Posters Presentations
202
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Day Three: Session X
J
iL2a
False Positives Associated with Enterococcus
Quantification Using EPA-Approved Methods for
Industrial Discharge
Presenter: Emelie Andersson
University of North Carolina, Chapel Hill, Institute of Marine Sciences
Authors: Emelie Andersson, Angelia Blackwood, Kellen Lauer, Rachel Noble
Abstract
Industrial wastewater discharged into
recreational waters is routinely monitored
for Enterococcus spv fecal coliforms, or E.
coli. The most commonly used culture-based,
U.S. Environmental Protection Agency- (EPA-)
approved methods for quantification of
Enterococcus sp. are EPA method 1600 and
Enterolert*. For this study discharge samples
were collected twice per month for 1 year
and quantified by EPA 1600 and Enterolert.
Additionally a suite of qPCR analyses were
conducted on the samples, including total
Enterococcus (EPA 1611), quantification of spe-
cific Enterococcus species, E.faecalis, E.faecium,
and E. casseliflavus, and quantification of three
human-associated Bacteroidales-based mark-
ers (Fecal Bacteroides, HF183, and BacHum).
Published guidelines were closely followed to
ensure quality quantitative data. Enterococcus
sp. concentrations ranged from 109-2,070
(mean=819) CFU/100 ml and 86,207-5,246,223
(mean =719,679) CE/100 ml as determined by EPA
1600 and EPA 1611, respectively. There was no
correlative relationship between EPA 1600 and
1611 (r=-0.10). Concentrations of E.faecalis and
E.faecium were below detection limit for 10 of 24
samples (42%). None of the human-associated
markers were quantifiable in any of the sam-
ples. Twenty isolates were selected from each
EPA 1600 plate for confirmation using Vitek-2
(Biomerieux, Inc.), and the results indicated that
the fecally associated Enterococcus sp. were not
strongly present, while 42% of the isolates were
identified as non-Enterococcus species. This
finding was supported by the molecular analy-
ses. Interestingly, plant and dairy-associated
species (such as Aerococcus viridans) dominated
the characterized species. The results indicate
that for certain types of industrial discharge,
Enterococcus sp. quantification is a questionable
proxy for presence of fecal contamination.
Biosketch
Ms. Emelie Andersson graduated in 2013
with a bachelor of science degree in biology
from Mount Olive College, North Carolina,
where she also was a student athlete. Since
March 2014, she has been working as a research
technician in Dr. Rachel Noble's laboratory
at the Institute of Marine Sciences of the
University of North Carolina at Chapel Hill in
Morehead City, North Carolina. Ms. Andersson
has an interest in microbial source tracking and
how it impacts public health.
203
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Application of Hydrodynamic Modeling to Predict
Viral Impacts from Wastewater Treatment Plant
Discharges Adjacent to Shellfish Growing Areas
Presenter: Yaping Ao
U.S. Food and Drug Administration
Authors: Yaping Ao, Gregory Goblick
U.S. EPA's 2016 Recreational Waters Conference
Abstract
Since the inception of the National
Shellfish Certification Program in 1925—now
the National Shellfish Sanitation Program-
dilution analysis has been used as a means to
minimize the presence of enteric pathogenic
microorganisms in shellfish growing areas.
Today, failures and bypasses at wastewater
treatment plants (WWTPs) and combined sewer
overflows (CSOs) that release untreated or
partially treated sewage into shellfish receiv-
ing waters are of particular interest to shellfish
control authorities and public health officials.
To assess the risk posed by wastewater efflu-
ents, the U.S. Food and Drug Administration
(FDA) has used several approaches, including
hydrodynamic modeling, to predict the extent
of sewage impacts on receiving waters. Models
used by FDA were calibrated and validated by
hydrographic dyes studies and microbiological
analyses. The simulated results are extremely
helpful to shellfish management authorities
for determining appropriate classification
and management for shellfish growing areas.
Hydrodynamic modeling also has proven to be
a valuable tool for a current joint United States/
Canada quantitative norovirus risk assessment.
FDA and the National Oceanic and Atmospheric
Administration will be collaborating on efforts
to develop ecoforecasting capabilities using
modeling and weather (precipitation) forecasts
to predict sewage impacts on shellfish grow-
ing areas that occur from WWTP bypasses and
CSOs during storm-related events.
Biosketch
Ms. Yaping Ao is a visiting associate
with the U.S. Food and Drug Administration,
Center for Food Safety and Applied Nutrition
in College Park, Maryland. She serves as a lead
modeler in the application of computer fate and
transport models to assess pollution source
impacts to shellfish growing areas. She assisted
with the development of dilution models to
support a joint U.S.-Canada norovirus risk
assessment and has provided training on and
led specialized field and hydrographic studies
to identify and assess pollution sources in the
environment. Ms. Ao's expertise includes sup-
porting the development of guidance for irriga-
tion water use for produce safety. She received
her master of science degree in civil engineering
from Marquette University in Wisconsin and
her bachelor of science degree in environmental
engineering from the Chengdu University of
Technology, China.
204
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Day Three: Session X
J
iL2a
Beach Sand and the Potential for Infectious Disease
Transmission: Observations and Recommendations
Presenter: Joao Brandao, PhD
National Institute of Health
Department of Environmental Health - Water and Soil Unit
Authors: Helena Solo-Gabriele, Joao Brandao
Abstract
Recent studies suggest that sand can serve
as a vehicle for exposure of humans to patho-
gens at beach sites, resulting in increased health
risks. Sampling for microorganisms in sand
should, therefore, be considered for inclusion in
regulatory programs aimed at protecting rec-
reational beach users from infectious disease.
We reviewed the literature on pathogen levels
in beach sand and their potential for affecting
human health. In an effort to provide specific
recommendations for sand sampling programs,
we outline published guidelines for beach
monitoring programs, which are currently
focused exclusively on measuring microbial
levels in water. We also provide background on
spatial distribution and temporal characteristics
of microbes in sand, as these factors influence
sampling programs. First steps toward estab-
lishing a sand sampling program include iden-
tifying appropriate beach sites and using initial
sanitary assessments to refine site selection. A
tiered approach is recommended for monitor-
ing, which would include analyzing samples
from many sites for fecal indicator organisms
and other conventional analytes, while test-
ing for specific pathogens and unconventional
indicators would be reserved for high-risk sites
and possible outbreaks. Given the diversity of
microbes found in sand, studies are urgently
needed to identify the most significant etiologi-
cal agent of disease and to relate microbial mea-
surements in sand to the risk to human health.
(J Marine Biol Ass UK, 2015
Biosketch
Dr. Joao Brandao is an applied chemist-
biotechnologist from the New University of
Lisbon, Portugal, specializing in clinical mycol-
ogy and environmental research. He has con-
ducted research in those two areas of expertise
since 2001 and lectured on them at Lusofona
University in Lisbon. Mr. Brandao is the envi-
ronmental microbiology research manager at
the Department of Environmental Health of the
National Institutes of Health, a national del-
egate of the Portuguese Association of Medical
Mycology to the European Confederation
of Medical Mycology, (council member) and
national member of the European Microbiology
Research Group for the European Commission
(Bathing Water Directive). Of special interest to
Dr. Brandao are microbial pathogen contami-
nants in sand, air, and water; nosocomial and
endemic fungal infections of environmental
origin; and prevention and early molecular
detection. Other special interests include water
contaminants and quantitative microbial risk
assessment in water quality and microbial
source tracking tools (for point-source pollution
detection and resolution).
205
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U.S. EPA's 2016 Recreational Waters Conference
Risk Levels of Toxic Cyanobacteria in
Portuguese Recreational Freshwaters
Presenter: Joao Brandao, PhD
National Institute of Health
Authors: Carina Menezes, Catarina Churro, Joao Brandao, Elsa Dias
Abstract
In this work, we present the results from
monitoring cyanobacteria and associated
toxins in several recreational freshwaters from
central and southern Portugal. We identified
two distinct scenarios: (a) reservoirs that never
exhibited potentially toxic cyanobacteria; and
(b) reservoirs that showed a persistent occur-
rence of cyanobacterial blooms, often associated
with the presence of microcystins. Comparing
our results with the guideline values for cya-
nobacterial cells in bathing waters and the
corresponding expected microcystin levels, we
determined that 30% of the reservoirs had high
risk for public health. However, the cyanobacte-
rial cell density criterion might overestimate
the risk because it includes nontoxic strains of
potentially toxic species. Also, this criterion is
based on the worst scenario for toxin cell pro-
duction quota, which might be higher than the
real toxin cell content detected in cyanobacterial
bloom samples. In fact, the risk level decreases
considerably when we compare the real micro-
cystin concentrations in water samples with
the expected values. The applicability of these
criteria in risk assessment of cyanobacteria
and cyanotoxins in recreational freshwaters is
discussed.
Biosketch
Dr. Joao Brandao is an applied chemist-
biotechnologist from the New University of
Lisbon, Portugal, specializing in clinical mycol-
ogy and environmental research. He has con-
ducted research in those two areas of expertise
since 2001 and lectured on them at Lusofona
University in Lisbon. Mr. Brandao is the envi-
ronmental microbiology research manager at
the Department of Environmental Health of the
National Institutes of Health, a national del-
egate of the Portuguese Association of Medical
Mycology to the European Confederation
of Medical Mycology, (council member) and
national member of the European Microbiology
Research Group for the European Commission
(Bathing Water Directive). Of special interest to
Dr. Brandao are microbial pathogen contami-
nants in sand, air, and water; nosocomial and
endemic fungal infections of environmental
origin; and prevention and early molecular
detection. Other special interests include water
contaminants and quantitative microbial risk
assessment in water quality and microbial
source tracking tools (for point-source pollution
detection and resolution).
206
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Day Three: Session X
J
iL2a
Stretching Budgets with Composite Sampling
Presenter: Shannon Briggs, PhD
Michigan Department of Environmental Quality
Author: Shannon Briggs
Abstract
The advantage of using composite samples
is that combining multiple samples provides
an indication of the water quality at the cost of
one sample. In general, results for composite
samples are equivalent to taking the arith-
metic mean of the analytical results of each
of the samples that comprise the composite.
Mathematically an arithmetic mean is never
less than and is generally greater than a geo-
metric mean, although this is not always true in
real-world sampling. A composite beach sample
could potentially result in more beach closings
and advisories than would a geometric mean
for the same beach.
Arithmetic and geometric means, however,
tend to be similar for samples from beaches
that have homogeneous historical monitoring
data. Homogeneity can be determined with the
Kruskal-Wallis test, which is a nonparametric
test equivalent to a one-way analysis of variance
that determines whether there are between-
site differences. Furthermore, an arithmetic or
geometric mean is less likely to exceed criteria
by selecting beaches that have homogeneity and
consistently low bacteria levels.
