Bioremediation Resource Guide


United States	Solid Waste	EPA/542-B-93-004
Environmental Protection	and Emergency Response September 1993
Aqency	Technology Innovation Office
Washington, DC 20460
&EFW Bioremediation
Resource Guide
Abstracts of Policy,
Guidance, and Technical
Assistance Documents
Summary of Regulatory Mechanisms that
Affect Bioremediation Technologies
Descriptions of Bioremediation-Related Databases, Hotlines, Catalogs, and Dockets
Easy to Use Matrix that Assists in Identification of Appropriate Documents
CJ-'J,, Recycled/Recyclable
~y V) Printed with Soy/Can oia Ink on paper that
Q \^J contains at least 60% recycled fiber

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BIOREMEDIATION
RESOURCE GUIDE
and
BIOREMEDIATION
RESOURCE MATRIX
U.S. Environmental Protection Agency
Office of Solid Waste and Emergency Response
Technology Innovation Office
Washington, DC 20460
September 1993

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EXECUTIVE SUMMARY
This Bioremediation Resource Guide is intended to support decision-making by Regional and State Corrective
Action permit writers, Remedial Project Managers (RPMs), On-Scene Coordinators, contractors, and others
involved in evaluating cleanup alternatives for Resource Conservation and Recovery Act of 1976 (RCRA),
Underground Storage Tank (UST), and Comprehensive Environmental Response, Compensation, and Liability
Act of 1980 (CERCLA) sites by directing readers to bioremediation resource documents, databases, hotlines, and
dockets as well as identifying regulatory mechanisms (e.g., Research Development and Demonstration Permits)
that have the potential to ease the implementation of bioremediation at hazardous waste sites.
This Guide provides abstracts of representative examples of over 80 bioremediation bibliographies, guidance,
workshop reports, overview documents, study/test results, and test designs/protocols. The Bioremediation
Resource Matrix, which accompanies this Guide, identifies the technology, media, and contaminants covered in
each abstracted document. The included documents focus for the most part on soil and ground water and on in-
situ, slurry phase, and land treatment bioremediation. Information contained in this Guide is not intended to be
all-inclusive, nor does it represent an endorsement by the U.S. Environmental Protection Agency (EPA).
TABLE OF CONTENTS
INTRODUCTION	2
HOW TO ORDER DOCUMENTS LISTED IN THIS GUIDE	3
SOURCES OF BIOREMEDIATION INFORMATION/TECHNICAL ASSISTANCE	4
FEDERAL REGULATIONS AND GUIDANCE RELEVANT
TO BIOREMEDIATION	5
ABSTRACTS OF BIOREMEDIATION RESOURCES	6
REQUEST FOR COMMENTS	27
ORDER FORMS	29
BIOREMEDIATION RESOURCE MATRIX	Center pull-out
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INTRODUCTION
EPA is committed to identifying the most effective and efficient means of addressing the thousands of hazardous
waste sites in the United States. Therefore, the Office of Solid Waste and Emergency Response's (OSWER's)
Technology Innovation Office (TIO) at EPA is working in conjunction with the EPA Regions and research
centers, and industry to identify and further the implementation of innovative treatment technologies. Currently,
bioremediation is a frequently selected innovative technology.
The goal of OSWER is to encourage the development and use of innovative hazardous waste treatment technolo-
gies. One way of enhancing the use of these technologies is to ensure that decision-makers can avail themselves
of the most current information on technologies, policies, and other sources of assistance. To this end, this Guide,
which identifies documents that can directly assist RPMs and permit writers in investigating existing information
on bioremediation remedial applications for contaminants usually found at RCRA, UST, and CERCLA sites, was
prepared.
HOW TO USE THIS GUIDE
When using this Guide to identify resource information on bioremediation, you may wish to take the following
steps:
1. Turn to the Bioremediation Resource Matrix located in the center of this Guide. This matrix lists over 80
bioremediation related documents and identifies the type of information provided by each document as well
as a document ordering number.
2. Select the document(s) that appear to fit your needs based on the content information in the matrix.
3. Check the abstract identification code. This number refers to an abstract for the document. The number
corresponds to a page number in the Guide and the letter corresponds to an abstract on that page.
For example:
Abstract

