United States
          Environmental Protection
          Agency
            Office of Research and
            Development
            Washington DC 20460
EPA/625/R-01/001A
February 2001
&EPA
US EFA Office o! Research jr-d Di«tojwipnt
Summary of Workshop on
Biodegradation of MTBE
          February 1-3,2000

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                                         EPA/625/R-01/001A
                                            February 2001
Summary of Workshop on
 Biodegradation of MTBE

           February 1-3,2000
       Workshop Sponsored by the
U.S. Environmental Protection Agency and
       American Petroleum Institute
                 Prepared by:

            Eastern Research Group, Inc.
            Lexington, MA 02421-3136


             Contract No. 68-D7-0001
             Work Assignment 3-11
       Technology Transfer and Support Division
      National Risk Management Research Laboratory
         Office of Research and Development
         U.S. Environmental Protection Agency
              Cincinnati, OH 45268

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                                        Notice

       The views expressed in these Proceedings are those of the individual authors and do not
necessarily reflect the views and policies of the U.S. Environmental Protection Agency (EPA).
Scientists in EPA's Office of Research and Development have prepared the EPA sections and
those sections have been reviewed in accordance with EPA's peer and administrative review
policies and approved for presentation and publication. Mention  of trade names or commercial
products does not constitute endorsement or recommendation for use.
                                          -11-

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                                        Foreword

       The U.S. Environmental Protection Agency is charged by Congress with protecting the
Nation's  land, air, and water resources. Under a mandate of national environmental laws, the
Agency strives to formulate  and implement  actions leading to a compatible balance between
human activities and the ability of natural systems to support and nurture  life. To meet this
mandate,  EPA's research program  is  providing data  and  technical support  for  solving
environmental problems today and building a science knowledge base necessary to manage our
ecological resources wisely, understand how pollutants affect our health, and prevent or reduce
environmental risks in the future.

       The National  Risk  Management  Research Laboratory  is  the  Agency's  center  for
investigation of technological and management approaches for reducing risks from threats to
human health and the environment.  The focus of the Laboratory's research  program  is on
methods for the prevention and control of pollution to air, land, water and subsurface resources;
protection of water quality in public water systems; remediation  of contaminated sites and ground
water; and prevention and control of  indoor air pollution. The goal  of this research effort is to
catalyze   development   and   implementation  of  innovative,   cost-effective   environmental
technologies; develop scientific and engineering information needed by EPA to support regulatory
and policy decisions; and provide technical support and information transfer  to ensure effective
implementation of environmental regulations and strategies.

       This  publication has  been  produced as part of the  Laboratory's  strategic long-term
research plan. It is published and made available by EPA's Office of Research and Development
to assist the user community and to link researchers with their clients.
                                     E. Timothy Oppelt, Director
                                     National Risk Management Research Laboratory
                                           -m-

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                                  Acknowledgments

       The workshop entitled Biodegradation ofMTBE was organized and sponsored by the
U.S. Environmental Protection Agency (EPA)  and the  American Petroleum Institute (API).
Appreciation is given to all those who contributed to the workshop through presentations and
participation in the discussions.

       This  workshop  report  was prepared by EPA's National Risk  Management Research
Laboratory (NRMRL) with support  from Eastern Research Group, Inc.  (ERG) and Battelle
Memorial Institute. Joan Colson, NRMRL, served as project officer. EPA wishes to acknowledge
the work performed by Lauren Lariviere of ERG and Andrea Leeson of Battelle Memorial
Institute for their contributions to the workshop and to the development of this report.

       A  special acknowledgment  is made to  the  following people who provided technical
consultation for the planning and organization of the workshop and for the editorial review of the
report:

Fran Kremer, US EPA
Bruce Bauman, API
Kirk O'Reilly, Chevron
Benjamin Blaney, US EPA
                                         -IV-

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                                 Table of Contents

Notice	  ii
Foreword 	 iii
Acknowledgments 	 iv

1.0             	  1

2.0 SCOPE OF THE PROBLEM	  2

3.0 CURRENT             	  4
       3.1 Overview of MTBE Biodegradation 	  4
             3.1.1  Aerobic Degradation of MTBE 	  4
             3.1.2  Anaerobic Degradation of MTBE 	  6
             3.1.3 MicrobialCometabolismofMTBE	  7
       3.2 Enhanced In Situ Bioremediation 	  8
             3.2.1 Bioaugmentation	  8
             3.2.2 Stimulation of Indigenous Microorganisms  	  9
       3.3 Natural Attenuation 	 10
       3.4 Ex Situ Bioremediation	 12

4.0                    	 16

5.0 COLLABORATIVE EFFORTS 	 22

6.0 LITERATURE CITED  	 23

Attachment A -
Attachment B -        List
Attachment C - Poster         List
Attachment D -        List


                                      Tables

Table 1.  State Drinking Water Regulations (Speth, 2000) 	  3
                                     Figures
Figure 1.  Proposed Degradation Pathway of MTBE by              PM1 (Church and
         Tratnyek, 2000)  	 5
                                       -v-

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                                      1.0 OVERVIEW






       A workshop on biodegradation of methyl  tert-butyl  ether (MTBE)-contaminated  soils  and



groundwater was held in Cincinnati, OH, on February  1-3, 2000, and was sponsored by the U.S.



Environmental Protection  Agency's  (EPA)  National   Risk Management  Research  Laboratory



(NRMRL) and the American Petroleum Institute  (API). Researchers in  academia,  industry,  and



government agencies were invited to attend and present current research. The goals of the workshop



were:



        !      To gain an understanding of the types of MTBE research that various organizations are



              conducting and of the conclusions that this research is generating.



              To identify the remaining research needs on MTBE biodegradability.



              To understand what research is being planned for the future and to identify potential



              opportunities for collaboration.



       The  following  sections present information  discussed during the  Biodegradation of  MTBE



Workshop and present  a  summary of the  authors' written and oral presentations. These sections



include:



        !      Scope of the problem (Section 2.0)



        !      Current research (Section 3.0)



        !      Research needs (Section 4.0)



        !      Collaborative efforts (Section 5.0).



       Numerous presentations are summarized in this report. The presentations referenced throughout



the report are those made at the February 2000 workshop.

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                               2.0 SCOPE OF THE PROBLEM




       MTBE has become the subject of significant attention in recent years due to public focus on several




sites where MTBE plumes are very large and are impacting drinking water sources. The attention has been




particularly acute in California where gasoline usage is the highest in the U.S. and the population density and




water usage results in increased potential  for contaminant migration into drinking water wells. MTBE




production and usage in the U.S. has risen steadily since 1982, resulting in potential contamination in many




more areas (Bauman, 2000).




       The National Water-Quality Assessment Program (NAWQ A) of the U. S. Geological Survey (USGS)




has assessed the extent of MTBE contamination in the U.S. and the role of non-point MTBE sources on




distribution. These studies have shown that MTBE is widely distributed in the hydrosphere, and that significant




regional patterns are present.  In a national study of 2,948 wells during the period 1985-1995, approximately




20% of mixed well types  located in areas using MTBE as the principal fuel oxygenate contained detectable




concentrations of MTBE (>0.2 |Jg/L). However, in areas where MTBE use was not widespread, less than




5% of wells contained measurable MTBE  concentrations. This general pattern of MTBE distribution in




groundwater was confirmed by more localized studies (Chapelle, 2000).




       At present, there is inadequate health effects data for the USEPA to set an oral reference dose for




MTBE. However, because MTBE has a very unpleasant taste and odor, the EPA has issued an Advisory




on MTBE in drinking water of 20-40 • g/L.  Table 1 shows the Standards, Guidelines, and Action Levels as




currently set by individual states. Four states have health-based Primary Drinking Water Standards. At the




time of this workshop, three states have enforceable guidelines, while twelve more have guidelines, or action




levels, in place. The levels range from 5 • g/L  (CA) to 240  • g/L (MI). The specifics of enforcement are




determined by each State (Speth, 2000).

