touched the lives of
many Alaskans. While the long-term
effects of the spill  are still being evalu-
ated, there is the potential for habitat
and food chain disruption, as well as
decreased survivability and  repro-
ductivity of animals exposed to the oil.
These effects, while perhaps not im-
mediately fatal to a given individual,
have a direct bearing on the survival of
the speciesasa whole and consequently
of the ecosystem of which it  is a
member.
    In the immediate aftermath of the
accident, attempts to clean up the
spilled  oil were hampered  by bad
weather and the remoteness of the lo-
cation. Ultimately, a massive cleanup
was organized that involved more than
10,000 individuals,  several  hundred
vessels and aircraft, and highly special-
ized equipment. Many  conventional
clean-up techniques (such as booms,
high- and low-pressure spraying, skim-
mers, and manual  scrubbing) were
employed to remove oil from the sur-
face of the rocks and beaches. These
techniques, however, were unable to
remove all of the oil from the beaches,
or oil trapped under rocks and in the
matrix of beach sediments.
                                     Glaciers have left their mark on the
                                     coastline of Prince William Sound. Coastal
                                     topography is often steep, and ranges from
                                     vertical cliffs to sand, pebble, and boulder
                                     beaches. In some areas, streams and snow
                                     melt also introduce large amounts of fresh
                                     water to the near-shore waters of the
                                     Sound.
Workers used several
techniques, including
high-pressure spraying, to
clean up the spilled oil in
1989.

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              The Effects of Oil on the Food Chain
A CHAIN OF EVENTS - Spilled crude oil has the potential to affect every level of the
marine food chain. Floating oil may contaminate plankton, which includes algae, fish
eggs, and the larvae of various invertebrates (such as oysters and shrimp). In turn, the
small fish that feed on these organisms can become contaminated. Larger animals in
the food chain  (including bigger fish, bears, and humans) may then eat these
contaminated fish. In addition, marine animals and birds may be exposed directly to
oil in the water column, which they can ingest or get on their fur or feathers. Spilled
oil may also preventthe germination and growth of marine plants and the reproduction
of invertebrates, either by smothering or by toxic effects.

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Enhancing the Cleanup with
Bioremediation

    To enhance the clean-up efforts,
the U.S.  Environmental Protection
Agency's (EPA's) Office of  Research
and Development (ORD) suggested that
hioremediation might  be useful. A
panel of national and international ex-
pert  scientists  in  the  field  of
bioremediation was convened, which
recommended that ORD conduct a field
demonstration project to evaluate the
feasibility of using bioremediation to
assist in clean-up operations.
    Bioremediation involves the use
of microorganisms (such as bacteria) to
enhance the "degradation" of oil and
other types  of chemicals. Scientists
have observed that biodegradation oc-
curs naturally in the environment af-
ter a spill of crude oil due to the presence
of indigenous microorganisms. These
microorganisms degrade the hydrocar-
bons found in the crude oil (which they
use as a food source) into a harmless
substance consisting primarily of car-
bon dioxide, water, and fatty acids.
    A few days after the Alaskan spill,
microorganisms began to  multiply
naturally in response to the presence of
oil. With such a bounty of hydrocar-
bons, however, the ability of these mi-
croorganisms to degrade the oil was
limited by the availability of nutrients
(nitrogen and  phosphorus). Without
these nutrients, the microorganisms
were unable to fully utilize the hydro-
carbons as a food source.
    Therefore, the panel of expert sci-
entists that was convened recom-
mended that ORD apply fertilizers to
designated test beaches  in Prince Wil-
liam Sound. These fertilizers would
help  the  microorganisms to degrade
the oil. The rationale behind this ap-
proach is that the greater the number
of microorganisms or the greater the
microbial activity, the greater the abil-
ity of the organisms to break down the
oil and the faster the rate of  degrada-
tion. Bioremediation has the potential
to clean  up the oil trapped  beneath
rocks and in the beach sediments, and
has the added advantage of being less
disruptive to the  environment  than
conventional clean-up techniques such
as pressure spraying.