Abstract A on
Identification
6 A —— page 6 of the
Code
I
Resource Guide

page 6 in the

Resource Guide

4. Review the abstract that corresponds to the document in which you are interested to confirm that the docu-
ment will fit your needs.
5. If the document appears to be appropriate, check the document number highlighted under the abstract. For
example:
' 1^ ADbcument tfcimbw:
6. Turn to the section entitled "How to Order Documents Listed in this Guide" on page 3 of this Guide and order
your document using the directions listed. You will find ordering forms identified in the section entitled
"Order Forms," which begins on page 29 of this Guide.
7. When seeking information on technical assistance sources, turn to page 4 of this Guide.
8. To identify information on Federal regulations and guidance relevant to bioremediation, turn to page 5 of this
Guide.
9. If you would like to comment on this Guide or would like additional information, turn to page 27 of this
Guide and follow the directions for mailing or faxing your comments/questions.
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HOW TO ORDER DOCUMENTS LISTED IN THIS GUIDE
Documents listed in this Guide are available through a variety of sources. When ordering documents listed in the
"Bioremediation Abstracts" section of this Guide, use the number listed in the shaded bar below the abstract. If
using the Bioremediation Resource Matrix in the center of the Guide to order documents, use the number listed
below the document title. If multiple document ordering numbers are identified, select the appropriate number
based on the directions below. EPA/540 and EPA/600 documents may be available through CERI and/or NCF.PI.
These document repositories provide in-stock documents free of charge, but document supplies may be limited.
Prior to purchasing a document through NTIS, you may also wish to review a copy at a technical library.
Document Type
Publications with the following numbers:
EPA/530
EPA/540
EPA/600
EPA/625
Document Source
Center for Environmental Research Information
(CERI)
Cincinnati, OH 45268
(513) 569-7562
Out of stock documents may be ordered from NCEP1 or may be purchased from NTIS.
Publications with the following numbers:
EPA/540
EPA/542
EPA/600
National Center for Environmental
Publications and Information (NCEPI)
11029 Kenwood Road, Building 5
Cincinnati, OH 45242
(513) 891-6561
fax requests to (513) 891-6685
A document title or number is needed to place an order with NCEPI. Out of stock documents may be ordered
from CERI or may be purchased from NTIS.
Publications with EPA/530 numbers RCRA Information Center (RIC)
401 M St., S.W. Mailcode: WI1-562
Washington, DC 20460
(202) 260-9327
"Order Form for Free Office of Solid Waste Publications," included in the "Order Forms" section of this Guide
can be used to order from the RIC.
OSWER Directives
(EPA personnel only)
Superfund Document Center
401 M St., S.W. Mailcode: OS-245
Washington, DC 20460
Attn: Superfund Directives
(202) 260-9760
Publications with NTIS numbers and Office of Solid
Waste and Environmental Response (OSWER)
Directives (for Non-EPA personnel)
NTIS
5285 Port Royal Road
Springfield, VA 22161
(703) 487-4650
NTIS provides documents for a fee. "NTIS Order Form," included in the "Order Forms" section of this Guide can
be used to order from NTIS.
If you have difficulty finding a document, call:
RCRA/Superfund/UST Hotline 1-800-424-9346
Operates Monday-Friday, 8:30 a.m. - 7:30 p.m., Eastern Time.
Hotline staff can help EPA staff or members of the public locate documents and assist callers with placing docu-
ment orders.
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SOURCES OF BIOREMEDIATION INFORMATION/
TECHNICAL ASSISTANCE
Numerous computer-based bulletin boards, regulatory hotlines, dockets, databases, catalogs, and periodicals are also avail-
able. These resources provide technical information on bioremediation and other innovative technologies and guide you to
additional valuable resources.
BULLETIN BOARDS:
• Alternative T reatment Technologies
Information Center (ATTIC) (data line) 301-670-3808
To obtain information on ATTIC 908-906-6828
A collection of hazardous waste databases accessed through a bulletin board
that provides hazardous waste abstracts, news bulletins, conference informa-
tion, and a message board.
• Cleanup Information Bulletin
(CLU-IN) (data line) 301-589-8366
A bulletin board for hazardous waste professionals that provides current
information on innovative technologies. Provides information bulletins, mes-
sage and on-fiie exchange, and on-line databases and directories.
Help Line 301-589-8368
Addresses questions about CLU-IN access and contents; addresses problems
with the service.
' Office of Research and
Development (ORD) (data line) 800-258-9605
Bulletin Board Service (BBS) (data line) 513-569-7610
Provides a bibliography of 18,000 documents, news excerpts from
"Bioremediation in the Field," and a message board.
Help Line 513-569-7272
Provides information on access to and contents of the ORD BBS.
CATALOGS:
• Catalog of Hazardous and Solid Waste Publications,
Sixth Edition EPA/530-B-92-001
Catalogs Office of Solid Waste policy directives, guidance documents, bro-
chures, Regulatory Development Branch memos, and other documents rel-
evant to hazardous and solid waste.
' Compendium of Superfund Program Publications
EPA/540/8-91/014, NTIS PR 881
Provides abstracts and ordering information for fact sheets, directives, publica-
tions, and computer materials on Superfund. Use the document ordering
directions to compendium.
DATABASES:
• DIALOG Database 800-3-DIALOG
A large database that contains files relevant to hazardous waste including:
Biotechnology Abstracts; Enviroiine; Corrective Action Search, Pollution Ab-
stracts; National Technical Information Services (NTIS); and others.
- NTIS Database
Contains abstracts of government-sponsored research, development, and
engineering analyses prepared by approximately 250 Federal agencies and
some State and local governments. Accessible via the DIALOG system.
• Records of Decision System (RODS)
(To get information on accessing RODs) 703-603-8881
Contains the full text of all signed Records of Decision for hazardous waste
clean-up sites nationwide. Direct access to RODS is available to EPA
personnel and organizations that have relevant EPA contracts.
' Vendor Information System for innovative
Treatment Technologies (VISITT) 800-245-4505
Contains current information on availability, performance, and cost of innova-
tive technologies to remedy hazardous waste sites.
DOCKETS:
• Federal Facilities Docket Hotline
.800-548-1016
Provides the name, address, NPL status, agency, and Region for the 1,930
Federal facilities listed on the Federal Facilities Docket. Facilities are on the
docket because they reported being a RCRA TSDF or having spilled or having
the potential to release CERCLA hazardous waste. Operates Monday-Friday,
8 a.m. - 6 p.m., Eastern Time.
• RCRA Information Center 202-260-9327
Indexes and provides public access to all regulatory materials supporting the
Agency's actions under RCRA, and disseminates current Office of Solid Waste
publications. Operates Monday Friday, 8:30 a.m. - 4 p.m., Eastern Time.
• Superfund Docket 202-260-3046
Provides access to Superfund regulatory documents, Superfund Federal
Register Notices, and Records of Decision. Operates Monday - Friday,
9 a.m. - 4 p.m., Eastern Time.
• UST Docket 202-260-9720
Provides documents and regulatory information pertinent to RCRA's Subtitle I
(the Underground Storage Tank program). Operates Monday - Friday,
9 a.m. - 4 p.m., Eastern Time.
HOTLINES/REGULATORY/TECHNICAL ASSISTANCE:
• EPA Headquarters Library 202-260-5921
Offers reports from various EPA offices and trade and environmental journals.
Features the "Hazardous Waste Collection" department. Operates Monday -
Friday, 10a.m. - 4 p.m., Eastern Time.
• RCRA/Superfund/UST Hotline 800-424-9346
Provides regulatory assistance related to RCRA, CERCLA, and UST programs.
Serves as a liaison between the regulated community and EPA personnel and
provides information on the availability of relevant documents. Operates
Monday - Friday, 8:30 a.m. - 7:30 p.m., Eastern Time.
• TSCA Hotline 202-554-1404
Answers public and private regulatory questions on TSCA. Refers callers to
appropriate EPA contacts, and takes TSCA-relevant document orders. Oper-
ates Monday - Friday, 8:30 a.m. - 5 p.m., Eastern Time.
• Superfund Health Risk Technical
Support Center 513-569-7300
Provides EPA Regional Superfund risk assessors, Slate agencies, and those
working under EPA contract with technical, typically chemical-specific, support
and risk assessment review. Operates Monday-Friday8a.m.-5p.m., Eastern
Time.
INFORMATION CENTER:
' National Center for Environmental
Publications and Information (NCEPI) 513-891-6561
(Fax requests) 513-891-6685
Stores and distributes to public and private callers a limited supply ol most
EPA publications, videos, posters, and other multi-media materials. Callers
should know document titles or numbers when calling. The following
periodicals can be obtained from NCEPt, as supplies last:
- Bioremediation in the Field
A periodical devoted to bioremediation that contains 140 potential applications
of bioremediation, including site type, name, and contact.
- Groundwater Currents
A newsletter that reports on innovative in-situ and ex-situ groundwater
remediation technologies to be applied in the field.
- Tech Trends
An applied technology journal that provides information on Superfund remov-
als, remedial actions, and RCRA corrective actions.
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FEDERAL REGULATIONS AND GUIDANCE RELEVANT TO
BIOREMEDIATION
This table lists pertinent RCRA regulations, with the Code of Federal Regulations (CFR) and Federal Register (FR)
citations. This table provides information on guidance documents relevant to these regulations. It is important to note
that many of these Federal regulations are considered optional. As such, States may choose not to become authorized
for these provisions. In addition, States may elect to have more stringent regulations than the Federal regulations
identified here. Contact your State environmental protection agency when considering the applicability of any of the
following Federal regulations.
CITATION
REGULATION
DESCRIPTION
GUIDANCE
40 CFR §261.4(e)-(f)
July 19, 1988
53 FR 27290
40 CFR §270.65
July 15, 1985
50 FR 28728
40 CFR §264.600
December 10, 1987
52	FR 461164
40 CFR §270.42(e)
March 7,1989
54	FR 9596
(Changes certain permit
modifications for hazardous
waste)
40 CFR §268.40
June 1,1990
55	FR 22686
(Presents third third
wastes)
40 CFR §268.45
August 18, 1992
57	FR 37279
40 CFR §268.44(h)
August 17, 1988
53	FR 31143, 31185, 31188,
31196,31199, 31202
(presents final rule on first third
wastes and national capacity
variances)
40 CFR §260, 264.552 et al
February 16, 1993
58	FR 8658
(Presents final CAMU rule)
July 27, 1990
55 FR 30842-30845
(Proposes corrective
action & CAMU)
Treatability Study
Exemption
Research Development and
Demonstration Permits
Subpart X Miscellaneous
Units
RCRA Permit Modification
Rule: Temporary
Authorization
Land Disposal Restrictions
(LDR) Subpart D -
Treatment
Standards
Treatment Standards lor
Hazardous Debris
Variance from an LDR
Treatment Standard
Corrective Action
Management
Unit (CAMU)
Allows for treatability studies
under RCRA
Allows the issuance of a RCRA
permit for a pilot scale study
pertaining to an innovative or
experimental technology
Allows the issuance of a RCRA
permit for a miscellaneous unit
Allows the permitting agency to
grant a facility a temporary
authorization to perform certain
activities (e.g., cleanups,
corrective action and closure
activities) for up to 180 days
Sets forth RCRA hazardous
waste treatment standards
Discusses biological destruc-
tion of hazardous constituents
from debris surface
Allows for a site-specific
treatability variance to be
issued as a nonrulemaking
procedure
Encourages treatment,
including use of innovative
treatment (specifically
bioremediation), instead of
containment
Conducting Treatability Studies
Under RCRA (7/92, OSWER
Directive 9380.3-09FS, NTIS
PB92-963-501)
EPA Guidance Manual for
Research Demonstration and
Design Permits
(7/86, EPA/530-SW-86- 008,
OSWER Directive 9527.00-1 A)
Modify RCRA Permits (9/89,
EPA/530-SW-89-050)
Land Disposal Restrictions
Summary of Requirements
(2/91, OSWER Directive
9934.0-1 A)
Regional Guide: Issuing
Site-Specific Treatability
Variances for Contaminated
Soils and Debris from LDRs
(1/92, OSWER Directive
9380.3-08FS)
Environmental Fact Sheet:
EPA Issues Final Rules for
Corrective Action
Management Units and
Temporary Units
(1/93, EPA/530-F-93-001)
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ABSTRACTS OF BIOREMEDIATION RESOURCES
The following abstracts describe the contents of pertinent bioremediation documents and are categorized by document type.
Document types included in this Section are:
Begins on Page
• Bibliographies 6
• Guidance/W orkshops 8
• Overview Documents 10
• Study/Test Results 13
• Test Designs/Protocols 24
• Examples of Other Relevant Documents 26
To quickly identify documents pertinent to your interest area, see the Bioremediation Resource Matrix in the cenLer of this
Guide. The documents in the matrix are categorized using the document types identified above and can be cross-referenced
with the abstracts using the code to the left of the document titles on the matrix. In an effort to limit the number of resources
listed here, documents on oil spills, waste minimization, natural process bioremediation, leaching/immobilization, ex-situ
bioremediation, and other media (e.g., air and surface water) as well as Records of Decision and proceedings are not in-
cluded. Samples of documents focused on ex-situ bioremediation of wastewater and leachate using a biofilter and a
bioreactor are included. However, those seeking information on these topics or other topics not addressed in this Guide may
wish to contact the hotlines, dockets, etc. listed on page 4 of this Guide. These abstracts were pulled from a TIO literature
survey under development and the NTIS Database.
BIBLIOGRAPHIES
6A 1
Biodegradation of Pesticides. (Latest citations from the
NTIS Database), (Published Search).
NERAC, Inc., Tolland, CT, November 1992
NTIS I>KuiiK;r.t Number PB97-854297/XAB
The bibliography contains citations concerning the
biodeterioration of pesticides in soil and water by naturally
occurring microbes, chemicals, and microbial innoculants. The
citations examine the kinetics and metabolic process of pesti-
cide biodegradation, and include means to enhance the
process. Mathematical models are also included. (Contains a
minimum of 61 citations and includes a subject term index and
title list.)
6B
Biodegradation of Toxic Wastes. (Latest citations from
the Energy Database), (Published Search).
NERAC, Inc., Tolland, CT, October 1992
NITS Document Number: PSP3>S51#9/LL„!
The bibliography contains citations concerning the use of
bacteria to decompose hydrocarbons and other hazardous
materials. Microbial breakdown of coal wastewater, jet fuel,
phenols, and cyanides is also discussed. Cost comparisons
between biodegradation and conventional physical and chemi-
cal methods are considered briefly. (Contains 250 citations
and includes a subject term index and title list.)
6C
Bioremediation, January 1988-March 1992 (Citations
from the NTIS Database).
National Technical Information Service, Springfield, VA,
February 1992
• WiiiM li ££&£$ ¦¦ ¦¦ -vM ;
The bibliography contains citations concerning the decomposi-
tion of toxic materials by biological means. Bacterial decompo-
sition of jet fuel, wood preservatives, explosives, crude oil,
halogenated organics, diesel fuel, aviation fuel, and creosote
are discussed. Enhancement of decomposition rates by addition
of nutrients is also included.
6D
Federal Publications on Alternative and Innovative
Treatment Technologies for Corrective Action and Site
Remediation: Second Edition.
Member Agencies of the Federal Remediation Technologies
Roundtable: U.S. Environmental Protection Agency, U.S.
Department of Defense, U.S. Air Force, U.S. Army, U.S.
Navy, U.S. Department of Energy. U.S. Department of
Interior, Washington, DC, August 1992
EPA Docum«itJNvHnben 54MMJ2/Q01 ,
NTIS Dcxftinient NnipBer. &B&3445696/LL
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The Federal Remediation Technologies Roundlablc developed
this bibliography to publicize the availability of Federal docu-
ments pertaining to innovative and alternative technologies to
treat hazardous waste. The bibliography addresses technolo-
gies that provide for the treatment of hazardous waste. It
emphasizes innovative technologies for which detailed cost and
performance data are not available. It includes citations for
documents addressing: international surveys and conferences;
technology survey reports; treatability studies; groundwater;
thermal processes; biological; physical/chemical; and commu-
nity relations. Citations provided here will supplement listings
in this Guide.
7A
Hazardous Materials: Microbiological Decomposition.
(Latest citations from the BioBusiness Database).
National Technical Information Service, Springfield, VA,
May 1992
N' FfM	Nisn''"u"i	AH
The bibliography contains citations concerning the decomposi-
tion of toxic materials by biological means. Bacteria, enzymes,
and bioluminescence are among the methods discussed. Bacte-
ria and enzymes that digest toluene, polychlorinated biphenyls
(PCBs), selenium wastes, oil shale waste, uranium, oil sludge,
pesticides, rubber wastes, and pentachlorophenol are discussed.
Flavobacterium and white rot fungus are among the biological
agents highlighted. (Contains 250 citations and includes a
subject term index and title list.)
7B
Literature Survey of Innovative Technologies for
Hazardous Waste Site Remediation, 1987-1991.
U.S. Environmental Protection Agency, Office of Solid
Waste and Emergency Response, Washington, DC, July
1992
EPA fisviJiTiPsrF NymlK!"	5
NTIS Di-coficul '"Viiiuly.r.	H.'5M AB
EPA's Office of Solid Waste and Emergency Response is
seeking to further the use of innovative hazardous waste
treatment technologies in its programs. In order to achieve
more permanent remedies, the Agency is encouraging the use
of new or innovative technologies that are capable of treating
contaminated soils/sludges and groundwater more effectively,
less expensively, and in a manner more acceptable to the public
than existing conventional methods. The bibliography is
intended to increase the efficiency of the technology
evaluation process. The document is not meant to be compre-
hensive in scope nor is it meant to convey an endorsement of
the citations. It is meant to provide a survey of publications
which could be useful when innovative technologies are inves-
tigated. As a research aid, the bibliography can help provide
insights into current developments and provide references
which may serve as a basis for further investigations.
7C
Selected Alternative and Innovative Treatment Technolo-
gies for Corrective Action and Site Remediation, (A
Bibliography of EPA Information Resources), Winter
Update.
U.S. Environmental Protection Agency, Washington, DC,
January 1993
This bibliography provides citations for documents relevant
to alternative and innovative treatment technologies appli-
cable at Superfund and RCRA sites. The bibliography also
includes: conferences and international surveys; technology
survey reports/guidance; treatability studies documents;
groundwater documents; technology support documents;
physical/chemical treatment documents; community rela-
tions documents; bulletin boards and databases; and technol-
ogy newsletters. Citations provided here will supplement
listings in this Guide.
7D
Subsurface Science Program Bibliography, 1985-1992,
(Bibliography).
U.S. Department of Energy, Office of Health and Environ-
mental Research, Washington, DC, August 1992
MIS OtfLiLiiiciii jSumbei; DE92-(WiH>42/XA3B
The Subsurface Science Program sponsors long-term basic
research on (1) the fundamental physical, chemical, and
biological mechanisms that control the reactivity, mobilization,
stability, and transport of chemical mixtures in subsoils and
groundwater; (2) hydrogeology, including the hydraulic, mi-
crobiological, and geochemical properties of the vadose and
saturated zones that control contaminant mobility and stability,
including predictive modeling of coupled hydraulic-geochemi-
cal-microbial processes; and (3) the microbiology of deep
sediments and groundwater. This research, focused as it is on
the natural subsurface environments that are most significantly
affected by the more than 40 years of waste generation and
disposal at DOE sites, is making important contributions to
cleanup of DOE sites. Past DOE waste-disposal practices have
resulted in subsurface contamination at DOE sites by unique
combinations of radioactive materials and organic and
inorganic chemicals (including heavy metals), which make
site cleanup particularly difficult. The long- term (10- to 30-
7