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Table 1. MTBE State Drinking Water Regulations (Speth, 2000)
Primary Drinking Water Standards
Maine
New Jersey
New York
South Carolina
                                    Concentration
                                        •g/L
 35
 70
 50
20-40
  Health Based
  Health Based
  Health Based
  Health Based
Enforceable Guidelines
California
Michigan
West Virginia

Guideline or Action Level
Arizona
California
Connecticut
Illinois
Kansas
Maryland
Massachusetts
New Hampshire
Pennsylvania
Rhode Island
Vermont
Wisconsin
  5
 240
20-40
 35
 13
 70
 70
20-40
 10
 70
 15
20-40
20-40
 40
 60
Aesthetically Based
   Health Based
  EPA Advisory
   Health Based
   Health Based
   Health Based
   Health Based
   EPA Advisory
Aesthetically Based
   Health Based
Aesthetically Based
   EPA Advisory
   EPA Advisory
   EPA Advisory
   Health Based

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                              3.0  CURRENT RESEARCH

       This section describes the current research on biodegradation  of MTBE as described
during the workshop presentations.  It has been divided into four general sections including an
overview of MTBE biodegradation, enhanced in situ bioremediation, natural attenuation, and ex
situ bioremediation.

3.1    Overview of MTBE Biodegradation
       MTBE has been  shown to biodegrade under  various  conditions including  aerobic,
anaerobic,  and  cometabolic  conditions,   however  it is not well  understood under  which
geochemical conditions degradation occurs.. A summary  of the  research in  these  areas is
provided in the following sections.

3.1.1  Aerobic Degradation of MTBE
       Several researchers described successful mineralization of MTBE in laboratory-scale
research  (Cowan,  2000;  Morales and Deshusses,  2000; Salanitro,  2000; Scow et al., 2000;
Venosa et al., 2000: Suidan et al, 2000). Microorganisms were isolated from a variety of sources,
generally from petroleum or chemical plant wastewater bioreactors.
       Scow et al.  (2000) and Salanitro (2000) have identified  pure cultures capable of utilizing
MTBE as a sole carbon and energy source. Salanitro (2000) and other researchers  (Cowan,
2000; Morales  and Deshusses, 2000; Venosa et al., 2000) have also developed microbial consortia
capable of mineralizing MTBE under aerobic  conditions. Microbial cell yields tend to be  lower on
MTBE than those observed for aromatic hydrocarbons (0.1-0.2 g cells/g MTBE).  In  addition,
biodegradation rates tend to be slower than those observed for the aromatic hydrocarbons.
       The microorganism, bacterial  strain  PM1,  isolated by Scow et al. (2000)  was further
studied by  Church and Tratnyek (2000) to determine  the degradation pathway.  This study
confirmed the mineralization of MTBE by strain PM1 and ascertained that the  degradation rates
of tert-amyl methyl  ether (TAME), ethyl tert-butyl ether (ETBE), di-isopropyl ether (DIPE), tert-
                                           -4-

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butyl alcohol (TEA), and tert-amy\ alcohol (TAA) were of the same order of magnitude as the
degradation rate of MTBE.  Together with a consistency in product formation,  these results
suggested that similar enzyme systems are responsible for all of the reactions. The proposed
aerobic degradation pathway for MTBE is shown in Figure 1.
       The  degradation pathway shown in Figure  1  contains some  hypothesized steps in the
pathway.  Clearly,  aerobic biodegradation  of MTBE is demonstrable. Additional research is
necessary to clarify the microorganisms involved in the process, factors that impact cell yield and
biodegradation rates, and the degradation pathway.
MTBE
       CH3
   H3C-C-O-CH3
       CH3
        1
        t
ETBE
   H3C-C-O-C-H
       CH3
 OPE  CH3   H
   H3C-C-O-C-CH3
        H    CH3
      CH3
  H3C-C-O-CH2CH3
      CH3
      I
TAME
H3CH2C-C-O-CH3
 CH3
-c-o
 CH3
3A
CH3
H3C-C-OH




IP

CH3
i
Y
y

CH3
H3C-C-OH =

H
™ CH3
H3CH2C-C-OH ^ 	
CH3
i
Y
y
AT
O
=> H3C-C-CH3 --->

TAF CH3 O
1 II
H3CH2C-C-O-C-H
CH3





Acetaldehyde,
Acetate, etc.

Figure 1. Proposed Degradation Pathway of MTBE by Bacterial strain PM1 (Church and Tratnyek, 2000)
                                          -5-

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3.1.2  Anerobic Degradation of MTBE
       The majority  of researchers have investigated MTBE biodegradation under aerobic
conditions, and in fact, Morales and Deshusses (2000) and other researchers were unable to
demonstrate any MTBE biodegradation under anaerobic conditions. However,  Finneran and
Lovley (2000) and Kropp  et  al.  (2000) have demonstrated biodegradation  of  MTBE under
anaerobic conditions.
       In the study by Finneran  and Lovley (2000),  several  sediments were investigated for
MTBE and TEA  biodegradation  potential.  Results varied among sediments, with the most
success occurring when Fe (III) oxide and humic substances were added to the serum bottles.
Radiolabeled [14C] MTBE was  added during  investigation of two of the sediments  and
conversion  to carbon  dioxide and methane  was observed,  although  at low levels. TEA  was
observed to biodegrade much more rapidly than MTBE under iron-reducing and  methanogenic
conditions. Anaerobic TEA degradation is relatively rapid and extensive. Rates are  comparable to
those seen for aerobic TEA degradation. Sediment adapted to degrade TEA converts 50% of the
added uniformly labeled [C-14]  TEA to both [C-14] CO2 and [C-14] CH4 in 45 days.
       Kropp et  al.  (2000)  conducted a  similar study  in which sediment  slurries  were
investigated for anaerobic biodegradation of MTBE and other alternative  gasoline oxygenates
such as methanol,  ethanol,  and isopropanol  as well as several of the ethers  such  as TAME,
ETBE, and DIPE. Kropp et al. (2000) found  that the simple alcohols  were  susceptible to
anaerobic  biodegradation, but the effect of increased branching,  as seen  with TEA,  was
increased recalcitrance to anaerobic  decay.  This same observation  (that  increased branching
tends to cause recalcitrance to anaerobic decay) was also seen with MTBE and its isomer butyl
methyl ether. In general, while Kropp  et al.  (2000)  found definite evidence for anaerobic
degradation  of MTBE  and other  ether oxygenates  under methanogenic  conditions,  the
phenomenon was not widespread. Kropp et al. concluded that MTBE should be considered as a
compound for which anaerobic biodegradation is extremely difficult.
       Information on  the pathway of anaerobic MTBE has not yet been  investigated.
Investigation of the anaerobic  biodegradation of MTBE is  still in the early  stages and more
research is necessary to fully understand this process.

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3.1.3  Microbial Cometabolism of MTBE
       Hyman (2000) provided a  review of microbial cometabolism of MTBE. A summary of
this review is provided in this section.
       Several aerobic microorganisms, including bacteria and fungi, have been identified that
are capable  of cometabolically-degrading MTBE. There are also several primary substrates that
have been identified that  can be used to  stimulate MTBE biodegradation, including alkanes,
aromatics, and cyclic compounds. In general, MTBE cometabolism appears to be associated most
strongly with microorganisms that grow aerobically on the short chain alkanes (
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with iso-pentane as a primary substrate in laboratory-scale bioreactors. Field applications of iso-
pentane-degrading bacteria  are  currently  being  implemented  by Stringfellow  (2000).  Field
evidence obtained by Butler et al. (2000) strongly suggests that cometabolism of MTBE was the
primary mechanism for MTBE removal from the aquifer.

3.2     Enhanced In Situ Bioremediation
        The information in this section discusses the enhanced in situ bioremediation techniques.
This  section  is  divided into  bioaugmentation  studies  and  studies  in  which indigenous
microorganisms were stimulated.