Wymer et al. (2005), Kinzelman et al.
(2006), Bertke (2007), Wymer and Wade 2007,
Reicherts and Emerson (2010), and USEPA (2010)
support the use of composite sampling to char-
acterize the mean indicator density and encour-
age more sampling while minimizing costs. In
Michigan, composite sampling is the cost-effec-
tive approach for implementing qPCR methods.
Results from culture and qPCR methods on
composite samples will be evaluated to establish
correlations between the two methods.
Biosketch
Dr. Shannon Briggs is a toxicologist for
the Water Resources Division of the Michigan
Department of Environmental Quality (DEQ).
She received her bachelor of science degree in
animal science and her doctorate in pharma-
cology and toxicology from Michigan State
University. She is a member of a planning
team that will host the 2016 Great Lakes Beach
Conference in Marquette, Michigan, October
5-7, 2016. Dr. Briggs assists local health depart-
ments with state and federal grants for monitor-
ing beaches across the State of Michigan. She is
leading a water quality initiative of the DEQ to
provide rapid testing equipment and training
for 10 new laboratories that will test beaches
using the U.S. Environmental Protection
Agency's draft Method C (i.e., qPCR method
for E. coli). Dr. Briggs is an active member, past
president, and cofounder of the Great Lakes
Beach Association.
207
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U.S. EPA's 2016 Recreational Waters Conference
Pollution Source Identification, Tracking, and
Sanitary Survey on Italian Beaches
Presenter: Annalaura Carducci
University of Pisa
Author: Annalaura Carducci, Marco Verani, lleana Federigi, Renato Iannelli
Abstract
In Italy, sea-bathing tourism is a very
important socioeconomic resource, but it is
being impacted by the rapid urbanization of the
coastal environment, which represents pollution
pressure on water quality with point and dif-
fuse sources of fecal contamination. This study
was carried out at Versilia, a popular bathing
destination northwest of Tuscany, where short-
term pollution posed the problem of possible
classification as "scarce" (according to the
current European Bathing Water Directive)
because of fecal contamination from drainage
ditches. Our goal was to understand the impact
of polluted streams on seawater contamina-
tion and the effect of meteorological conditions
on freshwater and seawater bacterial indicator
levels. The monitoring results from the 2012 to
2015 bathing seasons were analyzed and, only
for 2015, we used cultural and biomolecular
techniques to detect fecal-oral pathogens and
viral indicators at ditch mouths. Our results
demonstrate a relationship between bacterial
indicator levels and rainfall amount; however,
microbiological pollution also was highly vari-
able in dry weather, suggesting the presence
of undetected sources of fecal contamination.
Collected data from seawater and river mouths
show a time-dependent dilution effect of the
sea that varied between the two halves of each
bathing season. During 2015—the last year of
monitoring—besides bacteria indicators, we
found only the human adenovirus genome. This
analytical survey of the pollution sources in
Versilia recreational water could be used to cre-
ate larger monitoring data sets for developing
predictive models of microbial contamination in
relation to climatic conditions and disinfection
intervention
Biosketch
Ms. Carducci is a professor of general and
applied hygiene in the Department of Biology
at the University of Pisa, where she graduated
with bachelor's and master's degrees in biology,
specializing in hygiene and public health. Prof.
Carducci supervises the Biology Department's
Hygiene and Environmental Virology
Laboratory and Health Communication
Observatory. The focus of her teaching activity
as well as her research includes microbiological
monitoring of food and environmental matrices
and biological risk analysis for life and work-
ing settings and for foods, health, and risk
communication.
208
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Day Three: Session X
J
iL2a
Expanding EPA Method 544: Addition of Seven
Microcystin Congeners for Analysis of Lake Erie
Beach Samples, a Comparative Study with ELISA
Presenter: Mark Citriglia
Northeast Ohio Regional Sewer District
Authors: Cheryl Soltis-Muth, Deborah Schordock
Abstract
Thirty-seven Lake Erie beach water sam-
ples from the summer of 2015 were analyzed for
microcystins by U.S. Environmental Protection
Agency (EPA) method 544 using solid phase
extraction (SPE) and analysis on a Shimadzu
LCMS-8050 Liquid Chromatograph Tandem
Mass Spectrometer (LC/MS/MS). These same
samples were lysed by a series of three freeze/
thaw cycles and filtered. The resulting extracts
were analyzed by ELISA and also by directly
injecting them into the LC/MS/MS with the
same operating conditions as with EPA method
544. In 19 cases, the ELISA result was greater
than the sum of the individual microcystin
results from EPA method 544 (SPE & LC/MS/
MS). In all except one of the 19 instances, the
ELISA also was greater than the sum of the
individual microcystin results from the direct
inject LC/MS/MS method. This indicates that
there may be additional microcystin congeners
in the samples that are not being detected by
EPA method 544.
Seven additional microcystin congeners
were added to the EPA method 544 analysis.
The beach water extracts from 2015 will be
rerun to determine if the difference between
the ELISA and EPA method 544 results is due to
microcystin congeners previously untested by
the method.
Future work includes validating the SPE
extraction procedure in EPA method 544 with
the additional microcystin congeners and
continuing the ELISA and LC/MS/MS method
comparison study into 2016.
Biosketch
Mr. Mark Citriglia is currently the man-
ager of analytical services for the Northeast
Ohio Regional Sewer District (NEORSD). Mr.
Citriglia has a bachelor of science degree in biol-
ogy with a minor in chemistry from Cleveland
State University and is a certified State of Ohio
wastewater class III operator and a class IV
certified wastewater analyst. He has worked
for NEORSD for 27 years, beginning his career
there as a wastewater analyst in the Analytical
Services Department (laboratory), successfully
gaining the skills and knowledge necessary to
advance to managing the lab. As the laboratory
manager for the past 12 years, Mr. Citrigilia has
been committed to operating the lab in a safe,
professional, and proficient manner, generating
data of known and documented quality. He vol-
untarily adopted the quality policies and proce-
dures outlined by the National Environmental
Laboratory Accreditation Program, obtaining
accreditation in 2007. Currently, he is expanding
the lab's capabilities to include a research and
development department along with additional
accreditations. Mr. Citriglia is a member of the
Water Environment Federation (WEF), NELAC
Institute, and Lab Safety Institute. He was the
recipient of the 2007-2008 Northeast Section
Ohio Water Environment Association Lab
Analyst Award, 2009 Ohio Water Environment
Association Lab Analyst Award, and 2010 WEF
Laboratory Excellence Award.
209
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U.S. EPA's 2016 Recreational Waters Conference
Make It "Tough:" qPCR Master Mix Comparison
Presenter: Mark Citriglia
Northeast Ohio Regional Sewer District
Author: Nichole Schafer
Abstract
Within the 2012 Recreational
Water Quality Criteria guidelines, the
U.S. Environmental Protection Agency (EPA)
provides information for states that want to
adopt water quality standards based on rapid
methods that EPA has developed and validated
using qPCR. Currently EPA has validated
methods 1609 and 1611 for the quantification of
Enterococcus in environmental waters using
qPCR. The Northeast Ohio Regional Sewer
District laboratory collected samples from two
northeast Ohio beaches and analyzed them
using EPA method 1609 and a qPCR assay for E.
coli. The objectives of this study were to iden-
tify a master mix that (1) could be used for both
assays, (2) reduced inhibition, (3) is similar to
the Environmental Master Mix (EMM) recom-
mended by EPA, (4) uses an internal amplifica-
tion control allowing for analysis on the same
platform, and (5) contains a purified DNA
polymerase that is validated to be free of extra-
neous E. coli DNA. A total of 241 samples were
analyzed for both organisms using two differ-
ent chemistries, EMM as stated in method 1609
and Tough Mix. The data for each organism
and master mix combination were compared
for inhibition, method QC, level of quantifica-
tion, and difference between assay absolute and
relative quantitation. Accuracy, sensitivity, and
specificity were calculated as compared to the
conventional method. In conclusion, we have
determined that the Tough Mix yielded data
comparable in accuracy and reproducibility
to the EPA-recommended mix, but the level
of inhibition and instance of positive QC data
decreased with the Tough Mix.
Biosketch
Mr. Mark Citriglia is currently the man-
ager of analytical services for the Northeast
Ohio Regional Sewer District (NEORSD).
Mr. Citriglia has a bachelor of science degree
in biology with a minor in chemistry from
Cleveland State University and is a certified
State of Ohio wastewater class III operator and
a class IV certified wastewater analyst. He has
worked for NEORSD for 27 years, beginning
his career there as a wastewater analyst in the
Analytical Services Department (laboratory),
successfully gaining the skills and knowledge
necessary to advance to managing the lab. As
the laboratory manager for the past 12 years,
Mr. Citrigilia has been committed to operat-
ing the lab in a safe, professional, and profi-
cient manner, generating data of known and
documented quality. He voluntarily adopted
the quality policies and procedures outlined
by the National Environmental Laboratory
Accreditation Program, obtaining accredita-
tion in 2007. Currently, he is expanding the
lab's capabilities to include a research and
development department along with additional
accreditations. Mr. Citriglia is a member of the
Water Environment Federation (WEF), NELAC
Institute, and Lab Safety Institute. He was the
recipient of the 2007-2008 Northeast Section
Ohio Water Environment Association Lab
Analyst Award, 2009 Ohio Water Environment
Association Lab Analyst Award, and 2010 WEF
Laboratory Excellence Award.
210
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Day Three: Session X
J
iL2a
qPCR: A Screening Tool for Harmful Algal
Blooms
Presenter: Mark Citriglia
Northeast Ohio Regional Sewer District
Authors: Nichole Schafer, Mark Citriglia
Abstract
Lake Erie has seen an increase in the num-
ber and severity of harmful algal blooms (HAB)
caused by cyanobacteria (blue-green algae). The
cyanobacteria present in the HABs can poten-
tially produce toxins capable of causing illness
and/or death. Timely and accurate identification
and reporting of these toxins is critical for issu-
ing water quality advisories. The methods used
to analyze toxin are very expensive, and select-
ing the correct one can be difficult. Another
challenge is the necessity for a skilled analyst
to be available to identify the microscopic algae.
Furthermore, some cyanobacteria can produce
multiple toxins (e.g., the genus Anabaena can
produce anatoxin-a, saxitoxin, and microcystin;
the genus Aphanizomenon can produce saxi-
toxin, anatoxin-a, and clylindrospermopsin).