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year) goal of the Subsurface Science Program is to provide
a foundation of fundamental knowledge that can be used to
reduce environmental risks and to provide a sound scientific
basis for cost-effective cleanup strategies. The. Subsurface
Science Program is organized into nine interdisciplinary
subprograms, or areas of basic research emphasis. The subpro-
grams currently cover the areas of Co-Contaminant
Chemistry, Colloids/Biocolloids, Multiphase Fluid Flow,
Biodegradation/ Microbial Physiology, Deep Microbiology,
Coupled Processes, Field-Scale (Natural Heterogeneity and
Scale), and Environmental Science Research Center.
GUIDANCE/WORKSHOPS
Bioremediation of Hazardous Waste Sites Workshop;
Speaker Slide Copies and Supporting Information.
U.S. Environmental Protection Agency, Center for Environ-
mental Research Information, Cincinnati, OH, February
1989
NTiS Document Nux.iber PBS205/XAB
Basic requirements for implementing biological systems to
remediate hazardous wastes initial data requirements; example
site for bioremediation; reactor design; and in-situ design.
8B^ I
Biotechnology Workgroup for Department of Defense
Soil and Groundwater Decontamination Applications;
Final Report for Period Ending March 1989.
Reuter, R. H., Life. Systems Inc., Cleveland, OH, Naval Civil
Engineering Lab. Port Hueneme, CA, June 1991
^-TTS Document Ncrrh-.-r AD \2^7 9*>6WXAR
This report contains materials used in and generated by the
Department of Defense Biotechnology Workshop on Soil and
Groundwater Decontamination Applications. Various
bioremediation techniques for treating soil and water contami-
nated with sludges, solvents, toxins, acids, bases, and heavy
metals were discussed as well as the overall place ofbiotechnol-
ogy in Installation Restoration programs. Among the specific
applications discussed were: biochemical sensors to determine
environmental stress in organisms; in-situ detoxification and
biodecontamination of pollutants in soils and waste streams;
sequestration, removal, and recovery of metals in waste streams
with metal-binding proteins; and the use of vegetation to limit
the transport to sequester, and/or to remove contaminants from
soil or water.
80
Contaminants and Remedial Options at Wood Preserv-
ing Sites.
Selvakumar, A.; Sudell, G.; and Wolf, G., Foster Wheeler
Enviresponse, Inc., Edison, NJ, October 1992
NTIS Document Nuro'xrr FB92-232222/XAB
The report will assist Federal, State, or private, site removal
and remedial managers operating under the Comprehensive
Environmental Response, Compensation, and Liability Act
(CERCLA), the Resource Conservation and Recovery Act
(RCRA), or State rules. Tt provides information that
facilitates the selection of treatment technologies and services
at wood preserving sites, in order to meet the regulations'
acceptable levels of cleanliness. Within the context of the
United States wood preserving industry, the reference identi-
fies the sources and types of wood preserving contaminants,
characterizes them, and defines their behavior in the
environment. It addresses the goals in technology selection and
describes the principal remedial options for contaminated
wood preserving sites. It also considers ways to combine these
options to increase treatment efficiency. Finally, this
remedial aid provides a comprehensive bibliography,
organized by its relevance to each section, to complement the
information offered in these pages.
8D
Environmental Biotechnology of Hazardous Wastes
Research Planning Workshop, The National Science
Foundation.
Blackburn, J.W.; Donaldson, T.L.; and Sayler, G.S., and Oak
Ridge National Laboratory., Oak Ridge, TN, Tennessee
University, Knoxville, TN, Center for Environmental
Biotechnology, U.S. Department of Energy, Washington,
DC, August 1988
NTJS Docujiiwit Nombei. DES9 00I340/XAB
Environmental biotechnology is "the direct use of microorgan-
isms and their capabilities to solve environmental problems and
for in-situ agricultural applications and industrial waste treat-
ment." Environmental biotechnology is at the interface be-
tween responsible disciplines in engineering, molecular biol-
ogy, and ecological sciences. The purpose of this research
planning effort was to define limitations in the current knowl-
edge base and to develop a research agenda and strategy that
will lead to the successful practice of environmental biotech-
nology. The focus was specifically directed at environmental
protection. However, it was recognized that the research plan
8