3.2.1   Bioaugmentation
        Two studies  were  presented in which MTBE-degrading  microbial cultures  were
introduced into the subsurface (Salanitro, 2000; Scow et al., 2000). Scow et al. (2000) worked
with the bacterial strain PM1. The objectives of this study were to determine, both in laboratory
and field  experiments,  if strain PM1  was  effective  at removal  of MTBE from a  contaminated
groundwater aquifer at the Port Hueneme Naval Facility in Oxnard, CA. Microcosm studies were
first conducted to determine whether MTBE biodegradation by strain PM1 would occur in site
sediments. MTBE biodegradation was significantly higher in those microcosms inoculated with
strain PM1 than in those microcosms  without inoculation. Initial concentrations of MTBE were
removed  within 5 days, and subsequent concentration spikes were removed more rapidly.
Nutrient addition appeared to have no impact on biodegradation rates.
        The field study was initiated in November 1999 and currently, the  system  has not
operated for a sufficient period of time to determine the effectiveness of the process. The field
study consists of two test plots located 610 m downgradient from the source of MTBE. Both
plots are aerated using an oxygen generator from which oxygen is injected into  seven 20-gallon
tanks associated with each plot.  Plot A receives only oxygen and Plot B  receives oxygen and
was inoculated with strain PM1 (density of approximately 109 cells per ml in the final injection
solution).
        Salanitro (2000) has demonstrated the use of biobarriers, also at the Port Hueneme Naval
Facility in Oxnard, CA and at a  site in Tahoe  City, NV. Salanitro (2000) worked  with a mixed
culture, MC-100, and examined its MTBE biodegradation potential first in laboratory studies using

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site  groundwater.  MTBE  biodegradation  was  much more rapid  when  microcosms were
inoculated with MC-100 than in uninoculated microcosms.  MTBE concentrations  of 10 to  12
mg/L were degraded to below detection limits within two weeks. MTBE (70-80 mg/L) and BTEX
(45 mg/L) in groundwater with high concentrations of gasoline (700 mg/L) were also completely
degraded in microcosms inoculated with MC-100.
       The  field studies at Port Hueneme consisted of creating three test plots: one with oxygen
injection only; one with oxygen injection augmented with MC-100; and one control (no treatment).
The  experiment was conducted for one year. In the control test plot, no significant  decline in
MTBE  concentrations was  observed. In the oxygen-injection-only  plot, MTBE degradation
appeared to  occur after a lag time of approximately 260 days. However, TEA was not degraded
in this test plot. In the inoculated test plot, MTBE biodegradation occurred soon after inoculation
and was non-detectable after 260 days. TEA was not  detected in  this test plot. Similar results
were obtained at a different field site in Tahoe City.
       The  results from both  of these studies  indicate  that bioaugmentation has  merit and
warrants further research. Additional research is  needed to verify  results and to determine the
effectiveness of bioaugmentation  under different operating conditions  and  under different
hydrogeologies.

3.2.2  Stimulation of Indigenous Microorganisms
       Stimulation of indigenous microorganisms was investigated by Mackay et al. (2000).
Laboratory and field experiments were conducted at an MTBE plume at Vandenberg Air Force
Base, CA. Microcosm studies with site sediments suggested that native aerobic MTBE-degrading
bacteria were present  in the site sediments and could be stimulated to degrade MTBE solely  by
adding oxygen (Wilson et al., 1999). In two separate field tests,  dissolved oxygen was released
into the MTBE plume by diffusion through the walls of tubing pressurized with oxygen and in
contact with the  groundwater flowing through  unpumped well  screens  or permeable  walls.
Upgradient concentrations  of MTBE ranged from 100-400 ug/L. In both field tests,  significant
reductions in MTBE concentrations (<5-100 ug/L) were measured downgradient of the diffusive
oxygen release systems in repeated  sampling events, suggesting that oxygen release led to
stimulation of in situ biodegradation of MTBE. Appearance of TEA also indicated the  activity of
MTBE-degrading microorganisms.

                                          -9-

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       Results from this research indicate that oxygen addition alone may be sufficient at some
sites to effect the biodegradation of MTBE. Further research is needed to optimize the process
and to determine factors that may affect the process.

3.3    Natural Attenuation   Several field analyses of natural  attenuation of MTBE were
presented. Some of the studies showed that natural attenuation of MTBE was possible, but the
degree of attenuation varied greatly from site to site. Evidence of biodegradation in groundwater
was  demonstrated by Borden (2000),  Butler et al. (2000), Landmeyer (2000), and Wilson (2000);
and by Baehr et al. (2000) in  the vadose zone. In contrast, Hunter (2000) and Weaver (2000)
found no evidence of biodegradation; however, both  of these studies were conducted in areas
with high groundwater velocity and, at Weaver's sites, high recharge rates. Happel et al. (2000)
presented preliminary results that also indicated fairly slow attenuation of MTBE as compared to
BTEX (two orders of magnitude lower).
       Related to  natural attenuation is the  development of a new method for monitoring
petrochemical biodegradation  as described by Mills  and Haines  (2000). In this method,  the
isotopic composition of biodegradation products  was analyzed. This method may  allow  the
differentiation between degradation of gasoline components, MTBE, and natural organic matter,
thereby offering the potential for more conclusive evidence of MTBE biodegradation.
       Borden (2000) described an  extensive three-dimensional field characterization that was
conducted to define the horizontal and  vertical distribution of BTEX,  MTBE,  and indicator
parameters in a shallow coastal plain  aquifer. Field-scale degradation rates were highest near the
source and  declined  further  downgradient.  Laboratory  microcosm  studies conducted  under
aerobic and  denitrifying conditions  showed an  identical  pattern of  biodegradation  with high
biodegradation  rates near  the source and lower rates  further  downgradient.  Mathematical
modeling studies using BIOPLUME II  and a three-dimensional analytical solution showed that: 1)
the field data could not be adequately fitted using a spatially uniform first-order decay rate; and 2)
use of a spatial uniform first-order  decay  rate would substantially underestimate contaminant
concentrations  and risks  to  downgradient receptors. Therefore,  while biodegradation was
occurring in the aquifer, current models  were inadequate to predict  MTBE natural attenuation
accurately.
                                          -10-

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       Schirmer et al. (2000) conducted a natural gradient experiment in the Borden Aquifer,
CFB Borden, Ontario. MTBE was injected in 1988 in a 2,800 L slug at a concentration of 270
mg/L plus 19 mg/L BTEX and 515  mg/L  Cl. It was found that approximately 3% of the initial
MTBE mass remained after eight years. MTBE was found where expected based on modeling,
but it was found sporadically and at concentrations much lower than predicted. The nature of the
aquifer, the characteristics of the contaminant, and the fact that the slug was introduced 1.5  m
below the  water table,  indicated that the processes of sorption,  abiotic degradation,  and
volatilization were not significant contributors to the observed MTBE attenuation (Butler et al.,
2000). The most likely explanation appears to be biodegradation. Additional laboratory studies by
Butler et al. (2000) demonstrated biologically-catalyzed MTBE degradation in the Borden aquifer;
however, this result appeared to be incidental and difficult to predict.  Cometabolism was easily
initiated in laboratory microcosms and this may be  the more likely  mechanism for MTBE
biodegradation in the Borden aquifer.
       Landmeyer (2000) conducted  a  study  of the fate of  MTBE  in anaerobic  aquifer
sediments. Very little biodegradation was  observed under anaerobic conditions  over a 7-month
period. However, recent evidence indicates that complete  degradation of MTBE to carbon
dioxide is possible under mixed anaerobic/aerobic conditions, such as  those  present where
anaerobic groundwater  discharges to aerobic surface waters.  Other field evidence  indicates
significant uptake of MTBE by oak trees.
       Wilson (2000) is in the process of conducting  a survey of existing underground storage
tank (UST) sites in association with BP/Amoco. Groundwater samples  were analyzed for MTBE,
TEA, BTEX, naphthalene, methane, iron (II), total organic carbon (TOC), oxygen,  sulfate, and
sulfide. Results are still being analyzed, but  methane concentration doesn't appear to explain  TEA
or MTBE concentration. There is generally more TEA than MTBE, possibly as a biodegradation
product from MTBE or possibly due to the higher solubility of TEA.
       Baehr et al. (2000) conducted a study investigating the concentration of MTBE measured
in the unsaturated zone. Concentrations indicated that degradation of MTBE in the unsaturated
zone in southern NJ is sufficient to eliminate the atmosphere as a viable source of MTBE present
in shallow ground water. This may have some implication on  natural attenuation at gasoline-spill
sites.  Degradation  of BTEX compounds within the capillary zone has been shown to greatly
enhance  the transport of BTEX  mass from the water table to the  unsaturated zone due to
                                          -11-