The Northeast Ohio Regional Sewer District
(NEORSD) decided to experiment with a rapid
method using qPCR to screen for a total cya-
nobacteria gene and specific toxin-producing
genes (i.e., microcytins, saxitoxin, and cylindro-
spermopsin). The NEORSD laboratory experi-
mented with the Phytoxigene, CyanoDTec qPCR
assays to screen samples submitted for cyano-
toxin analysis. A portion of the sample submit-
ted for analysis was filtered, and the DNA was
extracted and analyzed on multiple qPCR plat-
forms. The results of our study indicate that this
method has the potential to eliminate the need
for microscopic examination and to simplify the
process of selecting the appropriate method for
toxin analysis.
Biosketch
Mr. Mark Citriglia is currently the man-
ager of analytical services for the Northeast
Ohio Regional Sewer District (NEORSD).
Mr. Citriglia has a bachelor of science degree
in biology with a minor in chemistry from
Cleveland State University and is a certified
State of Ohio wastewater class III operator and
a class IV certified wastewater analyst. He has
worked for NEORSD for 27 years, beginning
his career there as a wastewater analyst in the
Analytical Services Department (laboratory),
successfully gaining the skills and knowledge
necessary to advance to managing the lab. As
the laboratory manager for the past 12 years,
Mr. Citrigilia has been committed to operat-
ing the lab in a safe, professional, and profi-
cient manner, generating data of known and
documented quality. He voluntarily adopted
the quality policies and procedures outlined
by the National Environmental Laboratory
Accreditation Program, obtaining accredita-
tion in 2007. Currently, he is expanding the
lab's capabilities to include a research and
development department along with additional
accreditations. Mr. Citriglia is a member of the
Water Environment Federation (WEF), NELAC
Institute, and Lab Safety Institute. He was the
recipient of the 2007-2008 Northeast Section
Ohio Water Environment Association Lab
Analyst Award, 2009 Ohio Water Environment
Association Lab Analyst Award, and 2010 WEF
Laboratory Excellence Award.
211
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U.S. EPA's 2016 Recreational Waters Conference
Rapid and Efficient Extraction Method of
Chlorophyll by Bead Beating
Presenter: Mark Citriglia
Northeast Ohio Regional Sewer District
Authors: Debmalya Bhattacharyya, Mark Citriglia
Abstract
Estimation of chlorophyll a has been
routinely applied to determine algal and phy-
toplankton levels in marine and freshwater
to assess the eutrophic status and, thereby
monitor water quality The existing method
extracts chlorophyll a from glass fiber filters by
manual mastication in the presence of cold 90%
acetone. The manual mastication of the filters
is a prolonged and arduous process that allows
processing of only two samples (in triplicates)
in 20 minutes (i.e., about 3 minutes on average
for processing each glass fiber filter). An alter-
native method of bead beating the glass fiber
filters is using an Omni bead ruptor 24. The new
method significantly reduced the time of pro-
cessing of two samples analyzed in triplicates.
Comparative analyses of the new and existing
extraction methods were conducted keeping
all other processing parameters constant and
the chlorophyll a content was determined.
Comparison of the results from the two meth-
ods was statistically analyzed by Passing and
Bablok regression, which suggests that there
were no significant systematic or proportional
differences between the methods of extraction.
It also was noted that the standard deviations
among the data obtained by the bead beating
method were less than by the existing method,
which probably is due to less handling being
required. We conclude that the application of
bead beating of the glass fiber filters can be
used with better efficiency to routinely process
chlorophyll a samples.
Biosketch
Mr. Mark Citriglia is currently the man-
ager of analytical services for the Northeast
Ohio Regional Sewer District (NEORSD).
Mr. Citriglia has a bachelor of science degree
in biology with a minor in chemistry from
Cleveland State University and is a certified
State of Ohio wastewater class III operator and
a class IV certified wastewater analyst. He has
worked for NEORSD for 27 years, beginning
his career there as a wastewater analyst in the
Analytical Services Department (laboratory),
successfully gaining the skills and knowledge
necessary to advance to managing the lab. As
the laboratory manager for the past 12 years,
Mr. Citrigilia has been committed to operat-
ing the lab in a safe, professional, and profi-
cient manner, generating data of known and
documented quality. He voluntarily adopted
the quality policies and procedures outlined
by the National Environmental Laboratory
Accreditation Program, obtaining accredita-
tion in 2007. Currently, he is expanding the
lab's capabilities to include a research and
development department along with additional
accreditations. Mr. Citriglia is a member of the
Water Environment Federation (WEF), NELAC
Institute, and Lab Safety Institute. He was the
recipient of the 2007-2008 Northeast Section
Ohio Water Environment Association Lab
Analyst Award, 2009 Ohio Water Environment
Association Lab Analyst Award, and 2010 WEF
Laboratory Excellence Award.
212
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Day Three: Session X
J
iL2a
Child Exposure to Water and Sand at the Beach:
Findings from Studies of over 80,000 Subjects at
13 Beaches
Presenter: Stephanie DeFlorio-Barker, PhD
U.S. Environmental Protection Agency, Human Studies Facility
Authors: Stephanie DeFlorio-Barker, Benjamin F. Arnold, John M. Colford Jr.,
Steve Weinberg, Ken Schiff, Elizabeth Sams, Alfred Dufour, Timothy J. Wade
Abstract
Swimming and recreating in lakes, oceans,
and rivers are common activities among adults
and children; some studies have suggested that
children could be at greater risk of illness than
adults following such exposures. These effects
could be caused by differences in immunity
or differing behavioral factors such as poorer
hygiene, as well as longer exposures to and
greater ingestion of potentially contaminated
water and sand. We pooled data from 13 pro-
spective cohorts to examine exposures to poten-
tially contaminated media such as beach water
and sand among children compared to adults,
and summarized time spent in the water among
swimmers (n=48,573) by age category. Age cat-
egories were younger than 1 year, 1-3 years, 4-7
years, 8-12 years, 13-18 years, 19-34 years, and
35 years and older. Children aged 4-7 and 8-12
had the highest exposures at the beach of any
of the age groups. Among children aged 8-12,
27% swallowed water, 63% dug in the sand, and
17% got sand in their mouth, compared to 6%,
22%, and 5% of adults 35 and older, respectively.
Across all beach sites, children 8-12 spent 122
minutes in the water (SD= 85 minutes), while
adults aged 35 and over spent 75 minutes in the
water (SD=68 minutes). All exposures reported
among children were statistically significantly
greater than those reported among adults
(p < 0.001). These findings and estimates might
help inform quantitative microbial risk assess-
ments to better estimate health risks associated
with recreational water exposures.
Note: This abstract does not represent EPA
policy.
Biosketch
Dr. Stephanie DeFlorio-Barker is a postdoc-
toral research fellow at the U.S. Environmental
Protection Agency. She received her bachelor of
science degree in molecular and cellular biol-
ogy from the University of Illinois at Urbana-
Champaign, and her master of public health
degree in epidemiology and doctorate in envi-
ronmental and occupational epidemiology from
the School of Public Health at the University
of Illinois at Chicago. Dr. DeFlorio-Barker's
research interests include water quality, eco-
nomic burden of environmental exposures, risk
assessment, and communication of environ-
mental health risks. For her doctoral disserta-
tion, she conducted research on the severity and
economic burden associated with surface water
recreation in the United States.
213
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U.S. EPA's 2016 Recreational Waters Conference
Recreational Water Quality in Massachusetts:
Historical Inventory of Freshwater and Marine Beach
Surveillance from 2001-2015
Presenter: Irena Draksic
Massachusetts Department of Public Health, Bureau of Environmental Health
Authors: Michael Celona, Vanessa Curran, Irena Draksic, Michael Beattie,
Margaret Round, Marc Nascarella
Abstract
Water quality data in Massachusetts
represents a rich resource for understanding
recreational water quality dynamics. With
over 1,100 freshwater and marine beaches,
Massachusetts has the greatest number of
beaches in the northeastern United States. The
Massachusetts Department of Public Health
has been monitoring bacterial levels at these
beaches since 2001, where they have collected
a robust data set of over 200,000 individual
water quality samples. This data set represents
a robust historical inventory of laboratory
results from individual water quality samples,
coupled with field observations recorded by the
sample collector. For example, in addition to
data describing bacterial levels (bacterial levels
in colony-forming units per 100 ml), this inven-
tory includes parameters such as temperature,
bather density, and rainfall. These samples
were all collected during the meteorological
summer, at a frequency of approximately once
per week. Multiyear data analyses suggest that
exceedances of water quality standards occur
less frequently now, as compared to previous
years. For example, of 14,874 samples collected
at 1,086 beaches in 2014, approximately 3.5% of
beaches exceeded the Enterobacteriaceae bacte-
ria standard. Historically, about 5% of beaches
exceed the respective marine or freshwater
bacterial standard. Our field-collected data and
field observation suggest that the majority of
exceedances are directly related to rain events
and likely the result of increased surface runoff.
The data described here will be used in future
efforts to understand the relationship between
climate-related changes in rainfall and the
health impacts of potential increased exposure
to fecal bacteria.
Biosketch
Ms. Irena Draksic is a beach inspector
at the Massachusetts Department of Public
Health, Bureau of Environmental Health's
Environmental Toxicology Program (ETP). She
has been with the department for 11 years.
Ms. Draksic manages beach data and closures
during the bathing beach season, assists with
sanitary surveys and freshwater algae bloom
sampling, and provides training and technical
support to contract laboratories and local com-
munities. She has a bachelor of science degree in
biology from Denison University and a master
of arts degree in environmental science and
policy from Clark University.
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Day Three: Session X
J
iL2a
Preparing for New Water Quality Standards at NYC
Beaches: Retrospective Evaluation of Water Quality
Data and Updating Hydrodynamic Models
Presenter: Charles Dujardin
Great Lakes Environmental Center
Authors: Christopher Boyd, Anand Kumaraswamy, Charles Dujardin
Abstract
The New York City (NYC) Department
of Health and Mental Hygiene (DOHMH) is
responsible for monitoring and surveillance
of all of the city's permitted beaches. The U.S.
Environmental Protection Agency's (EPA's) 2012
Recreational Water Quality Criteria will impact
beach notification in NYC because of its more
conservative geometric mean, new statisti-
cal threshold value capturing long-term risk,
and new beach action value alerting the public
before water quality declines to unacceptable
levels. DOHMH conducted a retrospective
analysis of beach monitoring data to analyze
the potential increase of advisory and closure
days if the new 2012 criteria had been in effect.
The study found on average a 31-day increase in
the number of pollution advisory days per year
and a 6-day increase in the number of closure
days per year. These increases were not uni-
formly distributed across the city's beaches. The
implications of the new criteria include the need
for increased beach program staffing for more
frequent sampling and risk communication and
education outreach.