A Cooperative Agreement

     Because of the need for rapid re-
sponse, ORD quickly drafted a research
plan for the bioremediation field test
and submitted it to EPA's Science Ad-
visory Board (SAB). The SAB, which
Congress established in the late 1970s
to provide advice to EPA regarding the
scientific and technical aspects of en-
vironmental problems and issues, ap-
proved the  research plan with minor
modifications. The SAB also stated that
the project would be a significant con-
tribution to future environmental re-
search planning and technology.
     ORD then approached Exxon and
proposed a  cooperative effort to con-
duct the  bioremediation study under
the Federal Technology Transfer Act
of 1986. The Act encourages collabora-
tion between the  private and public
sectors for the economic, environmen-
tal, and social benefit of  the United
States.
     In early June of 1989, ORD en-
tered into a formal cooperative agree-
ment with Exxon to test the capability
of bioremediation in treating contami-
nated beaches in Prince William Sound.
To ensure the independence  of study
results,  EPA  provided the technical
expertise  to carry out the  bio-
remediation project, and was respon-
sible for oversight and management of
the study. EPA also agreed to provide
supplemental resources for any other

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Scientists used
backpack sprayers to
apply the liquid
oleophilic fertilizer to
a test beach in Snug
Harbor.
efforts that would be necessary to make
the technology useful in the cleanup of
future spills. Exxon paid for the logisti-
cal support directly applicable to the
study (such as lodging for the scientists
and transportation from Valdez, Alaska,
to the test sites) and for laboratory and
field support. In 1989, EPA's contribu-
tion  to  the  Alaskan  Oil  Spill
Bioremediation Project was approxi-
mately $1.6 million and Exxon's share
was about $3 million.

Snug Harbor

     After planning and  mobilizing
staff and facilities,  scientists surveyed
beaches to find a suitable test area for
          the project. Snug Harbor,
          which  is located on the
          southeastern side of Knight
          Island, was selected as the
          test site. The area is  sur-
          rounded by mountains with
          steep vertical ascents and
          peaks of up to 2,000 feet.
          The shoreline, which was
          moderately oiled, had a rea-
          sonable uniformity of beach
          material  (cobblestone,
          gravel,  or sand).  The test
          area was also sheltered from
          storms and subject to mini-
          mal  freshwater  runoff
          (streams and  snow melt),
          which could interfere with
          the field tests. The degree
          of  contamination at  Snug
          Harbor simulated those
conditions considered typical of a beach
following physical washing  (the pri-
mary  clean-up procedure used by
Exxon).
                            Slow-release, water-soluble fertilizer
                       briquettes were bagged in herring nets and
                        anchored in the tidal zone on a test beach
                                             in Snug Harbor.
    Nutrient application began on
June 8,  1989. Two types of nutrient-
rich fertilizers were applied to the test
beaches:

  A slow-release, water-soluble fertil-
  izer, in which nutrients were slowly
  released and distributed to the oil-
  contaminated beach surfaces by rain
  and tidal action. Fertilizer  "bri-
  quettes" (similar in size and weight
  to charcoal) were bagged in herring
  nets,  placed on the  beach surface,
  and anchored in the tidal zone with
  steel-reinforced rods.

• A liquid oleophilic fertilizer, in which
  the nutrients adhered to the oil cov-
  ering the rock and gravel surfaces,
  thereby making nitrogen and phos-
  phorus available at the site of micro-
  bial activity. This  fertilizer was
  sprayed over the contaminated test
  areas.

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Beach Name
Seal
Seal

Seal
Otter
Otter
Eagle
1 SnugHarbons
Snug Harbor Test Plots
I William Sound,
Beach Type Nutrient Application | Alaska. A small
Cobble
Cobble

Cobble
Mixed sand and gravel
Mixed sand and gravel
Mixed sand and gravel
None (reference)
Oleophilic

Water-soluble
Water-soluble
Oleophilic
None (reference)
promontory divides
Seal Beach from Otter
Beach, while a more
conspicuous
promontory divides
Eagle Beach from
OtterBeach.