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volatilization and upward diffusive transport, resulting in a significant natural attenuation pathway.
Given that MTBE is degraded in the vadose zone, a similar pathway may exist for MTBE natural
attenuation.
       Hunter (2000) presented data on  a small gasoline spill (7-12 gallons of reformulated
gasoline  [RFG]) that  contaminated bedrock drinking water wells. MTBE concentration in  the
reformulated gasoline was estimated to be  11% by volume. Contaminated soil was removed  and
households were provided with point-of-entry filtration. Otherwise, no other remedial efforts were
employed. Initial MTBE concentrations  were approximately  6,500 ug/L. Within two years, all
wells  were below  the 35  ug/L health standard.  It is  believed  that removal was due to rapid
groundwater flow and dispersion rather than biodegradation.
       Weaver (2000) characterized four plumes on Long Island, NY. The aquifers all had high
groundwater velocities and recharge rates.  In general, the MTBE plumes were thousands of feet
long. All plumes were documented to "dive" into the aquifer possibly due to recharge. Inadequate
site characterization would have missed the plumes if groundwater monitoring well screens were
only screened  across the water table.
       Happel et  al.  (2000) conducted an analysis of compliance data from over 500 Leaking
Underground  Fuel Tanks (LUFT) sites in CA.  Approximately  7,000 sampling events were
conducted  on  these  500 wells. Approximately  50% of the  sampling events detected  MTBE.
Preliminary data indicate that MTBE  attenuated  at a rate two orders of magnitude lower than
BTEX.
       These studies illustrate both the potential of MTBE natural  attenuation as well as  the
inadequacy of natural attenuation. Natural  attenuation  of MTBE  is highly sensitive to site
characteristics  and may simply not  be  feasible at some  sites. In  addition,  Borden  (2000)
demonstrated that existing models are not adequate to predict MTBE natural attenuation and may
significantly underestimate the plume size over time. In particular, these studies illustrate the need
for additional research into the factors that influence natural attenuation.

3.4    Ex Situ Bioremediation
       Several researchers are investigating the  potential for ex situ bioremediation of MTBE.
Ex situ bioremediation of MTBE  could be applied as part  of a pump-and-treat approach  for
                                          -12-

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remediation of contaminated groundwater,  or it may be used as part of the treatment train for
drinking water.  The  majority of the investigations  that were presented  are  currently  at  the
laboratory-scale; however, Chang et al. (2000) presented data from a pilot-scale test.
       Cowan et al.  (2000) examined the kinetics of an MTBE-degrading microbial culture. The
microbial growth rate was slower than for most heterotrophs, with maximum specific growth
rates ranging from 0.017 to 0.057 h-1 at 30°C. Consequently, the low growth rates limited the
types  of bioreactors that could be used for water treatment.  Reactors  that were examined
included a sequencing batch reactor (SBR), a submerged attached growth air-lift (SAGAL), and
a cyclically operated submerged attached growth bioreactor (COSAG). With the SBR, effluent
concentrations were sustained at <20 ug/L; however, two shocks occurred  to the system  during
the experiment and  recovery times  were quite long (1.5-2 months). The SAGAL reactor
performed well, also sustaining effluent MTBE concentrations <20 ug/L.  Variations in reactor
temperature impacted the reactor performance. The COSAG bioreactor is currently in operation
and data is currently being evaluated; however, results to date are promising, with no MTBE
detected in the reactor effluent at a hydraulic residence time of 4.5 hours.
       Venosa et al. (2000) described results from four  bioreactors operated for over one year
to determine  MTBE biodegradation  under  different substrate/co-substrate  conditions.  The
reactors used  were porous pot reactors. The reactor conditions were as follows:
        !       influent  MTBE concentration of 150 mg/L with MTBE the only organic  carbon
               source;
        !       influent  MTBE concentration  of 75  mg/L  with ethanol also  added at a
               concentration of 75 mg/L;
        !       influent  MTBE concentration of 75 mg/L with diethyl ether also  added at a
               concentration of 75 mg/L; and
        !       influent MTBE concentration of 75 mg/L with diisopropyl  ether also added at a
               concentration of 75 mg/L.
       Results showed  that at high  biomass concentrations, MTBE was biodegraded  in  the
presence or absence  of other carbon sources. Mineralization of MTBE occurred, as confirmed
through chemical  oxygen demand  (COD)  and carbon analysis. Little loss occurred from  the
control reactor, confirming  system integrity.  This bioreactor design was  useful for laboratory
                                          -13-

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situations, but would probably be ineffective in the field due to limited flow rates. Future research
includes pilot-scale evaluations using commercially available membrane bioreactors.
       In a related study by  Suidan et  al.  (2000), the kinetics of MTBE biodegradation of
cultures developed in the Venosa et al. (2000) studies were examined. Studies were conducted in
batch reactors  and several parameters were investigated including MTBE, TEA, total  and
inorganic carbon,  dissolved oxygen, pH, and gaseous carbon dioxide and oxygen. MTBE was
mineralized to 1 • g/L within 24 hours with initial concentrations at 5, 15 and 40 mg/L. Results
indicated that biotransformation of TEA was the rate-limiting step in the mineralization of MTBE
In addition, the presence  of ethanol  competed  with  TEA  biodegradation, but not MTBE
degradation.
       Morales and Deshusses (2000)  conducted microcosm  and column studies  with  an
MTBE-degrading  consortium.  Biodegradation was only observed under aerobic conditions and in
the presence of the MTBE-degrading consortium. No degradation was observed under anaerobic
conditions or with indigenous microorganisms. In the  presence of the consortium,  complete
degradation of 20-25 mg/L MTBE was observed in approximately 10 days. The degradation rate
decreased with successive MTBE spikes,  possibly due to toxic levels of nitrite.  Column studies
were operated for 6 months with an approximate 100% MTBE removal efficiency for loadings of
0.25 - 3 g/rrf-h for  columns packed with soil and 0.25 - 3 g/m?-h for columns packed with perlite.
Microcosm studies showed 70% conversion of MTBE to carbon dioxide, with a lower conversion
in columns. A new treatment technique Deshusses called "pump and trickle" where groundwater
is brought to the  surface and reinjected in an infiltration  trench seeded with MTBE degrading
micro-organisms was proposed.
       Chang et al. (2000) described the use of a mixed culture and bacterial  strain PM1 in a
biotrickling filter unit used to treat MTBE-contaminated  groundwater. The biotrickling filter unit
consisted of seven granular activated carbon (GAC) packed-bed columns with a diameter of 14
inches and a depth of six ft. Start-up and operation of the columns was not steady, but shut down
periods were unrelated to problems with the biotrickling filter.  One column was inoculated with
the strain PM-1 from a pure culture grown on ethanol. Influent concentrations of MTBE ranged
from 290-460 |Jg/L, with removal efficiencies of greater than 90%.
                                          -14-

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        A pilot-scale, compost-based biofilter for treatment  of MTBE vapor also has been
investigated.  To date, the removal efficiency approaches 100% for loading rates less than about
300 g/irf-d.
                                         -15-

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                                   4.0 RESEARCH NEEDS




       Historically, there has been concern regarding the recalcitrance of MTBE. While all researchers




presented data and agreed upon the biodegradability of MTBE, it was apparent that additional research was




necessary to more fully understand both the basic microbiology of MTBE biodegradation, as well as develop




effective technologies for remediation of MTBE-contaminated groundwater and soils and to understand the




environmental conditions under which MTBE is degraded. It is critically important that this research on




MTBE biodegradation be conducted to  improve the understanding and performance of MTBE remedial




technologies.




       The following areas appear key for further research into the microbiology of MTBE biodegradation:




        !       The influence of various environmental parameters on MTBE biodegradation, including




               geochemical factors and temperature should be investigated.




        !       The effect of BTEX on MTBE biodegradation in moderate to low BTEX concentrations and




               high BTEX concentrations needs to be understood.