Additionally, to better anticipate poor
water quality as a result of rain events,
DOHMH and the NYC Department of
Environmental Protection (DEP) revisited the
advisory "triggers" using a hydrodynamic
model developed by Stevens Institute and math-
ematical kinetics previously developed by DEP.
Regression curves were developed between
duration of hours the model exceeded 110
colony-forming units/100 milliliters and rainfall
volume. From these regressions, advisory trig-
gers and "durations" can be developed for each
beach. For a given rainfall volume, expected
time and duration of rainfall, and expected
duration of exceedance, an assessment can be
conducted to determine whether an advisory
should be issued for a particular day.
Biosketch
Mr. Charles Dujardin received his bache-
lor's degree in chemical engineering and mas-
ter's degree in environmental engineering from
Manhattan College. Mr. Dujardin has extensive
experience with water quality assessments,
including total maximum daily load develop-
ment for pathogens and dissolved oxygen,
particularly in urban settings. He has been the
project manager for many water quality model-
ing studies conducted for the City of New York,
which have evaluated engineering alternatives
for controlling combined sewer overflows and
stormwater discharges. Also included has been
the development of a state-of-the-art eutro-
phication model to assess the effectiveness of
nutrient abatement schemes. For Region 2 of the
U.S. Environmental Protection Agency, he was
project manager for the development of was-
teload allocations for toxic metals discharged
to the New York-New Jersey Harbor. He has
extensive experience with assimilative capac-
ity studies, site feasibility studies, and seasonal
treatment alternatives.
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On Rapid Assessment Methods Using Statistical
Modeling: Multiple Least Squares Regression vs.
Logistic Regression
Presenter: Jay Fleisher, PhD
NOVA Southeastern University, College of Medicine
Author: Jay Fleisher
U.S. EPA's 2016 Recreational Waters Conference
Abstract
Background: There is a need to develop
rapid assessment of bacterial water quality. To
this end many statistical models have been pub-
lished mostly using environmental variables to
predict concentrations of a particular FIO. The
majority of these statistical models have used
multiple least squares regression, in which the
major indicator of the goodness of fit of these
models have largely depended on the R2 value,
which to date has been quite low. Since beach
management decisions have to be dichotomous
in nature (i.e., open/close beach), we explored
the use of the multiple logistic model in relation
to the multiple least squares approach.
Methods: 668 samples were used in this
analysis. 10 major environmental variables and
several FIOs were collected on each sample date.
Both types of models were run on those data.
Results: Our best multiple least squares
regression was computed with an R2 value
of 0.26, while the multiple logistic regression
model yielded a maximum sensitivity of 72.9
percent and a maximum specificity of 65.9 per-
cent at a cut point = 0.1. A backward selection
routine was used in both the models.
Conclusions: Since the logistic regression
yields a much less nebulous goodness of fit
statistic, coupled with the fact that the beach
manager's decision is a dichotomous one, more
attention should be paid to research using the
multiple logistic model.
Biosketches
Dr. Jay Fleisher received his bachelor of
science degree in environmental health science
and master of science degree in environmental
science from the City University of New York,
his master of science degree in epidemiology
from Columbia University's School of Public
Health, and his doctorate in environmental
epidemiology/biostatistics from the Institute of
Environmental Medicine, New York University.
Dr. Fleisher holds faculty positions at Florida's
Nova Southeastern University and University
of Miami. Dr. Fleisher's research interests are in
the fields of chronic and infectious illnesses. He
has focused his research efforts on the health
effects of exposure to waters contaminated with
domestic sewage, indicator organism variabil-
ity, indicator organism-pathogen relationships,
risk assessment, statistical water quality sam-
pling protocols, assessing compliance, setting
of microbial water quality standards, popula-
tion health burden assessment, risk perception,
and risk vs. current standards. Dr. Fleisher
has advised numerous international commit-
tees, organizations, and government agencies
on various aspects of these recreational water
quality issues. In addition, he has authored over
70 peer-reviewed publications and six book
chapters.
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Day Three: Session X
J
iL2a
Wayne Creek: A Microbial Source Tracking Case
Study
Presenter: Raul Gonzalez, PhD
Hampton Roads Sanitation District
Authors: Raul Gonzalez, Kyle Curtis, Danny Barker
Abstract
Microbial source tracking (MST) meth-
ods were used by Hampton Roads Sanitation
District to characterize chronic bacterial con-
tamination during dry and wet weather in
the Lafayette River (Norfolk, Virginia). The
objectives of this study were to 1) partition
the sources of fecal contamination entering
the Lafayette River, and 2) define the origin
of contamination to work in a smaller, more
manageable area. During this study, the City of
Norfolk identified a leaking sewer pipe in the
subwatershed. Therefore, an additional objec-
tive was to evaluate water quality after the
repair. Sites were sampled on a downstream-
to-upstream transect, high-priority stormwater
segments were visited early in the sampling
campaign, and subsequent samples were col-
lected upstream in the stormwater network.
Septic systems, which were initially suspected
of causing the observed contamination, did not
influence the headwaters at the sample loca-
tions monitored. To quantify the magnitude of
human contamination relative to raw wastewa-
ter, MST data were calibrated using pump sta-
tion raw influent. Human fecal contamination,
measured by the HF183 marker, was confined to
one "main" channel, which reduced the prob-
lem area to a manageable size. After the leaking
force main was found and repaired, human con-
tamination in the headwaters disappeared. Prior
to this repair, water quality adjacent and down-
stream of the leak was similar to raw influent
levels. Costs and obstacles encountered when
starting an MST laboratory will be discussed.
Biosketch
Dr. Raul Gonzalez is an environmental
scientist at Hampton Roads Sanitation District
(HRSD), where he uses state-of-the-art source
tracking techniques to identify potential sources
of contamination in Hampton Roads. Since
joining HRSD, he has been working on vari-
ous water quality projects to quantify storm-
water loads and bacteria runoff. Dr. Gonzalez
is a native of California, where he graduated
from University of California, Los Angeles
with a degree in biology. After graduation, he
worked at the Los Angeles County Sanitation
Districts before returning to graduate school at
University of North Carolina- (UNC-) Chapel
Hill, where he earned his doctorate in envi-
ronmental science and engineering. While at
UNC, Dr. Gonzalez used molecular methods
to pinpoint causes of high bacteria levels in
surface waters. He now uses this microbial
source tracking experience to assist regional
stakeholders.
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Heal the Bay's Beach Report Card: Communicating
Complex Water Quality Issues and Improving Public
Health
Presenter: Leslie Griffin
Heal the Bay
Authors: Leslie Griffin, James Alamillo
U.S. EPA's 2016 Recreational Waters Conference
Abstract
Beach water quality often gains wide-
spread attention only when there is a substan-
tial sewage spill. Beachgoers typically do not
concern themselves with the potential health
effects associated with swimming in ocean
waters impacted by more everyday sources such
as polluted runoff, most likely due to a simple
lack of awareness. Historically it has been chal-
lenging for health agencies to effectively convey
the health risks associated with swimming at
these beaches. The focus of beach water quality
risk management is typically on epidemiologi-
cal research, risk assessment, and limitations
with conventional sampling and measurement;
yet few resources have been directed toward
ensuring that the messaging effectively reaches
the beachgoer. For 25 years, Heal the Bay has
provided the public with an easy-to-use tool
for deciding where to swim based on the most
recent bacteriological data. The Beach Report
Card® (BRC) provides the beachgoing public on
the West Coast with easy-to-understand water
quality information about their favorite beaches.
The BRC assigns a letter grade based on sample
results from local health agencies. Grades are
calculated using an algorithm that takes into
account frequency and magnitude of exceed-
ances of single sample and geometric mean rec-
reational water quality standards. The BRC has
been an incredibly successful and effective tool
for public notification. Today, the BRC provides
health information for more than 600 beaches
along the West Coast and has become part of
the public and regulatory vernacular. This past
summer, the BRC expanded its dissemination
of beach water quality information by posting
predictive model or "Nowcasting" results for
three southern California beaches. This effort
was the result of a Clean Beaches Initiative
grant from the state and will expand to include
18-25 beach locations over the next 3 years.
Biosketch
Ms. Leslie Griffin is the beach water
quality scientist at the Los Angeles-based
environmental organization, Heal the Bay.
Native to the East Coast, she relocated across
country to receive her bachelor and master
of science degrees in environmental science
with an emphasis in water quality from Loyola
Marymount University. She worked on passive
sampling of PAHs for 2 years while obtaining
her master's degree. While pursuing her educa-
tion, Ms. Griffin interned at Heal the Bay as an
aquarist and a watershed educator. In 2015, she
began working full time with the organization
as the data analyst for the Beach Report Card
program. Currently, Ms. Griffin manages the
Beach Report Card program—working to ensure
accurate and timely dissemination of weekly
beach water quality info for over 600 locations
along the West Coast, as well as implementing
a daily predictive modeling—or "nowcasting"—
program for five beaches in Southern California.
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Day Three: Session X
J
iL2a
The Next Generation PCR-Based Quantification
Method for Ambient Waters: Digital PCR
Presenter: John Griffith, PhD
Southern California Coastal Water Research Project
Authors: Yiping Cao, John Griffith, Stephen Weisberg
Abstract
Real-time quantitative PCR (qPCR) is
increasingly being used for ambient water mon-
itoring, but development of digital polymerase
chain reaction (digital PCR) has the potential
to further advance use of molecular techniques
in such applications. Digital PCR refines qPCR
by partitioning the sample into thousands to
millions of miniature reactions that are exam-
ined individually for binary end-point results,
with DNA density calculated from the fraction
of positives using Poisson statistics. This direct
quantification removes the need for standard
curves, eliminating the labor and materials
associated with creating and running standards
with each batch, and removing biases associ-
ated with standards variability and mismatch-
ing amplification efficiency between standards
and samples. Confining reactions and binary
end-point measurements to small partitions
also leads to other performance advantages,
including reduced susceptibility to inhibition,
increased repeatability and reproducibility,
and increased capacity to measure multiple
targets in one analysis. As such, digital PCR is
well suited to ambient water monitoring appli-
cations, and is particularly advantageous as
molecular methods move towards autonomous
field application.
Biosketch
Dr. John Griffith is principal scientist in the
Department of Microbiology as well as coordi-
nator of molecular technology at the Southern
California Coastal Water Research Project. He
received his doctorate in marine biology and
biological oceanography from the University
of Southern California. Dr. Griffith has been at
the forefront of research to develop, evaluate,
and implement methods for rapid assessment of
beach water quality, microbial source identifica-
tion, and detection of waterborne pathogens.
