United States
Environmental Protection Agency
REGIONAL/ORD WORKSHOP ON
EMERGING ISSUES ASSOCIATED WITH
AQUATIC ENVIRONMENTAL
PATHOGENS
SUMMARY REPORT
September 5 - September 7, 2001
Fort Meade, MD
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
TABLE OF CONTENTS
FOREWORD 1
EXECUTIVE SUMMARY 3
RESEARCH NEEDS IDENTIFIED AT THE WORKSHOP 5
ORD'S RESPONSE TO REGIONAL RESEARCH NEEDS 9
WORKSHOP AGENDA:
PRESENTATION TITLES, PRESENTERS 'NAMES, AND PAGE NUMBERS 15
APPENDIX A: LIST OF PARTICIPANTS A-l
APPENDIX B: SLIDES FROM PRESENTATIONS B-l
APPENDIX C: BREAKOUT GROUP SUMMARY C-l
APPENDIX D: FLIP CHART NOTES D-l
APPENDIX E: WORKSHOP PARTICIPANT EVALUATION SUMMARY E-l
US EPA ijiio of Research and Development
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
FOREWORD
The ORD/Regional Training Workshop on Emerging Issues Associated with Aquatic
Environmental Pathogens was the seventh in a series of Regional Science Topic Workshops
sponsored by the Office of Science Policy (OSP) in the Office of Research and Development
(ORD) at the U.S. Environmental Protection Agency (EPA). Other workshops in this series
included:
Asthma: The Regional Science Issues
Communicating Science: Waves of the Future Info Fair
Fully Integrated Environmental Location Decision Support (FIELDS)
Non-Indigenous Species
Pesticides
Endocrine Disrupters
The objectives of the National Regional Science Topic Workshops are to: 1) establish a better
cross-Agency understanding of the science applicable to specific region-selected human health
and/or ecological topics, and 2) develop a network of EPA scientists who will continue to
exchange information on these science topics as the Agency moves forward in planning
education, research, and risk management programs.
Each year, EPA regions, through the Regional Science Council, identify priority science topics
on which to conduct workshops. The workshops address the science issues of greatest interest to
the regions on the selected topic area. Each workshop is planned and conducted by a team of
regional, ORD, and interested program office scientists, and is led by a regional chairperson and
facilitated by one or more Regional Science Liaisons to ORD. Participants maintain the cross-
Agency science networks they establish at the workshops through planned post-workshop
projects and activities, such as the identification of collaborative research opportunities, creation
of information sharing mechanisms (e.g., interactive web sites), and development of technical
fact sheets for regional use.
For additional information on a specific workshop or on the Regional Science Topic Workshop
series, contact David Klauder in ORD's Office of Science Policy (202-564-6496).
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
EXECUTIVE SUMMARY
The ORD/Regional Training Workshop on Emerging Issues associated with Aquatic
Environmental Pathogens was held on September 5 - September 7, 2001, at Fort Meade,
Maryland. The workshop was chaired by Ron Landy (Region 3 Liaison) and facilitated by Mick
Kulik (Region 3).
The workshop was organized into six sessions:
I. Science Support to EPA Programs and Regions
II. Incidence and Risk Assessment of Emerging Pathogens
III. Assessing Exposures to Emerging Pathogens
IV. Emerging Pathogens Risk Management Challenges:
A. Emerging Pathogens in Drinking Water
B. Emerging Pathogens in Recreational Water
V. Pathogen TMDLs / CAFOs / Biosolids
VI. Over the Scientific Horizon - Meeting Regional Research Priorities
Regional, Office of Research and Development, and Office of Water scientists presented current
work on methodologies being developed, and new data on a variety of pathogens of concern;
EPA regulations or rules under development or in the process of revision, and new or improved
approaches for risk assessment and risk management. The need for new and better methods as
more pathogens are recognized was an often-repeated theme throughout the workshop; however,
a number of the presenters described such new methods currently being developed, or, in the
form of case studies, gave alternatives that have been used by regulators when a method was
unavailable. The meeting was limited primarily to EPA staff, with representation from all ten
regions, all 5 ORD National Labs and Centers, and all the relevant components of the Office of
Water. When dealing with topics where external views were considered critical, outside
speakers were invited.
Breakout sessions, and a panel discussion, were held with the express purpose of generating
detailed lists of the type of data, procedures, and information needed to help with the regulation
and control of different types of pathogens, in both drinking and recreational water.
Many participants expressed a wish to conduct workshops on this topic on a regular basis;
reasons given were that there will always be new pathogens emerging, and new methods and
data needed by regional scientists and regulators. The workshop provided an informal way of
keeping in contact with individuals in similar fields and situations, as well as a way of keeping
informed of new developments.
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
RESEARCH NEEDS IDENTIFIED AT THE WORKSHOP
The list of research needs below was initiated by a panel discussion held on the first day of the
workshop with the purpose of identifying high-priority regional needs. The list includes issues
brought up during discussions, presentations, question and answer sessions, and break-out group
sessions throughout the workshop.
METHODS - NEW/BETTER METHODS NEEDED
The most commonly and emphatically expressed need by the regions was the development of
better methods: new methods for pathogens not currently being monitored, simpler methods that
can be used reliably, and faster methods where time is an issue (e.g., beach closings). In
particular, the need mentioned most frequently was the need to develop methods for directly
measuring pathogens of concern so that we would no longer have to rely on using indicators.
The problems with using indicators were discussed at length during the workshop.
A method is needed for the detection of E. coli O157, which is not detected by routine E. coli
monitoring.
A better method for E. coli, specifically one that will not give as many false positive results due
to the presence of a different organism (eg., Colilert).
For coliform bacteria, a simpler method for determining assimilable organic carbon (AOC) is
needed; AOC levels are correlated with coliform and Mycobacterium avium complex presence.
Methods that give accurate results within the same day are especially valuable to regional
offices performing assessment of beaches, as decisions on whether to close a beach have
to be made quickly and often carry significant financial consequences.
Methods specifically intended for biosolids and groundwater need to be developed, and
existing methods standardized.
Methods need to be developed or adapted to be able to detect organisms associated with
biofilms
Improvements to existing methods for some pathogens were also suggested.
Specifically, a better method for measuring Aeromonas that will be easier to use for
routine, regulatory monitoring. In the case of Cryptosporidium, the existing method can
be misinterpreted.
Recovery methods for use in soil are needed for all pathogens.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
TECHNICAL ASSISTANCE. AND RESEARCH NEEDS HIGHLIGHTED DURING
WORKSHOP DISCUSSIONS:
Criteria need to be developed for selection of sampling sites, especially for coliform
bacteria testing, as false positive readings have been connected with sampling sites.
Better information on the significance of indicators used in water quality monitoring
would be helfpul. Indicators are sometimes the only available test, and as such will
continue to be used, but a clear explanation of their shortcomings is necessary.
Standard and detailed techniques for risk management are needed for pathogens in both
drinking water and recreational water. More detailed information is needed to determine
the efficacy of disinfection methods on the removal of various pathogens. Research is
also needed on sample flushing procedures.
New research is needed to develop a database on indicator organisms; the bulk of the
existing data on indicators deals with the same few organisms.
Consensus should be reached regarding the best indicator organisms for detecting
pathogens in biosolids.
DAT A NEEDED: PATHOGENS
All Pathogens:
Better data on pathogen characterization is needed for all pathogens, particularly
information on virulence determinants and indicator regrowth.
Determination of acceptable loads: some studies exist, but information on acceptable
loads should be available for all pathogens of major concern. Dose-response studies
would be useful in determining acceptable loads by defining the dose that results in
disease.
Antibiotic resistance should be considered as a potential concern in all bacterial
pathogens. Studies may need to be done to determine to what extent pathogens are
exhibiting resistance, and if so, whether it is increasing.
Development of models to predict die-off rates of pathogens in different environmental
conditions.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Specific Pathogens:
Studies are needed to show the impact of chloramines on Legionella.
More information is needed on pathogens associated with animal feeding operations, e.g.,
Campylobacter and Salmonella.
An assessment is needed on the potential public health impact of prions, which may be
associated with stream effluent near meat packing plants.
EPIDEMIOLOGY
In general, a better understanding of the epidemiology of aquatic pathogens was considered
necessary.
Endemic levels of pathogens should be monitored, to help recognize instances when a
pathogen reaches epidemic levels.
The characteristics of disease caused by each pathogen should be studied; this
information will be useful in tracing pathogens to the distribution system. Information
on the disease progression and secondary infection is also needed.
A system should be developed for better tracking of outbreaks, to help with regulatory
management. This would require the identification of new ways to assess disease
occurrence; hospital admissions, which is the indicator currently used, can give
unreliable estimates of disease occurrence.
Information is also needed on ways to deal with sensitive (young and elderly) and super-
sensitive (immuno-compromised) populations, as opposed to the general population,
when it comes to pathogens in drinking water.
DISTRIBUTION SYSTEMS
The characteristics of the system that could allow for intrusion should be studied in all
drinking water systems and all potential sources of pathogens external to the distribution
system identified (e.g., distance between water and sewer/utility pipes, pressure drops
that could allow pathogens to enter the system).
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
WATERSHED APPROACH
More information, as well as performance standards, are needed on the subject of
watershed protection.
Techniques are not clearly understood when it comes to bacterial source tracking in
watersheds. Some of the questions that still need to be answered are:
» When is bacterial source tracking feasible?
» What techniques work best for watersheds?
* What is the size limit (when is a watershed too big for bacterial source tracking to be
effective)?
» How far can the source library extend when performing these tests?
COMMUNICATION (ORD/REGIONS/LOCAL AGENCIES^
» Communication between ORD and the regions would allow newly developed science to
be a prominent part of the regulatory decision making.
» Communication with local agencies is also essential in reaching consensus regarding
aquatic pathogens and when/if they should be considered a public health priority.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
ORD'S RESPONSE TO REGIONAL RESEARCH NEEDS
On the last day of the workshop, a panel of ORD representatives addressed the research needs
identified by the regions throughout the workshop. The Office of Science Policy, as well as four
of the ORD National Laboratories and Centers were represented, and are listed below:
Office of Science Policy (OSP) - Molly Whitworth
National Risk Management Research Laboratory (NRMRL) - Gene Rice
National Health & Environmental Effects Research Laboratory (NHEERL) -
Rebecca Calderon
National Exposure Research Laboratory (NERL) - Gerry Stelma
National Center for Environmental Research (NCER) - Cynthia Nolt-Helms
OSP - Molly Whitworth - Introduction. Organizational Structure within ORD
The Office of Science Policy (OSP) coordinates the planning of ORD research. There are five
Research Coordination Teams (RCTs) within OSP - Water, Air, Waste, Pesticides/Toxics, and
Multi-Media - which are tasked with developing ORD research plans to support Agency science
needs. Review of the current research programs takes place from January to May of each year,
and recommendations are made on the priorities for future research funding.
Throughout the year, RCT members - Associate Lab and Center Directors, program office
representatives and regional representatives - meet regularly to share and discuss information.
Associate Lab Directors and Associate Center Directors are also available to give technical
assistance. Alternatively, regional needs and requests for assistance can be communicated via
the regional representative to each RCT.
Four of the ORD National Laboratories and Centers were represented in this discussion
addressing the needs identified by the regions (listed in the preceding section of this report).
Each representative directly addressed the research needs expressed by providing the answer to
three questions:
1. Which research topics and needs are currently being addressed by the laboratory?
2. Where are the gaps in data (as perceived by the ORD laboratory)?
3. In what ways can the regions communicate their research needs to ORD?
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
NRMRL - Gene Rice
Question 1: Which research topics and needs are currently being addressed by NRMRL?
The National Risk Management Laboratory is primarily oriented toward the engineering aspects
of water treatment. Ongoing research includes the following topics:
* Treatment strategies for the physical removal and chemical inactivation of
pathogens in drinking water and wastewater
* Microbiological methods for indicators and some pathogens, for measuring
occurrence
» Pathogen monitoring, tracing and watershed tracking
» Watershed management
* Models of:
Inactivation kinetics
Disinfection
Determining Ct (concentration over time) values
* Effects of biofilms on drinking water systems, such as:
Effect on total water quality
Presence of pathogens, especially opportunistic pathogens
* Regulation of pathogens from biosolids and Confined Animal Feeding Operations
Question 2: Where are the gaps in data (asperceived by NRMRL)?
* Data gaps exist in the removal efficiency for pathogens (efficiency is often
limited by the methods used).
Question 3: In what ways can the regions communicate their research needs to ORD?
* Meetings/workshops are important for continued interaction between ORD and
the regions.
* Often the research that addresses regional needs is not adequately communicated
to the regions - meetings are an effective way of doing so.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
NHEERL - Rebecca Calderon
Question 1: Which research topics and needs are currently being addressed by NHEERL?
Health Division of NHEERL:
* Maintaining a relationship with the Centers for Disease Control and Prevention,
especially in collecting data on outbreaks
* Data collection on endemic levels of disease, which can help deal with outbreak
situations as they arise
* Infectious dose studies are planned, especially for Cryptosporidium
* There is a new research initiative focusing on bathing beaches
Ecology Division of NHEERL:
* Programs involving landscape ecology, GIS and satellite imaging; NHEERL has
worked extensively with the regions on some of these projects
* Source tracking of enteric microbial organisms
* Database on Enterococci
* Methods development (especially DNA micro-arrays) in conjunction with NERL
* Microbial community structure and identification of the ecological niches of
pathogens
* "Coastal 2000" program: an extensive program dealing with marine environments
Question 2: Where are the gaps in data (as perceived by NHEERL)
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* Algal toxins - information was requested on whether this problem should be
considered in epidemiological studies
Question 3: In what ways can the regions communicate their research needs to ORD?
* Scientists and program directors are interested in communicating with the
regions, as regional problems can be incorporated into ORD studies
» Holding forums and meetings/workshops on a regular schedule
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
NERL - Gerry Stelma
Question 1: Which research topics and needs are currently being addressed by NERL?
* Occurrence data on hazardous organisms and chemicals
* Studies of human exposure to hazardous organisms and chemicals (a method is
still needed to measure human exposure)
» Development of monitoring tools, including:
Real-time PCR
DNA micro-arrays (using real-time PCR)
Flow cytometry using labeled antibodies (a new, bench-top unit has been
developed)
Biosensors to measure fecal indicators
Dose-response relationships, for both healthy and immuno-compromised
individuals
Better understanding of where indicators come from; this would be
especially useful for performing TMDLs and managing watersheds
Method for measuring Enterococci (which are more accurate indicators in
marine water than E. coif)
Biosensors to measure algal toxins; some technologies already exist but
cannot currently be used for water samples
Question 2: Where are the gaps in data (asperceived by NERL)?
* Better methods are needed for source tracking
» The best existing methods should be identified, and further research on those
methods supported
* Data is needed on the transport of microorganisms in manure, soil, sludge
Question 3: In what ways can the regions communicate their research needs to ORD?
* Holding workshops on a regular basis would facilitate communication -
especially after meeting in informal settings such as workshops
» Additional travel, both on the part of ORD and the regional labs would also
maintain the lines of communication
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
NCER - Cynthia Nolt-Helms
Question 1: Which research topics and needs are currently being addressed by NCER?
Current programs funded by NCER:
* Too many areas to present them all in this forum, check out our website and use
it's SEARCH feature to find specifics of interest to you: www.epa.gov/ncerqa/.
Some examples include:
* Drinking water program - pathogens research including:
Dose-response relationships for Cryptosporidium
Infectivity and virulence of Cryptosporidium in healthy adults
Development of indicators, such as a portable system for on-site detection
ofLegionella
* Recreational water program (a small program which may not continue)
» Hazardous algal blooms
* Water and watersheds
» Global climate change
Question 2: Where are the gaps in data (asperceived by NCER)?
New programs initiated by NCER in response to data gaps include:
* Risk from pathogens in drinking water
* Risk from pharmaceuticals
* Risk from tropical waters (recreational water)
* Risk from "swash zone" - interface zone of a beach where the waves and sand
meet (recreational water)
Question 3: In what ways can the regions communicate their research needs to ORD?
* Information on research funded is available on the NCER webpage:
www. epa. gov/ncerqa
Abstracts, annual progress reports, and final reports for all funded studies
are posted on the website
* Input (from both ORD and regions) can be communicated to NCER through
participation in relevancy reviews of grant proposals or by contacting any NCER
staff
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
* Outreach to regions to determine the best way of making research results known:
Progress Reviews where principal investigators provide presentations on
their research (annually/semi-annually, and at the end of the study)
Posting of publications on the website
Regional informational meetings being held at most regional offices by
NCER staff in FY01
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
WORKSHOP AGENDA
ORD/Regional Workshop on Emerging Issues
Associated with Aquatic Environmental Pathogens
September 5-7, 2001
Ft. Meade, Maryland
Wednesday, SEPTEMBER 5
Introduction
8:00 Registration
8:30 Welcome - Deborah Dietrich (ORD) and Rich Pepino (Region 3)
Workshop Goals/Logistics -Ron Landy (Region 3), Meeting Coordinator
Workshop Structure - Mick Kulik (Region 3), Meeting Facilitator
Science Support to EPA Programs and Regions
1. Where/how do emerging pathogens fit within the Agency mandates?
What are the scientific needs to support these mandates?