        !       The by-products of MTBE biodegradation, such as TEA, should be studied since they are




               often detected at sites.




        !       A better understanding of the cause for low growth rates and low cell yields on  MTBE




               should be developed. Adequate biomass must be maintained for efficient degradation of




               MTBE.




        !       Given that evidence has been shown for MTBE biodegradation under aerobic, anaerobic, and




               cometabolic conditions, identification of the microorganisms involved in these processes may




               provide a link between research conducted in different laboratories.
                                               16

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        !       Likewise, given the variety  of conditions under which MTBE biodegradation has been




               observed, more research is needed on the mechanism of MTBE biodegradation, including




               pathways and regulators of MTBE metabolism under aerobic, anaerobic, or cometabolic




               conditions. This may have an impact on ex situ   bioreactor performance and provide




               information on the potential for and predictability of in situ bioactivity.




       Prior to implementing MTBE remedial technologies, it is also apparent that there needs to be a better




understanding of the scope of the problem nationwide and the state-of-the-art for treatment technologies.




These research needs are summarized as follows:




        !       Develop a database containing  information  on MTBE-contaminated  sites nationwide




               representing various environmental conditions. Site data should ideally include contaminant




               concentrations and distribution, geochemical data, and hydrogeological information. The EPA




               and BP/Amoco have formed a collaborative effort to obtain this information from a number




               of petroleum-industry sites. Additional input from other sources would be beneficial.




        !       Assess ability of various technologies to achieve different target levels and associated costs




               to achieve the target level.




        !       Develop a database of technologies that are at pilot- or full-scale and may work for MTBE.




               As much cost and performance data as possible should be included.




       Detailed suggestions for research needs on specific  technologies  were discussed during the




workshop. The technologies under discussion could be broadly categorized by monitored natural attenuation,




enhanced in situ treatment, and ex situ treatment. In addition, a number of research needs were apparent




in the area of site characterization. In the following paragraphs, the research needs for these specific areas




are discussed.
                                                17

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       Site Characterization.  Site characterization is a critical component of the site cleanup. If site




characterization is not adequately performed, site cleanup may not be achieved and serious health and




environmental impacts  could occur later.  Conventional site characterization strategies that have  been




implemented at BTEX-contaminated sites may not be adequate to delineate the MTBE plume or to identify




and quantify MTBE biodegradation indicators. The following research needs have been identified:




        !       Sites must be more comprehensively characterized. Plumes may be deeper and longer than




               expected.




        !       Source mass should be better characterized since this impacts treatment.




        !       Understand the effect recharge has on the downward movement of an MTBE plume.




        !       Guidance in the form of a protocol should be developed on the proper site characterization




               methods and analytical methods.




       Monitored Natural Attenuation. Monitored natural attenuation may be applicable under specific site




conditions; however, a significant amount of research is still needed to  fully understand the processes that




impact natural attenuation of MTBE. The following research needs have been identified:




        !       Determine data needs beyond those obtained for BTEX assessments. Microbiological studies




               may help determine these data needs.




        !       Screen a large number of sites to better understand how prevalent MTBE biodegradation is




               and how significantly MTBE biodegradation contributes to natural attenuation of MTBE. This




               is also necessary to determine in situ MTBE biodegradation rates.




        !       Determine specific site conditions  conducive to or inhibiting biodegradation of MTBE. This




               data could come from a combination of microbiological studies and assessment of a database




               of site data.




        !       Understand the role of groundwater/surface water interfaces.






                                               18

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        !       Develop a protocol for conducting natural attenuation assessments of MTBE.




       Enhanced In Situ Biodegradation. Enhanced in situ biodegradation is being investigated in the field




and promising results have been demonstrated. Additional research needs are as follows:




Conduct additional pilot-scale field trials and assess the following parameters:




        !       Life cycle costs and reliability




        !       Achievable degradation rates




        !       Biomass required and maintained




        !       Electron acceptor delivery methods




        !       Adequate methods to evaluate performance




        !       Development of techniques for effective electron acceptor delivery




        !       Study of enhanced in situ  MTBE biodegradation under a variety of conditions including




               different  hydrogeological  conditions, different  contaminant concentrations,  and mixed




               contaminant systems




        !       Development of aggressive source area technologies.  It is unknown whether enhanced




               biodegradation will be effective for residual nonaqueous phase liquids.




        !       Compilation of case studies of enhanced in situ bioremediation to find  determinants of




               success or failure




        !       Development of techniques for determining the presence of MTBE-degrading bacteria and




               identify what factors may be limiting  their activity. Microbiological studies would provide




               information to assist in this  determination




        !       Development of protocols  for conducting and monitoring in situ MTBE bioremediation




               technologies
                                               19

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       Ex Situ Bioremediation. Ex situ bioremediation techniques have shown successful biodegradation




of MTBE under a variety of conditions. In addition to some additional research at the laboratory-scale level,




there are several areas of research to be explored at the field-scale. Additional research needs are as follows:









Conduct pilot-scale field trials and assess the following parameters:




        !       Life cycle costs and reliability




        !       Achievable degradation rates




        !       Biomass required and maintained




        !       Adequate methods to evaluate performance




        !       Long term performance data with  shock  loadings and other operational performance




               requirements




        !       Mechanisms and processes to control degradation in aboveground water treatment reactors




        !       Reactor performance and costs under different influent conditions, including varying MTBE




               concentrations, loadings, and mixed contaminants




        !       Existing GAC systems for biological activity and evaluate efficacy and cost of inoculating




               existing GAC reactors with MTBE-degrading cultures




        !       Biotreatment as cost competitive compared to existing technologies such as GAC treatment




        !       State-of-the-practice database providing operational information of various reactor types




        !       Protocols for conducting and evaluating ex situ bioremediation of MTBE




        Overall. A combination  of technologies is  likely  to  be the most  appropriate choice for site




remediation. As such, it is important to examine the best treatment train technologies that bring MTBE




concentrations to low levels (i.e. thermal destructive technologies combined with air sparging or SVE followed
                                               20

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by biodegradation). This is an important area of research since a treatment train approach may likely be




necessary at many sites.
                                                21

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                              5.0 COLLABORATIVE EFFORTS

       The need for collaborative efforts into investigating bioremediation of MTBE became evident during
the workshop. Different research groups have different strengths, and combining these strengths would bring
the most powerful approach to solving the problem of MTBE contamination. A work group comprised of
government agencies  (e.g., EPA and USGS), industry representatives (e.g.,  the American Petroleum
Institute), and academia would be the most productive. Additional suggestions are as follows:
       1.      Many different areas of expertise were evident during the workshop. These can be grouped
               into three broad  categories: microbiology, bioreactor design, and field expertise. The team
               could  include a combination of these areas of expertise. The  microbiology of MTBE
               bioremediation is not fully understood and  researchers working with MTBE-degrading
               microbialconsortia or those examining field biodegradation would benefit from the input from
               microbiologists.  Likewise, researchers involved with bioreactor design and implementation
               could create a strong team if working with researchers with significant field experience.
       2.      Lead organizations  should be aware of the need to create this combination of experts when
               developing new  programs. The best way to create this  awareness is through widespread
               dissemination of current research and existing research needs.
       3.      In  order to disseminate the current  information on MTBE bioremediation, workshops
               designed for lead organizations could be developed. Government agencies with experience
               conducting these types of workshops could collaborate with various experts in the field of
               MTBE bioremediation.
       4.      An organization is needed that would take the lead on disseminating information on MTBE
               remediation. A combination of government agencies, industry, and academia would provide
               an appropriate forum for this activity.
                                               22

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                                 6.0 LITERATURE CITED

Baehr, A.L., E.G. Charles, and R. J. Baker. 2000. Field Evidence for Methyl  tert-Butyl Ether (MTBE)
Degradation in the Unsaturated Zone at Low Concentrations. Presented at the MTBE Biodegradation
Workshop, Cincinnati, OH, February 1-3.