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typifies a central research mode! for the future development of
effective and ecologically sound biotechnology for other envi-
ronmental applications.
9A I
Guidance Manual on Hazardous Waste Land Treatment
Closure/Post-Closure, 40 CFR Part 265 Final Report.
Sims, J.L. and Sims, R.C., Utah State Univ., Logan, UT,
Department of Civil and Environmental Engineering,
Sponsored by the U.S. Environmental Protection Agency,
Office of Solid Waste, Washington, DC, April 1987
F f' V I >i H il- fVsr.TitN-*( aV:,.l M - 1
N'US	Xundiet: PB&7-1 S:Ui'I.Va AF-.
The guidance manual addresses closure/post-closure of hazard-
ous waste land treatment (HWLT) units under 40 CFR Part 265
Subpart G and Section 265.280 of Subpart M. The manual
specifically addresses five areas: (1) general information on
HWLT and methods of closure; (2) objectives of closure and
post-closure; (3) factors affecting closure and post-closure; (4)
methods for addressing closure and post-closure based on
migration potential; and (5) management during closure and
post-closure.
Innovative Technology: Slurry-Phase Biodegradation
Fact Sheet (Final).
U.S. Environmental Protection Agency, Office of Emer-
gency and Remedial Response, Washington, DC, November
1989
'JTIS	F»9fK£?43WX'A0
The fact sheet provides technology description, site character-
istics affecting treatment feasibility, technology considerations,
and technology status for Slurry Phase Biodegradation (SPB).
The sheet describes how SPB is potentially effective in treating
various organic contaminanLs.
9C
Methodologies for Evaluating In-Situ Bioremediation of
Chlorinated Solvents, Research Report August 21,1989 -
June 14,1991.
Grbic-Galic, D.; McCarty, P.L.; Roberts, P.V.; and Semprini,
L., Stanford University, CA, Department of Civil Engineer-
ing, Robert S. Kerr Environmental Research Laboratory,
Ada, OK, March 1992
l/PA t-Miiuwa! Number: MKV-R f?
MT1S	Number'
The report summarizes the behavior of and requisite conditions
for a class of natural biological processes that can transform
chlorinated aliphatic compounds. These compounds are among
the most prevalent hazardous chemical contaminants found in
municipal and industrial wastewaters, landfills and landfill
leachates, industrial disposal sites, and groundwater. Biologi-
cal degradation is one approach that has the potential for
destroying hazardous chemicals so that they can be rendered
harmless for all time. Methodologies are presented that are
useful for evaluating the potential for biorestoration of ground-
water contaminated with chlorinated aliphatic compounds.
Section 1 provides an introduction and an overview of the
problems with chlorinated aliphatic compounds in groundwa-
ter. Section 2 presents a review of the processes affecting the
movement and fate of chlorinated aliphatics in the subsurface,
including ad vaction, dispersion, sorption and relative mobility,
diffusional transport, and immiscible transport. Methodologies
and results are presented for evaluating the presence of a native
mcthantrophic community and its ability to degrade the con-
taminants of concern; determining the sorption of contaminants
to the aquifer material; and preliminary designing of an in-situ
treatment approach using the model previously described.
9D
"No Migration" Variances to the Hazardous Waste Land
Disposal Prohibitions: A Guidance Manual for Petition-
ers.
U.S. Environmental Protection Agency, Office of Solid
Waste, Washington, DC, March 1990
}M'A I iin'iimt'H N'lrnN-r. c> Vfp/SW
s w f 1-^ fU i i nr il-	i	A A V*
This manual is intended to assist hazardous waste management
facility owners and operators who may be considering petition-
ing the U.S. Environmental Protection Agency for variance
from land disposal prohibitions at specific sites for specific
wastes. The guidance manual also provides the EPA interpre-
tation and suggested procedures to implement the regulatory
standards and procedures set out in 40 CFR 268.6. This manual
also contains descriptions of additional requirements for "no
migration" petitions in the Land Disposal Restrictions First
Third Rule.
9E
Rotating Biological Contactors. Engineering Bulletin.
U.S. Environmental Protection Agency, Office of Emer-
gency and Remedial Response, Washington, DC, October
1992
F.PA	Kuinbrt
N l'IK tMvumcnr-Numlwr: Phv?	Ait
9

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Rotating biological contactors employ aerobic fixed-film
treatment to degrade either organic and/or nitrogenous (ammo-
nia-nitrogen) constituents present in aqueous waste streams.
Fixed-film systems provide a surface to which the biomass can
adhere. Treatment is achieved as the waste passes by the media,
enabling fixed-film systems to acclimate biomass capable of
degrading organic waste. Fixed-film rotating biological
contactor reactors provide a surface to which soil organisms
can adhere; many indigenous soil organisms are effective
degraders of hazardous wastes. The bulletin provides
information on the technology applicability, the technology
limitations, a description of the technology, the types of
residuals produced, site requirements, the latest performance
data, the status of the technology, and sources of further
information.
10A ||
Slurry Biodegradation Engineering Bulletin.
Science Applications International Corporation, Cincinnati,
OH, Sponsored by the U.S. Environmental Protection
Agency, Office of Emergency and Remedial Response,
Washington, DC, September 1990
EPA	htuutxM. oS-C'K
N'TUi Pe.auwfit Mowix-r. PB92• :2H<>IWXAH
In a slurry biodegradation system, an aqueous slurry is created
by combining soil or sludge with water. This slurry is then
biodegraded aerobically using a self-contained reactor or in a
lined lagoon. Thus, slurry biodegradation can be compared to
an activated sludge process or an aerated lagoon, depending on
the case. There are two main objectives for using the technol-
ogy: to destroy the organic conlaminanL and, equally important,
to reduce the volume of contaminated material. Slurry biodeg-
radation can be the sole treatment technology in a complete
cleanup system, or it can be used in conjunction with other
biological, chemical, and physical treatment. It may be demon-
strated in the Superfund Innovative Technology Evaluation
(SITE) program. Commercial-scale units are in operation.
Vendors should be contacted to determine the availability of a
unit for a particular site. The bulletin provides information on
the technology applicability, the types of residuals produced,
the latest performance data, site requirements, the status of the
technology, and sources for further information.
OVERVIEW DOCUMENTS
Bioprocessing Applications in the Management of
Nuclear and Chemical Wastes.
Genung, R.K., Oak Ridge National Laboratory, TN, U.S.
Department of Energy, Washington, DC, November 1988
N HS Dri-Luintin N.i'al>cj IiE8y-CV336N/.£A8
The projected requirements for waste management and envi-
ronmental restoration activities within the United States will
probably cost tens of billions of dollars annually during the next
two decades. Expenditures of this magnitude clearly have the
potential to affect the international competitiveness of many US
industries and the continued operation of many Federal facili-
ties. It is argued that the costs of implementing current technol-
ogy will be too high unless the standards and schedules for
compliance are relaxed. Since this is socially unacceptable,
efforts to improve the efficiency of existing technologies and to
develop new technologies should be pursued. A sizable re-
search, development, and demonstration effort can be easily
justified if the potential for reducing costs can be shown.
Bioprocessing systems for the treatment of nuclear and chemi-
cally hazardous wastes offer such promise.
Bioremediation: An Information Update on Applying
Bioremediation to Site Cleanup.
U.S. Environmental Protection Agency, Office of Solid
Waste and Emergency Response, Washington, DC, March
1992
tPA Doturncai. Kualar
NUN EHvcunsem Number.	! 26.1
The Bioremediation Field Initiative was established to provide
the U.S. Environmental Protection Agency (EPA) and State
Project Managers, consulting engineers, and industry with
timely information regarding new developments in the applica-
tion of bioremediation at hazardous waste sites. The initiative
provides evaluation of the performance of selected full-scale
field applications; provides technical assistance to Remedial
Project Managers (RPMs) and On-Scene Coordinators
(OSCs), through the Technical Support Centers; and is
developing a database on the field applications of
bioremediation, which is summarized in this bulletin.
Critical Review of In-Situ Bioremediation Topical
Report, January 1990 - March 1992.
Ray, C; Rittmann, B.E.; Seagren, E.; Valocchi, A.J.; and
Wrenn, B., Illinois University at Urbana-Champaign, TL,
Newmark Civil Engineering Laboratory, March 1992
NT1S Dt-'Uir.icm NiiniHri • fRQ |
In-situ bioremediation, which is the managed, in-place
cleanup of contaminated groundwater aquifers and surface
soils by microorganisms, is a promising technology because it
10