Chair. Jake Joyce (Region 7)
8:45 Overview of Emerging Pathogens - Jake Joyce (Region 7)
9:10 The Clean Water Act's Requirements for Pathogens -
Jim Pendergast (OW) IS]
9:30 Safe Drinking Water Act/Contaminants Pathogens Candidate List -
PaulBerger (OW) [PAGE 26]
9:50 Workgroup updates (15 minutes each)
Unregulated Contaminant Monitoring of Pathogens -Jim Sinclair (OW)
Update on EPA's Implementation Guidance of AWQC for Bacteria -
Jennifer Wigal (OW) [PAGE IS]
Developing a Strategy for Waterborne Microbial Disease Control -
Steve Schaub (OW) If]
10:35 Break
10:50 EPA Implementation of Beaches Environmental Assessment & Coastal Health
(BEACH) Act - Charles Kovatch (OW)
11:30 Q&A
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
2. Is our science on pathogens appropriate to support regulatory
management?
11:40 Summary of Major Regional Scientific/Technical Priorities -
Yield Binetti (Region 3) [PAGE 32]
11:50 Regional panel discussion - facilitated Q & A - Moderator: Vicki Binetti
Panel: Lisa Byron (Region 2), Joel Hansel (Region 4), Bob Benson (Region 8),
Howard Neukrug (Philadelphia Water Department) [PAGE 33]
12:30 Lunch
Incidence and Risk Assessment of Emerging Pathogens
1. What do we know about the nature and magnitude of waterborne disease
in the United States?
Co-chairs: Kim Ngo (Region 6) and Rebecca Calderon (ORD/NHEERL)
1:30 Waterborne Disease Outbreaks in the U.S. - Regional Issues -
Kim Ngo (Region 6) [PAGE 3?]
1:45 Waterborne Disease: EPA's Intramural Epidemiology Program -
Rebecca Calderon (ORD/NHEERL) [PAGE 38]
2:40 An Estimate of National Waterborne Disease Occurrence -
Susan Shaw (OW) 39]
3:00 Q&A
3:30 Break
2. Are our microbial risk assessment tools (epidemiological and
"toxicological") adequate to evaluate emerging pathogens?
Chair: Bruce Macler (Region 9)
3:45 Microbial Risk Assessment: State of the Art and Science -
Bruce Macler (Region 9) [PAGE 40]
Microbial Risk Assessment in other Agencies
4:00 A Dose-Response Envelope for E. coli O157:H7 -MarkPowell (USDA) [PAGE 41]
4:15 Vibrioparahaemolyticus Risk Assessment - Marianne Miliotis (FDA) [PAGE 41]
Microbial Risk Assessment at EPA
4:45 Pathogen Risk Assessment Protocol for Environmental Water Media -
Steve Schaub (OW) [PAGE 44]
5:00 Risk Assessment in Regulatory Development - StigRegli (OW) [PAGE 45]
5:15 Microbial Risk Assessment Activities in Cincinnati Division of EPA's National
Center for Environmental Assessment - John Lipscomb (ORD/NCEA) [PAGE 46]
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Thursday, SEPTEMBER 6
Incidence and Risk Assessment of Emerging Pathogens (continued)
8:30 (Two concurrent breakout group discussions) [PAGE 48]
1. What endpoints and factors should be considered in microbial risk
assessments? Moderator: Bruce Macler (Region 9)
2. Recognizing the uncertainties inherent in microbial risk assessment; what
data would provide the most value added? Moderator: Mike Messner (OW)
9:20 Breakout group reports - 10 minutes each
9:40 Q & A session with leads for breakout groups
10:00 Break
Assessing Exposures to Emerging Pathogens
1. How do we look for these organisms? What is the current status of
methods to monitor exposure?
Co-chairs: Bobbye Smith (Region 9), Gerry Stelma (ORD/NERL), and Fred
Genthner (ORD/NHEERL)
10:15 Assessing Exposures to Emerging Pathogens - What do the Regions Need?
Terry Fleming (Region 9) [PAGE 49]
10:35 Assessing Exposure to Bacterial Pathogens -Gerry Stelma (ORD/NERL)
10:55 Development of Molecular Methods to Detect Emerging Viruses -
Ann Grimm (ORD/NERL) SI]
11:15 Detection of Protozoan Pathogens in Water - Frank Schaefer (ORD/NERL)
11:25 Current Approaches to Identifying Sources of Fecal Contamination -
Gerry Stelma (ORD/NERL) S3]
11:45 Antimicrobial Resistance Among Enteric Bacteria Isolated from Human and Animal
Wastes and Impacted Surface Waters -
Mark Sobsey (University of North Carolina)
12:05 Q&A
12:20 Lunch
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
First of two concurrent sessions:
Emerging Pathogens in Drinking Water Risk Management Challenges
Co-chairs: Kim Harris (Region 5) and Gene Rice (ORD/NRMRL)
1:20 User Friendly Models for Evaluating Hydrogeologic Barriers to Viruses -
Bart Faulkner (ORD/NRMRL) 88]
1:40 Water Supply and Water Resources Division Response to Waterborne Disease
Outbreaks - Kim Fox (ORD/NRMRL) [PAGE 86]
1. What are the microorganisms of concern and how do they get into the
distribution system?
2:00 Disinfection of Emerging Pathogens - Gene Rice (ORD/NRMRL) [PAGE 87]
2:20 Biofilms in Drinking Water Distribution Systems -
MarkMeckes (ORD/NRMRL)
2. What is the drinking water distribution rule?
2:40 Update on Six-Year Review of TCR and Potential Distribution System Rule -
Ken Rotert (OW) 89]
3. How do we handle cross-contamination and biofilm problems in the field?
What are the inherent risks to distribution systems? Can we minimize the
risk? Case study example/panel discussion.
3:00 Case Study: Biofilm Problem at Greenville, South Carolina -
David Parker (Region 4) 60]
3:20 The View from the Field - Water Distributor's Perspectives -
Mark LeChevallier (American Water Works Service Company, Inc.) [PAGE 1]
3:40 Break
4:00 Panel Discussion -Moderator: Dave Parker (Region 4)
Panel: Kim Ngo (Region 6), Kim Harris (Region 5), Mark LeChevallier (American
Water Works Service Company), and Mark Meckes (ORD/NRMRL)
(Allparticipants reconvene)
4:45 ORD's Drinking Water Research Program Tracking System -
AlDufour (ORD/NERL) and Cheryl Itkin (ORD/NCEA) [PAGE 69]
5:00 Adjourn
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Second of two concurrent sessions:
Emerging Pathogens in Recreational Water Risk Management Challenges
Co-chairs: Nancy Grundahl (Region 3) andAl Dufour (ORD/NERL)
1:00 EPA's National BEACH Survey - Rick Hoffman (OW) [PAGE &\
1:40 Development of Improved Monitoring Approaches for Recreational Water -
Steve Schaub (OW) 4]
2:00 Use of Molecular Biology Techniques to Provide the Public with More Information
on Recreational Water Closures in Norfolk -
Alpha Diallo (Norfolk Public Health Lab) 6S]
2:20 Acceptable Goals and Thresholds for Pathogens from Recreational Water Exposures -
Al Dufour (ORD/NERL) [PAGE 66]
2:40 Determining Sources of Microbial Contamination - Regional Applied Research
Effort - Joel Hansel (Region 4) &}
3:00 Break
3:20 Rapid Methods for Measuring Recreational Water Quality: A Look to the Future -
Al Dufour (ORD/NERL) 68]
3:40 Facilitated Q & A with panel of previous speakers
(Allparticipants reconvene)
4:45 ORD's Drinking Water Research Program Tracking System -
Al Dufour (ORD/NERL) and Cheryl Itkin (ORD/NCEA) [PAGE 69]
5:00 Adjourn
5:00-7:00 Buffet Dinner and Poster Session *
Friday, SEPTEMBER 7
Pathogen TMDLs/CAFOs/Biosolids
Co-chairs: Stephanie Harris (Region 10) andJ. E. Smith, Jr. (ORD/NRMRL)
8:30 A Regional Perspective on CAFO/Biosolid Regulations: How ORD Can Help Us! -
Stephanie Harris (Region 10) [PAGE 71]
8:50 Overview of Watershed Features and Dynamics Associated with Drinking and
Recreational Water Protection - J. E. Smith, Jr. (ORD/NRMRL) [PAGE 72]
9:00 Potential Watershed Movement of Pathogens of Animal Origin -
John Cicmanec (ORD/NRMRL) [PAGE 73]
9:25 Emerging Infectious Disease Agents and Issues Associated with the Management of
Treated Sludges (Biosolids) - J. E. Smith, Jr. (ORD/NRMRL)
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
9:50 Risks from Pathogens in Biosolid Fertilizers/Land Application of Sludge - NRC
Report - Tom Burke (Johns Hopkins University) [PACs-E 78]
10:10 Q&A
10:30 Break
Over the Scientific Horizon Meeting Regional Research Priorities
Moderator: Paula Estornell
Presentations by ORD panelists based on previous days' discussions and questions, especially
those raised during first day session on "Is our science appropriate... "
10:45 Molly Whitworth - Office of Science Policy (OSP)
10:55 Gene Rice - National Risk Management Research Laboratory (NRMRL) [PACs-E 10]
11:05 Re becca Colder on - National Health and Environmental Effects Research
Laboratory (NHEERL) II]
11:15 Gerry Stelma/Al Dufour - National Exposure Research Laboratory (NERL)
11:25 Cynthia Nolt-Helms - National Center for Environmental Research (NCER)
13]
11:35 Q & A and Discussion with Panel
12:00 Closing Session - Rich Pepino (Region 3)
What have we learned from this workshop? Do we need new research initiatives?
Are there specific emerging pathogens or situations that merit regulation?
What science do we need to support this?
Were the goals of the workshop achieved? Where do we go next and how?
Discussion
12:30 Adjourn
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
*POSTERS
5:00 - 7:00 p.m. September 6
Ft Meade Golf Course Club House
"Identification of Sources of Fecal E. coli in Minnesota Watersheds Using rep-PCR DNA
Fingerprinting" - Priscilla Dombeck (University of Minnesota)
"Alpine E. coli O157 Outbreak" - Sandy Spence (Region 8)
"Calicivirus Outbreak at Big Horn Mountain Lodge" - Barbara Barron (Region 8)
"Enterococci in the Water Column and Shoreline Interstitial Waters at Beaches on the
Gulf of Mexico and Santa Rosa Sound, Pensacola Florida" - FredGenthner
(ORD/NHEERL)
"NARMS (National Antimicrobial Resistance Monitoring System) Relevance to
Environmental Pathogens" - Linda Silvers (FDA Center for Veterinary Medicine)
"Identifying Sources of Fecal Coliform Bacteria in Selected Streams on Virginia's
TMDL Priority List" - Ken Hyer (VA USGS District)
"Survey of U.S. Public Health Laboratories: Detection of Microbial Pathogens of the
Contaminant Candidate List"- Elizabeth Hilborn (ORD/NHEERL)
"Waterborne Pathogen Best Management Practices in Agricultural Watersheds" - Barry
Rosen (USDA/National Resources Conservation Service)
"Antimicrobial Resistance Among Bacteria Isolated from Human and Animal
Wastes and Impacted Waters: Comparison with NARMS Findings" - D. Cole, D.
Robinette, J. Bumgarner andM. Sobsey
""Biological Assessment of Beach, Leachate, and Effluent Sites at Lake Texoma"- Mary
E. Gonsoulin (ORD/NRMRL)
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Welcome: Deborah Dietrich (ORD) and Rich Pepino (Region 3)
Workshop Goals/Logistics: Ron Landy (Region 3), Meeting Coordinator
Workshop Structure: Mick Kulik (Region 3), Meeting Facilitator
Science Support to EPA Programs and Regions
Where/How do emerging pathogens fit within the Agency mandates? What are the
scientific needs to support these Mandates?
Chair: Jake Joyce (Region 7)
Overview of Emerging Pathogens - Jerome (Jake) Joyce (Region 7)
Emerging pathogens are microbes that have relatively recently come into the view of the health
sciences as a cause of disease. These pathogens can be a relatively newly-discovered agent of
infection such as Legionella bacteria causing Legionnaire's disease. An emerging pathogen may
also be a relatively well-known agent that changes in virulence such as the E. coli O157:H7
strain acquiring the Shiga toxin. Emerging pathogens can also result from the importation of a
new infectious agent such as West Nile Virus, which is well-known in the old world, but recently
introduced into the new world. Emerging pathogens can be re-emerging disease agents such as
yellow fever or malaria which have been eradicated in the continental U.S. but can make a
comeback due to global weather changes.
Emerging pathogens can be engendered by a complex mixture of factors, such as:
Social factors: e.g., changing moral values, migration of human populations;
Political factors: e.g., warfare or oppressive governments, resulting refugee
populations;
Economic factors: e.g., prioritization of health funds, access to primary health care
and modern sanitation;
Ecological factors: e.g., urbanization, land use changes, climate changes; and
Technological factors: e.g., rapid transportation, bio-engineered pathogens.
The factors above are not freestanding, but are all intertwined with each other.
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Finally, emerging pathogens can arise among an immuno-compromised population. An
opportunistic infectious agent is one that is not able to harm a healthy individual, but can
produce disease in a person with compromised immune status.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
The Clean Water Act's Requirements for Pathogens - James Pendergast (OW)
The Federal Water Pollution Control Act amendments of 1972 (commonly known as the Clean
Water Act) provide a comprehensive approach for protecting water quality of the nation's waters
from chemical, physical, and biological stresses, including pathogens. This presentation
described how the Act has been used to assess and control pathogens, and has served as the basis
for addressing emerging environmental issues, both in the area of human health and aquatic life.
The Clean Water Act (CWA) utilizes a series of steps in controlling pathogens. The Act states
that water quality criteria must be established by the EPA for pathogens and indicators in
recreational and coastal waters. The states then develop standards for water considering both
public health and recreation. The states also identify those waters which do not meet their
standards, in compliance with Section 303d of the CWA. A listing is assembled of these waters,
known as the "303d" list, and Total Maximum Daily Loads (TMDLs) must be determined for the
pathogens or pollutants which are determined to be causing the impairment. Once established,
the TMDLs are used to determine what actions are needed to return the body of water to the
purposes designated by the state.
The CWA National Pollutant Discharge Elimination System (NPDES) permit program exists to
improve water quality by regulating point-sources of pollution. Compliance with the Act is
monitored and enforced by EPA. Additionally, the states operate programs to abate nonpoint
sources of pollution using funds supplemented by EPA under Section 319 of the CWA.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Safe Drinking Water Act and Contaminants Pathogens Candidate List - Paul
S. Berger (OW)
The 1996 amendments to the Safe Drinking Water Act (SDWA) require EPA to regulate any
contaminant that: 1) causes an adverse health effect; 2) occurs (or probably occurs) in drinking
water at a frequency or concentration that warrants public health concern; and, 3) provides a
meaningful opportunity to improve public health protection. The SDWA also requires EPA to
publish a list every 5 years of non-regulated contaminants that occur, or probably occur, in
drinking water and may need to be regulated. This list, known as the Contaminant Candidate
List (CCL), becomes the focus for increased research and regulatory attention. According to the
SDWA, within 5 years of publication of each CCL, EPA must decide whether or not to regulate
at least 5 of these contaminants. In response to this Congressional mandate, EPA published a list
of contaminants (CCL) in the Federal Register on March 2, 1998 (63 FR 1027410287). The
pathogens on the CCL include:
Protozoa: Acanthamoeba, Microsporidia (Enterocytozoon, Encephalitozoori)
Viruses: Caliciviruses, adenoviruses, coxsackieviruses, echoviruses
Bacteria: Helicobacter pylori, Mycobacterium avium complex, Aeromonas
hydrophila
Algae: Algae and their toxins
Many, perhaps nearly all, of the contaminants on the CCL cannot be regulated in a cost-effective
manner without research into their health effects, occurrence in water, susceptibility to water
treatment processes, effect on sensitive subpopulations, risk assessment, and/or the utility of
inexpensive surrogates for their presence. Therefore, to allow the Agency to make a
scientifically defensible decision on whether to regulate, the contaminants on the CCL are being
given high priority with regard to drinking water research. (This situation, however, will not
foreclose on research or regulations for contaminants not on the CCL.)
