United States Office of Research and Office of Solid Waste and

Environmental Protection Development Emergency Response

Agency Washington, DC 20460 Washington, DC 20460

EPA/540/F-95/506G September 1995

Bioremediation Field

Initiative Site Profile:

Escambia Wood 

Preserving Site

Background

The Escambia Wood Preserving Site located in Brookhaven, Mississippi, is a

former wood preserving facility that used pentachlorophenol (PCP) and creosote to treat wooden poles. Located on the site are two pressure treatment

cylinders, a wastewater treatment system, five bulk product storage tanks,

seven condenser ponds (including a 3,000,000-gal., unlined primary surface

impoundment), and excavated soil materials containing process chemicals

from past spills. In April 1991, U.S. EPA Region 4 initiated a removal action to

eliminate all sources of potential releases to the environment.

In the fall of 1991, PCP-contaminated soil from previous spill events was

used to evaluate multiple soil treatment regimes involving three different

fungal strains during an 8-week treatability study. A follow-on field-scale

demonstration study using one specific strain of lignin-degrading fungi was

conducted over a 5-month period (June to November 1992). Both studies

were carried out by the U.S. EPA Risk Reduction Engineering Laboratory

(RREL) (now the National Risk Management Research Laboratory

[NRMRL]) and the U.S. Department of Agriculture Forest Products Laboratory (FPL) under the Superfund Innovative Technology Evaluation (SITE)

Program with support from the Bioremediation Field Initiative.

Site Characterization

In preparation for conducting a treatability study, candidate contaminated

source soils from the site were sampled at the surface in June 1991. Composite soil samples were analyzed for PCP and other volatile and semi-volatile organics. The composite samples contained elevated concentrations of

44 organic compounds, 12 of which are hazardous constituents of K001

waste. PCP concentrations were found to range from 25 to 342 mg/kg, with

an average of 143 mg/kg. Contaminant concentrations varied greatly

within the sampled soil. Subsequent excavation of these soils to provide

sufficient quantities of material for the treatability and demonstration studies showed increasing concentrations of PCP with depth, eventually encountering concentrations in excess of 5,000 mg/kg.

Treatability Study

The treatability study compared 10 treatments, combining three fungal

species, three inoculum loading levels, and the appropriate controls. The

experimental method combined a randomized complete block (RCB) designed without replication and a balanced incomplete block (BIB) design

with treatment replicated four times. Eleven 10-ft by 10-ft plots, each

holding about 4 tons of soil, were constructed. In the RCB design, six of the

plots each received a separate treatment. In the BIB design, each of the five

remaining plots was divided by interior borders into four 2.5-ft. by 2.5-ft

SITE FACTS

Location: Brookhaven,

Mississippi

Laboratories/Agencies: U.S.

EPA National Risk Management

Research Laboratory (NRMRL),

U.S. Department of Agriculture

Forest Products Laboratory

(FPL), Superfund Innovative

Technology Evaluation (SITE)

Program, U.S. EPA Region 4

Media and Contaminants:

Pentachlorophenol (PCP) and

creosote in soil

Treatment: Lignin-degrading

fungi

Date of Initiative Selection:

Spring 1991

Objective: To evaluate the

effectiveness of lignin-degrading

fungi treatment for wood

preserving wastes

Bioremediation Field Initiative

Contacts: John Glaser and

Richard Brenner, U.S. EPA

NRMRL, 26 West Martin Luther

King Drive, Cincinnati, OH 45268

Regional Contact:

De’Lyntoneus Moore, U.S. EPA

Region 4, Waste Management

Division, 345 Courtland Street,

Atlanta, GA 30365

Printed on paper that contains at least

50 percent recycled fiber.
split plots. These plots were used to evaluate one of the

treatments from the RCB design and four additional

treatments.

Excavated soil was mechanically sieved to pass through

a 1-in. screen, mixed, and placed in the plots to a depth

of 10-in. The plots were then inoculated with the fungi.

After inoculation, each plot periodically was irrigated

and tilled with a garden rototiller. Wood chips were

added to each plot to provide a substrate to sustain

growth of the fungi. Figure 1 is a schematic of the soil

preparation, showing the treatment plots. Treatment

with Phanerochaete chrysosporium resulted in an average

69 percent reduction in PCP for initial concentrations in

excess of 1,000 mg/kg, while an average reduction of 89

percent was achieved with Phanerochaete sordida with

initial PCP concentrations of 600-700 mg/kg.