Borden, R.C. 2000. Transport and Fate of a BTEX and MTBE Plume - What Do We Know? Presented at
the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Butler, B.J., M. Schirmer, and J.F. Barker. 2000. The Fate of MTBE in the Borden Aquifer. Presented at
the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Chang, D.P.Y., E.D. Schroeder, K.M. Scow, B.M. Converse, J. Scarano, N. Watanabe, and K. Romstad.
2000. Experience with Laboratory and Fi eld-Scale Ex Situ Biodegradation of MTBE. Presented at the MTBE
Biodegradation Workshop, Cincinnati, OH, February 1-3.

Chapelle, F.H. 2000. The Distribution and Environmental Fate of MTBE in the Hydrosphere: The Approach
of the U.S. Geological Survey. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February
1-3.

Church, C.D. and P.G Tratnyek. 2000. Process Level Investigations of the In Situ Degradation of MTBE.
Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Cowan, R.M., J.K. Truskowski, and K. Park. 2000. MTBE Biodegradation Research at Rutgers, The State
University of New Jersey. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Da Silva, M.B., N. Lovanh, C.S. Hunt, and P.J.J. Alvarez. 2000. The Effects of Ethanol on BTEX Natural
Attenuation:  Pure Culture and  Aquifer Column Experiments. Presented at the MTBE  Biodegradation
Workshop, Cincinnati, OH, February 1-3.

Finneran, K.T. and D.R Lovley. 2000. Anaerobic Degradation of Methyl terf-Butyl Ether (MTBE) and tert-
Butyl Alcohol (TEA). Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February  1-3.

Hanson, J.R., C.E. Ackerman,  and K.M.  Scow. 1999. Biodegradation of Methyl  tert-butyl ether by a
Bacterial  Pure Culture. Appl Environ. Microbiol 65:4788-4792.

Happel, A.M., E.H. Beckenbach, and K.N. Emerson. 2000. Evaluating Attenuation of MTBE: What We
Have Learned From LUFT Data and Laboratory Studies. Presented at the MTBE Biodegradation Workshop,
Cincinnati, OH, February 1-3.
                                             23

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Hunter, B. 2000. Natural Attenuation of MTBE at a Site Where 24 Bedrock Wells Were Contaminated by
a 10-Gallon Gasoline Spill. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Hyman, M. 2000. Microbial Cometabolism of MTBE. Presented at the MTBE Biodegradation Workshop,
Cincinnati, OH, February 1-3.

Kropp, K.G., M.R. Mormile, and J.M. Suflita. 2000. Anaerobic Biodegradation of MTBE and Alternative
Gasoline Oxygenates. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Landmeyer, IE. 2000. MTBE Attenuation Processes: Ambient and Enhanced Redox Conditions, Stream Bed:
Groundwater Interactions, and Plant Uptake. Presented at the MTBE Biodegradation Workshop, Cincinnati,
OH, February 1-3.

Mills, M.A. and J.R. Haines. 2000. Monitoring Petrochemical Biodegradation by Continuous-Flow Isotope
Ratio Mass Spectrometry. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Morales, M. and M. Deshusses. 2000. Research in Bioremediation of MTBE at UC Riverside: Lessons from
Laboratory Experiments. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Salanitro, J. 2000. In Situ Control of MTBE Plumes with Inoculated  Biobarriers. Presented at the MTBE
Biodegradation Workshop, Cincinnati, OH, February 1-3.

Schirmer, M., C. Hubbard, B. Butler, R Devlin, and J. Barker. 2000. The Borden Field Experiment - Where
Has the MTBE Gone? Demonstrating In Situ Remediation - The  Borden Aquifer Research Facility.
Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Scow, K.M., A. Smith, J. Leung, D. Mackay, and E. Lory. 2000. Bioaugmentation of MTBE-Contaminated
Groundwater with Bacterial strain PM1. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH,
February  1-3.

Speth, T.  2000. Drinking Water Issues. Presented at the MTBE Biodegradation Workshop, Cincinnati, OH,
February  1-3.

Stringfellow, W.T. 2000. Using iso-Pentane to Stimulate Biodegradation in Groundwater Treatment Systems.
Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Suidan, M.T., GJ. Wilson, AP. Richter,  and A.D. Venosa. 2000.  Kinetics of MTBE Biodegradation.
Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Venosa, A.D., M.T. Suidan, G.J. Wilson, and A.P. Richter. 2000.  Aqueous Mineralization of MTBE.
Presented at the MTBE Biodegradation Workshop, Cincinnati, OH, February 1-3.

Wilson, R.D. et al. 1999. Laboratory-Scale Evaluation of In Situ Aerobic MTBE Biodegradation Options for
Vandenberg Air Force Base,  CA. Proceedings of the Conference  "Petroleum Hydrocarbons and
Organic Chemicals in Ground Water: Prevention, Detection and Remediation ", cosponsored by the API
and NGWA, November 17-19, Houston, TX.

                                             24

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            Attachment A




MTBE Biodegradation Workshop Agenda
                 -25-

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vvEPA
United States
Environmental Protection Agency
Office of Research and Development
L
) American
Petroleum

MTBE  Biodegradation Workshop
Marriott Kingsgate Conference Center
Cincinnati, Ohio
February 1-3, 2000
                                Agenda
Meeting Goals
     Ben Blaney, US EPA
OVERVIEW OF ISSUES
MTBE and Underground Storage Tanks
     Sammy Ng, US EPA
EPA's ORD Current and Future Research on MTBE Bioremediation
     Fran Kremerand Stephen Schmelling, U.S. EPA
MTBE Biodegradation: API and Industry Perspectives
     Bruce Bauman API
The Distribution and Environmental Fate of MTBE in the Hydrosphere: The Approach of the U.S.
Geological Survey
     Francis Chapelle, USGS
EPA Region 9 Perspective on MTBE Response
     Steve Under, US EPA
PRESENTATIONS
I n Situ Treatment
Transport and Fate of a BTEX and MTBE Plume: What do we know?
     Bob Borden, NCSU
Bioaugmentation of MTBE-Contaminated Groundwater with Bacterial Strain PM1
     Doug Mackay, University of Waterloo
Microbial Cometabolism of MTBE
     Michael Hyman, North Carolina State University
                                  -26-

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Anaerobic Biodegradation of MTBE and TEA
      Kevin Finneran, University of Massachusetts-Amherst

In Situ Control of MTBE Plumes with Inoculated Biobarriers
      Joseph Salanitro, Equilon Enterprises

In Situ Treatment of MTBE by Biostimulation of Native Aerobic Microorganisms
      Doug Mackay
Monitored Natural Attenuation

Evaluating Attenuation of MTBE: What we have Learned from LUFT Data and Laboratory Studies

      Anne Happel, Lawrence Livermore Laboratory

The Fate of MTBE in the Borden
      Aquifer/Barbara Butler, University of Waterloo

MTBE Attenuation Processes: Ambient and Enhanced Redox Conditions, Stream-Bed-Ground-Water
Interactions, and Plant Uptake
      James Landmeyer, USGS

Field Evidence for MTBE Degradation in the Unsaturated Zone at Low Concentrations
      Arthur Baehr, USGS

Natural Attenuation of MTBE in the Subsurface Under Methanogenic Conditions
      John  Wilson, US EPA

Comparative Evaluation of MTBE Sites on Long Island
      Jim Weaver, US EPA


Ex Situ  Treatment

Drinking Water Issues
      Thomas Speth, US EPA

MTBE Biodegradation: Kinetics, Reactor Engineering, and the Potential for Ex-Situ Treatment of
Groundwater
      Robert Cowan, Rutgers University

Mineralization of MTBE in Continuous Flow High Biomass Bioreactors
      Albert Venosa, US EPA

Kinetics  of MTBE Biodegradation
      Makram Suidan, University of Cincinnati

Research in Bioremediation of MTBE at UC Riverside: Lessons from Laboratory Experiments
      Marc Deshusses,  University of Califormia-Riverside

Experience with Laboratory and Field Scale Ex-Situ Biodegradation of MTBE
      Daniel Chang, University of California-Riverside

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POSTERS

The Effects of Ethanol on BTEX Natural Attenuation: Pure Culture and Aquifer Column Experiments
      Pedro Alvarez, US EPA

The Borden Field Experiment - Where Has the MTBE Gone? Demonstrating In Situ Remediation - the
Borden Aquifer Research Facility
      Jim Barker, University of Waterloo