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Unregulated Contaminant Monitoring of Pathogens - James L Sinclair (OW)
The 1996 amendments to the Safe Drinking Water Act authorize monitoring of unregulated
contaminants in drinking water. This provision is being implemented as the Unregulated
Contaminant Monitoring Rule (UCMR). Monitoring will be done for contaminants that lack
sufficient occurrence information. The occurrence data from unregulated contaminant
monitoring will be used with other information to make regulatory determinations for
contaminants in drinking water. Contaminants to be monitored will be selected from the
occurrence priorities list of the Contaminant Candidate List (CCL). Of the 10 CCL pathogens,
eight (8) are listed as occurrence priorities:
Algae and toxins Echoviruses
Microsporidia Coxsackieviruses
Aeromonas hydrophila Adenoviruses
Helicobacter pylori Caliciviruses
There are three types of unregulated contaminant monitoring based on analytical methods'
availability for contaminants. The first monitoring option is assessment monitoring and will
include all water systems serving more than 10,000 people plus a representative sample of 800
small systems (serving fewer than 10,000 people). Assessment monitoring will be done for
contaminants with methods that were well-developed and validated when the UCMR was
promulgated in September 1999. Sampling will be four times a year for surface water systems
and two times a year for ground water systems. The next monitoring option is the screening
survey. It includes a representative sample of 120 large systems and 180 small systems for a
total of 300 systems. Screening surveys will be done for contaminants that will have methods
ready by the time of screening survey monitoring. Sampling times and frequency are flexible.
The third monitoring option is the prescreen survey. It includes up to 200 systems which are
vulnerable to contaminant occurrence. Samples will be taken at vulnerable times, if possible.
The prescreen survey is for contaminants that don't have well developed methods, or have
methods that are expensive.
No CCL microbial contaminant has methods that could be used for assessment monitoring.
Aeromonas has a method that can be used with a screening survey. The 7 remaining CCL
microorganisms will be monitored with prescreen surveys when methods that can be used with
these surveys are available. A lack of contaminant occurrence information may hinder designing
a prescreen survey for vulnerable times and locations. Future work needed for UCMR
monitoring of CCL pathogens is method development and preliminary surveys of source water
and treated water. Preliminary surveys of pathogens will assist in the design of UCMR surveys.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Update on EPA's Implementation Guidance for Ambient Water Quality Criteria
for Bacteria - Jennifer Wigal (OW)
The presentation provided an update on EPA's progress toward finalizing the Implementation
Guidance for Ambient Water Quality Criteria for Bacteria - a document developed in response
to commitments made in the Beach Action Plan (1999). The draft of the Implementation
Guidance was distributed for public comment on February 14th, 2000. Key topics included the
transition from fecal coliform bacteria as indicators to E. coli in freshwater and Enterococci in
marine water, as well as a new recommendation to apply the criteria to waterbodies, rather than
sources of contaminants. Additionally, the application of the criteria to tropical waters was
recommended by EPA unless an alternate indicator is developed. A policy recommendation for
secondary contact waters (used for activities such as boating) was also included in the document.
Approximately fifty comment letters were received from states, industry, and environmental
groups, addressing all areas of the guidance, but focused especially in the following areas:
Science underlying the criteria;
Application of criteria to non-human sources;
Application of criteria during high flows;
Monitoring; and
Extent of state flexibility in implementation.
The EPA workgroup is still in the process of revising the guidance - which is scheduled to be
completed in the Fall of 2001 - based on discussions of the comments received. Additional
revisions will include the relationship of the criteria implementation in WQS and BEACH
programs, and considering the relative risk from non-human sources (i.e., how much is coming
from each source?).
Finally, EPA's recommended Water Quality Criteria for Bacteria were presented for fresh and
marine water, for designated beach areas, as well as moderately to infrequently used full-body
contact recreation areas.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Developing a Strategy for Waterborne Microbial Disease Control - Stephen
Schaub (OW)
In 1998, pathogens were the second most frequent cause of impairments to waters under Section
303(d) of the Clean Water Act. These impairments are primarily due to the increasing
populations of humans and livestock, and their wastes. EPA has a number of existing mandates,
regulations and initiatives such as the Interim Enhanced Surface Water Treatment Rule and
EPA's Beach Action Plan which are important in reducing health risks from pathogens. EPA is
adopting a strategy for future actions to integrate the protection of public health under both the
Safe Drinking Water Act and the Clean Water Act. The strategy will guide decisions on the
most effective use of available resources, support communication and partnerships with
stakeholders, and enhance public communication.
The specific goals of the strategy include identification of priority activities for:
1) The recognition of significant known pathogens and emerging pathogens,
2) Identification and control of pollutant sources so that waters will meet protective use
criteria,
3) Coordination of regulatory and research activities, regulatory approaches and R&D
program support, and
4) Participation of public agencies and stakeholders.
The approaches to be used in meeting these goals were outlined as well:
Develop Ambient Water Quality Criteria, as well as monitoring protocols, and
indicators for pathogens.
Establish risk assessment guidelines for the Agency.
Establish treatment requirements or discharge criteria, as well as monitoring
requirements for reused water and unregulated industrial water.
Determine effectiveness and establish model management practices for
contamination sources.
Further areas of concern not immediately addressed by the strategy included pathogens found in
sediments, ecosystem microbiology, risks associated with animal-borne pathogens, and algal
toxins in both marine and fresh water.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
EPA Implementation of the Beaches Environmental Assessment and Coastal
Health (BEACH) Act - Charles Kovatch (OW)
Swimmers need to know about microbial contamination at beaches to minimize their exposure
and reduce swimming-related illness. Recreational water quality monitoring and public
notification programs vary from state to state with respect to indicator organisms, sampling
location and frequency, and beach postings and advisories. In an effort to protect public health
and improve beach monitoring and public notification, Congress passed the Beaches
Environmental Assessment and Coastal Health (BEACH) Act in October 2000 to amend the
Clean Water Act; the BEACH Act assigned specific duties to EPA and the states, which were
outlined in this presentation. The provisions of the BEACH Act apply to coastal recreation
waters located in coastal and Great Lakes states.
As required by the BEACH Act, states must adopt EPA's current recreational water quality
criteria by April 2004. Also, EPA (specifically ORD) is conducting research on health risks
associated with exposure to pathogens in recreational waters, and improved and rapid detection
methods for pathogen indicators. This research is scheduled to be completed by October 2003,
and EPA will promptly publish revised water quality criteria based on the research by October
2005. Once EPA publishes the new recreational water quality criteria, states will have three
years to adopt the criteria.
In order to address the inconsistency in beach monitoring and notification approaches among and
within states, the EPA must publish performance criteria by April 2002 for:
Monitoring and assessment procedures for pathogens and pathogen indicators, and
Notification of exceeding or likelihood of exceeding recreation water quality
standards.
To be eligible for BEACH Act grants states must be consistent with the criteria when developing
and implementing their beach monitoring and notification programs. Currently, 34 out of 35
eligible states have applied and been awarded FY01 Development grants.
As part of its role in the implementation of the BEACH Act, EPA is also required to "establish,
maintain, and make available to the public a national coastal recreational water pollution
occurrence database" by October 2003. The database will list the coastal recreation waters
subject to monitoring and notification under the BEACH Act, and their status.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Additional provisions of the BEACH Act include:
State development and implementation of a program by 2003 for jurisdiction over
coastal recreational waters;
EPA provides technical assistance for assessment and monitoring of floatable
material; and
EPA implementation of monitoring and notification program when a state program is
inconsistent with performance criteria.
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Is our science on pathogens appropriate and adequate to support regulatory
management?
Summary of Major Regional Scientific/Technical Priorities - Vicki Binetti
(Region 3)
EPA regional offices, in conjunction with state regulators and state health departments, are
responsible for implementing legal mandates to keep pathogens out of the drinking water
system. At the regional level, the goal of regulation is to protect public health - guidelines are
always being developed to be used for that purpose.
Regional needs that always exist include:
Identification of sources of contamination, and guidelines on how to measure and
monitor their impact;
Information and guidance on enforcement and correcting of violations; and
Guidelines on dealing with potential outbreaks / contamination / boil-water alerts.
At the end of this presentation, the panel of speakers participating in the discussion to follow was
introduced. The panel members were Joel Hansel (Region 4), Lisa Byron (Region 2), Bob
Benson, (Region 8) and Howard Neukrug (Philadelphia Water Commission). Each panel
member briefly addressed the question: "Is our science on pathogens appropriate and adequate
to support regulatory management? "
A summary of the scientific needs expressed during the panel presentations, and the
ensuing discussion, can be found in Appendix D.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Regional Panel Discussion and Facilitated Q & A
Joel Hansel - Region 4
A clear distinction between recreation and consumption or ingestion (drinking water) would
facilitate regulatory management of aquatic pathogens. Disease endpoints other than the
gastrointestinal symptoms traditionally considered should be taken into account; Pfiesteria is an
example of a pathogen (in recreational water) whose health effects include skin rashes, as well as
neurological symptoms.
In the case of Total Maximum Daily Loads, the cause of the impairment has to be determined;
bacterial source tracking can sometimes be used to make that determination. A case study was
presented documenting the contamination with fecal coliform bacteria of a small watershed in a
rural area. The surrounding land was used primarily for small-scale agriculture and keeping
livestock. Leaking septic tanks were initially assumed to be the source of the contamination.
Bacterial source tracing was conducted after a library of potential sources was prepared. It was
determined, as a result, that the source of the contamination was the presence of livestock.
Particularly, in light of the fact that the livestock were not contained and had free access to water
bodies.
Fecal coliform levels decreased soon after the livestock were fenced and prevented from
accessing the watershed. A new problem was encountered, however, when the increased
presence of wildlife caused fecal coliform levels to increase again. As wildlife was not
perceived to be a health concern - coliform levels were still lower than they had been - no steps
were taken to keep wildlife away from the watershed.
Research needs mentioned, particularly with respect to bacterial source tracing, included:
When is a watershed too big for bacterial source tracing to be effective?
How far can a source library extend, and how many isolates are needed for
each source?
What technique works best, and can it be done faster?
What is the risk associated with wildlife?
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Lisa Gray Byron - Region 2
The New York City watershed was presented as an example of the special concerns associated
with an unfiltered water supply, particularly the need to protect the entire watershed.
Cryptosporidium and Giardia are two pathogens that the New York City watershed team is
seeking to control, although at present there is a lack of science that can be used to evaluate the
watershed for the presence of these two pathogens. New York City is currently concentrating
efforts in performing a risk assessment on its watershed, and risk can be better assessed if the
bacterial load can be characterized.
Science support needed for these purposes includes:
Studies on the die-off rates of pathogens, and on the distance they are able to travel;
to be used possibly in constructing models;
Acceptable load information for the entire watershed (i.e., from all sources
combined);
Recovery methods to detect pathogens in soil;
Evaluation of best management practices (BMPs), e.g., buffer strips; and
Better detection methods. An example that illustrates this need is the existing method
for total coliform bacteria which can give false positive results in the presence of
Aeromonas.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Bob Benson - Region 8
Inadequate science is a problem often encountered in regulatory management, and is
compounded by the fact that waiting for a method to be validated by science is not always
feasible for the regions. Examples used to illustrate this point included:
A case where data were misinterpreted because of a problem with the methodology
for Cryptosporidium monitoring;
A biofilm problem traceable to problems with methods used in routine monitoring;
An E. coli O157 outbreak which occurred eleven years ago, likely because E. coll
O157 is not detected by routine E. coli monitoring;
An outbreak of calicivirus at a snowmobile park was traced to drinking water
contamination using a research method (not a validated method); and
In the case ofAeromonas, it is not always clear whether it is due to drinking water
contamination or an isolated case. There is a method for measuring Aeromonas, but
it will not be a part of routine, regulated monitoring until 2003.
Regardless of whether the science behind the methods has been validated, the regions frequently
have to make regulatory decisions (e.g., issuing boil orders, or performing disinfection) without
the benefit of the most advanced scientific knowledge.
Information that is helpful in making those decisions includes:
Characteristics of the disease, especially for tracing a potential outbreak to the
distribution system;
Information on any highly susceptible population (e.g., immuno-compromised
individuals); and
Information on disinfection, especially for systems that do not disinfect on a regular
basis.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Howard Neukrug - Philadelphia Water Commission
Along with the common goals of EPA and state regulators - safe drinking and recreational water
- consideration must also be given to the public perception of the quality of drinking water. It is
not realistic that good methods will always be available (once a method is perfected, another
pathogen may then emerge as a disease-causing organism). Watersheds can be protected to an
extent, but the sources of pathogens cannot be completely eliminated. It is important to know
the endemic, as opposed to the epidemic (outbreak), levels of pathogens to be able to inform the
public about potential problems. Additional ways of disease transmission, other than
waterborne, should also be considered.
A multi-sided approach to the problem of pathogens in drinking water is essential, paying
particular attention to source water protection.
Specific science needs include:
Monitoring and tracking the incidence of pathogens at endemic levels;
More research on the effects of disinfection; and
Operator training, especially in dealing with outbreak levels.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Incidence and Risk Assessment of Emerging Pathogens
What do we know about the nature and magnitude ofwaterborne disease in the United
States?
Co-Chairs: Kim Ngo (Region 6) and Rebecca Calderon (ORD/NHEERL)
Waterborne Disease Outbreaks in the U.S. - Regional Issues - Kim Ngo
(Region 6)
Statistics kept by the Centers for Disease Control and Prevention (CDC) over the past twenty
five years show that most waterborne disease outbreaks (86% of 371 outbreaks) can be
associated with source water contamination. The source water was usually ground water. Some
outbreaks (12%) can be traced to the distribution system. All of the water systems involved
were in compliance with the Safe Drinking Water Act, and half of them (50%) were practicing
disinfection. Protozoa, particularly Giardia and Cryptosporidium, bacteria (such as E. coif), and
viruses were the most common pathogens.
Following a storm event in July 1998, lighting struck a wastewater lift station causing 167,000
gallons of raw sewage to spill into Brushy Creek. Samples were taken from the distribution
system and tested negative for total coliform bacteria. When ground water was tested, four out
of five wells were found to be positive for E. coli. The Texas Department of Health was notified
of a potential outbreak situation. Following further studies by the Health Department it was
determined that 6,000 people were exposed, 25% of which became ill. Eighty five confirmed
cases of cryptosporidiosis were reported during the outbreak, and another forty five from
September to December of 1998.
The multiple barrier approach is a multi-faceted approach in protecting drinking water, which
involves identifying all the "barriers" - components of the drinking water system where
pathogens can enter - and taking steps to protect each one. Brushy Creek and other recent
outbreak cases can be attributed to one component or "barrier" failing to prevent pathogens from
entering.
Better reporting of outbreaks is one of the foremost regional needs mentioned - currently,
reporting is mandated only by the states, and each state's list of reportable diseases is different.
Keeping such data, especially on endemic levels of disease, will allow occurrence trends to be
monitored, and potential outbreak situations to be recognized. One suggestion made to improve
reporting was to possibly include an enforcement team within the EPA's investigative team.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Waterborne Disease: EPA's Intramural Epidemiology Program - Rebecca
Calderon (ORD/NHEERL)
The current epidemiology program is focused primarily on evaluating endemic (a high
background prevalence of disease) and epidemic (a temporal excess of illness over some
background level) disease associated with drinking water microbes. Waterborne disease
surveillance has been a joint activity between the EPA and the Centers for Disease Control.
Information is collected on outbreaks or epidemics associated with drinking water, recreational
water, and outbreaks aboard ocean-going vessels. However, using a self-reporting, or passive
surveillance system, can result in under-reporting and inconsistency in reporting. New etiologic
agents that have emerged in studies of outbreaks include Giardia, Cryptosporidium, Legionella,
andE. coliO157.
In addition to the epidemic work, there are two basic approaches to evaluating endemic
gastrointestinal illnesses associated with exposure to drinking water microbes. The first
approach is an evaluation of "acute gastrointestinal illness", in which illness is measured by
study participants' self-reported symptoms. In order to evaluate the risk attributable to water,
community intervention studies have been conducted or are in progress in which illness rates are
measured before and after a community upgrades community drinking water treatment to reduce
exposure to microbes in drinking water. The second approach is to evaluate health effects for
specific organisms such as Cryptosporidium. Serological surveys in paired cities involve
systematic collection and antibody-testing of blood samples from representative populations in
the exposed and unexposed cities. The Cryptosporidium serological work suggests that some
exposure may be protective - 2 and 1/4 times higher portion of the population is antibody
positive - and that efforts to reduce exposures through drinking water treatment may lead to
more disease and outbreaks. Other organism-specific work is being conducted on some of the
Candidate Contaminant List microorganisms, specifically by testing stool samples of volunteers
before, during, and after illness.
Intervention studies are conducted on the same population (before and after intervention) and
one such study has shown that up to 35% of microbial gastroenteritis can be linked to drinking
water. After intervention at the community level, gastrointestinal disease rates decreased.