He is the author of more than 60 articles on the
use of Foraminifera as bioindicators, prokary-
otic and viral diversity in marine plankton,
microbial source tracking, rapid water qual-
ity measurement methods, and relationships
between microbial water quality indicators and
swimming-related illness.
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U.S. EPA's 2016 Recreational Waters Conference
Comparison of Agar- and Pectin-Based Methods for
the Detection of Male-Specific Coliphage
Presenter: Joseph Guzman
Orange County Public Health Laboratory
Authors: J.A. Guzman, T.T. Chiem, R.C. Alexander
Abstract
Water quality is an important public
health concern affecting millions of lives. Many
waterborne illnesses are caused by enteric virus
contamination in the water. Current test meth-
ods involve the detection of bacterial indicators,
which is inadequate in predicting the patho-
genic levels of enteric virus. Due to similarities
to enteric virus in structure, composition, and
resistance to environmental extremes, coli-
phages have been considered as a surrogate for
determining enteric virus levels in water quality
assessment. This study focused on the detection
of male-specific (F+) coliphage using agar-based
U.S. Environmental Protection Agency (EPA)
method 1602—single agar layer (SAL)—in com-
parison to commercially available pectin-based
Easyphage. Each method involved combin-
ing surface water, sewage, or spiked samples
with log-phase Escherichia coli on the associated
media. After appropriate incubation, detection
of coliphage was identified by plaque forma-
tion in a lawn of E. coli host growth. In spiking
experiments, samples with a predetermined
concentration of F+ coliphage corresponding to
80 plaque-forming units (PFU) per 100 mil-
liliters (mL) were tested by both methods to
determine recovery rates. In spiked samples, the
highest recovery rate for SAL was 25% (20/80)
and 98% (78/80) for Easyphage with a mean
recovery rate of 7% and 69%, respectively. In
surface water samples, Easyphage consistently
showed more recovery of coliphage than the
SAL method with a mean recovery of 123.6
PFU/lOOml for Easyphage and 53.8 PFU/lOOml
for SAL. In one sample, Easyphage recov-
ered 25 times more coliphage than SAL. The
pectin-based Easyphage method is less labor-
intensive and more effective in the recovery of
F+ coliphage in comparison to the agar-based
SAL method. Incorporating the Easyphage
method with traditional bacterial indicators
would provide a more complete assessment of
water quality.
Biosketch
Mr. Joseph Guzman is the supervis-
ing public health microbiologist oversee-
ing the Orange County Public Health Water
Quality Laboratory located in Newport Beach,
California. He received his bachelor of science
degree in medical microbiology from California
State University, Long Beach, and, for the last 25
years, has been with the Orange County Public
Health Laboratory working in all areas of public
health microbiology. Mr. Guzman first started to
focus on water quality testing in 1998, becoming
involved in the bacterial monitoring of Orange
County's beaches, harbors, and estuaries. The
laboratory continues to do routine beach moni-
toring while also participating in research proj-
ects in the development of rapid indicator meth-
ods, development of alternative fecal indicators,
predictive modeling, and determining sources
of bacterial pollution in receiving waters.
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Day Three: Session X
J
iL2a
Beachgoer Behavior During a Retrospectively
Detected Algal Bloom at a Great Lakes Beach
Presenter: Elizabeth D. Hilborn, PhD
U.S. Environmental Protection Agency, Environmental Public Health Division
Authors: Elizabeth D. Hilborn, Whitney S. Krueger, Blake A. Schaeffer, Richard P. Stumpf,
Elizabeth A. Sams, Timothy ). Wade
Abstract
Algal blooms occur among nutrient rich,
warm surface waters and can adversely impact
recreational beaches. During July-September
2003, a prospective study of beachgoers was
conducted on weekends at a public beach on
one of the Great Lakes in the United States.
We measured each beachgoer's activity at the
start and end of their beach visit and the envi-
ronmental factors—water and air temperature,
wind speed, and wave height—at the study site
each day. At the time, there was no notification
of algal blooms; however, we retrospectively
evaluated the presence of algal blooms using
MERIS data from the Envisat-1 satellite.
A total of 2,840 people participated in the
study over 16 study days. The majority (55%)
were female, and 751 (26%) were < 18 years of
age. An algal bloom was detected retrospec-
tively by remotely sensed satellite imagery dur-
ing August 16-24. This peak bloom period (PB)
included 4 study days. During PB study days,
more study participants 226/742 (31%) reported
body contact with the water compared to con-
tact 531/2098 (25%) on nonpeak days. During the
4 PB days, of the environmental factors, only
mean water temperature was significantly dif-
ferent—25 degrees Centigrade (°C) vs. 23 °C (p <
0.05)—from other days.
These results suggest that beachgoer body
contact with water was not deterred by the pres-
ence of an algal bloom and that interventions
to actively discourage water contact during a
bloom are needed to reduce exposure to blooms.
Note: This is an abstract of a proposed pre-
sentation and does not necessarily reflect EPA
policy.
Biosketch
Dr. Elizabeth D. Hilborn is a senior health
scientist (epidemiologist) in the Environmental
Public Health Division in the Office of Research
and Development of the U.S. Environmental
Protection Agency (EPA). Dr. Hilborn's expertise
is in the human health effects of waterborne
contaminants including toxic cyanobacteria. She
currently serves on the Interagency Working
Group for the Harmful Algal Blooms, Hypoxia,
Research and Control Act; on the Scientific
Committee for the 10th International Conference
on Toxic Cyanobacteria; and on multiple
cyanobacteria-focused EPA committees and
workgroups. Dr. Hilborn earned her bachelor of
science degree in biology from the University of
North Carolina (UNC) at Chapel Hill, a doctor
of veterinary medicine degree at North Carolina
State University College of Veterinary Medicine,
and a master of public health degree at the
UNC at Chapel Hill. Dr. Hilborn served in the
Centers for Disease Control and Prevention's
Epidemic Intelligence Service and is board-cer-
tified with the American College of Veterinary
Preventive Medicine.
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U.S. EPA's 2016 Recreational Waters Conference
Washington County, Minnesota, Lake Surveillance
for Naegleria fowleri from 2011-2014
Presenter: Vincent Hill, PhD
Centers for Disease Control and Prevention
Authors: Bonnie Mull, Jessica L. Collin-Pilarski, Amy Kahler, Erik Anderson, Matt Downing,
Fred Anderson, Vincent Hill
Abstract
The presence of the thermophilic free-
living ameba, Naegleria fowleri, was investigated
in lakes in Washington County, Minnesota,
including one lake associated with the deaths
of two children in 2010 and 2012 from primary
amebic meningoencephalitis (PAM). Samples of
lake water and sediment were collected from 10
lakes in the summer months annually from 2011
to 2014. In addition to culture and PCR analyses
for N. fowleri, samples were analyzed for a suite
of physical, chemical, and biological parameters
potentially related to its presence and growth.
Environmental parameters such as rainfall and
air temperature also were investigated as poten-
tial factors related to detecting the pathogen.
N. fowleri was detected in five of the 10 lakes
in 2011, in one lake in 2012, and in none of the
lakes in 2013 or 2014. It was detected in 2011 and
2012 in the lake associated with the two cases of
PAM. Average air temperatures prior to sample
collection were higher in 2011 and 2012 than in
2013 and 2014, suggesting that the hot weather
during those summer periods might have been
associated with the growth and detection of
N. fowleri. The amount of precipitation within
13 days of sample collection was positively asso-
ciated with detection of N. fowleri. The results
of this study provide environmental and water
quality data that will help increase understand-
ing of N. fowleri ecology and potential environ-
mental and water quality factors that could be
associated with the detection of this pathogen
in recreational water bodies.
Biosketch
Dr. Vincent Hill is a research environ-
mental engineer in the Waterborne Disease
Prevention Branch at the Centers for Disease
Control and Prevention, National Center for
Emerging and Zoonotic Infectious Diseases.
Dr. Hill's work focuses on the development and
application of environmental sampling meth-
ods, microbial detection methods, and treatment
technologies for water and wastewater sys-
tems. He is an author of over 90 peer-reviewed
journal articles and scientific reports focused
on environmental microbiology and engineer-
ing. In addition to directing the Waterborne
Disease Prevention Branch's Environmental
Microbiology Laboratory, Dr. Hill also is team
lead for the Water, Sanitation, and Hygiene
Laboratory Team.
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Day Three: Session X
J
iL2a
An Economic Evaluation of the Demand for
Beach Safety Information
Presenter: Abigail Kaminski
University of Maine, School of Economics
Authors: Abigail Kaminski, Kathleen Bell, Caroline Noblet, Keith Evans
Abstract
Coastal beaches and the recreation oppor-
tunities they provide are important economic
assets. It is vital that the health and safety of
users be considered in determining how these
resources are managed. The health and safety
of coastal waters is impacted by a diverse and
changing set of problems; large-scale nutrient
runoff, land use change, heavy precipitation
events, and failing or aging infrastructure can
all have negative impacts on coastal water qual-
ity. State and local organizations use a variety
of means to communicate this information
to beach users, but little is known about the
diversity of beach users and how they seek out
information on beach safety.
In this study, we analyze beach safety
information-seeking behavior using data from
a survey of Maine and New Hampshire beach
users. We estimate a series of discrete regres-
sion models to explain search behaviors. Our
sample seek out surf condition information at a
higher rate than water quality information, and
results to date show differences in the factors
that influence demand for these two types of
information, suggesting that users might regard
the risks associated with each differently. These
findings provide valuable insight for coastal
resource managers and public health officials
as they develop effective plans to reach diverse
beach users.
Biosketch
Ms. Abigail Kaminski is a graduate student
in the School of Economics at the University of
Maine, pursuing a master of science degree in
resource economics and policy. She received
her bachelor of arts degree in economics and
geography from Clark University in Worcester,
Massachusetts. After graduating from Clark,
Ms. Kaminski spent 2 years teaching middle
school math with Teach for America in Miami,
Florida, and 1 year as an assistant director of
admissions at Clark University. She is currently
working as a research assistant on an interdisci-
plinary research team focused on strengthening
decision-making around the management of
coupled coastal systems. Ms. Kaminski's thesis
research focuses on the decision-making behav-
ior of coastal beach users, and she has a broad
interest in the ways diverse individuals interact
with and value natural resources.