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is versatile and can have significant economic advantages.
Many common contaminants are biodegradable, and new
microbial capabilities for degradation are being discovered all
the time. Success in the field and in laboratory studies point
out the promise. On the other hand, the promises are not yet
fulfilled, mainly because of the complexity of the subsurface
situation. The report provides a comprehensive and in-depth
critical review of in-situ bioremediation. It is organized to
evaluate the possibilities and restrictions inherent in all facets
of in-situ bioremediation, including microbiology, hydrody-
namics, engineering, and its legal and other nontechnical
aspects. Several of the key conclusions are illustrated by case
studies of successful field projects. Finally, the research
needed to advance in-situ bioremediation to become a reliable
and acceptable tool is outlined.
11A
Hazardous Waste Land Treatment: A Technology and
Regulatory Assessment.
Brown, K.W.; Evans, G.B.; and Overcash, M., Argonne
National Laboratory, IL, U.S. Department of Energy,
Washington, DC, September 1987
NTIS	VuiuFu-j: Of*MM557i/XAE
Land treatment is a waste management technology that pro-
vides a high level of treatment for some hazardous wastes by
using the soil as a treatment medium to degrade, transform, or
immobilize waste constituents. It is characterized by very long
retention times, and its costs are comparable to or lower than
those of other waste treatment or disposal technologies. Also,
most land treatment sites can be returned to unrestricted use
after closure. Research and field experience for hazardous
waste land treatment (HWLT) has demonstrated that a wide
range of elements and compounds are treatable in a soil-based
system. Also, studies show that heavy metals and organics are
unlikely to migrate under HWLT conditions; preliminary stud-
ies of emission levels of volatile compounds show that they
comply with regulatory requirements. However, because HWLT
is regulated as "land disposal" (a term that incorrectly describes
it), its widespread adoption is constrained. If hazardous waste
regulations are changed to regulate HWLT as a separate treat-
ment technology, it could be further developed and integrated
into the US hazardous waste management system.
In-Situ/On-Site Biodegradation of Refined Oils and
Fuels. (A Technology Review).
Riser-Roberts, E„ Naval Civil Engineering Laboratory, Port
Hueneme, C A, June 1992
Hl'lS CfcitismeiH MyntHfj-; ADAJJ2MJ1/J/XAli
This extensive literature review covers all aspects of in situ and
on-site bioremediation of gasoline, middle distillate fuels, and
other refined petroleum products, using information available
through 1988. Topics covered include: composition of differ-
ent fuels, factors affecting biodegradation rates, responsible
microorganisms, degradation pathways, factors enhancing bio-
degradation, and potential limitations. This report is in three
volumes. Volume 1 covers general information of most interest
to managers and non-specialists. Volume 2 contains technical
information required for in-depth coverage. Volume 3 covers
related information, including detailed applied information on
screening and microbial enumberation methods, laboratory
microorganism studies, bioreclamation site preparation,
bioreactor use and selection, cost data, and competing technolo-
gies.
Literature Review and Preliminary Assessment of
Biological Transformations and Biotreatment Technol-
ogy for Petroleum Hydrocarbons and Chlorinated
Solvents.
Korte, N.E., Oak Ridge National Laboratory, TN, Washing-
ton, DC, Sponsored by the U.S. Department of Energy,
Washington, DC, December 1990
NT1-S Crauficcfit Sutnter: OF.1*J -WW !/>AB	;
Chlorinated solvents and petroleum hydrocarbons may un-
dergo a number of natural degradation processes when applied
Lo soil or groundwater. Indeed, the existence of these reactions
has led to extensive research and the development of biodegra-
dation as a remedial action technique. Unfortunately, the
scientific literature demonstrates that there is considerable
controversy concerning many aspects of the field. For example,
different investigators are often unable to agree on relative rates
of biodegradation or even whether certain compounds are
biodegradable. This report examines the recent scientific
literature, describes the biodegradation reactions that are known
to occur, and discusses some of the controversies. The potential
value of biodegradation for remedial action of soils and ground-
water is also presented both from a review of the literature and
from interviews with remedial action contractors.
j lip f
Low-Temperature Effects on Systems for Composting of
Explosives-Contaminated Soils, Part 1. Literature
Review Special Report.
Ayorinde, O.A. and Reynolds, C.M., Cold Regions Research
and Engineering Laboratory, Hanover, NH, December 1989
M1IS IVmhiWBI Niirol.*r
11

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This report reviews literature on the influence of major param-
eters on composting, with emphasis on temperature and explo-
sives. Heat energy is produced by composting as a result of a
microbial conversion of chemical energy to thermal energy.
Hence, heat production and transfer, the influence of engineer-
ing design on compost pile temperatures, and the control and
measurement of compost pile temperature are also examined.
In addition, the report includes a general discussion on
composting, fundamental composting principles, available types
of composting systems, applications of composting technol-
ogy, and the established parameters that influence composting
under various environmental conditions that may be applicable
to cold regions' treatment of hazardous waste.
12A ||
Overview of In-Situ Waste Treatment Technologies.
Hyde, R.A.; Piper, R.B.; Roy; M.W.,Walker, S., EG and G
Idaho, Inc., Idaho Falls, ID, 1992
NTI5 Dt< unKi'* dumber !}E92-QI "i 2fX/\ B
In-situ technologies are becoming an attractive remedial alter-
native for eliminating environmental problems. In-situ treat-
ments typically reduce risks and costs associated with
retrieving, packaging, and storing or disposing-waste and are
generally preferred over ex-situ treatments. Each in-situ tech-
nology has specific applications, and, in order to provide the
most economical and practical solution to a waste problem,
these applications must be understood. This paper presents an
overview of thirty different in-situ remedial technologies for
buried wastes or contaminated soil areas. The objective of this
paper is to familiarize those involved in waste remediation
activities with available and emerging in-situ technologies so
that they may consider these options in the remediation of
hazardous and/or radioactive waste sites. Several types of in-
situ technologies are discussed, including biological treat-
ments, containment technologies, physical/chemical treatments,
solidification/stabilization technologies, and thermal treatments.
Each category of in-situ technology is briefly examined in this
paper. Specific treatments belonging to these categories are
also reviewed. Much of the information on in-situ treatment
technologies in this paper was obtained directly from vendors
and universities and this information has not been verified.
12B
Reductive Dehalogenation: A Subsurface
Bioremediation Process, Journal Article.
Sims, J.L.; Suflita, J.M.; and Russell, H.H., Robert S. Kerr
Environmental Research Laboratory, Ada, OK, Utah Water
Research Lab, Logan, UT, Oklahoma University, Norman
Department of Botany and Microbiology, 1990
NTTS I^tih^tNeitiber: *%9t-M4£73/XAB . ,V
Introduction and large-scale production of synthetic haloge-
nated organic chemicals over the last 50 years has resulted in a
group of contaminants that tend to persist in the environment
and resist both biotic and abiotic degradation. The low solubil-
ity of these types of contaminants, along with their toxicity and
tendency to accumulate in food chains, make them particularly
relevant targets for remediation activities. Among the mecha-
nisms that result in dehalogenation of some classes of organic
contaminants are stimulation of metabolic sequences through
introduction of electron donor and acceptor combinations;
addition of nutrients to meet the needs of dehalogenating
microorganisms; possible use of engineered microorganisms;
and use of enzyme systems capable of catalyzing reductive
dehalogenation. The current state of research and development
in the area of reductive dehalogenation is discussed along with
possible technological application of relevant processes and
mechanisms to remediation of soil and groundwater contami-
nated with chlorinated organics. In addition, an overview of
research needs is suggested, which might be of interest for
development of in-situ systems to reduce the mass of haloge-
nated organic contaminants in soil and groundwater.
12C
Reductive Dehalogenation of Organic Contaminants in
Soils and Groundwater. Groundwater Issue.
Sims, J.L.; Suflita, J.M.; and Russell, H.H., Robert S. Kerr
Environmental Research Lab, Ada, OK, January 1991
NTIS Dtifctaksit Nurabee PB9H91056/3SAB
Introduction and large scale production of synthetic haloge-
nated organic chemicals over the last 50 years has resulted in a
group of contaminants which tend to persist in the environment
and resist both biotic and abiotic degradation. The low solubil-
ity of these types of contaminants, along with their toxicity and
tendency to accumulate in food chains, make them particularly
relevant targets for remediation activities. Although the pro-
cesses involved in dechlorination of many of these organic
compounds are well understood in the fields of chemistry and
microbiology, technological applications of these processes to
environmental remediation are relatively new—particularly at
pilot or field scale. It is well established, however, that there are
several mechanisms which result in dehalogenation of some
classes of organic contaminants, often rendering them less
offensive environmentally. These include: stimulation of
metabolic sequences through introduction of electron donor
and acceptor combinations; addition of nutrients to meet the
needs of dehalogenating microorganisms; possible use of engi-
neered micro-organisms; and use of enzyme systems capable of
catalyzing reductive dehalogenation.
12