Future work planned includes continued monitoring of outbreaks, as well as serology (paired-
city) and intervention studies. Microbe-specific studies will focus on the Contaminants
Candidates List pathogens. Finally, epidemiology studies are planned in response to the October
2000 passage of the BEACH Act; and NHEERL has begun discussion on the formulation of a
pathogen strategy.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
An Estimate of National Waterborne Disease Occurrence- Susan Shaw (OW)
Section 1458(d)(l) of the Safe Drinking Water Act requires EPA and CDC to conduct
epidemiology studies in five communities, and to prepare a report on the findings of the studies
and on the national occurrence of waterborne disease. In response, EPA and CDC will conduct
disease occurrence studies focusing on infectious disease, specifically gastrointestinal (GI)
illness and specific pathogens. The National Estimate will focus on drinking water related
gastrointestinal illness.
Several studies are ongoing or have recently been completed, including:
Household Intervention Studies, including a "special population" intervention study;
Community Intervention Studies;
A national survey study (FoodNet); and
A study on childhood cryptosporidiosis and drinking water.
In order to estimate national occurrence of waterborne disease, the gastrointestinal disease rate
(from all causes) is needed, as well as an approximation of the fraction of disease that can be
attributed to drinking water. Several studies (Payment '91, Colford '01, Calderon '01) estimate
that 30% of all GI disease is drinking water attributable - that fraction being most accurate for
highly challenged drinking water systems. A national telephone survey study (FoodNet) is being
conducted to determine the total rate of GI disease. A recent estimate, based on findings of the
studies mentioned above, predicts the national rate of waterborne GI disease to be 211 million
cases per year, or 0.79 cases per person per year. Similar preliminary estimates have been
developed using an attributable fraction of 15% and 5% to account for better drinking water
systems.
As ongoing epidemiology studies are completed and more data is gathered through FoodNet, the
National Estimate will become more representative of the entire population; an updated report on
the National Estimate is scheduled for 2003. Analysis of the data will also be done by
categories, such as the presence or absence of a Public Water System, the type of source water,
and the region.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Are our microbial risk assessment tools (epidemiological and "toxicological") adequate
to evaluate emerging pathogens?
Microbial Risk Assessment: State of the Art and Science - Bruce Macler
(Region 9)
Microbial Risk Assessments are used to collect relevant information that may be used in
management decisions involving the control of aquatic pathogens; information such as
estimating disease incidence rates, predicting levels of specific diseases, and evaluating the
effect of regulatory measures can be obtained using microbial risk assessments.
Approaches used in risk assessment can be based on epidemiology, to provide direct,
quantitative data; on system analysis, for qualitative assessments of vulnerability; or on dose-
response or exposure modeling.
Experimental epidemiology studies can include clinical studies, such as determining the
infective dose for a specific pathogen; or population intervention studies, to estimate the
endemic waterborne disease risk from drinking water. Other epidemiology studies are
observational, such as disease surveillance studies. Even though they provide quantitative data,
epidemiology studies are limited by confounding factors (human variation, number of
participants, other factors), and do not always show a clear connection between cause and effect.
System analysis methods, specifically Hazard Assessment Critical Control Point (HACCP)
methods, use a non-quantitative approach to assess vulnerabilities in a system. Once hazards are
analyzed, control points are established at key points in the system and monitored, taking
corrective actions when necessary. This procedure is currently being used widely in the food
industry to comply with the Food and Drug Administration's (FDA) regulations.
Dose-response modeling is based on the toxicological risk assessment approach for chemicals.
Dose-response data for infectivity are typically derived from healthy, adult populations, and
therefore would have to be corrected to include sensitive populations. Relevant disease
endpoints could be used to that effect. Mixtures of organisms may also need to be considered in
dose-response modeling.
Microbial exposure assessments take into account the routes of exposure, and the levels and
distribution of the organisms of concern, as well as the relative risk posed by each pathogen.
Estimates of occurrence are needed for these studies, as well as some information on pathogen
viability.
All of the approaches mentioned above are used in making regulatory decisions; each can be
used to clarify certain risk questions, but not others. Further research is needed to improve all
approaches currently used in microbial risk assessment.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
A Dose-Response Envelope for E. coli O157:H7 - Mark Powell (USDA)
The objective of the research conducted was to develop a dose-response model for illness by E.
coli O157:H7 that bounds the uncertainty in the dose-response relationship for the organism.
No human clinical trial data are available for E. coli O157:H7 dose-response; however, data
exist for two surrogate pathogens: enteropathic E. coli (EPEC) and Shigella dysenteriae. Using
that data, and E. coli O157 outbreak data, a most likely value of the dose-response for E. coli
O157:H7 can be estimated - and was inferred to be somewhat higher than S. dysenteriae and
somewhat lower than EPEC. The predicted exposure distribution of E. coli O157:H7 was
determined (by another model) for the approximate number of ground beef servings consumed
annually in the United States. Using both the predictive exposure distribution model, and the
dose-response data, an estimated distribution can be derived for the annual number of cases of
E. coli O157:H7 that can be attributed to ground beef.
Some recognized sources of under-reporting include ill persons not seeking medical care,
physicians not obtaining cultures from all patients thought to be infected, and laboratories not
culturing all stool samples for E. coli O157:H7. The uncertainty due to each of these sources of
under-reporting was characterized (using a variety of epidemiological data) and the estimates
were corrected accordingly to take under-reporting into account.
The dose-response envelope modeling concluded that at a dose level of 100 colony forming units
(cfu), fewer than fifteen persons exposed would become ill. The median response rate was
predicted at six percent, meaning that six percent of persons exposed are likely to become ill.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Vibrio parahaemolyticus Risk Assessment - Marianne Miliotis (FDA/OFSAN)
Vibrioparahaemolyticus (Vp) is an important agent of diarrhea associated with consumption of
seafood. It has been isolated from clinical sources, as well as nonclinical sources including
estuarine and marine environments in many regions of the world. Since it was first isolated in
1950, it has accounted for about 40-60% of all foodborne illnesses in Japan. Up until recently,
the allowable numbers of Vp associated with oysters was 104 Vp/m\. Studies following recent
outbreaks in 1997 and 1998 estimated infectious doses to be as low as 100 to 1000 organisms.
The FDA conducted a risk assessment on the potential public health impact of Vibrio
parahaemolyticus infections resulting from the consumption of raw molluscan shellfish, in
response to these outbreaks. The objectives of the risk assessment were twofold: 1) to produce
a mathematical model of the risk of illness incurred by consumers of raw oysters containing
pathogenic Vp; and 2) to provide FDA with information that will assist the agency with the
review of current programs relating to the regulation of Vp in raw molluscan shellfish to ensure
that such programs protect the public health.
The risk assessment was conducted according to the conceptual framework recommended by
CODEX, which involves four steps:
1) Hazard Identification: the identification of pathogenic Vp as the hazard;
2) Exposure assessment: the likelihood and levels of ingesting Vp by eating raw
molluscan shellfish harboring this organism;
3) Hazard Characterization/Dose-Response: the relationship between the levels of Vp
ingested, and the frequency and magnitude of illness; and
4) Risk Characterization: the integration of dose-response and exposure assessments to
assess illness.
This risk assessment is a product pathway analysis and is divided into three modules: Harvest,
Post Harvest, and Public Health. Because of significant differences in oyster harvesting
methods, handling practices, and climates within the United States, five geographic regions
(Northeast Atlantic, Mid-Atlantic, Pacific Northwest, Louisiana Gulf Coast, and the remaining
Gulf Coast) for each of the four seasons were modeled separately for the three modules. Like all
risk assessments, because of data gaps and uncertainty, assumptions had to be made in order to
conduct the risk assessment.
The risk assessment provided estimates of risk for illness among consumers of raw oysters of
4,750 cases per year, ranging from 1,000 to 16,000 cases. Risks increase with increasing levels
of total Vp and, therefore, increasing levels of pathogenic strains of Vp.
Water and air temperatures at the time of harvest were found to be the major factors influencing
the initial levels of this pathogen in oysters. Air temperature was also found to influence the
growth of Vp in oysters after harvest and, thus, the levels in oysters at the time of consumption.
The single most important factor related to the risk of illness caused by this organism is the level
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of Vp in oysters at the time of harvest. Intervention measures aimed at controlling or reducing
the levels of Vp in oysters such as mild heat treatment (5 min at 50°C) of oysters, practically
eliminates the likelihood of illness occurring. Quick-freezing and frozen storage of oysters, which
causes a one to two log decrease in viable Vp oyster levels, also substantially reduces the
probability of illness. Rapid refrigeration after harvest also significantly reduces risk of illness.
The risk assessment also suggested that approximately 15% of the illnesses were associated with
the consumption of oysters containing greater than 10,000 Vp/g at the time of harvest.
The model was validated using "real-world" data not incorporated in the risk assessment, and
comparing model predicted levels with actual survey levels recorded by a 1998-1999 FDA study.
The model predictions were in good agreement with the data recorded in the study, confirming the
validity of the model.
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Pathogen Risk Assessment Protocol for Environmental Water Media - Stephen
Schaub (OW)
EPA's Office of Science and Technology (OST) has been working through the process of
developing a microbiological risk assessment protocol for water that accounts for all of the unique
features of microbial pathogens and human infectious disease, while remaining consistent with
common threads of risk assessment from the existing chemical and ecological risk assessment
protocols. In microbial assessments for water, human infection attributes to be considered are
specific to microorganisms (growth and die-off, strain variability in infectivity, occurrence and
risk) and to human exposure (immunity, variable infection endpoints, secondary spread). The
pathogen risk assessment protocol has adopted the approach of ecological risk assessment protocol,
and is consistent with major food microbiology components of the Codex Alimentarius.
Model development is the first step in the process and serves the purpose of formulating the
problem. Risk characterization in this model would include characterization of exposure
(pathogen) and characterization of human health effects (host).
The goal of the protocol, once completed, is to be used as a non-regulatory agency protocol for
conducting risk assessments, as are the models currently used for providing ecological and
chemical guidance.
In order for the protocol to be validated, additional research is needed to fine-tune the model, such
as:
Testing to validate protocol for different types of pathogens (e.g., Contaminants
Candidate List pathogens) and in all media;
Risk analysis tools such as models on human susceptibility, dose-response, animal and
in-vitro models and mechanisms of infection;
EPA's risk characterization procedures incorporated into the model;
Risk assessment procedures that are in a form useful to risk managers and the public;
Procedures to assess environmental factors' impact to infectivity; and
Approach to deal with exposure doses and duration.
EPA is also involved in refining the existing system for implementing microbial risk assessment.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Risk Assessment in Regulatory Development - Stig Regli (OW)
The presentation was a comparison of risk assessment under various regulatory rules: the 1989
Surface Water Treatment Rule (SWTR), the 1996 Safe Drinking Water Act (SOWA), the final
Interim Enhanced Surface Water Treatment Rule (IESWTR), and the Long-Term 2 Enhanced
Surface Water Treatment Rule (LT2ESWTR).
The Maximum Contaminant Level Goals (MCLGs) were promulgated by EPA to be zero for
Giardia, Legionella, and viruses; for regulatory purposes, the 1989 SWTR set the limits as close to
the MCLGs as feasibly possible. SWTR was then characterized as reducing risk to less than one
infection per 10,000 people per year from giardiasis, assuming that in controlling Giardia, the rule
would also protect the public from most other pathogens. In reality, infection was a conservative
indicator of illness from Giardia. The risk model used suggested that most systems using the rule
would be protected to an acceptable risk (less than one infection per 10,000 people per year). A
provision made for systems with poor source water quality recommended removal or inactivation
to a set level, greater than the SWTR. Source water quality was difficult to quantitatively evaluate,
however, and the recommendation was one that could not be implemented.
In 1996, the Safe Drinking Water Act (SDWA) changed the existing rule by stating that the
MCGLs should first be recommended at levels at which no health effects occur; actual levels of
pathogens were promulgated to be as close to MCLGs as (economically) feasible; sensitive sub-
populations were to be taken into consideration in the cost-benefit analysis.
The regulations were amended next in 1998 with the publication of the Interim Enhanced Surface
Water Treatment Rule (IESWT). In this case, an MCLG of zero was established for
Cryptosporidium, and recommendations were to control Cryptosporidium and other pathogens to
the extent that was feasible both economically and technically. Risk assessment could be
performed to support the cost benefit analysis, and for most models more costs were avoided than
incurred. Risk assessment carried large uncertainties, however, due to the limited occurrence and
dose-response data available.
The Long Term 2 Enhanced Surface Water Treatment Rule (LT2ESWTR) reflects the evolving
role of risk assessment in regulatory management. It is different from IESWT in that it is not
based on a specific risk goal; rather, risk characterization is incorporated into the proposed rule.
Taking into account the uncertainties inherent in characterizing occurrence, treatment, and dose-
response data for different pathogens, the rule recommends performance of a risk analysis, taking
into account the level of uncertainty, in order to evaluate the costs and benefits as required.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Microbial Risk Assessment Activities in Cincinnati Division of EPA's National
Center for Environmental Assessment - John Lipscomb (ORD/NCEA)
The microbial risk assessment group of NCEA-Cincinnati has several ongoing projects within a
Pathogen Risk Assessment Program. These activities can be separated into four general areas:
(1) Methods assessment and development, including the use of alternative statistical
methods in the presence of limited dose-response data;
(2) Modeling the impact of microbial contamination of waterways and systems on human
exposure;
(3) Assessment of the risk contribution of variance in strain virulence; and
(4) Human population susceptibilities and contribution of differentially susceptible sub-
populations to endemic and epidemic infectious waterborne disease.
The ultimate goal of the research efforts is to develop guidance on the conduct of quantitative
assessments for waterborne pathogens. Within the first area, the evaluation and development of
methods to quantify microbial risk is being addressed through the development of draft
assessments for three of the pathogens on EPA's Contaminant Candidate List (Coxsackievims,
Calicivims, andMycobacterium avium complex). Limited by the availability of dose-response
information, NCEA is investigating alternative approaches towards estimating the dose-response
relationship for pathogens.
Within the second area, the development of a model to integrate the exposure to microbial
contamination from watershed through treatment to drinking water delivery will provide additional
information to more realistically weigh the risk of contamination, and evaluate the contribution of
environmental factors ultimately contributing to pathogen survival and human exposure.
Within the third area, the estimation and comparison of the human disease burdens associated
with specific pathogens and with exposure to specific water sources is being examined. This
project is evaluating the usefulness of three different measures of health impact: Natural Units
(cases, deaths, etc.), Disability Adjusted Life Years, and Cost of Illness. A joint EPA/FDA project
has been established to quantify infective foodborne and waterborne doses of microbial strains
with different virulence in animal models of susceptibility.
Within the fourth area, NCEA is addressing the importance of the secondary spread of disease
and the contribution of differentially susceptible segments of the population. Cooperative
Agreements are producing: 1) an infectious disease framework based on disease transmission
theory; and 2) a deterministic differential equation incorporating three modes of transmission -
endogenous pathogen-to-human, human-to-human, and human-to-water to human - and sensitivity
analysis to determine the relative contributions of each. An Inter-Agency Agreement with
FDA/NCTR has been established to evaluate the usefulness of animal models of susceptibility as
surrogates for human susceptible segments of the population (infants, the elderly, diabetic,
immuno-compromised, and malnourished individuals). The isolation of pathogenic organisms
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shed in feces from these models will provide some information useful in estimating the
contribution of these segments of the population to recirculation of pathogens, and the secondary
spread of disease via waterborne and foodborne routes.
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Breakout Group Discussions
See APPENDIX C: Breakout Group Summaries
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Assessing Exposures to Emerging Pathogens
How do we look for these organisms? What is the current status of methods to monitor
exposure?
Co-Chairs: Bobbye Smith (Region 9), Gerry Stelma (ORD/NERL) and Fred Genthner
(ORD/NHEERL)
Assessing Exposures to Emerging Pathogens - What the Regions Need - Terry
Fleming (Region 9)
In implementing the goals of the Clean Water Act, to protect the public against exposures to
disease causing pathogens in drinking and recreational waters, regions and states must be able to
monitor for these pathogens.
The Regions have identified five major research needs to improve monitoring for pathogens:
1) Rapid detection methods for indicator bacteria and pathogens in recreational waters;
current methods for testing recreational water may take three days, when contamination
may be present for one day.
2) Better indicators, including an approved source-tracking method. Indicators need to be
appropriate for non-point source problems, and yield results quickly.
3) Dose-response relationships (relative risk) for these pathogens, alone and/or in
combinations, so that standards can be set.
4) Predictive models or other tools for evaluating exposures due to storm events or sewage
overflows; such models would allow the state to protect public health (e.g., close
beaches after a storm).
5) Approved "alternative" methods for detection of pathogens in drinking and recreational
waters. New, faster, less expensive methods are being developed, but need to be
approved by the EPA quickly so they can be used.