Process Level Investigations of the In Situ Degradation of MTBE
      Clinton Church, Oregon Graduate Institute

Research in Bioremdediation of MTBE at UC Riverside: Lessons from Laboratory Experiments
      Marc Deshusses

Biodegradation of MTBE in Soil Monitored by I RMS
      John Haines,  US EPA

Natural Attenuation of MTBE at a Site where 24 Bedrock Wells were Contaminated by a 10-Gallon
Gasoline Spill
      Bruce Hunter, Maine Department of Environmental Protection

Anaerobic Biodegradation of MTBE and Alternative Gasoline Oxygenates
      Kevin Kropp,  University of Oklahoma

Using Iso-Pentane to Stimulate MTBE Biodegradation In Groundwater Treatment Systems
      William Stringfellow, Lawrence Berkely National Laboratory

Structure and Behavior of the MTBE Plume at Port Hueneme, CA
      John  Wilson
                                          -28-

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             Attachment B




MTBE Biodegradation Workshop Speaker List
                   29

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x>EPA
United States
Environmental Protection Agency
Office of Research and Development
American
Petroleum
        MTBE Biodegradation Workshop
        Marriott Kingsgate Conference Center
        Cincinnati, Ohio
        February 1-3, 2000
        Speaker List

        Arthur Baehr
        Water Resources
        U.S. Geological Survey
        Mount View Office Park
        810 Bear Tavern Road
        West Trenton, NJ 08628
        609-771-3978
        Fax:609-771-3915
        E-mail: abaehr@usgs.gov

        Michael Barcelona
        Research Professor
        Department of Civil &
        Environmental Engineering
        University of Michigan
        1221 1st Building
        Ann Arbor, Ml 45109
        734-763-6512
        Fax:734-763-6513
        E-mail: mikebar@engin.umich.edu

        Bruce Bauman
        American Petroleum Institute (API)
        1220 L Street, NW
        Washington, DC 20005
        202-682-8000
        E-mail: bauman@api.org
                             Ben Blaney
                             Assistant Laboratory Director
                             for Waste Research
                             U.S. Environmental Protection Agency
                             26 West Martin Luther King Drive
                             Cincinnati, OH 45268
                             513-569-7852
                             Fax:513-569-7680
                             E-mail: blaney.ben@epa.gov

                             Bob Borden
                             Department of Civil Engineering
                             North Carolina State University
                             Box 7908
                             Raleigh, NC 27695-7908
                             919-515-1625
                             Fax:919-515-7908
                             E-mail: rcborden@eos.ncsu.edu

                             Barbara Butler
                             Research Associate & Adjunct
                             Assistant Professor
                             Department of Biology
                             University of Waterloo
                             200 University Avenue
                             Waterloo, ON N21 3G1
                             CANADA
                             519-885-1211
                             Fax:519-746-0614
                             E-mail: bjbutler@uwaterloo.ca
                                            30

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Daniel Chang
Professor
Department of Civil &
Environmental Engineering
University of California, Davis
One Shields Avenue
Davis, CA 95616-5294
530-752-2537
Fax: 530-752-7872
E-mail: dpchang@ucdavis.edu

Francis Chapelle
U.S. Geological Survey
720 Gracern Road - Suite 129
Columbia, SC 29210
803-750-6116
Fax:803-750-6181
E-mail: chapelle@usgs.gov

Joan Colson
U.S. Environmental Protection Agency
26 West Martin Luther King Drive (G75)
Cincinnati, OH 45268
513-569-7501
Fax:513-569-7585
E-mail: colson.joan@epa.gov

Robert Cowan
Department of Environmental Sciences
Rutgers University
14 College Farm Road
New Brunswick, NJ 08901-8551
732-932-8750
Fax: 732-932-8644
E-mail: cowan@envsci.rutgers.edu

Marc Deshusses
Assistant Professor
Department of Chemical and
Environmental Engineering
University of California, Riverside
Bourns Hall B321
Riverside, CA 92521
909-787-2477
Fax: 909-787-2425
E-mail: mdeshuss@engr.ucr.edu
Carl Enfield
U.S. Environmental Protection Agency
26 West Martin Luther King Drive
Cincinnati, OH 45268
513-569-7489
E-mail: enfield.carl@epa.gov

Kevin Finneran
Department of Microbiology
University of Massachusetts - Amherst
Morrill Science Center
Amherst,  MA 01003
413-545-9649
Fax:413-545-1578
E-mail: finneran@microbio.umass.edu

Anne Happel
Lawrence Livermore National Laboratory
700 East  Avenue L-542
Livermore, CA 94550-9234
925-422-1425
Fax: 925-423-7998
E-mail: happel1@llnl.gov

Michael Hyman
Department of Microbiology
North  Carolina State University
Raleigh, NC 27695-7615
919-515-7814
Fax:919-515-7867
E-mail: hymanm@mbio.ncsu.edu

Fran Kremer
U.S. Environmental Protection Agncy
26 West Martin Luther King Drive (481)
Cincinnati, OH 45268
513-569-7346
E-mail: kremer.fran@epa.gov
James Landmeyer
U.S. Geological Survey
720 Gracern Road - Suite 129
                                          31

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Columbia, SC 29210-7651
803-750-6128
Fax:803-750-6181
E-mail: jlandmey@usgs.gov

Steve Linder
U.S. Environmental Protection Agency
75 Hawthorne Street (WST-8)
San Francisco, CA 94105
415-744-2036
Fax:415-744-1026
E-mail: linder.steven@epa.gov

Doug Mackay
University of Waterloo
744 Frenchman's Road
Stanford, CA 94305
650-324-2809
Fax: 650-324-2259
E-mail: d4mackay@uwaterloo.ca

Hugh McKinnon
Associate Lab Director
National Risk Management Risk
Research Laboratory
U.S. Environmental Protection Agency
26 West Martin Luther King Drive (225)
Cincinnati, OH 45268
513-569-7689
E-mail: mckinnon.hugh@epa.gov

Sammy Ng
Acting Director, Office of
Underground Storage Tanks
U.S. Environmental Protection Agency
401 M Street, SW (5401G)
Washington,  DC 20460
703-603-9900
Fax:703-603-0175
E-mail: ng.sammy@epa.gov
P.O. Box1627
Richmond, CA 94802-0627
510-242-5365
Fax:510-242-1954
E-mail: kito@chevron.com

Joseph Salanitro
Equilon Enterprises
3333 South Highway 6 - P.O. 8ox 1380
Houston, TX 77251-1380
281-544-7552
Fax:281-544-8727
E-mail: jpsalanitro@equilon.com

Stephen Schmelling
National Risk Management
Research Laboratory
U.S. Environmental Protection Agency
919 Kerr Research Drive - P.O. Box 1198
Ada, OK 74821
580-436-8540
Fax: 580-436-8581
E-mail: schmelling.steve@epa.gov

Thomas Speth
U.S. Environmental Protection Agency
26 West Martin Luther King Drive (B24)
Cincinnati, OH 45268
513-569-7208
E-mail: speth.thomas@epamail.epa.gov

Makram Suidan
Water Quality Processes Program
Department of Civil and
Environmental Engineering
University of Cincinnati
P.O. Box210071
Cincinnati, OH 45221-0071
513-556-3695
Fax:513-556-4003
E-mail: makram.suidan@uc.edu
Kirk O'Reilly
Senior Environmental Specialist
Chevron Research & Technology
Albert Venosa
U.S. Environmental Protection Agency
26 West Martin Luther King Drive
Cincinnati, OH 45268
                                          32

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513-569-7668
Fax:513-569-7105
E-mail: venosa.albert@epa.gov

Jim Weaver
National Exposure Research Laboratory
Ecosystems Research Division
U.S. Environmental Protection Agency
960 College Station Road
Athens, GA 30605-2700
706-355-8329
Fax: 706-355-8302
E-mail: weaver.jim@epa.gov

John Wilson
Research Microbiologist
Office of Reserch and Development
National Risk Management
Research Laboratory
U.S. Environmental Protection Agency
Kerr Research Laboratory
919 Kerr Research Drive
Ada, OK 74820
580-436-8534
Fax: 580-436-8703
E-mail: wilson.johnt@epa.gov
                                           33