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A test plot in Snug
Harbor where the
oleophilic fertilizer
was applied looked
dramatically cleaner
than an adjacent
untreated plot.
    Each fertilizer was applied to two
types of beaches—one  comprised of
mixed sand and gravel; the other made
up of cobblestone. Two "reference"
test plots,  where no nutrients  were
added, also were set up for comparison
against the treatment plots.
    Approximately 2 weeks after the
oleophilic fertilizer was applied to the
cobblestone beach plot, scientists ob-
served visible reductions in the amount
of oil on rock surfaces.  This was par-
ticularly evident from the air, where
the contrast with oiled areas surround-
ing the plot was dramatic. To the sci-
entists who surveyed the test plot by
helicopter, it looked as if a clean rec-
tangle had been etched on the beach's
surface.  Close examination of this
treated  cobblestone plot verified that
much of the oil on the rocks' surfaces
was gone, although oil remained in the
mixed gravel below the rocks.
     EPA scientists also observed re-
duced amounts of oil in the mixed sand
and  gravel beach  plot  treated with
oleophilic fertilizer within 2 weeks,
though the difference  between  the
treated area and its reference plot was
not as striking as that observed on the
cobblestone beach. This is because tides
mixed up the sand and gravel, whereas
the  cobblestone remained relatively
stationary. Therefore, the visual disap-
pearance of the oil was less apparent in
the sand and gravel plot. The oil below
the beach surface was disappearing as
well. All other plots (including those
treated  with only  the  fertilizer  bri-
quettes) appeared as oiled  as they had
been at the beginning of the field study.
     Over the next 2 to 3 weeks, the

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cleaned rectangle on the cobblestone
beach remained clearly visible. The oil
in the  sand and gravel below the
cobblestone persisted, but became less
apparent over the course of the sum-
mer. The oleophilic-treated mixed sand
and gravel plot also appeared increas-
ingly cleaner than its untreated refer-
ence plot.  Beaches  treated with the
fertilizer briquettes were relatively
unchanged.
    Toward the end of the summer
season,  the entire test  area became
steadily cleaner. Most of the areas sur-
rounding the test  plots  were also
cleaner,- scientists attributed this to
several  storms and  frequent rainfall,
which helped replenish nutrients in
this area and enhance the natural bio-
degradation processes.  However, a
heavily oiled area south of Snug Harbor
that was never treated remained con-
siderably  contaminated, suggesting
that nature alone could not account for
the dramatic reduction of oil observed
in the test area.

Passage Cove

    Based on the promising results of
the initial field test at Snug Harbor and
the absence of any adverse effects on
the area's  ecosystems,  EPA recom-
mended to Exxon  in July that the
bioremediation efforts be scaled up
during the remainder of the summer.
Passage Cove served as the main refer-
ence beach for a large-scale application
of nutrients by Exxon clean-up crews,
which commenced on August 1,1989.
    Passage Cove, which is located on
the northwestern side of Knight Island,
had been heavily oiled by the spill.
Even though the site had been physi-
cally washed by Exxon clean-up crews,
considerable oil remained  on the
shoreline and in the beach sediments.
In fact, contamination was discovered
as far as 2 feet below the surface. Scien-
tists did find, however, that the physi-
cal washing had spread the oil into a
very thin layer over a large surface area
of rock and gravel, which made it easier
for the microorganisms to gain access
to and break down the oil.
    Scientists set up three beach plots
for research at Passage Cove. All of the
plots  were comprised of cobblestone
overlaying sand  and gravel.  The
oleophilic fertilizer and a granular form
of a slow-release, water-soluble fertil-
izer were applied in tandem to one test
plot. These fertilizers were applied to-
gether because subsurface oil contami-
nation was a concern at Passage Cove
and there  were some questions about
how deep the oleophilic fertilizer could
penetrate  the beach's subsurface. Un-
like the water-soluble fertilizer, the
oleophilic fertilizer has a syrup-like
consistency which could hinder its
ability to permeate the subsurface.
    A third type of fertilizer, a fertil-
izer solution containing inorganic ni-
trogen and  phosphorus dissolved  in
seawater,  was sprayed across the sec-
ond test plot by fixed sprinkler systems
(similar  to lawn  sprinklers). An
untreated  reference plot was also set
up for comparison purposes.
    Within 2 weeks following  the ap-
plication of the oleophilic and water-
soluble fertilizers, the treated beaches
were considerably cleaner than the ref-
erence plot. Not only did the rock sur-
faces  look cleaner, but the oil beneath
the cobblestone was also disappearing.
The beach plot treated with the fertil-
izer solution from the sprinkler system
behaved in a similar manner, and be-
came steadily cleaner. The reference
plot showed no sign of oil loss. By the
end of August, the treated plots looked
equally clean. In contrast, the refer-
ence plot appeared very much as it did
in the beginning of the field study. Oil
in the subsurface still remained in all
the test plots.
                                               9