Regions are often compromised due to the lack of adequate methods. When new methods are
developed, the approval process is a crucial bottleneck.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Assessing Exposure to Bacterial Pathogens - Gerard Stelma (ORD/NERL)
Human exposure to bacterial pathogens can be measured by looking for specific biomarkers of
exposure. Examples include antibodies, reactions to bacterial antigens to which prior exposures
have occurred, or measurement of specific products of the infecting agent in bodily fluids.
Exposure studies generate information on the sources and geographic distribution of exposures,
and on the number of people exposed to various pathogens.
Three pathogens of importance were discussed in the presentation: 1) Helicobacter pylori; 2)
Mycobacterium avium; and 3) Legionellapneumophila.
Serum antibodies are the biomarker currently used for the detection of//, pylori exposure; this
method works well even for asymptomatic individuals. Another method considered is testing for
salivary antibodies - a less invasive method, which, however, is not as effective at detecting
exposure even in individuals with clinical symptoms. H. pylori is suspected of being potentially
waterborne, although that is not its usual mode of transmission. If it is determined that the
pathogen is waterborne, H. pylori exposure and its association with untreated drinking water could
be studied.
Detection of exposure toM avium has so far been done using a skin test method; non-specific
reactions, however, led to false positive and/or negative results. Another disadvantage of the skin
test is the requirement that patients return to have the test read. NERL microbiologists are
currently examining the efficacy of interferon induction byM avium antigens as a potential
replacement of the skin test method for monitoring exposure. A pilot study is being conducted in
collaboration with a Los Angeles hospital - where M. avium occurs commonly in the water.
Specifically, interferon assays are being conducted to determine whether it is possible to obtain
historical exposure data. A full scale study using interferon assays would provide nationwide
exposure data, identifying areas of high and low exposures, and examine the relationship between
M. avium occurrence and its presence in potable water.
Legionellapneumophila exposures are also being studied, using a urinary antigen for a biomarker;
the antigen is present only in patients currently infected. The study is intended to generate accurate
data on the incidence of legionellosis in the United States. Only a small number of cases are
currently reported annually to the Centers for Disease Control (400-500), most likely because,
while patients are often hospitalized, they are simply treated for pneumonia symptoms. In most
cases, no cultures are taken to determine the organism responsible. A realistic nationwide estimate
of the incidence of legionellosis is likely to be much higher than the current number of reported
cases. The study will also attempt to relate data on incidence with the occurrence of L.
pneumophila in potable water.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Development of Molecular Methods to Detect Emerging Viruses - Ann Grimm
(ORD/NERL)
Enteric viruses are believed to cause a large number of gastroenteritis cases in the United States;
caliciviruses specifically can be a major cause of waterborne, viral outbreaks. Methods are being
developed by NERL for the detection of calicivirus: primers and probes have been designed to
detect the virus in water samples. Polymerase chain reaction (PCR) techniques are used to amplify
many of the caliciviruses, since cell culture methods are not generally available. Different primer
sets and real time PCR (RT-PCR) conditions are being examined, and clinical samples are
screened for new types of calicivirus to test primer conditions. A combination of different methods
can also be used, and has the advantage of allowing testing for several viruses at once. Current
methods were used to confirm that a calicivirus outbreak in Wyoming was waterborne, when
nucleic acid in two viruses - from a water sample and fecal samples - was found to be identical.
The Hepatitis E virus (HEV) is another emerging virus of concern; although it has a fecal-oral
transmission route, an outbreak in India in 1959 appears to have been waterborne. HEV causes
infectious hepatitis in adults, and has a fatality rate of 15-25% in pregnant women. Sporadic cases
have appeared in the United States, but a recent estimate using a new assay showed that up to 15%
of the population could have antibodies to the virus - a fraction too high to be attributable solely to
infections related to foreign travel. In addition, a recent United States-acquired infection was 97%
similar to the (endemic) swine strain of the virus, raising the possibility that this might be a
zoonotic virus.
Assays have been developed to detect the presence of Hepatitis E virus, including:
A general assay, which detects all strains except the "Mexico" strain;
An assay specific to the "Mexico" strain; and
And a United States assay, to specifically match the swine strain.
In order to test the assays, the infectious dose (rD50) was determined for several strains of the virus,
and it was concluded that assays could detect a single infectious dose present in a sample. Further
testing showed that all assays were able to detect HEV in spiked environmental water samples.
The assays can now be used for exposure studies, and future research will involve the testing of
natural waters for the Hepatitis E virus.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Detection of Protozoan Pathogens in Water - Frank Schaefer (ORD/NERL)
Parasites, by their nature, do not replicate like bacteria, outside a host. With rare exception, the
waterborne parasites of significance to the Agency are enterics which are transmitted by food,
person-to-person contact, fomites, or water. Unlike clinical fecal specimens in which parasites are
numerous, water samples, due to the tremendous dilution of the contaminant fecal material, contain
very few parasites. Consequently, large amounts of water must be concentrated in order to detect
them. Once the particulates are concentrated, the transmission forms of the parasites must be
processed to selectively remove them from other particulates. This processing step is critical for
both microscopic and molecular methods, so the transmission forms are not masked by the
particulates in the case of microscopy, or inhibited by the particulates and other residual chemicals
in the case of molecular methods. It is only when these first 2 steps have been completed that an
assay can be performed successfully.
The current approach to detecting protozoan parasites like Giardia, Cryptosporidium,
Microsporidians, Cyclospora, and Toxoplasma in water involves concentrating 10 liters of water
using either a flat plate membrane filter or a capsule filter with an absolute porosity smaller than
the organism. The parasites are then separated from the concentrated particulates either by
buoyant density centrifugation or by immunomagnetic separation. Assays to detect the parasites
include fluorescence microscopy, fluorescent in situ hybridization molecular probes, and real-time
PCR.
Once a complete method is developed and published, it still is considered a "research" method.
For regulatory purposes the Agency needs validated methods which have appropriate positive and
negative controls associated with them, as well as performance characteristics. Validation is a
multi-laboratory exercise which determines the precision, accuracy, and robustness of the method.
Presently, only Methods 1622 and 1623 for the detection ofGiardia cysts and Cryptosporidium
oocysts are validated.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Current Approaches to Identifying Sources of Fecal Contamination - Gerard
Stelma (ORD/NERL)
A number of approaches are being tested for their ability to trace sources of fecal contamination:
DNA-based approaches: ribotyping, PCR and RT-PCR, pulse field gel electrophoresis
(PFGE), and toxin biomarkers;
Antibiotic resistance patterns;
Bacteriophage analyses;
Chemical markers;
Neural networking.
Ribotyping, PCR and PFGE methods assume that bacterial population structure is clonal,
therefore E. coli from different animal species should be genetically divergent. If the assumption is
correct, then isolates from a contaminated site can be divided into groups of clonal origin and
matched to their sources; the proportion of isolate coming from each source could also be
determined.
The same assumption applies to toxin biomarker methods, however these methods require a
larger sample size. Toxin biomarkers have a high animal species specificity, although only three
species-specific probes exist so far.
Antibiotic resistance methods assume that a different set of antibiotics would be used to treat
humans than animals; furthermore, different domestic animal species would be treated with
different combinations. This approach could distinguish between contamination from wild and
domestic animals, as wild animals should receive little exposure to antibiotics. These methods are
easy to perform, and their results are comparable to those of molecular methods; they do require
numerous isolates, and the use of a wide battery of antibiotics in different concentrations.
Bacteriophages - male-specific coliphages - can be used if all that is required is to distinguish
between human and animal contamination, with no need for animal species specificity. Chemical
markers can also be used for this purpose.
Neural networking utilizes several different measures of fecal contamination, and is easy to
perform and inexpensive. However, it currently detects total coliform bacteria - a disadvantage
since many coliform bacteria are not necessarily of fecal origin. Additionally, some of the
methods can be difficult to interpret. The use of fecal Streptococcus bacteria is being considered
for this method, as they tend to persist longer than coliform bacteria.
The approaches outlined above and others show promise for use in the detection of fecal
contamination and source tracking. The best methods will not be found, however, until a multi-lab
comparative study is done using the same environmental samples.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Antimicrobial Resistance Among Enteric Bacteria Isolated from Human and
Animal Wastes and Impacted Surface Waters - Mark Sobsey (University of North
Carolina)
Human infection with antimicrobial resistant (AR) pathogens has been steadily increasing since the
late 1960's. Prevalent AR pathogens include Salmonella spp., Enterococcus spp., and Escherichia
coli (E. coli). These bacteria have livestock as well as human reservoirs and some, notably
Salmonella spp. and pathogenic E. coli, have been associated with waterborne disease.
Furthermore, E. coli and Enterococci are microbial indicators of fecal contamination used by the
EPA as quality criteria for recreational and/or drinking source water. AR microbes and
transferable genetic elements containing resistance genes may be transmitted via fecal
contamination, and human acquisition of Multiple Antimicrobial Resistant (MAR) pathogens or
transferable genetic elements from fecally contaminated water can occur either through direct
ingestion of drinking and recreational waters or through consumption of fecally contaminated
foods (e.g., edible bivalve molluscan shellfish and uncooked, irrigated produce).
Recent studies have been determining the occurrence and properties of AR enteric bacteria in
human and animal waste and impacted surface waters in North Carolina. Results reported are from
ongoing studies of cattle and swine farms and wastewater treatment plants (WWTPs) that are
supported in part by the EPA. Findings to date indicate that humans and animals shed multiple
antibiotic resistant bacteria in high proportions and these bacteria enter surface waters. Salmonella
typhimurium and other Salmonella in swine wastes and impacted waters show MAR S.
typhimurium DT-104, which is a major human pathogen. Enterococci isolates downstream from a
swine farm exhibited resistance to Vancomycin, which has not been previously reported in the
literature. Vancomycin-Resistant Enterococci (VRE) are a major public health problem, especially
among vulnerable populations in health care settings. Rural, background sites of surface water
often exhibited high levels of AR bacteria (more than 40% in one case). This is likely due to
human or animal fecal waste impacts because these sites are not true "background" stations.
Fligher prevalence of AR bacteria are found downstream than upstream from swine farms, and
their AR profiles are similar. The results of these studies indicate high prevalence of MAR
bacteria in surface waters impacted by animal agriculture and human waste sources. The high
concentrations and prevalence of these MAR bacteria suggest that waterborne exposure may pose a
risk to people and animals downstream.
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Emerging Pathogens in Drinking Water- Risk Management Challenges
First of two concurrent sessions
Co-Chairs: Kim Harris (Region 5) and Gene Rice (ORD/NRMRL)
User-Friendly Models for Evaluating Hydrogeologic Barriers to Viruses - Bart
Faulkner (ORD/NRMRL/Kerr Environmental Research Center)
The EPA Office of Water requested the Subsurface Protection and Remediation Division of
NRMRL to develop a groundwater "vulnerability" model for predicting virus attenuation. The
Office of Water's Draft Ground Water Rule states that aquifers - other than ones known to be
highly vulnerable - are subject to hydrogeologic sensitivity assessment. In order to be considered
a barrier to viruses, a layer between the virus and the water supply must provide 4-log (or 99.99%)
attenuation of active viruses. The computer model constructed reports the probability of log-4
attenuation considering the parameters entered, such as type of soil and species of virus. The
model was designed with the premise that limited information would be available to the users. A
literature review was performed in order to be able to incorporate in the model default parameters
for previously-studied viruses and the twelve United States Department of Agriculture (USD A)
soil categories. In addition, the model is designed to be expandable, and to report uncertainty.
The user interface runs on a Java application, and fields are filled in for the users once they supply
the information available. Should more parameters be known, users can change the information in
the fields as necessary. The model then performs Monte Carlo Analyses and reports the results in
graphical and text format, as well as the probability of failure.
The user interface is available on the EPA intranet at the following address:
http://intranet.ada.epa.gov/research/virmod.html
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Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Water Supply and Water Resources Division Response to Waterborne Disease
Outbreaks - Kim Fox (ORD/NRMRL)
The Water Supply & Water Resources Division (WSWRD) in NRMRL/ORD has had a successful
collaborative relationship with the Centers for Disease Control & Prevention (CDC) for over
twenty years. When invited, EPA has supplied technical assistance and advice on tracking
causative events, evaluation of drinking water problems, and on how to ensure drinking water was
safe to consume. These requests for assistance have come from CDC, EPA regional offices, states,
municipalities and foreign countries. The most publicized recent waterborne disease outbreak was
the Cryptosporidium event in Milwaukee, Wisconsin. Another well publicized drinking water
problem that WSWRD assisted with was the boil water order in Washington, DC. In both cases,
an on-site investigation was conducted.
Some organisms that have caused recent problems or outbreak situations include Giardia,
Cryptosporidium, Vibrio, Shigella, E.coli O157:H7, Salmonella, Entamoeba histolytica, and others.
Due to the fact that there is a time lag effect - from ingestion to illness - each outbreak must be
investigated based on what evidence is available at the time. The majority of outbreaks have
occurred because of failure in one of the barriers keeping pathogens out of the drinking water
supply, the likely sources of contamination being pathogens coming from the watershed, treatment
plant, distribution system, or a system in the affected building. Some of the outbreaks investigated
by WSWRD are outlined below:
In Gideon, Missouri, a Salmonella outbreak involved birds contaminating an
unprotected storage tank; when the storage tanks were flushed, Salmonella bacteria
from the storage tanks flowed through the drinking water system, and illness followed
the pattern of the water flow from the tanks flushing event.
In Karl Meyer Hall, storage tanks on the roof of the building had a zone of stagnant
water at the bottom; the system shut down overnight, and when it was turned back on,
water from the stagnant zone entered the system.
In the Milwaukee, Wisconsin Cryptosporidium outbreak, the onset of disease correlated
with the onset of high turbidity in the water. In Washington, DC, no outbreak occurred
but a boil-water order was issued after turbid water passed through the treatment plant.
In Alpine, Wyoming, an E. coll O157:H7 outbreak was linked to fecal contamination in
the watershed, when bacteria in fecal samples and in water samples were tested.
In order to more successfully prevent future waterborne disease outbreaks, the multiple-barrier
system is the best approach. Raw source water cannot be presumed to be free of pathogens,
therefore each barrier is equally important. Removal of particulate matter is especially important,
given the link between turbidity fluctuations and the passage of parti culate material through the
treatment process. Treatment goals for water systems are also needed, as an additional barrier to
pathogens.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
What are the microorganisms of concern and how do they get into the distribution
system?
Disinfection of Emerging Pathogens - Eugene Rice (ORD/NRMRL)
There is a growing awareness of the need to control waterborne microbial pathogens. The
presentation concentrated on the role of chemical inactivation, using chlorine, chloramines and
ozone as a means of controlling bacterial and protozoan species. Some of the pathogens discussed
were organisms on the Contaminant Candidate List including Aeromonas, Helicobacterpylori and
Microsporidia; as well as other pathogens of concern, e.g., E. coli O157:H7, Giardia and
Cryptosporidium.
The effects of chlorine and chloramine were tested for a variety of exposure times on different
Aeromonas species; chlorine was more effective and faster at disinfection at similar temperature
and pH levels.
Helicobacter pylori were found to be more resistant than Aeromonas and slightly more resistant
thanE1. coli O157:H7 to disinfection with chlorine, possibly due to aggregation with particulates.
Giardia was one of the organisms tested that is known to be resistant to disinfection with chlorine;
in this case, ultraviolet light was used and found to effectively inactivate Giardia.
Better characterization of the effects of agents used in disinfection was suggested as a topic for
further research, particularly of the effects of disinfection techniques at low temperatures.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Biofilms in Drinking Water Distribution Systems - Mark Meckes (ORD/NRMRL)
Biofilm forms in drinking water distribution systems when bacteria and other organisms adhere to
wetted surfaces and begin producing an extracellular polysaccharide envelope. That glue-like
substance can anchor the bacteria to surfaces, where they can reproduce and grow. Increased
temperature and rainfall (and the resulting increase in turbidity) are factors which contribute to the
growth of biofilms, as are structural features of pipes and the velocity of the water running through
them. Biofilm growing on surfaces which are normally in contact with water under low flow
velocities can be released with a sudden increase in flow. Biofilm growth is also influenced by the
availability of nutrients, the surface materials, and the concentration of residual disinfecting agents
present in the water. Furthermore, the presence of biofilm as pioneer organisms provides an
environment which could influence the colonization or entrapment of pathogenic or indicator
organisms.
Studies currently being conducted to understand the problem include the development of rapid,
cost-effective methods for detecting groups of bacteria specifically associated with biofilm, and for
obtaining quantitative information regarding their occurrence. One group of bacteria, the non
tuberculosis mycobacteria, are currently being studied to determine if baiting techniques can be
used to concentrate them, so that they may be more easily detected.
Studies have been completed or are ongoing to determine the effects of the following factors on
biofilms:
pH;
Disinfecting agents;
Nutrients; and
Cross-connection.
Another study, in the planning stage, will look at the survival and growth ofAeromonas spp. in
biofilm. This work will be designed to determine if unique control strategies are needed for these
organisms.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
What is the Drinking Water Distribution Rule?