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                Attachment C




MTBE Biodegradation Workshop Poster Presenters
                     34

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x>EPA
United States
Environmental Protection Agency
Office of Research and Development
American
Petroleum
        MTBE Biodegradation Workshop

        Marriott Kingsgate Conference Center
        Cincinnati, Ohio
        February 1-3, 2000
        Poster Presenter List
        Pedro Alvarez
        Civil and Environmental Engineering
        University of Iowa
        4116 Seamans Center
        Iowa City, IA 52242-1527
        319-335-5065
        Fax: 319-335-5660
        E-mail: pedro-alvarez@uiowa.edu

        Jim Barker
        Department of Earth Sciences
        University of Waterloo
        200 University Avenue
        Waterloo, ON N2L 3G1
        Canada
        519-885-1211
        Fax: 519-746-7484
        E-mail: barker@cgrnserc. uwaterloo. ca

        Clinton Church
        Oregon Graduate Institute
        20000 Northwest Walker Road
        Beaverton, OR 97006
        503-690-1651
        Fax: 503-690-1273
        E-mail: church@ese.ogi.edu
                           Marc Deshusses
                           Assistant Professor
                           Department of Chemical and Environmental
                           Engineering
                           University of California, Riverside
                           Bourns Hall B321
                           Riverside, CA 92521
                           909-787-2477
                           Fax: 909-787-242
                           E-mail: mdeshuss@engr.ucr.edu

                           John Haines
                           U.S. Environmental Protection Agency
                           26 West Martin Luther King Drive (420)
                           Cincinnati, OH 45268
                           513-569-7446
                           E-mail: haines.john@epa.gov

                           Bruce Hunter
                           Hydrogeologist
                           Maine Department of Environmental
                           Protection
                           17 State House Station
                           Augusta, ME 04333-0017
                           207-287-7672
                           Fax: 207-287-7826
                           E-mail:
                           bruce.e.hunterstate.me.us
                                           35

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 Kevin Kropp
 Post Doctoral Research Associate
 Department of Botany & Microbiology
 University of Oklahoma
 770 Van Vleet Oval
 Norman, OK 73019
 405-325-3771
 Fax: 405-325-7619
 E-mail:  kevinkropp@ou.edu

 William Stringfellow
 Research Engineer
 Center for Environmental Biotechnology
 Lawrence Berkeley National Laboratory
 (MS-70A-3317)
 Berkeley, CA 94720
 510-486-7903
 Fax: 510-486-7152
 E-mail: wstringfellow@lbl.gov

John Wilson
Research Microbiologist
U.S. Environmental Protection Agency
National Risk Management Research
Laboratory
Kerr Research Laboratory
919 Kerr Research Drive
Ada, OK 74820
580-436-8534
Fax: 580-436-8703
E-mail: wilson.johnt@epa.gov
                                           36

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                Attachment D




MTBE Biodegradation Workshop List of Attendees
                      37

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x>EPA
United States
Environmental Protection
Agency
Office of Research and
Development
American
Petroleum
        MTBE Biodegradation
        Workshop
        Marriott  Kingsgate Conference
        Center
        Cincinnati, Ohio
        February 1-3,  2000

        Attendee List
        Steven Acree
        Office of Research & Development
        Subsurface Protection & Remediation Division
        U.S. Environmental Protection Agency
        Robert Kerr Environmental Research Center
        P.O. Box1198
        Ada, OK 74821-1198
        580-436-8609
        E-mail: acree.steven@epa.gov

        Gilberto Alvarez
        Environmental Engineer
        U.S. Environmental Protection Agency
        77 West Jackson Boulevard (DU-7J)
        Chicago, IL 60604-3507
        312-886-6143
        Fax:312-353-3159
        E-mail: alvarez.gilberto@epa.gov

        David Ariail
        Environmental Engineer
        U.S. Environmental Protection Agency
        Sam Nunn Federal Center
        61 Forsyth Street, SW
                      Atlanta, GA 30303-8960
                             404-562-9464
                             Fax: 404-562-9439
                             E-mail: ariail.david@epa.gov


                             John Brophy
                             Office of Air and Radiation
                             U.S. Environmental Protection Agency
                             401 M Street, SW (6406-J)
                             Washington, DC 20460
                             202-564-9068
                             E-mail: brophy.john@epa.gov

                             Tom Conrardy
                             Bureau  of Petroleum Storage Systems
                             Florida Department of Environmental Protection
                             2600 Blair Stone Road (MS-4530)
                             Tallahassee, FL 32399-2400
                             E-mail: tom.conrardy@dep.state.fl.us

                             Linda Fiedler
                             U.S. Environmental Protection Agency
                             401 M Street, SW (5102-G)
                             Washington, DC 20460
                             703-603-7194
                             E-mail:
                             fiedler.linda@epa.gov
                                              38

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Annette Gatchett
U.S. Environmental Protection Agency
26 West Martin Luther King Drive (481)
Cincinnati, OH 45268
513-569-7697
E-mail: gatchett.annette@epa.gov

Joe Haas
Engineering Geologist
New York State Department
of Environmental Conservation
Building 40-SUNY
Stony Brook, NY 11790-2356
631-444-0332
Fax:631-444-0373
E-mail:jehaas@gw.dec.state.ny.us


Douglas Heath
Hydrogeologist
U.S. Environmental Protection Agency
One Congress Street - Suite  1100 (CNH)
Boston, MA  02114-2023
617-918-1585
Fax:617-918-1505
E-mail: heath.doug@epa.gov

Ravi Kolhatkar
BP-Amoco
150 West Warrenville Road (H-7)
Naperville, IL 60563
630-420-3824
Fax: 630-420-5016
E-mail: kolhatrv@bp.com

William Kramer
Principal Hydrogeologist
Handex Environmental
P.O. Box 451
500 Campus Drive
Morganville,  NJ 07751
732-536-8667
Fax: 732-536-7751
E-mail: bkramer@handexmail.com
   Andrea Leeson
   Research Leader
   Battelle
   505 King Avenue
   Columbus, OH 43201
   614-424-6424
   E-mail: leeson@battelle.org

   Ernie Lory
   National Environmental
   Technology Test Site Manager
   U.S. Navy
   NFESC-ESC411
   Port Hueneme, CA 93043
   805-982-1299
   Fax: 805-982-4304
   E-mail: loryee@nfesc.navy.mil

   Norm Novick
   Technology Assessment
   and Enhancement Coordination
   Exxon Mobil
   8280 Willow Oaks II - Room 6W117
   3225 Gallows Road
   Fairfax, VA  22037
   703-849-4968
   Fax:703-849-5217
   E-mail: norman_j_novick@email.mobil.com

   Rey Rodriguez
   Subcommittee Chair, MTBE Partnership
   California MTBE Research Partnership
   653 Michelle Street
   West Covina, CA 91790
   626-917-7747
   Fax:626-917-7847
   E-mail: mapper3d@aol.com

   Laurel Staley
   Chief
   Treatment & Destruction Branch
   U.S. Environmental Protection Agency
   26 West Martin Luther King Drive (420)
   Cincinnati, OH  45268
   513-569-7863
   Fax:513-569-7105
   E-mail: staley.laurel@epa.gov
39

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Curt Stanley
Senior Staff Hydrogeologist
Equilon Services
West Hollow Technology Center
P.O. Box1380
Houston, TX 77251-1380
281-544-7675
Fax:281-544-8727
E-mail: ccstanley@equilon.com

Hal White
Office of Underground Storage Tanks
Office of Solid Waste and Emergency
Response
U.S. Environmental Protection Agency
401 M Street, SW(5403-G)
Washington, DC 20460
703-603-7177
Fax:703-603-0175
E-mail: white.hal@epa.gov
Richard Willey
Hydrologist
Region 1
U.S. Environmental Protection Agency
One Congress Street (HBS)
Suite 1100
Boston, MA 02114-2023
617-918-1266
Fax:617-918-1291
E-mail: willey.dick@epa.gov
                                          40

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