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                                     Passage Cove
    Passage Cove is
  located on Knight
    Island in Prince
    William Sound,
Alaska. The test plot
  in Tern Beach was
       divided by a
       promontory.
           Passage Cove Test Plots
Beach Name    Beach Type
Raven
                             Tern
             Cobble over mixed sand
             and gravel
                                          Cobble over mixed sand
                                          and gravel
                              Kittiwake      Cobble over mixed sand
                                          and gravel
                                     Nutrient Application

                                     None (reference)
                                     Oleophilic and water-
                                     soluble
                                     Nutrient solution
                                     sprinkler system
                                              10

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Hydrocarbon Analyses

    To confirm that  biodegradation
was indeed taking place, scientists per-
formed a variety of chemical analyses
in the laboratory. These analyses indi-
cated that over the summer, smaller
and smaller concentrations of hydro-
carbons were present in the oil samples
taken at the test sites, thereby con-
firming that biodegradation was oc-
curring  (see  page 12). Scientists
measure hydrocarbon  concentrations
through the use of gas chromatogra-
phy. Oil is really a mixture of many
different hydrocarbons, each  with a
specific boiling point.  A boiling point
is the temperature at which  a com-
pound will "volatilize" or turn to vapor.
Gas chromatography capitalizes on the
differences in boiling points among
different hydrocarbons to separate,
identify, and indicate the relative con-
centration of each of these components
in crude oil.

Ecological Monitoring

     Ecological monitoring  studies
were conducted concurrently with the
fertilizer application tests at both Snug
Harbor  and Passage Cove. Although
dilution and tidal mixing should mini-
mize the potential for adverse ecologi-
cal effects, scientists were concerned
that  certain  components  of the
oleophilic fertilizer could be toxic to
some marine species. In addition, algal
blooms (excessive growth of algae in a
body of water) could occur as a result of
the sudden availability of nitrogen and
phosphorus. Too many nutrients in a
water body reduce the amount of oxy-
gen present, thereby favoring plant life
over animal life.
     To determine the potential toxic-
ity of oleophilic fertilizer to native or-
ganisms, a wide range  of  species
(including  stickleback fish,  Pacific
herring, silver salmon, mussels, oys-
ters, shrimp, and mysids) was tested.
EPA scientists collected samples of
seawater directly over the beaches that
had just been treated with a combina-
tion of the oleophilic and water-soluble
fertilizer (a "worst-case" scenario).
     Laboratory studies  with these
samples showed that certain compo-
nents of oleophilic fertilizer are mildly
toxic to the most sensitive marine spe-
cies  (oyster larvae) where there is no
dilution by tidal action. Oyster larvae
are two orders of magnitude more sen-
sitive than salmon. The potential tox-
icity of the fertilizer to salmon is a key
concern since these fish spawn in Prince
William Sound. The circumstances of
fertilizer application, however, are such
that the potential to adversely affect
marine and terrestrial life is very un-
likely. Scientists also found that add-
ing nutrients to oiled shorelines  did
not cause any increases in algae, or any
measurable nutrient accumulation in
adjacent embayments.
     EPA scientists also  placed mus-
sels in cages  just offshore from  the
fertilizer-treated beaches and moni-
tored them to determine if any toxic
substances were accumulating in their
tissues due to the release or breakdown
of the oil. No  oil was detected in  the
mussel tissues, and no oil was observed
in the water  offshore from the test
areas.

Microcosms

     Microcosms were constructed on
board a fishing vessel to provide
supplemental information to the field
demonstration project. Microcosms are
designed to simulate naturally occur-
ring processes on a smaller scale. They
ha ve the advantage of providing backup
information in the event some unfore-
                                               11

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a 2
           Snug Harbor-Mixed Sand and Gravel-Untreated Beach
    Jl
   c c c c  c
                Sampling Date: 6/8/89
                                      S 2
 111
                        Sampling Date: 7/29/89
..lllllllllllllll  h
   HHHHHH PHPHHHHHHHHHH
                       222222
                    Hydrc
                                                          Hydrocarbons
                Snug Harbor-Cobble-Water Soluble Fertilizer
               Sampling Date: 6/8/89
                         c c c c c
                        Sampling Date: 7/29/89
                                              .llllill
                                llilllll  ll
                                         HHHHHH PHPHHH
                                         CCCCCCRCHCCC
                                     H H  H H  H
                                     C C  C C  C
                                                            23456
             Snug Harbor-Mixed Sand and Gravel-Oleophilic Fertilizer
                Sampling Date: 6/8/89
         ll
III!
                        Sampling Date: 7/29/89
• 111
Illllllllllll  I.
           H PHPHHHHHHH
           CRCHCCCCCCC
                                         HHHHHHPH
                                                       HHHHHHHHHH
                                                          Hydrocarbons
 HYDROCARBON ANALYSES - These figures illustrate how the composition of oil
 extracted from Snug Harbor test plots changed from early June to late July of 1989.
 Crude oil is a complex mixture of many compounds, including numerous hydrocarbons.
 Hydrocarbons, as the name implies, are made up of chains of carbon and hydrogen
 atoms. The numbers along the bottom of the graphs refer to the number of carbons in
 each hydrocarbon chain in the samples of crude oil. The height of the bar indicates the
 relative concentration of each hydrocarbon in the sample.