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U.S. EPA's 2016 Recreational Waters Conference
Prolonged Survival of Viable Fecal Bacteroidales
in Marine Sediments as Assessed by Propidium
Monoazide Real-time PCR
Presenter: Minji Kim, PhD
Department of Civil and Environmental Engineering, University of California—Davis
Authors: Minji Kim and Stefan Wuertz
Abstract
Members of the order Bacteroidales are the
most widely used fecal identifiers in microbial
source tracking (MST). While their decay in
natural waters has been investigated at length
and rate constants have been determined under
various conditions, fate and transport studies
in sediments are few and far between. Yet this
knowledge is critical when using MST to evalu-
ate recreational water quality.
To estimate the decay of fecal Bacteroidales
in oxygen-free sediments, we constructed
anaerobic sediment microcosms in which
human, cow, and dog feces were completely
mixed with sediments and incubated them at
6 degrees Centigrade (°C) and 20 °C. In addi-
tion, intact sediment core microcosms in which
fecal slurry was spiked in the overlying water
were established to investigate the rate of
transport of Bacteroidales to the sediment sur-
face. Microcosms were sacrificed in triplicate
on each sampling date up to 42 d followed by
qPCR analysis with or without addition of the
DNA-modifying dye propidium monoazide to
differentiate genetic markers from viable cells
and total intracellular as well as extracellular
marker DNA.
In anaerobic microcosms, the survival and
persistence of host-associated Bacteroidales cells
and DNA were considerably extended, espe-
cially at the lower temperature of 6SC; cells and
DNA showed two-log reduction times (T99) of
109 d or higher at 6°C and 14-25 d (cells) as well
as 27-47 d (DNA) at 20°C. In core microcosms,
Bacteroidales cells in sediments decayed up to
ten times more slowly than cells in the overly-
ing water column. In both microcosms, various
host markers decayed at similar rates under
each condition tested. Cells decayed signifi-
cantly faster than total DNA in marine sedi-
ments at 20°C while their decay was comparable
at 6°C, suggesting seasonal variation should
be considered in application of MST in marine
sediments.
The prolonged persistence of fecal
Bacteroidales in sediments indicated in the study
implies that sediments can act as nonpoint
sources of fecal markers, causing false positive
results when monitoring beach water quality
because the bacteria do not reflect recent fecal
pollution events. This study demonstrates the
need to consider the contribution of MST mark-
ers in sediments in field monitoring approaches.
Biosketch
Dr. Minji Kim received her doctorate from
the University of California, Davis (UCD) and
has conducted postdoctoral research in Stefan
Wuertz's laboratory at UCD. In her graduate
studies, she investigated the decay of host-asso-
ciated microbial source tracking (MST) genetic
markers in marine and freshwater sediments. In
addition, she performed two MST water qual-
ity monitoring projects to identify the sources
of fecal contamination in a northern California
coastal area. As a postdoctoral scholar, Dr. Kim
continues her interest in integrating engineer-
ing with microbiology to address public health
concerns. She has worked to identify patho-
genic bacteria and viruses in aerosols and to
optimize a propidium monoazide quantitative
polymerase chain reaction (qPCR) method for
analysis of viable parasite oocysts on produce.
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Day Three: Session X
J
iL2a
How EPA's Source ID Technology and Digital
PCR Enhance Watershed Management
Presenter: Mauricio Larenas
Source Molecular Corporation
Author: Mauricio Larenas
Abstract
Identifying sources of fecal pollution is
important for effective watershed management.
The need to more accurately identify where the
contamination is coming from and how much
bacteria is present has spurred the development
of the U.S. Environmental Protection Agency's
(EPA's) source ID and Digital Polymerase Chain
Reaction (PCR) technology.
EPA's patented genetic testing methods
developed specifically to detect human, cattle,
chicken, and dog fecal pollution have under-
gone rigorous review and been proven to be
more sensitive and more accurate. The upcom-
ing EPA standard for microbial source tracking
(MST) will enable watershed managers to use
genetic-based methods for broader applications.
It also will increase the credibility of MST as a
useful source of information in assessing poten-
tial sources of fecal contamination and possible
public health risks.
Digital PCR is an advanced technology
that provides absolute quantification of the
target DNA, allowing water managers to know
the source of fecal pollution as well as exactly
how much fecal bacteria is in the water. It also
improves sensitivity, allowing watershed man-
agers to find the fecal source even if it has been
some time since the pollution event, and creates
the possibility of direct pathogen detection.
The presentation will provide case studies
and practical insight on how DNA-based test
methods have enabled water quality managers
to more effectively craft remediation plans.
Biosketch
Mr. Mauricio Larenas is a project man-
ager as well as the chief operating officer at
Source Molecular Corporation. In the past
10 years, he has provided advice and assis-
tance to hundreds of water managers across
the country on source identification projects.
He has helped many more to understand the
importance of identifying the sources of fecal
pollution and the cost-saving that comes with
applying microbial source tracking technology.
Mr. Larenas leads the company's development
of breakthrough technologies (e.g., Digital PCR)
as well as its participation in California's Source
Identification Protocol Project. He has worked
with the U.S. Environmental Protection Agency
in obtaining a license for Source Molecular's
laboratory to use the regulator's newly devel-
oped and patented genetic testing markers.
Mr. Larenas is currently overseeing the devel-
opment of Source Molecular's analytical data
management system, which will be in full com-
pliance with the laboratory's ISO/IEC 17025:2005
accreditation requirements. He holds a bach-
elor of science degree in biology from Florida
International University in Miami, Florida.
225
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U.S. EPA's 2016 Recreational Waters Conference
A Customized DNA Microarray for Microbial
Source Tracking in Environmental Systems
Presenter: Xiang Li, PhD
U.S. Environmental Protection Agency, Office of Research and Development
Authors: Xiang Li, Valerie Harwood, Bina Nayak, Christopher Staley, Michael Sadowsky,
Jennifer Weidhaas
Abstract
It is estimated that more than 160,000 miles
of rivers and streams in the United States are
impaired due to the presence of waterborne
pathogens. These pathogens typically originate
from human and other animal fecal pollution
sources; therefore, a rapid microbial source
tracking (MST) method is needed to facili-
tate water quality assessment and impaired
water remediation. We report a novel qualita-
tive deoxyribonucleic acid (DNA) microarray
technology consisting of 453 probes that can
detect general fecal and host-associated bacteria,
viruses, antibiotic resistance, and other envi-
ronmentally relevant genetic indicators. A novel
data normalization and reduction approach
also is presented to help alleviate false positives
often associated with high-density microarray
applications. To evaluate the performance of
the approach, DNA and complementary DNA
(cDNA) were isolated from swine, cattle, duck,
goose, and gull fecal reference samples, as well
as soiled poultry liter and raw municipal sew-
age. Based on nonmetric multidimensional scal-
ing analysis of the results, findings suggest that
the new microarray approach might be useful
for detecting pathogens and identifying fecal
contamination in recreational waters. The abil-
ity to simultaneously detect a large collection of
environmentally important genetic indicators in
a single test has the potential to provide water
quality managers with a wide range of informa-
tion in a short period of time. Future research is
warranted to measure microarray performance
in different geographic regions and water types.
Biosketch
Dr. Xiang Li is an Oak Ridge Institute for
Science and Education (ORISE) postdoctoral
fellow at the U.S. Environmental Protection
Agency in Cincinnati, Ohio. He received his
bachelor and master of science degrees from
Liaoning Normal University in China, and
his doctorate in environmental engineering
from West Virginia University. Prior to that,
Dr. Xiang Li worked at the Center of Disease
Control and Prevention in Dandong, China. His
main research interests are microbial source
tracking in environmental waters, microbio-
logical water quality modeling, and molecular
microorganism detection methods assessment.
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Day Three: Session X
J
iL2a
Citizen Science on the Bronx River: An Analysis
of Water Quality Data for an Urban Recreational
Resource
Presenter: Diane Mas, PhD
Fuss O'Neill, Inc.
Authors: Diane Mas, Kathalene Lamboy
Abstract
Citizen science not only offers the oppor-
tunity to leverage scarce resources for collecting
data, but also encourages a tangible connection
between the citizen scientists and the waters
they monitor. Adapting monitoring programs
based on insight gained from the evaluation
of data collected and changing priorities in a
watershed is a critical step in the continued
success and relevance of a volunteer monitoring
program.
Since 1990, over 1,000 water quality
samples have been collected by Bronx River
stewards at 19 locations along the 23-mile Bronx
River in New York. The number of samples
collected at each location varied from 10 to 200,
and 15 water quality parameters were included
in the data set. Analysis of this large data set
was undertaken to summarize the data, identify
water quality trends and data gaps, and make
recommendations for the water quality monitor-
ing program to improve the quality, usefulness,
and educational value of future monitoring data
for the Bronx River. The analysis consisted of
a graphical and tabular statistical summary to
provide a comprehensive characterization of the
data collected, analysis of correlation with other
environmental parameters, and spatial and tem-
poral trend analysis. The review and analysis
of the water quality data resulted in several rec-
ommendations for ongoing and future citizen
scientist monitoring efforts for this important
urban recreational resource, including adding
visual monitoring for algal blooms and estab-
lishing sites for long-term data collection.
Biosketch
Dr. Diane Mas is an associate in the Water
Environment and Natural Resources group at
Fuss & O'Neill, Inc. She received her bachelor of
arts degree in geology from Amherst College,
her master of science in engineering degree
from the Water Resources Program at Princeton
University, and her doctorate in civil engi-
neering from the University of Massachusetts
Amherst. Dr. Mas was formerly a hydrologist
for the National Weather Service and has held
teaching appointments at Smith College and
Hampshire College. She has spent nearly 20
years working in the areas of water quality
modeling, watershed management, and envi-
ronmental impact assessment. Dr. Mas's cur-
rent areas of practice focus on climate change
resiliency and adaptation for water resources,
water quality assessment and modeling and
watershed management, harmful algal bloom
impacts to drinking and recreational waters,
and the relationship between water quality and
public health.
227
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U.S. EPA's 2016 Recreational Waters Conference
Microbial Source Tracking Toolbox Approach in
Florida's Surface Waters
Presenter: Daisys Matthews
Florida Department of Environmental Protection
Authors: Daisys Matthews, David Whiting, Puja Jasrotia, Loretta Wolfe
Abstract
Microbial source tracking is a set of tech-
niques used to investigate and identify poten-
tial sources of elevated levels of fecal indicator
bacteria in a water body. Indicator bacteria such
as fecal coliforms, E. coli, and enterococci are
commonly found in the feces of humans and
other warm-blooded animals, but they also can
grow as free-living, non-enteric environmen-
tal strains that are not indicative of increased
pathogen risk. Standard microbiological cul-
ture-based methods cannot distinguish enteric
bacteria (i.e., from the gut of a host animal) from
non-enteric environmental bacteria. Listing
a water body as impaired when no increased
risk to human health exists can create sig-
nificant economic burdens. To focus time and
money on eliminating or reducing fecal pollu-
tion more quickly, the Florida Department of
Environmental Protection has devised a mul-
tipronged approach that uses molecular fecal
source markers and chemical tracers of human
fecal wastewater. Commonly used human
wastewater tracers include artificial sweeteners
(sucralose), drugs (carbamazepine and pri-
madone), pain relievers (acetaminophen), and
fragrances (tonalide). In addition, the laboratory
recently implemented a method to distinguish
between DNA from live bacteria and DNA from
dead bacteria in a water sample using the dye
propidium monoazide when performing qPCR
analyses.