Update on 6-Year Review of TCR and Potential Distribution System Rule - Ken
Rotert (OW)
The Total Coliform Rule (TCR) and other rules related to drinking water, must be reviewed, and
appropriate revisions made at least every 6 years according to the 1996 Amendments to the Safe
Drinking Water Act (SOWA). Since the TCR was promulgated in 1989, the TCR is currently
under review. Part of this review includes the examination of unsolicited comments received since
the Rule's promulgation, as well as comments from the EPA Regions, the States, ASDWA,
AWWA, AMWA and ASM received as a result of a January, 2001 solicitation by EPA's Office of
Ground Water and Drinking Water (OGWDW). Some of the comments received addressed the
topics of waiving total coliform (TC) monitoring in undisinfected supplies, statistically
determining the number of samples to collect each month, using the stakeholder process to review
monitoring requirements, eliminating fecal coliforms as an indicator, using fewer than five routine
samples, and allowing flexibility in the sample collection location. Additional comments
addressed dropping the Maximum Contaminant Level (MCL) for TC, providing a clarification
regarding whether two positive E. coli samples at different sites constitutes a violation and
allowing the use of dedicated sampling taps.
EPA intends to couple the review and potential revisions of the TCR with a broader distribution
system rulemaking effort intended to address the potential public health risks associated with
finished water in the distribution system. This follows the recommendation of the M/DBP Federal
Advisory Committee (FACA) Agreement in Principle. To initiate this process, an expert workshop
was convened to discuss the health risks associated with distribution systems. Workshop attendees
included representatives from EPA, American Water Works Service Company, academia, utility
personnel, State regulators and private consultants. Nine white papers are under preparation as a
result of this workshop. First drafts of all of the white papers are expected to be completed in
December, 2001.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
How do we handle cross-contamination and biofllm problems in the field? What are the
inherent risks to distribution systems? Can we minimize the risk? Case study
example/panel discussion.
Case Study: Biofilm Problem at Greenville, South Carolina - David Parker
(Region 4)
The presentation focused on the problems encountered in a drinking water system in North
Carolina. The system was not filtered, under the assumption that the source water was protected -
high turbidity and raw coliforms were, however, present in the source. The filtration plant did not
operate continuously ("start-stop" operation), and the finished water storage areas were not covered
(until 1992 when a floating reservoir cover was installed). The disinfection agent - chloramine -
was also thought to be inadequate.
Initially avoiding filtration on the grounds that the watershed (source) was tightly controlled to
limit human access, Greenville was unable to meet the filtration avoidance criteria due to high
turbidity, and entered into an agreement which mandated construction of a filter plant by 2000.
Events that preceded that agreement included a series of Total Coliform Rule violations, and boil
water notices when fecal coliform bacteria were found. An analysis made of the species of
coliform bacteria present, coupled with the observation that their occurrence was correlated with
higher temperatures (above 17°C) led to the realization that the water contamination was related to
a biofilm problem. In this case, sediment in the water system was the source of the biofilm
organisms, and the unfiltered lakes (source) provided nutrients for their growth.
Since biofllm organisms can be pathogenic, or mask the presence of other pathogenic forms, an
action plan was developed to remedy the problem. Total coliform samples were to be taken twice
as often as required per month, and species analysis conducted on any positive samples. An
annual, system-wide flushing program was implemented, with additional instructions to switch the
disinfecting agent from chloramine to free chlorine with each flushing; and the filtration plant was
instructed to run continuously, rather than starting and stopping. Finally some public education
measures were undertaken (for immuno-compromised individuals, in particular) and a public
health surveillance program was initiated.
As a result of the action plan - initiated in 1995 - Greenville has had no Total Coliform Rule
violations; the new filtration plant (mandated by the agreement mentioned above) began operations
in the summer of 2000, and improvements were made to the existing filtration plant.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
The View from the Field - Water Distributor's Perspectives - Mark LeChevallier
(American Water Works Service Company)
Distribution system problems were outlined in this presentation, including control of pathogens
through disinfection, pathogen monitoring, and problems associated with pressure changes in the
system.
Studies onMycobacterium avium complex (MAC) have shown that it can grow in biofilms, and
has been found both in water and biofilm samples from distribution systems. MAC is a pathogen
of concern in drinking water since it can affect immuno-compromised individuals. Disinfection
studies have shown resistance to chlorine, but inactivation at high temperature (60°C); in addition,
low nutrients and turbidity levels in raw water reduce its occurrence. Assimilable Organic Carbon
(AOC) and Biodegradable Organic Carbon (BDOC) levels were correlated with MAC occurrence
in water; the development of simpler AOC and BDOC methods, and improved MAC detection
were suggested areas where research was needed.
Literature on the disinfection ofLegionella suggests that chloramines may be more effective for
biofilms ofLegionella than free chlorine. Conversion of a hospital water system from free chlorine
to chloramines reduced Legionella levels 1,000-fold.
Monitoring of pressure in water systems was also mentioned as a way of preventing pathogens
from entering the drinking water system. Specifically, a pressure wave passing through the system
can result in negative pressure (briefly) in some areas; negative pressure would have the effect of
drawing material - potentially including pathogens - through a pipe connection. Start and stop
operations of plants are a major cause of pressure changes; however, they can not always be
avoided (e.g., in one case, lightning caused a plant to shut down and resulted in negative pressure).
Shutting down pumps during a pump test was shown to cause brief moments of low pressure.
High-speed pressure monitoring is essential in identifying potential areas where the system might
have been breached.
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Emerging Pathogens in Recreational Water- Risk Management Challenges
EPA's National BEACH Survey - Rick Hoffmann (OW)
Each year since 1997, EPA has conducted its National Health Protection Survey of Beaches
(Beach Survey). The annual Beach Survey is sent to more than 300 states and local governments.
It is designed to gather information about beach water quality, standards, monitoring, and beach
health advisories or closings issued during the previous year's bathing season. The survey's goal is
to provide comprehensive, reliable beach information to the public via the Internet.
For each beach, information is collected in three areas: 1) programmatic information, including
water quality standards, 2) monitoring procedures and costs, and 3) closing/advisory information;
beach-specific information, such as physical characteristics, usage, possible pollution sources; and
advisory/closing details, including the number of days, length of beach affected, reason and
source(s) of contamination and indicator used.
All coastal states and the Great Lakes states are sent surveys. Most (86%) returned the surveys for
the 2000 swimming season; however, incomplete information was often submitted. According to
survey results for the 2000 season:
The majority (96%) of beaches have a monitoring program, with more than half of
those testing samples at least one per week.
A quarter of the beaches (26%) had at least one advisory or closing in the past season.
Past surveys indicate that the percentage of beaches with at least one closing has
remained at or near 25% for the past four years.
The sources identified as causing the pollution were unknown much of the time; of the
known causes, storm water and other runoff, and wildlife, were the most frequently
reported.
The survey showed significant differences in the kinds of indicators used by different
states - most using one or more of the following: total or fecal coliform bacteria, E. coli
or Enterococci.
As mentioned above, the results of each national survey are posted on the Internet for purposes of
informing the public and can be viewed at:
http ://www. epa. gov/ost/beaches
Many of the states and territories also maintain their own web sites where the information on state
beaches can be found.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Development of Improved Monitoring Approaches for Recreational Water -
Stephen Schaub (OW)
The EPA Office of Research and Development and Office of Water, under the Environmental
Monitoring for Public Access and Community Tracking (EMPACT) program, jointly sponsored a
study during the summer of 2000 to better define the temporal and spatial sampling requirements at
recreational beaches, and more accurately determine potential public health risks for acute
gastrointestinal disease when monitoring for bacterial indicators of fecal pollution. To establish the
monitoring protocols for the study a data quality objectives process was conducted to insure that
there was a statistically valid approach and protocol to sampling and data gathering to answer the
questions regarding the minimum amounts of sampling data (frequency, location, sample number,
etc.) to be gathered for a beach at any given time to accurately assess water quality. The study
utilized five sites: a river, a lake, an estuary, and east and west coast ocean beaches. These beaches
were monitored daily over a period of two consecutive summer months. Sampling grids were
utilized to provide a common set of data points for characterization of indicator levels at the
beaches and included three distances from shore, multiple sampling depths, and distances along the
shoreline. Samples were taken daily: routine morning and afternoon samples, intensively, at
hourly sample intervals over a day, or intensively during/after significant rainfall events. A
statistical analysis of the study results revealed that in at least certain instances, wind, wave height,
tides, sun intensity, the presence of bathers, and rainfall events, in addition to sampling time and
sample location, could have an impact on levels of fecal bacteria indicators detected.
This fall, 2001, the EPA will sponsor an expert workshop to evaluate the EMPACT program beach
monitoring data, and statistical representation of critical elements impacting on fecal indicator
values at beaches. The workshop will determine the minimum sampling procedures required to
gain various levels of confidence regarding indicator levels at a beach for beach managers,
regulators or operators who are responsible for assessing recreational exposure risks and their
acceptability. This monitoring guidance will be used in later updates of the EPA's recreational
water criteria and guidelines.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Use of Molecular Techniques to Provide the Public with More Information on
Recreational Water Closures in Norfolk - Alpha Diallo (Norfolk Department of Public
Health)
The Norfolk Department of Public Health, in cooperation with the School of Public Health,
University of Washington has established the relative significance of E. coli isolates from avian,
domestic animal and urban wildlife origins over the past two years.
Testing is performed once a week at 34 sites for fecal Streptococcus., fecal coliform and E. coli
during the recreational season. This is reduced to twice monthly during the winter months.
Standards are a geometric mean standard of 35 Enterococcus bacteria per 100 ml. For fecal
coliform a geometric mean of 200 fecal coliform/100 ml for two or more samples over a 30 day
period or a fecal coliform bacteria level of 1,000/100 ml at any time. Secondary contact water
fecal coliform shall not exceed a log mean of 1,000 per 100 ml during a 30 day period. Exceeding
the above may trigger notification or advisories.
Of 714 isolates that were submitted for microbial source tracking by ribotyping 34% were from
avian origin, 17.9% from domestic animals (cats and dogs), 14.8% from humans, and only 6.2%
from urban wildlife. Although the weekly microbial counts were not high enough to warrant
action in most cases, these results make a case for the screening and testing for specific pathogens
in beaches and recreational waters.
Specific pathogen testing by PCR was initiated using primers supplied by Dr. Samadpour.
Subsequently these have been purchased from Sigma Genosys. Validation of the protocol was
performed using water samples known to be free of E. coli and testing for presence of the toxins
stxl and stx2. Validation for other pathogens is in progress and will be available if the need arises.
A web site has been developed, but needs to be set up on to the network of either the City of
Norfolk or the Virginia Department of Health. This action is in progress. Data generated in the
laboratory is maintained in an Access database file. Results are recorded and sent to the
Environmental Health Manager and Health Director at the same time as posting occurs. If any
results are found to be actionable, the Environmental Health (EH) Team is dispatched to resample
and more rigorous testing is effected.
The EH Director plays a key role in this monitoring program. He is the principal link with other
City Departments concerned with either regulating or alleviating risks to the community.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Acceptable Goals and Thresholds for Pathogens from Recreational Water
Exposures - Al Dufour (ORD/NERL)
The debate about what is an acceptable risk regarding exposure to recreational waters has been
ongoing for many years. Risk managers must make decisions based on the benefits of a popular
leisure time activity and the potential health risks that may be suffered because of that activity. In
addition to using risk-benefit data, risk managers are frequently required to examine the cost-
benefit paradigm. Neither the risk-benefit nor the cost-benefit models have been followed in the
past. This presentation reviewed the historical approaches used in the United States to set
thresholds for water quality safety and discussed the current direction of U.S. EPA research with
regard to how it may provide valuable information for making acceptable risk decisions.
The Office of Water has made great efforts in the past and present toward making the risk
assessment data available to the public, however, some feedback is necessary in order to determine
whether the meaning behind the data is understood.
A rationale for using coliform bacteria for testing was developed from some early studies (e.g., in
1951, shoreline in Connecticut was sampled for coliform bacteria and found 93% of samples tested
negative). EPA recommended threshold values for coliform bacteria in 1976, and used a ratio to
later convert those thresholds for fecal coliform bacteria densities. More recent studies indicated
that extrapolation of the total coliform to fecal coliform values was not always accurate. New
guidelines were developed for testing, taking into account that total or fecal coliform bacteria may
not be the best indicators of fecal contamination.
New EPA-recommended guidelines specify the use of either E. coll or Enterococci for fresh water
beaches, and the use of Enterococci for marine waters as indicators.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Determining Sources of Microbial Contamination - A Regional Applied
Research Effort - Joel Hansel (Region 4)
The research proposal presented was based on the premise that Total Maximum Daily Loads that
are submitted by the states must include load allocations to the sources of contamination. Bacterial
source tracing methods are needed, therefore, for the states to be able to identify sources of
pathogens.
The proposed research will consider the techniques of ribotyping, multiple antibiotic resistance,
and pulsed field gel electrophoresis and determine ways in which they can be used for performing
bacterial source tracing.
Fecal coliform bacteria, E. coli, and Enterococci from multiple sampling sites will be analyzed
using antibiotic resistance and ribotyping. Information on these bacteria from existing databases
will be incorporated into developing the new bacterial source tracing methods. Soils and
sediments will also be examined as possible sources of contamination.
The proposal is currently undergoing peer review, and research will start as soon as that process
has been completed. The project is expected to be completed by December 2003.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Rapid Methods for Measuring Recreational Water Quality: A Look to the
Future - Al Dufour (ORD/NERL)
Traditional methods used to monitor the quality of recreational waters require 24 to 48 hours
before results are available. This delay provides a situation where the potential health risk
associated with positive findings of poor water quality are identified long after recreationists are
exposed. Since adverse health effects have been associated with swimming in recreational waters
of poor quality, it is in the public's interest to eliminate this shortcoming in current monitoring
practices. A solution to this problem is to develop an inexpensive, fast, easy, specific, and accurate
test, which will give near real-time results regarding the quality of bathing waters. There is a small
number of biosensor technologies that may meet the characteristics listed above and that may be
available in the near future. Biosensors are analytical devices that integrate microorganisms,
enzymes, antibodies or nucleic acids into optical, electrochemical or other forms of electronic
devices to yield a signal which is proportional to the concentration of the analyte. The technologies
described fall into three categories: 1) those that make use of enzyme reactions, 2) those that use
antibodies for immobilizing or labeling proteins, and 3) those that identify specific nucleic acids.
One such method utilizes luciferin and luciferase to measure production of bioluminescent
adenosine triphosphate (ATP) by the bacteria. The amount of ATP can then be measured by
measuring the bioluminescence.
A second method uses a fluorophore labeled antibody which attaches to the bacterium; this process
can be completed in 10 minutes using a portable instrument, and is currently being tested.
Molecular methods are also being developed using non-viable cells, as is a method using a
Charged-Coupled Device (CCD) camera - which, however, may not be feasible to use due to cost.
New technologies were summarized at the end of the presentation with some of the advantages and
disadvantages of each.
Advantages and Disadvantages of High-technolog)'
Methods for Rapid Measurement of Water Quali
Technology
Limit of Viable
Approach Sensitivity Cells Cost
Flow Cytometry
Bioluminometer
Fiber Optics
Taqman
CCD camera
Antibodies
Antibodies
Molecular
100-1000
No Medium
No Medium
No Medium
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
ORD's Drinking Water Research Program Tracking System - Al Dufour
(ORD/NERL) and Cheryl Itkin (ORD/NCEA)
In response to the need to organize, manage, and disseminate information about environmental
research areas shared across EPA's Office of Research and Development (ORD), ORD
Laboratories and Centers together piloted the "ORD Drinking Water Tracking System." This is a
new approach that supports a single inventory of all the drinking water research projects across
ORD Laboratories and Centers and utilizes the latest web technology. The Environmental
Information Management System (EEVIS), the EPA's repository of descriptive information (a.k.a.,
metadata) about projects, models, data, and documents, is being used as the foundation for the
Drinking Water Tracking System. The EEVIS stores the project descriptions in a Web-enabled
Oracle database and generates the dynamic interface web pages including search forms, data entry
forms and dynamic reports organized by drinking water category. Scientists can access the
tracking system to create, view, maintain, and deliver information about drinking water projects in
ORD. Currently, the tracking system is only available to EPA, but may be made publicly available
in the future.
EPA access to the Drinking Water Research Tracking System
http://cfint.rtpnc.epa.gov/dwportal
For additional information on the ORD Drinking Water Tracking System contact:
Al Dufour 513-569-7330 or Clyde Dempsey 513-569-7842
For information about EEVIS visit http://www.epa.gov/eims or contact:
Robert Shepanek 202-564-3348 or Cheryl Itkin 202-564-3357
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Pathogen TMDLs / CAFOs / Biosolids
Co-Chairs: Stephanie Harris (Region 10) and I.E. Smith, Jr. (ORD/NRMRL)
A Regional Perspective on CAFO/Biosolid Regulations: How ORD Can Help
Us! Stephanie Harris (Region 10)
The presentation offered a regional perspective on several topics of concern which would be best
addressed by ORD; the topics presented have been raised by microbiologists and project officers,
legal counsel, and field inspectors.