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13A I
Report on Decontamination of PCB-Bearing Sediments,
Final Report.
Wilson, D.L., U.S. Environmental Protection Agency,
Hazardous Waste Engineering Research Laboratory,
Cincinnati, OH, October 1987
NTSS Oi«.LitTu::u	FBlvivi 13221Afj
The EPA has initiated a research program to identify chemical/
biological methods as alternatives to incineration and to chemi-
cal land disposal for cleanup of polychlorinated biphenyls
(PCB)-contaminated sediments. The overall objective of the
program is U> identify, validate, and demonstrate effective and
economical chemical/biological processes for removal/destruc-
tion of PCBs in sediments. The report summarizes research
progress on chemical/biological methods development for the
detoxification/destruction of PCBs in sediments.
13B
Status of Land Treatment as a Hazardous Waste Man-
agement Alternative in the United States.
Matthews, J.; McFarland, M.; and Sims, R., et al., U.S.
Environmental Protection Agency, Risk Reduction Engineer-
ing Laboratory, Cincinnati, OH, 1989
N'TO Uacurooiai Hnjuixitacr FB9! • 1 -to 1
Land treatment systems are widely used in the United States for
treating petroleum refinery waste. Many of the petroleum
compounds are degradable in bench scale studies. Proper
operation of the treaLmenL is critical for successful perfor-
mance.
Use of Innoculation in Bioremediation, Journal Article.
Pritchard, P.H., Environmental Research Laboratory, Gulf
Breeze, FL, 1992
EPA IVwmwr.t-NunilMt:
N'lls I)*h uirtrn1: Ntuntor. }3i J nf.l-'XAU
The potenlial for inoculating chemically-polluted sites with
microorganisms to foster the removal or degradation of con-
taminating organic materials (and some inorganic materials)
has been recognized. The purpose of this review is to address
and discuss several important aspects that may help define
problems associated with inoculation in bioremediation and to
thereby provide an indication of the research needed to allow
this process to become a meaningful and productive element of
bioremediaLion technologies.
STUDY/TEST RESULTS
13D
Alternative Biological Treatment Processes for
Remediation of Creosote- and PCP-Contaminated
Materials: Bench-Scale Treatability Studies.
Blattman, B.O.; Lantz, S.E.; and Mueller, J.G., et al.,
Southern BioProducts, Inc., Pendleton, SC, Prepared in
cooperation with Technical Resources, Inc., Gulf Breeze, FL,
Sponsored by the Environmental Research Laboratory. Gulf
Breeze, FL, March 1991.
fc't'A lJocumeiH Nuuihei
vns n«ai.Tf.m Niiniher PHI J -1 T'WKSA Aft
Bench-scale biotreatability studies were performed to deter-
mine the most effective of two bioremediation application
strategies to ameliorate creosote- and pentachlorophenol (PCP)-
conLaminated soils present at the American Creosote Works
Superfund site, Pensacola, Florida: solid-phase bioremediation
or slurry-phase bioremediation. When indigenous microorgan-
isms were employed as biocatalysts, solid-phase bioremediation
was slow and ineffective (8-12 weeks required to biodegrade
>50% of resident organics). Biodegradation was limited to
lower-molecular-weight constituents rather than the more haz-
ardous, higher-molecular-weight (HMW) compounds; PCP
and HMW polycyclic aromatic hydrocarbons (PAHs) contain-
ing four or more fused rings resisted biological attach. More-
over, supplementation with aqueous solution of inorganic nu-
trients had little effect on the overall effectiveness of the
treatment strategy. Alternatively, slurry-phase bioremediation
was much more effective: >50% of LargeLed organics were
biodegraded in 14 days. Again, however, more persistent con-
taminants, such as PCP and HMW PAHs, were not extensively
degraded when subjected to the action of indigenous microor-
ganisms.
Approach to Bioremediation of Contaminated Soil,
Journal Article: Published in Hazardous Waste and
Hazardous Materials, v26n2,1990.
Matthews, J. E.; Sims, J.L.; Sims, R.C.; and Utah State
University, Logan, UT, Robert S. Kerr Environmental
Research Laboratory, Ada, OK
NTTTSr	^JHn'tfc'r: PH'.-M - 1 |(tJ 'J\ Ah
Biological processes, including microbial degradation, have
been identified as critical mechanisms for attenuating organic
contaminants during transit through the vadose zone to the
groundwater. On-site soil remedial measures using biological
processes can reduce or eliminate groundwater contamination,
13

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thus reducing the need for extensive groundwater monitoring
and treatment requirements. On-site remedial systems that
utilize the soil as the treatment system accomplish treatment by
using naturally occurring microorganisms to treat the contami-
nants. Treatment often may be enhanced by a variety of
physical/chemical methods, such as fertilization, tilling, soil
pH adjustment, moisture control, etc. The development of a
bioremediation program for a specific contaminated soil sys-
tem includes: (1) a thorough site/soil/waste characterization;
(2) treatability studies; and (3) design and implementation of
the bioremediation plan.
Assessing Detoxification and Degradation of Wood
Preserving and Petroleum Wastes in Contaminated Soil,
Journal Article: Published in Waste Management and
Research, v8nl, February 1990.
Aprill, W.; Sims, JLL.; and Sims, R.C., et al., Robert S. Kerr
Environmental Research Laboratory, Ada, J.L.; OK, Pre-
pared in cooperation with Utah State University, Logan, UT,
Department of Civil and Environmental Engineering, 1990
*TJh Document Nombei. • W3yy	%\
The study was undertaken to evaluate in-situ soil bioremediation
processes, including degradation and detoxification, for wood
preserving and petroleum refining wastes at high concentra-
tions in an unacclimated soil. The soil solid phase, water soluble
fractions of soil, and column leachates were evaluated. A
mutagenic potential assay (Ames assay) and an aqueous toxic-
ity assay (Microtox(TM) assay) were used to evaluate detoxi-
fication; high performance liquid chromatography was used to
evaluate chemical concentration and degradation for eight
polynuclear aromatic hydrocarbons (PAHs). The group of
noncarcinogenic PAHs studied demonstrated greater degrada-
tion, ranging from 54-90% of mass added for the wastes; the
carcinogenic group of PAHs studied exhibited degradation
ranging from 24-53% of mass added. Although no mutagenic-
ity was observed in waste/soil mixtures after one year,
Microtox(TM) toxicity was observed in water soluble fractions
and in leachate samples. Integration of information concerning
degradation of hazardous constituents with bioassay informa-
tion represents an approach for designing treatability studies
and for evaluating effectiveness of in-situ soil bioremediation.
When combined with information from waste, site, and soil
characterization studies, data generated in treatability studies
may be used in predictive models to: evaluate effectiveness of
on-site soil bioremediation; develop appropriate containment
structures to prevent unacceptable waste transport from the
treatment zone; and design performance monitoring strategies.
Assessment of Problems Associated with Landfilling or
Land Application of Pesticide Waste and Feasibility of
Cleanup by Microbiological Degradation.
Case, L.; Dzantor, E.K.; and Felsot, A., et al., Illinois
Department of Energy and Natural Resources, Champaign,
IL, Hazardous Waste Research and Information Center,
Illinois Natural History Survey, Champaign, IL; Department
of Agronomy, Illinois University at Urbana-Champaign, IL.
Prepared in cooperation with Illinois Natural History Survey,
Champaign, IL, U.S. Department of Agronomy, Illinois
University at Urbana-Champaign, IL, October 1990
N m IJ»ku motif Camber	• ..
To improve the prospects for use of land application as a
remediation tool and the possibility of augmenting biodegrada-
lion of pesticide wastes in soil, a series of laboratory experi-
ments were designed to explore microbiological aspects of the
persistence of high concentrations of herbicides in soil. Most of
the studies focused on the alachlor, which is an acetanilide
herbicide used in corn and soybeans. In laboratory experiments,
alachlor was not degraded after simulated spills of 1000 ppm or
10,000ppm. In contrast, lOppmand lOOppm doses of alachlor
were partially degraded into water-soluble metabolites. Tech-
nical-grade alachlor and an emulsitlable concentrate formula-
tion were metabolized similarly. Microbial populations and
dehydrogenase activity in soil were reduced upon exposure to
1000 ppm or 10,000 ppm alachlor, and lack of degradation of
these high concentrations was attributed to microbial toxicity
from alachlor itself rather than additives in its formulation.
14C I
Biodegradation of Hydrocarbons as a Remediation
Method for Petroleum Contaminants in the Environment
or as a Treatment Method for Petroleum Wastes. (A
Review and Analysis of Recent Field Study Literature)
(Master's Thesis).
Lubbers, J.E., Kansas University, Lawrence, KS, U.S.
Department of Chemical and Petroleum Engineering,
December 1989
N ilS LVvti'ii^tir Number. AliA2J07 i >/l/XAB
The U.S. Navy Petroleum Office (NAVPETOFF) is develop-
ing future Navy petroleum sludge disposal and soil decontami-
nation procedures. This project was conducted for NAVPETOFF
to aid that development by evaluating the use of bacteria to
eliminate petroleum hydrocarbons as a disposal or decontami-
nation option. Electronic database searchers, interviews with
bioremediation researchers, and manual literature searches
were conducted to collect information about microbial
bioremediation from sources which postdate the 1984 amend-
ments to RCR A. From that body of information, reports of field
applications of microbial bioremediation on petroleum wastes
or contaminants were set apart as the primary references for
evaluation development. Summaries of reported microbial
bioremediation methods were developed are presented. These
14