Biosketch
Ms. Daisys Matthews is the environmental
manager of the molecular biology laboratory
at the Florida Department of Environmental
Protection (DEP). She received her bachelor
of science degree in biological sciences from
Florida International University in Miami,
Florida. For the past 15 years, Ms. Matthews has
been a part of the DEP's biology laboratory. Her
expertise is in microbiology and molecular biol-
ogy, and she currently is focused on microbial
source tracking.
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Day Three: Session X
J
iL2a
Deadend Hollow-Fiber Ultrafiltration for the
Concentration of Coliphage from Freshwater
Presenter: Brian McMinn
U.S. Environmental Protection Agency
Authors: Brian McMinn, Asja Korajkic, Emma Huff, Eric Rhodes
Abstract
To more accurately signify the pres-
ence of pathogenic viruses in ambient waters,
the U.S. Environmental Protection Agency is
investigating the potential use of coliphage for
assimilation into new recreational water quality
guidelines. Culturable coliphage are typically
present in lower densities than fecal indicator
bacteria, requiring more sensitive and robust
concentration methods for their detection. Here,
we evaluated deadend hollow-fiber ultrafiltra-
tion (D-HFUF) in conjunction with single agar
overlay (SAL) to concentrate and detect F+ and
somatic coliphage from ambient freshwater.
Two volumes (1 liter [L] and 10L) of river and
lake water were evaluated to determine if either
impacted coliphage recovery. For 1L samples,
recoveries ranged from 62.2% to 78.6%, while
higher volumes resulted in lowered recoveries
(36.4-51.2%) of both coliphage types. Sample
volume significantly impacted recovery in river
water (P< 0.001), but not in lake water, while
there was no significant difference in recover-
ies between phage types irrespective of the
water type or volume. Method sensitivity was
assessed by serially diluting sewage in river
water. Both coliphage types were reliably and
consistently detected in highly diluted sewage
(10,000-fold). The ability of the D-HFUF-SAL
method to perform in highly turbid waters
(> 100 NTU) also was assessed and there was
generally no significant difference in recoveries
at turbidities ranging from 38.4 to 118.7 NTU.
Our study indicates that the D-HFUF-SAL
method is a robust and sensitive method for
recovery of coliphage from difficult environ-
mental matrices and its simple design makes
it a good candidate for routine water quality
monitoring.
Biosketch
Mr. Brian McMinn is a microbiologist in
the U.S. Environmental Protection Agency's
Office of Research and Development, National
Exposure Research Laboratory in Cincinnati,
Ohio. He received his bachelor of science
degree in environmental health from Missouri
Southern State University and his master of
science degree in environmental microbiology
from Missouri State University. Mr. McMinn's
research interests include development and
optimization of methods to concentrate adeno-
viruses from drinking and ambient waters.
More recently, the focus of his research has been
on investigating the utility of different bacterio-
phage as indicators of human fecal pollution as
well as developing improved methods for their
concentration and detection.
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U.S. EPA's 2016 Recreational Waters Conference
Microbial Source Tracking at Two Beaches
Presenter: Heather Merritt
Maryland Department of the Environment
Author: Mark Frana
Abstract
The Maryland Department of the
Environment and Delaware Department of
Natural Resources and Environmental Control
are working with the University of Maryland
Salisbury to identify sources of enteric bacte-
ria at Maryland and Delaware beaches. They
are using microbial source tracking (MST), a
subspecialty of microbiology that incorporates
various techniques into identifying sources of
fecal contamination in water samples. Recent
studies using genetic markers for pollution
source tracking in coastal waters have proven
successful in identifying nonpoint sources of
pollution. Over the past 15 years, a variety of
MST methods have been developed, each with
its own advantages and limitations. One of the
most promising techniques includes the use
of quantitative PCR (qPCR) using DNA probes
specific for fecal bacteria (e.g., Bacteroides spp.)
originating from a specific source. This project
involves collecting water samples at one site
in Maryland and one site in Delaware where
previous monitoring studies have shown high
levels of fecal indicator organisms, particularly
after a rain event. At each location, an inten-
sive survey was conducted to determine the
likely sources of fecal indicator bacteria. The
samples are analyzed using qPCR and four
genetic markers (i.e., human, gull, poultry, and
ruminants) specific for bacteria associated with
those sources of contamination. The results of
this project will help guide remediation efforts
designed to reduce the levels of fecal contami-
nation in the study areas.
Biosketch
Ms. Heather Merritt is the beaches coordi-
nator in the Science Services Administration of
the Maryland Department of the Environment
(MDE), a position she has held for nearly 10
years. Ms. Merritt has a bachelor of science
degree in wildlife science from Virginia Tech
in Blacksburg, Virginia, and master of science
degree in environmental science from Marshall
University in Huntington, West Virginia.
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Day Three: Session X
J
iL2a
Hydraulic Connectivity Between Wastewater
Injection Wells and Submarine Springs (Seeps) in
Lahaina, Maui
Presenter: Scott Murakawa
Hawaii Department of Health, Clean Water Branch
Authors: Scott Murakawa, Wataru Kumagai
Abstract
The Lahaina Groundwater Tracer Study
identified a definite hydraulic connection
between the injection wells at the Lahaina
Wastewater Reclamation Facility (WWRF) and
the nearshore coastal waters off of Lahaina,
Maui (Glenn, Whittier, Dailer, et al., 2013).
Beginning in October 2011, the Hawaii State
Department of Health-Clean Water Branch
(DOH-CWB) began characterizing the subma-
rine spring (seep) water entering nearshore sur-
face waters to determine the potential impacts
of injection well effluent on coastal waters.
This presentation will focus on the
hydraulic connections identified in the Lahaina
Groundwater Tracer Study, the methods used
to sample seep water, and the fecal indicator
bacteria and nutrient results from the ongoing
nearshore seep coastal monitoring conducted in
Lahaina, Maui. The data suggest that there is an
underground microbial community in under-
ground aquifers that can affect bacteria and
nutrient levels of the effluent from the Lahaina
WWRF. Chlorination of treated effluent injected
into the injection wells between January 2012
and July 2014 resulted in an increase in total
nitrogen at nearshore seep groups, yet showed
no effects on fecal indicator bacteria levels. This
suggests that chlorination has resulted in the
die-off of the denitrifying bacteria population
residing in the complex groundwater networks
of West Maui.
Biosketch
Mr. Scott Murakawa and Mr. Wataru
Kumagai are environmental health specialists
with the State of Hawaii Department of Health,
Clean Water Branch. Mr. Murakawa has been
with the Clean Water Branch for 11 years, and
Mr. Kumagai has been with the Clean Water
Branch for 2 years. Both Mr. Murakawa and
Mr. Kumagai have worked on various water
quality projects throughout the State of Hawaii
during their time with the Department of
Health, including Hawaii's recreational beach
monitoring, Lahaina seep monitoring, and
national aquatic resource surveys.
231
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U.S. EPA's 2016 Recreational Waters Conference
Utilizing Virtual Beach for Bacteria Forecast
Modeling in Sarasota, FL
Presenter: Matthew Neet, PhD
University of South Carolina
Authors: Matthew Neet, Dan Ramage, Dwayne Porter, Heath Kelsey
Abstract
Bacteria level forecasts were developed
for beach waters around Sarasota, Florida.
Specifically, daily forecast procedures were
automated for 12 beach areas. The Virtual
Beach (VB) software package, developed by the
U.S. Environmental Protection Agency (EPA),
provided a set of statistical tools to develop the
robust models created in the study. VB was
utilized because it requires limited statistical
or programming skill to understand and oper-
ate. As data records were incomplete for all
variables of interest, multiple model runs were
made and various techniques were tested to
increase the number of records available for
modeling and ultimately adjusted r-square
values. Once predictive (statistical) bacterial
models were created and analyzed, a website
application was developed to show each beach
location and its associated forecast. The website
application was utilized to provide daily recom-
mendations about swimming in beach waters
at each of the 12 study sites. Model results and
daily Web application recommendations will be
presented to local stakeholders for buy-in and
critique.
Biosketch
Dr. Matthew Neet is a temporary research
professor in the Belle W. Baruch Institute for
Marine and Coastal Sciences at the University of
South Carolina. He received his bachelor of sci-
ence degree in biological sciences from Clemson
University and his master of earth and environ-
mental resource management degree, master of
public health degree in environmental health
sciences, and doctorate in environmental health
sciences from the University of South Carolina.
Dr. Neet has more than 15 years of experience
in the geographic information system (GIS),
global positioning system, remote sensing, and
image processing fields. He also has geospatial
experience at a variety of institutions, including
higher learning, state and county government,
consulting, defense contracting, and com-
mercial business institutions. Having recently
completed his doctorate, Dr. Neet currently is
studying bacterial modeling along multiple
beach habitats in the southeast. His research
interests include environmental justice, GIS,
remote sensing, bacterial forecasting, and eco-
logical modeling.
232
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Day Three: Session X
Validation of Host-Associated Bacteroidales Gene
Marker Assays for Microbial Source Tracking in an
Urban Tropical Environment of Singapore
Presenter: Jean Pierre Nshimyimana
Singapore-MIT Alliance for Research and Technology - CENSAM
Singapore Centre for Environmental Life Sciences Engineering
Authors: Jean Pierre Nshimyimana, Mercedes Cecilia Cruz, Janelle Thompson,
Stefan Wuertz
Abstract
Microbial source tracking (MST) is now a
widely applied tool in many temperate regions
of Europe and North America as well as
Australia, New Zealand, and Japan, but it has
not seen much use yet in tropical urban envi-
ronments. This study applied qPCR to validate
the use of host-associated Bacteroidales marker
assays based on the 16S rRNA gene to identify
human fecal pollution in the urban tropical
environment of Singapore. We collected a total
of 295 animal and human stool and sewage
samples. Animals tested included cats, dogs,
rabbits, chickens, birds, monkeys, and wild
boars. Following DNA extraction, samples were
analyzed by qPCR using seven assays targeting
human-associated Bacteroidales—HF183-SYBR,
HF183, BacHum, BacH and B. thetaiotaomicron
(B. theta), dog-associated Bacteroidales (BacCan),
and total Bacteroidales (BacUni). The qPCR
results were used to compute and compare sen-
sitivity and specificity among assays.
