5>EPA
     United States
     Environmental Protection
     Agency
RESEARCH  PROJECT COLLABORATIO
          National Exposure Research Laboratory
     Microbiological & Chemical Exposure Assessment Research Division &
     National Risk Management Research Laboratory
                Water Supply and Water Resource Division
INFLUENCE OF DISINFECTANT RESIDUAL ON BIOFILM DEVELOPMENT, MICROBIAL
ECOLOGY, AND PATHOGEN FATE AND TRANSPORT IN DRINKING WATER INFRASTRUCTURE
                                      1 /urn
 Legionella cells growing within an amoeba that may colonize piped water biofilms
                                 IMPACT STATEMENT
                     One of the U.S. Environmental Protection Agency's (EPA)
                     primary roles is that of an advocate for sustainable water
                     infrastructure. Biofilms, which form on interior surfaces
                     of  the drinking water infrastructure, can  sequester,
                     accumulate and protect  viruses, bacteria, fungi, and
                     parasitic  protozoa. Release  of biofilms may  therefore
                     release a more infectious dose back into drinking water.
                     The experimental studies performed through this project
                     are designed to evaluate risks to human health posed by
                     pathogens  associated  with these  biofilms, initially
                     focusing  on bacterial  pathogens  that may grow  in
                     association  with amoebae in biofilms. By providing the
                     science and engineering information that we and our
                     partners  need,  EPA  research  contributes  measurable
                     results that advance our efforts to ensure safe drinking
                     water.
BACKGROUND:
Biofilms form on interior surfaces of the drinking water infrastructure, including storage facilities and distribution
systems.  Pathogens may enter drinking water systems in the bulk water or through intrusions or physical breaches.
Biofilms can sorb pathogens (viruses, bacteria and parasitic protozoa) and serve as a protective niche for their survival.
In addition, opportunistic pathogens, principally respiratory pathogens such as nontuberculous mycobacteria [NTM] and
Legionella-\\ke bacterial  pathogens, may grow within biofilms in storage facilities and distribution systems, including
premise plumbing.  Recent data suggests that disinfection practices may selectively control opportunistic pathogens.
For example, chloramination is reputed to control Legionella, but may be less effective for NTM bacteria. However,
there are limited data  on  how disinfection strategies impact the biofilm  community structure that  may support
opportunistic pathogens and that may also provide a niche for sequestering fecal pathogens. Current models assessing
risks from drinking water are limited by not accounting for pathogen biofilm effects.

DESCRIPTION:
This project focuses on  providing basic data to bound risk estimates resulting from  pathogens associated with pipe
biofilms. Researchers will compare biofilm pathogen effects under two different disinfection scenarios (free chlorine or
chloramines) for a conventionally treated source water  (Potomac River).   Samples will  be collected from storage
facilities, dedicated sampling taps used for TCR compliance monitoring, and biofilm swabs  from household cold water
taps and shower heads (premise plumbing). Microbial community structures, NTM, and Legionella-\\ke bacteria and their
amoebae hosts will be characterized.  Potential novel biomarkers of exposure will be collected and used to explore  the
      National Exposure Research Laboratory &
      National Risk Management Research Laboratory

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potential for salivary and urine antibody tests.  Water  quality data, operations information,  and system condition
information will also be collected to aid in interpretation of microbial data.  In addition to disinfectant residual, key
water quality variables including nitrogen species (ammonia, nitrite, nitrate, and organic-nitrogen), phosphorus, metals
(lead, copper, arsenic), fluoride, organic carbon, pH, alkalinity, temperature, sulfate, chloride, and other inorganics will
be assessed. All potential pathogens will be genotyped and stored for future reference. In addition, if a water main
break or other ingress event occurs during the project-sampling phase, material  will  also be stored for molecular
identification of fecal pathogens and indicators.
EPA's Office of Research and Development has funded this research project in support of its  Aging Water Infrastructure
Research Program.  Phase I of this project will run from  March 2010 through March 2011,  and dependent on project
success, Phase II will run for a second year.
EPA GOAL: Goal #2 - Clean & Safe Water; Objective 2.1.1- Water Safe to Drink
ORD MULTI YEAR PLAN: Drinking Water (DW), Long Term Goal (LTG) - DW-1 Characterize risks associated with DW sources,
distribution, treatment, and use
RESEARCH PARTNERS:
•   EPA's National Exposure Research Laboratory:  Stacy Pfaller (NTM); Shay Fout (salivary antibodies)
•   EPA's National Risk Management Research Laboratory: Laura Boczek (biofilm disinfection); Randy Revetta (biofilm community
    structure)
•   EPA's National Health and Environmental Effects Research  Laboratory: Elizabeth Hilborn & Andrey Egorov (human health
    effects)
•   EPA's Office of Water:  Susan Shaw (coordination of EPA staff/home access)
•   Washington Aqueduct & WSSC contacts

EXPECTED OUTCOMES AND IMPACTS:
It is expected that this study will have  a  direct impact on public health by providing utilities information on pathogen
risks associated with biofilm growth  in  storage facilities and water distribution systems. This information will facilitate
better management decisions, which will result in a  decrease in the risk of waterborne illness.

OUTPUTS:
Primary outputs from  this project  will be journal and technical papers describing key  parameters that will enable
quantitative microbial risk assessments of opportunistic and fecal pathogens associated with biofilms in drinking water
systems. This information will enable updates to EPA's hydraulic models for drinking water distribution systems, so that
the role of pathogen-biofilm interactions can be included.

RESOURCES:
Aging Water Infrastructure Research Program: http://www.epa.Rov/awi/
National Exposure Research Laboratory: http://www.epa.Rov/nerl/
National Risk Management Research Laboratory: http://www.epa.Rov/nrmrl/

CONTACTS:
Nicholas Ashbolt, Principal Investigator - (513) 569-7318 or ashbolt.nick@epa.gov
Steven Doub, Media Relations - (513) 569-7503 ordoub.steven@epa.gov
Michelle Latham, Communications - (513) 569-7601 orlatham.michelle@epa.gov
      National Exposure Research Laboratory &
      National Risk Management Research Laboratory
EPA/600/F-10/012
February 2010

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