     While all of the July chromatographs show reduced amounts of hydrocarbons
 compared to the June graphs (indicating that degradation was taking place), the treated
 plots show more pronounced reductions relative to the untreated reference plot,
 indicating that degradation was enhanced in the treated plots.
                                     12

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seen complication results in the field
data. The microcosms also allow sci-
entists to test bioremediation concepts
under idealized conditions to better
understand what is happening in the
field.
     Tanks representing the test plots
were set up on the fishing vessel.
Perforated containers filled with con-
taminated cobblestone and contami-
nated mixed sand and gravel were
placed in the tanks.  Fertilizers were
then applied to simulate  the actual
test  applications. The  initial micro-
cosm results indicated that if suffi-
cient nutrients were supplied to the
microorganisms, enhanced biodegra-
dation of the oil would occur. Because
microcosms represent the test systems
that best reflect  field conditions, a
similar response could be expected in
the field.

The Culmination of the
Project

     By the end of September 1989,
Exxon had treated 74 miles of shore-
line in the largest  bioremediation
project ever conducted. Pontoon boats
and  other  small  craft  were used to
access the  shoreline, and crews used
airless spray pumps to apply fertilizer
to the oiled beaches. This large-scale
application of fertilizer was the culmi-
nation of the knowledge and experi-
ence gained  during  the  previous
months.

Winter Research

     Overall, the initial findings from
last summer's field and laboratory tests
indicate that using nutrients to  en-
hance biodegradation is effective and
environmentally safe. To  further
strengthen the success  of this
bioremediation approach, a variety of
questions still must be addressed about
the fertilizers, the application meth-
ods, the potential for adverse effects,
and other  details. Answering these
questions is critical to both the present
and  future  application  of bio-
remediation techniques to oil-con-
taminated  beaches.  Thus,  EPA and
                                     Researchers set up
                                     microcosms on board a
                                     fishing vessel to simulate
                                     test plots in the field and
                                     provide backup data.
During the large-scale
application of fertilizers
in 1989, Exxon crew
members usedpontoons
and other small craft to
apply the fertilizer to
contaminated beaches.
                                               13

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Exxon initiated additional research in
the winter following the spill  to ad-
dress questions remaining in the fol-
lowing key areas:

1. The mechanism by which the
oleophilic fertilizer works.
    While the summer study demon-
strated that the oleophilic fertilizer ef-
fectively cleaned  oiled beaches, the
precise mechanism by which this fer-
tilizer works needed further clarifica-
tion. Because of the dramatic effect of
this fertilizer in the field, there were
concerns that it was possibly washing
the oil from the surface of rocks. Labo-
ratory studies during the winter, how-
ever, confirmed  that the oleophilic
fertilizer enhanced the extent and rate
of oil degradation through the addition
of inorganic nutrients. The fertilizer
may also have enhanced biodegrada-
tion by indirectly increasing the num-
ber of  oil-degrading microorganisms
present in the beach sediments.

2. The optimization of fertilizer
application.
    The oleophilic and water-soluble
fertilizers were applied in combination
at Passage Cove in 1989  to provide
maximal distribution of nutrients to
oil-contaminated areas. Yet, there are
still some questions regarding the opti-
mal use  of this combined treatment.
For instance, could any reactions occur
that might reduce the effectiveness of
one or the other fertilizer? What appli-
cation sequence should be considered
for these two  fertilizers? Such ques-
tions must be  answered to ensure the
optimal application of nutrients in the
field.

3. The potential for adverse
ecological effects.
    During the summer field study,
questions were raised concerning the
potential for enhanced biodegradation
to produce by-products that may be
harmful to the environment.  Acute
and chronic tests with crustacean and
fish species were conducted during the
winter program that indicated no rea-
son for concerns in this area.