Region 10 contains a large number of cattle pastures - a likely source of human pathogens if the
manure is directly deposited or washed into the water source. Human pathogens which have been
identified in cattle manure include Salmonella, Cryptosporidium, Giardia, andE1. coli O157:H7.
Methods for measuring these pathogens exist, however, in the case of Salmonella and E. coli they
have been designed for use in food and drinking water; their application on source water has not
been evaluated. Another area where information is needed is the effectiveness of buffer zones in
reducing pathogen runoff, particularly for Cryptosporidium. The holding time regulation for
microbiological samples (6 hours) presents logistical problems and, if exceeded, puts the data into
question. The problem would be eliminated by using a mobile lab or portable instrument, thereby
performing the tests in the field; or, a way of preserving the sample could be tested.
Biosolids were also discussed, particularly the discrepancy between Class A and Class B biosolids;
Class A biosolids meet exceptionally stringent regulations and can have more applications than
Class B; more information is needed on whether these regulations are enforced and, if so, whether
they are protective of public health. Methods for testing biosolids were also suggested as an area
for more research, specifically coming up with simpler methods; some of the methods currently
accepted - e.g., two methods for Salmonella and virus assays - are complicated methods that
cannot be performed by all laboratories.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Overview of Watershed Features and Dynamics Associated with Drinking and
Recreational Water Protection - J E Smith, Jr. (ORD/NRMRL)
In considering a surface body of water it is difficult not to think that the water may at some time be
used either for recreational purposes or as a source of drinking water. In either case it is good
practice to minimize the entrance of pathogenic organisms. Based on the characteristics of the
watershed, and its use, the likely sources of pathogens can be identified and management plans put
in place.
Drinking water is the main concern where pathogens are involved, and recent waterborne disease
outbreaks have shown that compliance with the Surface Water Treatment Rule (SWTR) is not
always easy. While drinking water treatment requirements have become more stringent, treatment
can be even more efficient and effective when the sources of contamination in the watershed are
controlled. Sources are varied, and include onsite wastewater treatment systems; wet weather
flows, as from combined sewer and storm sewer overflows; and the application of treated
wastewater, treated sludge and/or animal manure to farm land and other areas.
Land application of treated sludge (biosolids) is a beneficial use. However, various human
pathogens have been identified in both raw sludges and animal manure, and some of these can
persist in the soil for long periods of time. Buffer zones between farm (application) areas and the
surface water may prevent transport of the pathogens - some data shows that organisms remain
viable for significantly shorter periods when plants are present. Other ways of removing pathogens
from sludge and/or manure include treatment by composting, aerobic digestion, alkaline
stabilization, anaerobic digestion and long-term storage.
Treatment of sludge has had as a goal reducing the levels of pathogens to below the number
required to cause human disease; better and cheaper methods are still needed to accomplish this
task. This is especially true since the trend is to reduce the number of pathogens to below the
detection level of current analytical methods. More sensitive analytical methods are under
development.
A workshop on emerging infectious disease agents associated with biosolids was completed in
June of 2000. State-of-the-science reports are being produced from information developed at the
workshop and will assist the National Academy of Science review of the science behind the United
States Sewage Sludge Regulations, as well as helping to define the need for controls on pathogens
associated with CAFOs.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Potential Watershed Management of Pathogens of Animal Origin - John
Cicmanec (ORD/NRMRL)
Microbial diseases/agents of concern typically include cryptosporidiosis, Escherichia coli
O157:H7, giardiasis, salmonellosis, and the caliciviruses - i.e., Norwalk agent. Animal microbial
diseases transmitted to humans account for up to 136 diseases. To give some feel for the
magnitude of the animal wastes problem, it is estimated that 50 times as much animal manure is
produced as are human wastes, and one calf sheds 10,000,000 Cryptosporidium oocysts per day.
Several of the waterborne disease outbreaks in recent years have been connected to animal
pathogens being transmitted through drinking water. E. coli O157:H7 is rare in most cattle, yet can
be present in 25% of animals in some herds; it remains viable in the soil for four months. Sheep
are the most common animal carriers of E. coli in Europe. Cryptosporidium parvum is most often
ingested with recreational water, and has a human and bovine genotype. Salmonella typhimurium
exhibits multiple antibiotic resistance, causes forty percent mortality in cattle and can be found in
many other mammals and birds. Risk assessment studies have been performed for all three
pathogens named above to estimate the number of infections per person, per year, from drinking
water. Infectivity assays have helped determine the infective dose for each organism.
The primary factors that affect transport of the pathogens to the watershed include:
Percolation Factor (pathogens leach through the soil);
Annual Runoff Factor (pathogens in water are washed off into the watershed); and
Soil Erosion Factor (pathogens are carried into the watershed while adsorbed to soil
particles).
By studying each of these factors, the watersheds most at risk for pathogen contamination have
been determined. These watersheds pose a higher risk of infection, either by accidental ingestion
of recreational water, or by drinking water, if pathogens have passed through the barriers of the
treatment plant. This information is important to risk managers, so that a potential outbreak
situation can be controlled more effectively.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Emerging Infectious Disease Agents and Issues Associated with the
Management of Treated Sludges (Biosolids) - J E Smith, Jr. (ORD/NRMRL)
Human pathogens sometimes found in sludge cause uncertainty about the practice of applying it to
land. EPA regulations published in 1993 regulate the treatment and testing for biosolids, and
define the areas where different classes can be applied. Class A biosolids are treated using
processes like composting, heat drying, and/or alkaline treatment before land application to control
pathogens. Sludge can also be considered class A based solely on testing for enteric viruses and
helminth ova. This alternative is now coming into question, since such testing does not provide
information about other pathogenic organisms that may be present. Class B biosolids meet less
stringent requirements and rely more heavily on the natural attenuation of pathogens after
treatment; most of the sludge applied to land is Class B.
Concerns of the public and some regulatory bodies relate to the fact that sludge (especially Class
B) being applied to land may still contain some pathogens. Research is ongoing to develop
improved analytical methods for measuring pathogens in biosolids, as well as for treating them to
reduce pathogen levels. Modifications of existing processes (heat treatment, lime stabilization) are
under consideration, and some new and innovative processes have been proposed (microwave
processes, solar drying, "gold coast" stabilization). In addition, how best to restrict access to land
treated with biosolids needs evaluation, since the possibility for pathogens to be transported by
animals (wildlife) or wind still exists.
Major research needs include: the standardization and validation of methods for both monitoring
and treatment of sludge; the development of a simple procedure for achieving a Class A sludge;
and investigation of some of the concerns that have been raised regarding worker safety at sludge
treatment and application sites.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Risks from Pathogens in Biosolid Fertilizers/Land Application of Sludge -
National Research Council Report - Tom Burke (Johns Hopkins University)
The purpose of this research study was to review the risks and risk assessment methods used by
EPA for establishing standards for chemical pollutants and pathogens in biosolids, and to consider
whether chemical and pathogen risks should be integrated. In 1993, EPA published Standards for
the Use or Disposal of Sewage Sludge (40 CFR Part 503). The rule set limits for chemical
pollutants and operations standards for eliminating pathogens in sludge. The standards set for
pathogens were based on the technology available for their detection and treatment.
The Clean Water Act requires EPA to periodically re-assess the scientific basis for the Part 503
rule. In addition, public concern about health risks and advances in risk assessment methods led to
a review of the current rule. The previous review by the Water Science and Technology Board, in
1996, concluded that properly treated and managed sludge can be safely used in food crop
production; however, the definitions of "treated and managed sludge" were vague, leading to
inconsistencies. The 1996 report recommended the development of methods for monitoring
specific pathogens in sludge, and a re-evaluation of the adequacy of a thirty-day waiting period
following applications of Class B sludge to pastures used for grazing.
The Committee on Toxicants and Pathogens in Biosolids Applied to Land is conducting a study
funded by the Office of Water, to be completed in the spring of 2002. The study plans to examine
new information on risks, and evaluate current practices (the thirty-day limited access period, for
example, does not seem to prevent wind-blown soil particles from transporting pathogens). Data
on chemicals and pathogens will also be examined, including data on organisms recently identified
as pathogens, and those that persist in soil. Infectious dose studies will be incorporated in the
study, to determine if current approaches are adequate or need to be revised. Information and
comments will finally be sought from a broad range of stakeholders.
Formulation of new approaches and identification of critical gaps are some of the major
implications expected from the study. The report should also provide an incentive for
harmonization in the way agencies approach the issue of risk management in biosolids.
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Over the Scientific Horizon - Meeting Regional Research Priorities
Moderator: Paula Estornell (Region 3)
See page 9: ORD's RESPONSE TO REGIONAL RESEARCH NEEDS
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US Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues associated with Aquatic Environmental Pathogens September 5 - September 7, 2001
Appendix A: List of Participants
Appendix B: Slides from Presentations
Appendix C: Breakout Group Summary
Appendix D: Flip Chart Notes
Appendix E: Workshop Participant Evaluation Summary
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U.S. Environmental Protection Agency
Regional/ORD Workshop:
Last Name
Regional Offices
Higgins
OO
McClelland
Reilly
Brignoni
Cabrera
Galizia
Gray Byron
Hemmett
McChesney
McKenna
Torrey
Binetti
Estornell
Gambatese
Grundahl
Hilliard
Kulik
Landy
Emerging Issues Associated with Aquatic Environmental
Appendix A:
First Name
Elizabeth
Maureen
J. Kevin
Rosa
Elvira
Audrey
Lisa
Roland
Dennis
Douglas
Leonard
Victoria
Paula
Jason
Nancy
Annie M.
Mick
Ronald
Pathogens September 5-September 7, 2001
List of Participants
Affiliation
Region 1
Region 1
Region 1
Region 2
Region 2
Region 2
Region 2
Region 2
Region 2
Region 2
Region 2
Region 3
Region 3
Region 3
Region 3
Region 3
Region 3
Region 3
Email Address
higgins .beth@epa.gov
mcclelland.maureen@epa.gov
reilly .kevin@epa.gov
brignoni.rosa@epa.gov
cabrera.elvira@epa.gov
galizia.audrey@epa.gov
byron.lisa@epa.gov
hemmett.roland@epa.gov
mcchesney.dennis@epa.gov
mckenna.douglas@epa.gov
torrey.leonard@epa.gov
binetti.victoria@epa.gov
estornell.paula@epa.gov
gambatese.jason@epa.gov
grundahl .nancy@epa.gov
hilliard.annie@epa.gov
kulik.michael@epa.gov
landy.ronald@epa.gov
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U.S. Environmental Protection Agency
Regional/ORD Workshop:
Pepino
Russell
Hansel
Johnson
Parker
Harris
Wirick
Carney
Gonsoulin
Lipka
Ngo
Schaub
Helvig
Joyce
Benson
Spence
Fleming
Macler
Roser
Smith
Emerging Issues Associated
Richard
David
Joel
Bonita
David
Kimberly
Holiday
Gerald
MaryE.
Douglas
Dzung Kim
Mike
John
Jake
Robert
Sandy
Terrence
Bruce
Sara
Barbara
(Bobbye)
with Aquatic Environmental
Region 3
Region 3
Region 4
Region 4
Region 4
Region 5
Region 5
Region 6
Region 6
Region 6
Region 6
Region 6
Region 7
Region 7
Region 8
Region 8
Region 9
Region 9
Region 9
Region 9
Pathogens September 5-September 7, 2001
pepino.richard@epa.gov
russell .dave@epa.gov
hansel j oel@epa.gov
johnson.bonita@epa.gov
parker.david@epa.gov
hams .kimberly@epa.gov
wirick.holiday@epa.gov
carney .gerald@epa.gov
gonsoulin .mary@epa.gov
lipka.douglas@epa.gov
ngo .kim@epa.gov
schaub.mike@epa.gov
helvig j ohn@epa.gov
j oyce j ake@epa.gov
benson.bob@epa.gov
spence . sandra@epa.gov
fleming.terrence@epa.gov
macler.bruce@epa.gov
roser.sara@epa.gov
smith.bobbye@epa.gov
Harris
Stephanie
Region 10
harris.stephanie@epa.gov
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated
Office of Research and Development
Dietrich
Marsh
Walton
ORD Laboratories and
Itkin
Lipscomb
Shepanek
Harrison
Levinson
Nolt-Helms
Barnwell, Jr.
Dufour
Grimm
Kryak
Mintz
Schaefer
Stelma Jr.
Bradley
Calderon
Genthner
Hilborn
Deborah
Marsha
Barbara
Centers
Cheryl
John
Robert
Bronda
Barbara
Cynthia
Thomas O.
Alfred P.
Ann
David
Bruce
Frank
Gerard
Patricia
Rebecca
Fred
Elizabeth
with Aquatic Environmental
ORD
ORD
ORD
ORD/NCEA
ORD/NCEA
ORD/NCEA
ORD/NCER
ORD/NCER
ORD/NCER
ORD/NERL
ORD/NERL
ORD/NERL
ORD/NERL
ORD/NERL
ORD/NERL
ORD/NERL
ORD/NHEERL
ORD/NHEERL
ORD/NHEERL
ORD/NHEERL
Pathogens September 5-September 7, 2001
dietrich.debbie@epa.gov
marsh.marsha@epa.gov
walton.barbarat@epa.gov
itkin.cheryl@epa.gov
lipscomb j ohn@epa.gov
shepanek.robert@epa.gov
harrison.bronda@epa.gov
levinson .barbara@epa.gov
nolt-helms.cynthia@epa.gov
barnwell.thomas@epa.gov
dufour.alfred@epa.gov
grimm.ann@epa.gov
kryak.davidd@epa.gov
mintz.bruce@epa.gov
schaefer.frank@epa.gov
stelma.gerard@epa.gov
bradley .patricia@epa.gov
calderon.rebecca@epa.gov
genthner.fred@epa.gov
hilborn.e@epa.gov
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U.S. Environmental Protection Agency
Regional/ORD Workshop:
Walton
Bauer
Cicmanec
Faulkner
Fox
Jacobs
Meckes
Olivas
Rice
Smith, Jr.
Klauder
Noyes
Whitworth
Program Offices
Hecht
Messner
Berger
Regli
Rotert
Shaw
Sinclair
Emerging Issues Associated
Barbara
Sarah
John
Bart
Kim
Scott
Mark
Yolanda
Eugene
James E.
David
Pamela
Molly
Judith
Mike
Paul
Stig
Ken
Susan
James
with Aquatic Environmental
ORD/NHEERL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/NRMRL
ORD/OSP
ORD/OSP
ORD/OSP
OW/IO
OW/OGWDW
OW/OGWDW
OW/OGWDW
OW/OGWDW
OW/OGWDW
OW/OGWDW
Pathogens September 5-September 7, 2001
walton.barbarat@epa.gov
bauer.sarah@epa.gov
cicmanec j ohn@epa.gov
faulkner.bart@epa.gov
fox.kim@epa.gov
j acobs . scott@epa.gov
meckes.mark@epa.gov
Yolanda Olivas/
ADA/USEPA/US@EPA
rice .gene@epa.gov
smith.james@epa.gov
klauder.david@epa.gov
noyes.pam@epa.gov
whitworth.molly@epa.gov
hecht.judy@epa.gov
mjmessner@aol .com
berger.paul@epa.gov
regli . stig@epa.gov
rotert.kenneth@epa.gov
shaw.susan@epa.gov
sinclair.james@epa.gov
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Verma
Dannel
Hoffmann
Kovatch
Kuzmack
Pendergast
Schaub
Wigal
Invited Guests
Burke
Diallo
Hyer
LeChevallier
Miliotis
Neukrug
Powell
Rosen
Silvers
Sobsey
Kiran
Mimi
Rick
Charles
Arnold
James
Stephen
Jennifer
Tom
Alpha A.
Ken
Mark
Marianne
Howard
Mark
Barry
Linda
Mark
OW/OPPTS
OW/OST
OW/OST
OW/OST
OW/OST
OW/OST
OW/OST
OW/OST
Johns Hopkins
University
Norfolk Department of
Public Health
VA USGS District
American Water Works
Service
Company, Inc.