Demonstration Study

A field-scale demonstration study was conducted in 1992

using the lignin-degrading fungus P. sordida to treat PCP

and 13 polycyclic aromatic hydrocarbons (PAHs) found in

the contaminated soil. P. sordida was selected based on the

results of the treatability study and its natural occurrence

in soil. The soil excavated for this study had much higher

PCP concentrations (up to 5,200 mg/kg) than that used in

the treatability study, although the average concentration

was decreased to approximately 1,000 mg/kg by dilution

with clean soil. The contaminated soil was placed in a 70-ft

by 100-ft soil treatment bed and two 25-ft by 50-ft control

beds. The treatment bed was inoculated with 10 percent

dry (w/w) fungal hyphae and inoculum substrate/soil.

No amendments were added to one of the control beds,

while the other was formed by blending in 10 percent dry

(w/w) sterile inoculum.

Depletion of targeted compounds in the treatment bed

and the two control beds was measured over a 20-week

period from June to November. A 64 percent reduction

in PCP concentration was achieved in the fungal treatment bed compared to 26 percent in the sterile inoculum-amended control bed and 18 percent in the

non-amended control bed. Substantial reductions in the

concentrations of 3- and 4-ring PAHs were noted in all

three beds, although depletion of 4-ring PAHs was greater

in the treatment bed than in the control beds. The 5- and

6-ring PAHs were not significantly diminished in either

the treatment or the control beds. The extent of treatment was controlled by two factors: low biological

activity of the fungal inoculum as determined by ergosterol measurements at the beginning of the study and

unfavorable weather conditions that severely curtailed

the desired tilling schedule for the soil beds.

Status

Although the treatment efficiencies achieved in the 5-

month field-scale demonstration study at Brookhaven

were somewhat disappointing after the highly promising treatability study, the demonstrated ability of lignindegrading fungi to remove significant quantities of PCP

at an initial concentration of approximately 1,000

mg/kg is considered encouraging. This concentration

is roughly 10 times greater than concentrations typically

attempted with bacterial-based treatments. The excessive rainfall encountered at Brookhaven, which frequently interfered with regularly scheduled tilling, may

have resulted in oxygen depletion in the treatment bed,

leading to loss of fungal mass and activity. This environmental consequence suggests that future applications of fungal treatment technology may need to avoid

soil bed configurations to prevent weather-related constraints. Current costs of fungal treatment operations

are estimated at $150-200/ton of soil. Reductions in

inoculum mass requirements and costs are anticipated

as new inoculum formulations and application techniques are developed.

The Bioremediation Field Initiative was established in 1990 to expand the nation’s field experience in bioremediation technologies.

The Initiative’s objectives are to more fully document the performance of full-scale applications of bioremediation; provide

technical assistance to regional and state site managers; and provide information on treatability studies, design, and operation of

bioremediation projects. The Initiative currently is performing field evaluations of bioremediation at eight other hazardous waste

sites: Libby Ground Water Superfund site, Libby, MT; Park City Pipeline, Park City, KS; Bendix Corporation/Allied Automotive

Superfund site, St. Joseph, MI; West KL Avenue Landfill Superfund site, Kalamazoo, MI; Eielson Air Force Base Superfund site,

Fairbanks, AK; Hill Air Force Base Superfund site, Salt Lake City, UT; Reilly Tar and Chemical Corporation Superfund site, St.

Louis Park, MN; and Public Service Company, Denver, CO. To obtain profiles on these additional sites or to be added to the

Initiative’s mailing list, call 513-569-7562. For further information on the Bioremediation Field Initiative, contact Fran Kremer,

Coordinator, Bioremediation Field Initiative, U.S. EPA, Office of Research and Development, 26 West Martin Luther King Drive,

Cincinnati, OH 45268; or Michael Forlini, U.S. EPA, Technology Innovation Office, Office of Solid Waste and Emergency Response,

401 M Street, SW., Washington, DC 20460.

Figure 1. Schematic of soil preparation, from excavation to screening,

mixing, placement in treatment plots, and inoculation with fungi.

2