4. The relationship between
nutrient application and algal
blooms.
    Scientists are conducting math-
ematical studies to determine the rela-
tionship between nutrient application
and enhanced algae growth. Through
these studies, scientists will be able to
predict what will happen to nutrients
in the environment, including how the
nutrients are likely to be transported
and mixed, and how they will be uti-
lized  by the microorganisms. The
studies will help scientists determine
the effects that may be expected from
any new experiments involving nutri-
ents (and other chemicals).

5. The analytical procedure for
measuring oil degradation.
    Oil  degradation is  commonly
measured by extracting oil from beach
material and then analyzing its com-
position in the laboratory to determine
the number and type of hydrocarbons
present. Oil degradation has been ex-
tensively studied over the last 20 years,
and scientists know  that certain hy-
drocarbons in crude oil degrade quickly,
while   others   are   slow   to
degrade. Scientists frequently use some
of the slower degrading hydrocarbons
as "internal markers," against which
the degradation rate of more quickly
degrading hydrocarbons can be mea-
sured.  In Alaska,  however,  scientists
discovered that the common internal
marker hydrocarbons were also rapidly
degraded in some cases. This made the
established procedure for measuring
oil degradation of limited use, and so
new analytical procedures are needed.
                        14

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If scientists can develop a new internal
marker technique, it will improve their
ability to assess oil degradation in the
field.

6. The statistical verification of
field data and the modification, if
necessary, of sampling
procedures.
    EPA is developing statistical pro-
cedures and computer programs that
will allow scientists to further analyze
the collected data, examine important
trends, and modify sampling proce-
dures, if necessary. Scientists are also
reevaluating  selected chemical  and
biological measurements used to study
oil degradation to focus^on more sensi-
tive, less variable approaches.

    Although these six studies were
begun in the winter, many of them will
continue throughout the summer of
1990 in conjunction with field research
activities.

Activitiesforthe
Summer of 1990

    A Bioremediation Monitoring
Program will be conducted early in the
summer of  1990. The program will
supplement many of the studies from
the winter research  and enhance
bioremediation application activities
planned for the remainder of the sum-
mer. The program is designed to moni-
tor Exxon's large-scale application of
nutrients. It will be a joint undertaking
by EPA, Exxon, the National Oceanic
and Atmospheric Administration
(NOAA), and the Alaskan Department
This clump of sand and
gravel shows the
consistency of oil in the
summer of 1989. By the
summer of 1990, much of
this oil had weathered
into a thicker, more glue-
like consistency.
                 The Character of
                   Crude in 1990
        The character of crude oil found on the shorelines
    of Prince William Sound in 1990 will differ from the oil
    that was present in 1989. After an oil spill occurs in a
    marine environment, winds and waves help spread
    and disperse the oil. Some of this oil will evaporate. Oil that mixes with the seawater
    produces  an oil-in-water emulsion  (globules of oil suspended in water), commonly
    referred to as "mousse." Although mousse was prevalent in 1989, cleaning activities
    and weathering have significantly reduced its occurrence in 1990.

        As time goes by, oil that has washed ashore becomes more glue-like in character
    and may eventually form into a hard layer of weathered oil or weathered oil mixed with
    fine sediments. This covering, which has the look and consistency of asphalt, is called
    a tarmat. Tarmats are found on some  shorelines in Prince William Sound. Some of
    these sites will be considered for bioremediation after the tarmats are removed.

        Oil may also penetrate a beach surface by seeping into the matrix of sediments or
    by  being  buried  by clean  sediments  that  have washed  over the  area. On some
    shorelines, pockets of subsurface oil may persist. Bioremediation holds great promise
    for cleaning up these areas in the summer of 1990.
                                           15

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This sheltered "low-
   energy" beachis
exposed to minimal
   wave action and
  consists of poorly
  sortedgravel and
          cobble.
                  of  Environmental  Conservation
                  (ADEC). The monitoring program will
                  focus on assessing three key effects of
                  bioremediation:

                    The amount of enhanced microbial
                    degradation of surface and subsurface
                    oil that can be achieved by nutrient
                    addition.

                    The potential toxicity associated
                    with nutrient addition.

                   • The amount of nutrients present in
                    the  water  off  treated beaches.