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summaries are introduced by a review of the biologic limits and
processes of the microbes commonly used for bioremediation.
The body of these summaries describes and illustrates their
techniques. Each summary concludes with an evaluation in the
form of a report of the method's effectiveness.
' 	"!i
Biodegradation of JP-5 Aviation Fuel by Subsurface
Microbial Communities, Progress Report January 1,
1987 - March 15,1987.
Swindoll, C. M., Naval Engineering Laboratory, Port
Hueneme, CA, 1988
MIS Docun <£•!;¦ NufhOer ADM?4.?/.VXAR
Leakage of the aviation fuels JP-5, JP-4 and AVGAS from
storage tanks has resulted in a severe environmental insult to a
Naval fuel farm and adjacent area. As part of the reclamation
effort, the indigenous microorganisms are being characterized.
This information will be used to optimize the bioreclamation of
the site. Approximately 60 aerobic microorganisms, including
more than 40 bacteria, 6 actinomycetes, and 10 fungi have been
isolated from soil contaminated with aviation fuels and adjacent
non-contaminated with aviation fuels and adjacent non-con-
taminated soil. All isolated bacteria were able to grow on JP-
5 as their sole carbon source. Most of the bacteria from the
contaminated sites were small, gram-negative rods, while most
bacteria from the non-contaminated site were gram-positive
rods. All of these microorganisms would be expected to
contribute to the bioremediation of the contaminated site.
15B
Biological Treatment of Soils Containing Manufactured
Gas Plant Residues, Topical Report, June 1988 -
February 1990.
Cushey, M. A. and Morgan, D.J., Remediation Technologies,
Inc., Pittsburgh, PA, Sponsored by Gas Research Institute,
Chicago, XL, May 1990
N'HS Oifcuiiunt N'.jmhsrr PR40-XH'9tW>h/KAH
The report summarizes the results of an on-going laboratory
research program directed by GRI to investigate the feasibility
of using biological treatment to remediate soils containing
residues from manufactured gas plant (MGP) sites, and to
develop and verify a rapid assessment protocol (the GRI Accel-
erated Treatability Protocol) for determining the potential for
biologically treating specific MGP site soils. This protocol,
which features soil characterization, desorption testing, and
slurry reactor testing, is cost competitive with traditional
treatability studies and can be completed in approximately half
the time. For the first two soils evaluated, total polynuclear
aromatic hydrocarbon (PAH) concentrations (a critical class of
chemicals at MGP sites) were reduced by 95 and 80% to
residual 'plateau' concentrations of less than 10 and 5,000 mg/
Kg, respectively. It is believed that the magnitude of the
'plateau' concentration is not primarily dictated by the avail-
ability of nutrients, oxygen, and adequate microbial popula-
tions but rather by limitations of mass transfer from the soil-
waste matrix to the bulk aqueous phase. Further tests with 13
other MGP soils are in-progress and the results of these will be
reported at a later time.
15C
Bioremediated Soil Venting of Light Hydrocarbons,
Journal Article: Published in Hazardous Waste and
Hazardous Materials, v7n4,1990.
Kampbell, D.H. and Ostendorf, D.W., University of Massa-
chusetts, Amherst, MA, Sponsored by Robert S. Kerr
Environmental Research Laboratory, Ada, OK, 1990
JhTTS OmiHRisnr Ntinibtr	H15?K/X.AK
The effectiveness and feasibility of bioremediated soil venting
of light hydrocarbons in the unsaturated zone was investigated.
Degradation mechanics were considered as a one dimensional
balance of storage, linear sorption, vertical advection, and
Michaelis-Menton kinetics. The resulting analytical solution
was tested successfully against field performance data of an
unsaturated clay soil bioreactor for a pollpellant waste gas
mixture of propane, n-butane, and isobutane. A series of
venting simulations was run to assess the biodegradation of
vapors above an aviation gasoline spill in sandy soil at Traverse
City, Michigan, based on field and microcosm estimates of the
kinetic parameters. Acclimated, nutrient rich soil effectively
and feasibly reduced effluent vapor concentration from the
strong influent concentration associated with dispersed re-
sidual gasoline in the contaminated capillary fringe. Aggre-
gated residual contamination required a stronger airflow for a
longer duration while natural kinetics were too slow for feasible
and effective treatment by bioremediated soil venting at Tra-
verse City.
15D f
Bioremediation Case Studies: Abstracts.
Devine, K., U.S. Environmental Protection Agency, Office
of Environmental Engineering and Technology Demonstra-
tion, Washington, DC, DEVO Enterprises, Inc., Washington,
DC, March 1992
£PA D'Xuiiient Nuujtvi (iOO/R
NI LS DHK.ijinctn Numfisr PBH2-2?2>WV-\b
The report contains abstracts of 132 case studies of
bioremediation technology applied to hazardous waste clean-
up. It was prepared to compile bioremediation studies in a
variety of locations and treating diverse contaminants, most of
15