FDA/CFSAN
Philadelphia Water
Commission
U.S. Department of
Agriculture
U.S. Department of
Agriculture
FDA Center for
Veterinary
Medicine
University of North
Carolina
verma.kiran@epa.gov
dannel .mimi@epa.gov
hoffrnann.rick@epa.gov
kovatch.charles@epa.gov
kuzmack.arnold@epa.gov
pendergast.j im@epa.gov
schaub . stephen@epa.gov
wigal.jennifer@epa.gov
adiallo@vdh.state.va.us
kenhyer@usgs.gov
mlecheva@amwater.com
mmilioti@cfsan.fda.gov
howard.neukrug@phila.gov
mpowell@oce .usda.gov
barry.rosen@ftw.nrcs.usda.gov
Lsilvers@cvm.fda.gov
sobsey@email.unc.edu
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Appendix B: Slides from Presentations
These slides can be found at
http://intranet.epa.gov/ospintra/regsci/pathogen.html
1. Overview of Emerging Pathogens
2. The Clean Water Act's Requirements for
Pathogens
3. Safe Drinking Water Act/Contaminants
Pathogens Candidate List
4. Unregulated Contaminant Monitoring of
Pathogens
5. Update onEPA's "Implementation Guidance for
Ambient Water Quality Criteria for Bacteria"
6. Developing a Strategy for Waterborne Microbial
Disease Control
1. EPA Implementation of Beaches Environmental
Assessment and Coastal Health (BEACH) Act
8. Summary of Major Regional Scientific/Technical
Priorities
9. Waterborne Disease Outbreaks in the U.S.
Regional Issues
10. Waterborne Disease: EPA's Intramural
Epidemiology Program
11. An Estimate of National Waterborne Disease
Occurrence
12. Microbial Risk Assessment: State of the Art and
Science
13. A Dose-Response Envelope for E. coli O15 7:H7
14. Vibrio parahaemolyticus Risk Assessment
Jake Joyce
James F. Pendergast
Paul S. Berger
James Sinclair
Jennifer Wigal
Stephen Schaub
Charles Kovatch
Vicky Binetti
Kim Ngo
Rebecca L. Calderon
Susan Shaw
Bruce A. Macler
Mark Powell
Marianne Miliotis
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
15. Pathogen Risk Assessment Protocol for
Environmental Water Media
16. Risk Assessment in Regulatory Development
17. Microbial Risk Assessment Activities in
Cincinnati Division of EPA's National Center for
Environmental Assessment
18. Assessing Exposures to Emerging Pathogens
What the Regions Need
19. Assessing Exposure to Bacterial Pathogens
20. Development of Molecular Methods to Detect
Emerging Viruses
21. Detection of Protozoan Pathogens in Water
22. Current Approaches to Identifying Sources of
Fecal Contamination
23. Antimicrobial Resistance Among Enteric Bacteria
Isolated from Human and Animal Wastes and
Impacted Surface Waters
24. User-Friendly Models for Evaluating
Hydrogeologic Barriers to Viruses
25. Water Supply and Water Resources Division
Response to Waterborne Disease Outbreaks
26. Disinfection of Emerging Pathogens
27. Biofilms in Drinking Water Distribution Systems
28. Update on 6-year Review ofTCR and Potential
Distribution System Rule
29. Case Study: Biofilm Problem at Greenville South
Carolina
3 0. EPA's National Beach Survey
31. Development of Improved Monitoring Approaches
for Recreational Water
Stephen Schaub
Stig Regli
John C. Lipscomb/Brenda Boutin
Terry Fleming
Gerard N. Stelma
Ann C. Grimm/Christina Newport/G.
Shay Fout
Frank W. Schaefer
Gerard N. Stelma
Mark Sobsey
Bart Faulkner et. al
Kim R. Fox
Eugene W. Rice
Mark C. Meckes
Ken Rotert
David Parker
Rick Hoffman
Stephen Schaub
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
32. Use of Molecular Techniques to Provide the
Public with more Information on Recreational
Water Closures in Norfolk
33. Acceptable Goals and Thresholds for Pathogens
from Recreational Water Exposures
34. Determining Sources ofMicrobial Contamination
35. Rapid Methods for Measuring Recreational Water
Quality: A Look to the Future
3 6. ORD 's Drinking Water Research Program
Tracking System
1>1. A Regional Perspective on CAFO/Biosolid
Regulations: How ORD Can Help Us!
3 8. Overview of Watershed Features and Dynamics
Associated with Drinking and Recreational Water
Protection
39. Potential Watershed Movement of Pathogens of
Animal Origin
40. Emerging Infectious Disease Agents and Issues
Associated with the Management of Treated
Sludges (Biosolids)
41. Risks from Pathogens in Biosolid Fertilizers/Land
Application of Sludge NRC Report
Alpha Diallo
Alfred P. Dufour
Joel Hansel
Alfred P. Dufour
Alfred P. Dufour/Cheryl Itkin
Stephanie Harris
J. E. Smith
John Cicmanec
J. E. Smith
Tom Burke
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U.S. Environmental Protection Agency
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Appendix C: Breakout Group Summary
DISCLAIMER: The thoughts below were collected during a brief interactive session
at the workshop. They do not represent Agency policy.
GROUP 1: What endpoints and factors should be considered in microbial risk
assessments?
Moderator - Bruce Macler (Region 9)
A. How do we account for mixtures of pathogens in sources in risk assessment?
Premise:
It is typical to have more than one pathogen present in the source
Only fecal pathogens are considered
There is no indicator or single pathogen that can predict the kind or number of other
pathogens that may be present
Suggestions:
* Consider the worst-case situation for the source
» Evaluate sources for general types of pathogens
B. How should we consider more sensitive people in risk assessment and management?
Premise:
Approximately twenty percent (20%) of the general population can be classified as
sensitive
Approximately one percent (1%) of the general population is considered super-
sensitive
Suggestions:
» Use a modifying factor in dose-response assessment to account for sensitivity (as is
done for Reference Dose)
» Focus on illness endpoints rather than infection
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
» Focus risk management on sensitive and supersensitive populations:
Education/Outreach to limit exposures
Guidance Materials
Vaccination programs
Minimize pathogens in sources
Redirect sources
» Identify all sensitive and super-sensitive populations
* Determine if sensitivity varies with pathogen type
» Consider the acute, subacute, and chronic effects of pathogens
» The cumulative impact of multiple pathogens must also be considered
GROUP 2: Recognizing the uncertainties inherent in microbial risk assessment:
what data would provide the most value added?
Moderator - Mike Messner (OW)
Participants in this second breakout group were asked to break into smaller groups in order to
consider separately the data needed for each of three groups of organisms, both in drinking and
recreational water, and the needs for total microbial risk in drinking water for all organisms:
1. Viruses in Drinking and Recreational Water (Subgroup 1)
2. Bacteria in Drinking and Recreational Water (Subgroup 2)
3. Protozoa in Drinking and Recreational Water (Subgroup 3)
4. Total Microbial Risk attributable to Microbes in Drinking Water (Subgroup 4)
The four sub-groups above each focused on four major areas where data is needed:
1) Occurrence - Where are they and in what quantities before they enter the treatment or
distribution system?
* Organisms that originate in the source water and pass through the treatment barriers
* Organisms that enter through pipe connections (not originating in source water)
and/or grow in the distribution system
2. Treatment and Distribution (Drinking Wateityeafffihat hfe|ipt@nlrt(lihhdi]sitgjinitai»$ systten?
3. Control (Recreational Water) - How effectively do watershed and other controls
reduce risk?
4. Infectivity/Illness - How infectious is the organism? (How is the probability of infection
related to dose? Given infection, what is the probability of illness / severity of illness?
How protective is immunity?
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
At the end of the breakout sessions the four subgroups reported their findings to the attendees.
Summaries of each subgroup's conclusions follow, below.
SUBGROUP 1'. Viruses in Drinking Water and Recreational Water
Moderator - Ann Grimm (ORD/NERL)
Research priorities were decided to be the same for all viruses, with no distinction made between
specific types of viruses. Additionally, needs were deemed to be similar in both drinking water
and recreational water.
The research needs were ranked in order of importance, and the justification for these rankings
was reported.
Priorities:
1. Development of methods
2. Data on occurrence
3. Research to identify an indicator
Justification:
1. Consideration of acute effects, especially on sensitive sub-populations
2. Consideration of potential long-term, insidious effects
SUBGROUP 2: Bacteria in Drinking Water and Recreational Water
Moderator - Gerard Stelma (ORD/NERL)
Bacteria were listed by genus in order of their importance to drinking water and recreational water.
For each bacterium, the data needs were ranked: one (1) was assigned to the type of data most
needed for estimating risk, and three (3) to the type of data least needed or already available.
In drinking water the most important bacteria were:
(I) Legionella;
(2) NT Mycobacteria; and
(3)E. coli O157:H7 and Cyanobacteria.
Data on treatment was the type most needed for both Legionella and NT Mycobacteria, while data
on occurrence was ranked most important for Cyanobacteria and E. coli O157:H7.
In the case of recreational water, the most important bacteria were:
(l)E. co//O157:H7
(2) Shigella and Campylobacter
Several other bacteria were considered equally important to be included under #3
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
(See Recreational Water table below)
Data was needed most in the area of control for E. coll O157:H7. Data on occurrence was
considered most important for Shigella, whereas data needs were not ranked in the case of
Campy lobacter.
The information on research needs for all the bacteria considered by Subgroup 2 is outlined in the
two tables, below.
Bacteria in Drinking Water
Bacterium (Rank)
(1) Legionella
(2) NT Mycobacteria
(3)E. co//O157:H7
(3) Cyanobacteria
Helicobacter
Aeromonas
HPC
Occurrence (Rank)
2
2
1
1
Treatment (Rank)
1
1
2
2
Infectivity (Rank)
O
2
O
3 - toxicity
Bacteria in Recreational Water
Bacterium (Rank)
(1)E. co//O157:H7
(2) Shigella
(2) Campylobacter
(3) Vibrio
(3) Aeromonas
(3) Pseudomonas
(3) Streptococcus
(3) Enterococcus
(3) Leptospira
(4) Cyanobacteria
Occurrence (Rank)
2
1
2
2
Control (Rank)
1
2
1
1
Infectivity (Rank)
3
O
O
O
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
SUBGROUP 3: Protozoa in Drinking Water and Recreational Water
Moderator - Mark LeChevallier (American Water Works Services Company)
Protozoa were first categorized by this subgroup according to the risk level each species presents
in drinking water and recreational water. The same three protozoan species were listed as having a
high risk level in both recreational and drinking water:
Cryptosporidium, Giardia, and Acanthamoeba.
Two additional protists, Entamoeba and Naegleria were identified as carrying a high-risk level in
recreational water only
With regard to data needs, the group focused on the data needed for drinking water treatment.
These needs were considered to be the same for all protozoa, regardless of species, and were
ranked in order of importance as follows:
(1) Exposure data
(2) Viability and infectivity data
(3) Data on strain variations
The full results reported by this subgroup are presented in the two tables on the following page.
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Risk Level in Drinking Water and Recreational Water
PATHOGEN
Cryptosporidium
Giardia
Acanthamoeba
Microsporidia
Entamoeba
Naegleria
Cyclospora
Toxoplasma
Isospora
RISK LEVEL
Drinking Water
H
H
H
M
L
L
L
L
L
Recreational water
H
H
H
?
H
H
L
L
L
[H = High; M = Medium; L = Low]
Data Needs for Protozoa in Drinking Water
SOURCE
WATER
Occurrence
t
Outbreaks
t
Reservoir and
transport hosts
GOAL:
TREATMENT
1
Variation
1
No surrogates
1
Difference in
design and
operation
T
NO MODEL
1.
EXPOSURE
High volume
methods
Frequency
monitoring
Attributable risk
from water
Relate to
antibody levels
2.
VIABILITY
INFECTIVITY
Cell culture=?
Animal
infectivity
=
Human
infectivity
3.
STRAIN
VARIATION
Infectivity
Variation
Genotypes
Pathogenicity
Role of
immunity
To improve treatment estimates it is necessary to improve exposure, viability and strain
variation.
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Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
SUBGROUP 4: Studies of TotalMicrobial Risk Attributable to Drinking Water
Moderator - Rebecca Calderon (ORD/NHEERL)
This last subgroup considered the data most needed to assess total microbial risk in drinking
water
The types of data needs they considered crucial are listed in order of importance below:
1. Intervention studies
2. Drinking water system classification strategy
3. Increased FoodNet coverage to correlate with the system
4. Serology data
5. GIS [Geographic information systems] data
6. Investigation of alternative disease surveillance
7. Probabilistic risk analysis
8. Medical outreach and education
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Appendix D: Flip Chart Notes
PART 1: WEDNESDAY, SEPTEMBER 5:
The notes included in this appendix represent the main points brought up during a Regional panel
discussion and facilitated question and answer session intended to develop a list of some of the
highest priority science issues to be addressed. These issues, and others brought up throughout the
meeting, were addressed by ORD representatives at the end of the workshop.
The discussion centered around two posed questions:
Question 1: Is our science on pathogens appropriate and adequate to support regulatory
management?
Question 2: What can be done today to address some of the microbial problems that are just
now emerging?
QUESTION 1:
Is our science on pathogens appropriate and adequate to support regulatory management?
Discussion of this question focused on the areas where more and/or better science is needed
to support regulatory management. The needs discussed can be divided into two general
areas: waterborne disease characterization and indicators.
WATERBORNE DISEASE CHARACTERIZATION NEEDS/GAPS
For recreational water, characterization of disease endpoints other than gastro-
intestinal symptoms
Analytic and disease incidence monitoring tools, such as:
* Quick, unambiguous methods for pathogen screening
» Non-traditional methods: expansion of FoodNet; monitoring of the sales of
over the counter anti-diarrheal medications
Definition of an outbreak and better ways to monitor and characterize outbreaks
Linking of outbreak/disease to source (whether drinking or recreational water)
Dose-response relationships
A better understanding of disease progression and its secondary spread
Understanding of the special susceptibilities of "at-risk" populations
Improved characterization of the incidence of waterborne disease. Specifically:
» Endemic vs. epidemic levels of disease incidence
EM
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
» Disease pathways, other than waterborne
Bridge the gap between drinking water, environmental, and health agencies
INDICATORS
Faster and better indicators are needed for the detection of pathogens, especially
bacterial pathogens.
A better understanding is needed of what indicators really mean, as well as what
information they cannot give.
QUESTION 2:
What can be done today to address some of the microbial problems that are just now emerging?
Discussion of this question centered around risk assessment and risk management.
Emphasize multi-barrier approach to risk management:
* source protection
* treatment
» disinfection
Information on how to approach risk management on a watershed basis, and for
source water assessment and protection
More information on the effectiveness of controls such as disinfection and best
management practices (BMPs)
Importance (to risk assessment) of factors such as die-off/survivability of pathogens
in varying environmental conditions
Better knowledge of the risks associated with recreational water use (ambient water
quality)
Data on bacteria source tracking (especially important for TMDLs)
» Methods
» Dimensions of investigation area
* Size of library
» Number of isolates
Assessment of risks from various pathogen sources; some examples include:
* Combined sewage overflows (CSOs)
» Storm water
* Biosolids
» Publicly owned treatment works (POTWs)
» Algal toxins
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U.S. Environmental Protection Agency
Regional/ORD Workshop: Emerging Issues Associated with Aquatic Environmental Pathogens September 5-September 7, 2001
Appendix E: Emerging Aquatic Pathogens Workshop
Participant Evaluation Summary
Most participants found that the workshop gave them a better overall understanding of the issues
associated with Emerging Aquatic Pathogens. Several participants were of the opinion that the
workshop was useful, but covered too broad a topic, and could have been longer, or divided into
separate, more specific workshops1. Attendees from the regional offices also liked the fact that the
sessions began by addressing regional needs from the perspective of regional presenters. Some
commended the inclusion of speakers from outside the EPA.
The responses regarding the most interesting and least interesting topic varied widely, with most
attendees explaining they were most interested in topics related to their own field (e.g., drinking
water vs. recreational water, microbiology vs. epidemiology.). The breakout sessions were thought
to be a good opportunity to delve deeper into specific topics, and participants especially
appreciated the reporting of each group's conclusions at the end of the sessions. Several attendees
did caution that there was not enough time to come up with meaningful, well-thought-out results.
The majority of participants found the format of the workshop to be a good balance of
presentations, discussions, and small group sessions; some, however, thought the time for
questions should be more flexible, to accomodate presentations that elicit longer discussions.
Many participants appreciated the opportunity to establish contacts between ORD and the regional
offices. Suggestions for continuing this interaction included the creation of an email listserv and
an arbitrated intranet discussion forum divided by topic. In general, the feedback for this workshop
was very favorable. The most common suggestion was that this workshop should be held regularly
(e.g. on an annual basis) to continue the interaction between ORD and the regions.
Note from the Organizers: Topics for the workshop were solicited from all 10 regions, leading to a fairly extensive
list including bioterrorism, airborne pathogens, resistant pathogens, and cyanobacteria. Because this broad list
covered far too much material for one workshop, the organizing team for the workshop selected two major topics on
which to focus: drinking water and recreational water. It was decided that the other topics would be covered in the
future, using the Placeware remote conferencing system. At one point there was discussion on having parallel
presentations for those interested in the two topics selected by the organizing team, however, it was decided that
there was more benefit in having the two groups interact.
E-l
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