                       In the  spring of  1990, shoreline
                   conditions were surveyed to determine
                   the extent of contamination in Prince
                   William Sound. Heavy winter storms,
                   along  with the natural processes of
                   weathering, had decreased the amount
                   of oil present on the shorelines. Never-
                   theless, oil remained  on many shore-
                   lines and in the subsurfaces of some
beaches.  Tarmats, thick asphalt-like
coverings of oil, were also scattered at
sites throughout the Sound.
    Based on these surveys, three types
of beaches were chosen to serve as sites
for the monitoring program: 1) a low-
energy beach with surface and subsur-
face  contamination,-  2) a  moderate/
high-energy beach with surface  and
subsurface contamination; 3) a moder-
ate/high-energy beach with subsurface
contamination. Terms such as  "high-
energy" and "low-energy" refer to the
degree of wave energy to which a beach
is exposed. Over 80 percent of the
coastline in Prince William Sound ex-
periences high or moderate wave en-
ergy levels.
    The selected beaches are  uni-
formly oiled and large enough to be
divided into two areas: one will be
treated by fertilizer, and the other will
remain untreated to  serve as a refer-
ence plot. Water and sediment samples
will  be taken on specific days after
                                           16

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           Clean-up Techniques for the Summer of 1990
In addition to bioremediation of approximately 400 sites, five other techniques will be
used to clean up the oil that persists in Prince William Sound and the Gulf of Alaska. These
techniques, which were chosen because they are the least disruptive to the environment.
                   Sorbent Booms. Sorbent booms are physical barriersthat intercept
                   and absorb oil. They will be used in areas where oil sheens persist.
                   The booms will be anchored near shore and replaced as necessary.
                   Manual Pickup. Small beach crews will use hand tools to pick up and
                   bag oily materials. Manual pickup will help improve the aesthetic
                   appearance of the beaches and remove potential sources of oily
                   debris that could foul fishing gear.
                   Tarmat Breakup/Removal. Tarmats, thick asphalt-like coverings of
                   oil which are slow to degrade, will be broken up with hand tools and
                   either scattered (to facilitate natural degradation or bioremediation)
                   or removed.
                   Tilling/Raking. In some areas, sediments will be raked or tilled in
                   order to expose  subsurface oil  to  natural  degradation or
                   bioremediation.
                   Spot Washing. Crews will use hand-held washing devices to remove
                   small accumulations of oil. The water and removed oil will then be
                   collected on the shoreline with sorbents (such as booms or "pom-
                   poms" that are designed to  absorb oil).
                                     17

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nutrient application and analyzed for
microbial activity and the amount of
nutrients and  oil present.  Scientists
will also use time-lapse photography
to characterize the visual changes in
the extent of surface oil.

The Research Component

    In addition to the Bioremediation
Monitoring Program,  EPA, Exxon,
ADEC, and the University of Alaska,
Fairbanks, will conduct research dur-
ing the summer of 1990.  Scientists
from all four organizations will par-
ticipate in designing and performing
experiments, and in collecting and in-
terpreting the data.
    The research planned for the
summer of 1990 is designed to address
certain questions about the effective-
ness and environmental safety  of
bioremediation that remain only partly
answered. The research program will
also  address questions  that will  be
important for  future applications of
bioremediation to oil-contaminated
beaches.
    EPA will conduct experiments to
answer the following key questions:

• How much  total oil removal at a
  given site can be expected from
  bioremediation?

   Scientists will   conduct   an
  experiment to determine the rate and
  extent  of degradation that can  be
  anticipated   when   different
  concentrations of fertilizer are applied
  to  contaminated beaches. Samples
  will be taken and analyzed in the
  laboratory. From these analyses, the
  scientists will  determine  what
  nutrient  concentrations  must  be
  maintained on the beaches to remove
  a given amount of oil.
What is the best fertilizer application
method for the  bioremediation of
subsurface oil?

  Scientists  believe  that  oil
degradation occurs at the top and/or
the bottom of a subsurface oil layer
(as opposed to throughout the layer).
Therefore,  nutrients must  be
consistently applied to these  areas
in  order  to enhance microbial
degradation  of oil.  Fertilizer
application will be tested  on  short
stretches of beaches with a distinct
layer of subsurface oil contamination..
The sprinkler application system
used in the summer of 1989 at Passage
Cove will be tested on one beach
plot.  Bathing  techniques will be
tested on another plot. These
techniques will work by saturating a
test area with a fertilizer solution so
that nutrients can seep through beach
sediments into the oiled subsurface.
An untreated  plot will serve as a
reference  plot  for  comparison
purposes.