-------
which were previously undocumented. All data are based on
vendor-supplied inf ormation and there was no opportunity to
independently confirm its accuracy. These 132 case studies,
from 10 different biotechnology companies, provide users with
reference information about on-going and/or completed field
applications and studies. About two-thirds of the cases were at
full-scale clean-up level with the remainder at pilot or labora-
tory scale. In 74% of the cases, soil was at least one of the media
treated. Soil alone accounts for 46% of the cases. Petroleum-
related wastes account for the largest contaminant with 82
cases. Thirty-one States are represented in the case studies.
Bioremediation Case Studies: An Analysis of Vendor
Supplied Data.
Devine, K., Environmental Protection Agency, Office of
U.S. Environmental Engineering and Technology Demon-
stration, Washington, D.C., DEVO Enterprises, Inc.,
Washington, DC, March 1992
tl'A Dcvnmi'su Numbei MWK viVM
NHS u mem Sumter P> 2 - J .O9/X A R
The report provides users with reference information about 132
on-going and/or completed field applications and studies from
10 different biotechnology companies. About two-thirds of the
cases were at full-scale clean-up level with the remainder at
pilot or laboratory scale. The report does not contain informa-
tion from all companies involved in bioremediation, only those
companies who responded to a request for voluntary submis-
sion.
16B
Bioremediation of Contaminated Surface Soils.
Matthews, J.E.; Sims, J.L.; Sims, R.C.; and Robert S. Ken-
Environmental Research Laboratory, Ada, OK, Dynamac
Corp., Ada, OK, Utah State University, Ixigan, IJT. Pre-
pared in cooperation with Dynamac Corporation, Ada, OK,
and Utah State University, Logan, UT, August 1989
N HS h»»cuni«*Mi Nuinbrr
Biological processes, including microbial degradation, have
been identified as critical mechanisms for attenuating organic
contaminants during transit through the vadose zone to the
groundwater. On-site soil remedial measures using biological
processes can reduce or eliminate groundwater contamination,
thus reducing the need for extensive groundwater monitoring
and treatment requirements. On-site remedial systems that
utilize the soil as the treatment system accomplish treatment by
using naturally occurring microorganisms to treat the contami-
nants. Treatment often may be enhanced by a variety of
physical/chemical methods, such as fertilization, tilling, soil
pH adjustment, moisture control, etc. The development of a
bioremediation program for a specific contaminated soil sys-
tem includes:. (1) a thorough site/soil/waste characterization;
(2) treatability studies; and (3) design and implementation of
the bioremediation plan. Biological remediation of soils con-
taminated with organic chemicals has been demonstrated to be
an alternative treatment technology that can often meet the goal
of achieving a permanent clean up remedy at hazardous waste
sites.
16C
Bioremediation of Explosives.
Alvarez, M.A.; Hanners, J.L.; and Unkefer, P.J.; et al, Los
Alamos National Laboratory, Los Alamos, NM, Sponsored
by the U.S. Department of Energy, Washington, DC, 1990
ivm nw = ..iv.u N,.rt.tior DEi'-'ij-U ! 11 Afi
The extensive manufacture, packing, and the use of explosives
has often resulted in significant contamination of soils and
groundwaters near these activities. Congressional mandate has
now required that such sites be remediated. An especially
promising technology for this explosives problem is biotech-
nology. When applicable, biotechnology is cheap and provides
complete conversion of hazardous compounds to harmless
biomass or carbon dioxide. The focus of this paper will be on
our present understanding of the microbial metabolism of the
explosives, TNT and RDX, which have been used most exten-
sively in the United States. To assure that an efficient process
is developed for TNT biodegradation, we are conducting appro-
priate lab scale tests with TNT contaminated soil. First, we are
testing their efficiency in soil/water slurries; we are also testing
their efficiency in a column system designed to simulate
composting conditions. A pilot scale test of this bacteria)
degradation will be conducted as soon as weather permits.
16D
Bioremediation of Hazardous Wastes.
Biosystems Technology Development Program, Office of
Research and Development, U.S. Environmental Protection
Agency, Ada, OK; Athens, GA; Cincinnati, OH; Gulf
Breeze, FL; and Research Triangle Park, NC, August 1992
i.l'.A i'iiruimeMi .Nuim-r hiKtfk 'O/I.'N* ,	,,
In 1987, the U.S. Environmental Protection Agency's Office of
Research and Development (ORD) initiated the Biosystems
Technology Development Program to anticipate and address
research needs in managing our nation's hazardous waste. This
document contains papers and posters presented at the fifth
annual Symposium on Bioremediation of Hazardous Wastes:
U.S. EPA's Biosystems Technology Development Program
held in Chicago in 1992. The five research and program areas
16

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SOURCES OF BIOREMEDIATION INFORMATION/
TECHNICAL ASSISTANCE
Numerous computer-based bulletin boards, regulatory hotlines, dockets, databases, catalogs, and periodicals are also avail-
able. These resources provide technical information on bioremediation and other innovative technologies and guide you to
additional valuable resources.
BULLETIN BOARDS:
• Alternative Treatment Technologies
Information Center (ATTIC) (data line) 301-670-3808
To obtain information on ATTIC 908-906-6828
A collection of hazardous waste databases accessed through a bulletin board
that provides hazardous waste abstracts, news bulletins, conference informa-
tion, and a message board.
• Cleanup Information Bulletin
(CLIMN) (data line) 301-589-8366
A bulletin board for hazardous waste professionals that provides current
information on innovative technologies. Provides information bulletins, mes-
sage and on-file exchange, and on-line databases and directories.
Help Line 301-589-8368
Addresses questions about CLU-IN access and contents; addresses problems
with the service.
' Office of Research and
Development (ORD) (data line) 800-258-9605
Bulletin Board Service (BBS) (data line) 513-569-7610
Provides a bibliography of 18,000 documents, news excerpts from
"Bioremediation in the Field," and a message board.
Help Line 513-569-7272
Provides information on access to and contents of the ORD BBS.
CATALOGS:
• Catalog of Hazardous and Solid Waste Publications,
Sixth Edition EPA/530-B-92-001
Catalogs Office ot Solid Waste policy directives, guidance documents, bro-
chures, Regulatory Development Branch memos, and other documents rel-
evant to hazardous and solid waste.
• Compendium of Superfund Program Publications
EPA/540/8-91/014, NTIS PR 881
Provides abstracts and ordering information for fact sheets, directives, publica-
tions, and computer materials on Superfund. Use the document ordering
directions to compendium.
DATABASES:
• DIALOG Database 800-3-DIALOG
A large database that contains files relevant to hazardous waste including:
Biotechnology Abstracts; Enviroline; Corrective Action Search; Pollution Ab-
stracts; National Technical Information Services (NTIS); and others.
- NTIS Database
Contains abstracts of government-sponsored research, development, and
engineering analyses prepared by approximately 250 Federal agencies and
some State and local governments. Accessible via the DIALOG system.
• Records of Decision System (RODS)
(To get information on accessing RODs) 703-603-8881
Contains the full text of all signed Records of Decision for hazardous waste
clean-up sites nationwide. Direct access to RODS is available to EPA
personnel and organizations that have relevant EPA contracts.
' Vendor Information System for Innovative
Treatment Technologies (VISITT) 800-245-4505
Contains current information on availability, performance, and cost of innova-
tive technologies to remedy hazardous waste sites.
DOCKETS:
• Federal Facilities Docket Hotline.
Provides the name, address, NPL status, agency, and Region for the 1,930
Federal facilities listed on the Federal Facilities Docket. Facilities are on the
docket because they reported being a RCRA TSDF or having spilled or ha ving
the potential to release CERCLA hazardous waste. Operates Monday - Friday,
8 a.m.-6p.m., Eastern Time.
• RCRA Information Center 202-260-9327
indexes and provides public access to all regulatory materials supporting the
Agency's actions under RCRA, and disseminates current Office of Solid Waste
publications. Operates Monday - Friday, 8:30 a.m. - 4 p.m., Eastern Time.
• Superfund Docket 202-260-3046
Provides access to Superfund regulatory documents, Superfund Federal
Register Notices, and Records ot Decision. Operates Monday - Friday,
9 a.m. -4p.m., Eastern Time.
• UST Docket 202-260-9720
Provides documents and regulatory information pertinent to RCRA's Subtitle I
(the Underground Storage Tank program). Operates Monday - Friday,
9 a.m. ¦ 4 p.m., Eastern Time.
HOTUNES/REGULATORY/TECHNICAL ASSISTANCE:
• EPA Headquarters Library 202-260-5921
Offers reports from various EPA offices and trade and environmental journals.
Features the "Hazardous Waste Collection" department. Operates Monday -
Friday, 10 a.m. -4 p.m., Eastern Time.
• RCRA/Superfund/UST Hotline 800-424-9346
Provides regulatory assistance related to RCRA, CERCLA, and USTprograms.
Serves as a liaison between the regulated community and EPA personnel and
provides information on the availability of relevant documents. Operates
Monday - Friday, 8:30 a.m. - 7:30 p.m., Eastern Time.
• TSCA Hotline 202-554-1404
Answers public and private regulatory questions on TSCA. Refers callers to
appropriate EPA contacts, and takes TSCA-relevant document orders. Oper-
ates Monday - Friday, 8:30 a.m. - 5 p.m., Eastern Time.
• Superfund Health Risk Technical
Support Center 513-569-7300
Provides EPA Regional Superfund risk assessors, State agencies, and those
working under EPA contract with technical, typically chemical-specific, support
and risk assessment review. Operates Monday - Friday 8a.m.-5p.m., Eastern
Time.
INFORMATION CENTER:
• National Center for Environmental
Publications and Information (NCEPI) 513-891-6561
(Fax requests) 513-891-6685
Stores and distributes to public and private callers a limited supply of most
EPA publications, videos, posters, and other multi-media materials. Callers
should know document titles or numbers when calling. The following
periodicals can be obtained from NCEPI, as supplies last:
- Bioremediation in the Field
A periodical devoted to bioremediation thatcontains 140 potential applications
of bioremediation, including site type, name, and contact.
- Groundwater Currents
A newsletter that reports on innovative in-situ and ex-situ groundwater
remediation technologies to be applied in the field.
- Tech Trends
An applied technology journal that provides information on Superfund remov-
als, remedial actions, and RCRA corrective actions.
.800-548-1016