Can the fertilizer application strategy
for the combined use  of oleophilic
and granular water-soluble fertilizer
be  further  optimized to assure
maximal degradation?

  This summer, bioremediation will
be   tested  on  beaches   with
considerably heavier concentrations
of oil than those tested last year. To
optimize  the effects of  these
fertilizers, it is important to remove
as much oil as possible in the shortest
timeframe. Therefore, scientists will
explore applying the oleophilic and
water-soluble fertilizers in different
combinations using laboratory
microcosms with fresh oiled beach
material.  Microcosms will permit
scientists to determine the optimum

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  application  strategy  without
  involving costly field operations.

• Can  additional information on
  biodegradation activity be obtained
  using new experimental measure-
  ments and analyses?

  To gain the most benefit from the
monitoring program, as much evidence
for enhanced hiodegradation must be
derived as possible. This means taking
advantage of new analytical techniques
that are not quite ready for routine use,
but that can betestedon samples taken
as part of the monitoring program. Sev-
eral state-of-the-art  techniques for
measuring oil degradation will  be
tested.

    The research data gathered during
the summer  of 1990 will be used to
supplement findings  from  the moni-
toring program to provide comprehen-
sive assessments on the effectiveness
of bioremediation.

Testing New Products

    ORD is also evaluating the ability
of several commercial products to en-
hance bioremediation in Alaska. In
February of 1990, ORD announced in
the Commerce Business Daily that it
was seeking  organizations  or compa-
nies that could offer commercial
methods capable of enhancing the bio-
degradation of crude oil residues in
Alaska. The Agency requested that
these proposals be submitted to the
National  Environmental Technology
Applications Corporation  (NETAC).
NET AC is an organization established
through a cooperative venture between
EPA and the University of Pittsburgh.
    As requested by EPA, NET AC as-
sembled  a Bioremediation Products
Evaluation Panel that met in March of

     ORD/NETAC: Bringing Innovative
         Technologies to the Market
     EPA and the University of Pittsburgh Trust have
     entered intoarnulti-yearcooperativeagreement
     to  establish  the National  Environmental
     Technology Applications Corporation (NETAC).
     NETAC's  purpose  is  to  facilitate  the
     commercialization  of technologies being
     developed  by the government and the private
     sector  that will  positively affect the  nation's
     most  pressing  environmental problems.
     NETAC's efforts encompass encouraging new
     technologies with promising commercialization
     potential, as well as innovations aimed solely at
     modifying and improving existing technologies
     or processes.
1990 to evaluate each of the 39 propos-
als submitted. The panel used detailed
screening criteria to evaluate the pro-
posals. Eleven proposals (two nutri-
ents,  one  dispersant, and eight
microbial cultures) were recommended
for further testing, along with a proto-
col for performing this testing.
    Ten vendors supplied products for
further testing. To evaluate these prod-
ucts, EPA scientists placed cleanbeach
material, weathered crude oil, seawa-
ter, and the commercial product in
glass flasks. The scientists performed
three different tests on these products.
The tests measured the degradation of
the oil, the change in the numbers of
oil-degrading microorganisms, and the
amount of oxygen used by microorgan-
isms while degrading oil.  (Increased
                                            19

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oxygen consumption is often an indi-
cator of increased microbial activity.)
     The Bioremediation  Products
Evaluation Panel will evaluate the re-
sults of these tests. It appears likely
that products having successful test
results will be approved for field tests
during the summer of 1990.
The Wave of the Future

    Bioremediation is a technology
that holds enormous promise for the
future. The successful field and labora-
tory tests already completed indicate
that bioremediation by nutrient addi-
tion offers a safe and effective way to
          ameliorate surface  and
          subsurface  oil. The activi-
          ties planned for the summer
          of 1990 are expected to
          verify these conclusions,
          and further expand our
          knowledge.
              While  prevention is
          the best defense, it is im-
          portant that technologies
          also be developed to  com-
          bat those oil spills that do
          occur.  For this reason,
          research efforts like the
          Alaskan Oil Spill Bioreme-
          diation Project are crucial.
          By understanding the sci-
          ence of those processes that
          can mitigate the potentially
          devastating effects of oil
          spills, we can help ensure
          the preservation of our rich
          and diverse natural  envi-
          ronment.
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