The five human-associated assays (HF183-
SYBR, HF183, BacHum, BacH, and B. theta) and
the total Bacteroidales assay BacUni had 100%
sensitivity for sewage. The overall sensitivity
for human stool samples ranged from 50 to 70%
and B. theta and BacHum displayed the high-
est human stool sensitivity of 68.6% and 65.7%,
respectively. In addition, these two assays had
the highest specificity (98.6% for B. theta and
91.4% for BacHum). The HF183 assay recently
recommended for MST in California, USA,
had a specificity of 90% and sensitivity of 60%
and 100%, respectively, to human stool and
sewage samples. BacCan assay sensitivity and
specificity to dog fecal samples were 80% and
97.3%, respectively. We recommend that B.theta,
BacHum, and BacCan be used for MST studies
in Singapore.
Biosketch
Mr. Jean Pierre Nshimyimana is cur-
rently pursuing doctoral studies in civil and
environmental engineering jointly at Nanyang
Technological University in Singapore and
Massachusetts Institute of Technology (MIT).
He will complete his studies later this year.
Mr. Nshimyimana received his master of
science degree in civil and environmental
engineering from MIT and an undergraduate
degree in environmental health sciences from
the School of Medicine and Health Sciences at
the University of Rwanda. In his research at
the Singapore-MIT Alliance for Research and
Technology (SMART) and Singapore Centre
for Environmental Life Sciences Engineering
(SCELSE), Mr. Nshimyimana is applying
molecular biology tools (e.g., qPCR, digital PCR,
and next generation sequencing) to address
(1) how bacterial communities and subgroups
of sewage-associated taxa and pathogen-like
bacteria vary as a function of land use, sites,
and water quality in tropical urban catch-
ments; (2) whether 16S rRNA host-associated
Bacteroidales assays can be used to effectively
identify and distinguish nonpoint sources
of fecal microbial pollution in urban tropical
environments; and (3) what the effect of pre-
dation is on persistence and decay of human
Bacteroidales-associated genetic markers in a
tropical sediments-rich environment.
233
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U.S. EPA's 2016 Recreational Waters Conference
Rapid Concentration to Support Improved Detection
of Indicator Bacteria in Recreational Waters
Presenter: Andy Page
InnovaPrep, LLC
Authors: Andy Page, Michael Hornback, David Alburty
Abstract
Rapid detection of fecal indicator bacteria
in recreational waters is needed to ensure public
safety. Molecular and other rapid microbio-
logical detection technologies have progressed
significantly in the last several decades and pro-
vide the greatest potential for overcoming these
challenges. However, their development has
significantly outpaced development of sample
concentration techniques, which are necessary
for rapid detection of low concentrations of
bacteria.
InnovaPrep has developed a suite of sys-
tems for concentration of bacteria and other bio-
logical particles from liquid samples. Volumes
of water from a few milliliters to tens of liters of
water are processed through flat membrane fil-
ters or hollow-fiber membrane filters to capture
any biological particles that are present. The
biological particles are then efficiently recovered
from the membrane surface with a tangential
flush using a carbonated "wet foam." The wet
foam is expanded up to six times the original
liquid volume and becomes highly viscous,
allowing it to act as the membrane surface and
recover the particles into volumes significantly
smaller than can be attained with traditional
liquid elutions. The process is scalable, efficient,
and typically results in concentration factors of
approximately 1000X per concentration stage.
The InnovaPrep concentration and wet
foam elution processes will be presented along
with developmental and commercially avail-
able InnovaPrep concentration systems. Data
from use of the Concentrating Pipette system for
concentration of indicator bacteria from recre-
ational waters and detection by quantitative
polymerase chain reaction will be presented,
including concentration efficiencies, concentra-
tion factors, and detection limits.
Biosketch
Mr. Andy Page, president and CTO of
InnovaPrep LLC, is an expert in the field of
collection and concentration of microorgan-
isms from air, surfaces, and liquids. He holds
multiple patents for biological collection and
concentration, including patents for membrane
concentration processes using wet foam elu-
tion. Mr. Page's patents underlie the InnovaPrep
Concentrating Pipette instrument, Large
Volume Concentrator, and other fractionation
and concentration technologies developed
by InnovaPrep for enabling improved rapid
detection of microorganisms in liquid samples,
including pathogens in environmental waters.
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Day Three: Session X
J
iL2a
Moving from the Shore to the Water: Advancements
in Local Health Department Beach Water Quality
Monitoring within the City of Milwaukee
Presenter: Lindsey Page
City of Milwaukee Health Department
Authors: Paul Biedrzycki, Lindsey Page, Todd Miller, Nicholas Tomaro
Abstract
Recreational water monitoring by local
health departments (LHDs) at public beaches
represents a core environmental health com-
petency In the City of Milwaukee, three public
beaches are monitored by the Milwaukee Health
Department (MHD) on a seasonal basis for lev-
els of Escherichia coli, the indicator organism for
pathogens of human health concern. Recently
water sampling and sanitary survey data col-
lection has been conducted in partnership with
the University of Wisconsin-Milwaukee (UWM)
Zilber School of Public Health (ZSPH). This col-
laboration has resulted in leveraging analytical
capabilities for developing beach water qual-
ity predictive models and investing in student
workforce development.
For the 2016 beach season, MHD and
faculty and students from ZSPH and the UWM-
School of Freshwater Sciences will develop and
deploy offshore buoys configured with sensors
to monitor real-time water quality conditions.
These advancements in LHD monitoring infra-
structure will ultimately enhance the qual-
ity, type, and amount of data collected at city
beaches to improve the accuracy of daily water
quality advisories issued by MHD.
Beach water quality monitoring programs
at many LHDs have been compromised by
funding shortfalls in the past decade, impact-
ing capacity and capabilities necessary to
adequately meet the public health mission. LHD
collaboration with academic and community
partners through the development of cost-effec-
tive and scientifically valid beach monitoring
outcomes is foundational to future sustain-
ability of this important function. The MHD
program is an example of how cross-sectoral
and coordinated partnerships can meaningfully
benefit community health, meet educational
outreach objectives and satisfy relevant research
interests.
Biosketch
Ms. Lindsey Page is the emergency
preparedness coordinator in the Division of
Disease Control and Environmental Health
at the City of Milwaukee Health Department.
She received her master of public health degree
from Northern Illinois University and, prior to
working for the City of Milwaukee, she was an
infectious disease epidemiologist for the North
Dakota Department of Health. Ms. Page cur-
rently is the cochair of the Wisconsin Coastal
Beaches Workgroup and manages the beach
monitoring program for three beaches in the
City of Milwaukee.
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U.S. EPA's 2016 Recreational Waters Conference
Beyond Swimming: Playing in the Cold Waters
of Alaska
Presenter: Gretchen Pikul
State of Alaska, Department of Environmental Conservation
Author: Gretchen Pikul
Abstract
The Kenai River is located on the Kenai
Peninsula in south central Alaska, and is con-
sidered the most popular sport fishing destina-
tion in Alaska. Due to a nearby bird rookery
and poor fish waste management by anglers,
bacteria levels exceed water quality standards
during the salmon fisheries in July and might
pose human health risks. The purpose of the
multi-year monitoring study was to determine
bacteria levels and sources, and develop data
trends and actions needed to improve the
effectiveness of public outreach and notifica-
tions. Water conditions in Alaska are differ-
ent from typical beaches used for swimming.
Recreational use of Alaskan beaches is highly
seasonal, with low temperature waters even
during summer. Recreational water use peri-
ods are limited, the fisheries are short (usually
3 weeks), and migratory birds are plentiful.
Swimming is not the primary activity on Alaska
beaches, but fishing, wading, dog walking,
kayaking, and boating are. Alaska needs a sec-
ondary recreation criteria to properly address
our specific bacteria concerns.
We are searching for better, more quantita-
tive risk assessment tools and more progressive
ways to improve beach sanitation resulting
in long-term solutions, beyond issuing beach
notices. Alaska is also searching for more
transparent ways to connect the BEACH Act
with other Clean Water Act programs. How
do bacteria exceedances lead to 303(d) listings,
Total Maximum Daily Loads, or other commu-
nity planning and state regulations? The study
results and implementation challenges will be
discussed during this presentation.
Biosketch
Ms. Gretchen Pikul is an environmental
program specialist at the Alaska Department
of Environmental Conservation (ADEC), where
she works in the Non-point Source section of
the Division of Water. She received her bach-
elor of science degree in geological sciences
from Salem State College in Massachusetts and
worked in the environmental private sector
before joining ADEC, where she has worked
in the Contaminated Sites DoD Oversight and
Water Quality sections. Ms. Pikul's main areas
of interests are total maximum daily loads,
restoration of impaired waters, and green infra-
structure to manage urban stormwater.
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Day Three: Session X
J
iL2a
Measuring the Impact of Harmful Algal Blooms: The
One Health Harmful Algal Bloom System (OHHABS)
Presenter: Virginia Roberts
Centers for Disease Control and Prevention
National Center for Emerging and Zoonotic Infectious Diseases
Authors: Virginia Roberts, Joana Yu, Irina Pyrkh, Lorraine Backer, Kathleen Fullerton,
Jonathan Yoder, Michael Beach
Abstract
Harmful cyanobacteria and algae blooms
(harmful algal blooms, or HABs) are an emerg-
ing public health issue in both coastal and
inland waters. Waterborne and foodborne
disease outbreaks associated with HAB events
or algal toxin exposures are reported to the U.S.
Centers for Disease Control and Prevention
(CDC) by state health departments via the Web-
based National Outbreak Reporting System
(NORS). However, these outbreak reports do
not include the individual-level data on harm-
ful algal bloom-related illnesses or exposures
needed to improve case definitions or prioritize
other HAB-related activities related to human
or animal health. In 2014, CDC established a
working group of state and federal partners
with expertise in HABs and illness surveillance
to design the One Health Harmful Algal Bloom
System (OHHABS), a Web-based reporting
system able to receive data about HAB-related
human illnesses, animal illnesses, and HAB
events using the same technological platform
as NORS. During 2014 and 2015, the OHHABS
working group drafted reporting forms; devel-
oped initial reporting definitions for human
illnesses, animal illnesses, and HAB events; and
launched the OHHABS pilot system. Following
the full OHHABS launch in 2016, maintenance
of the system will require ongoing efforts to
support state activities and demonstrate the
utility of the system through use of the data.
Continued and expanded collaboration with
human health, animal health, and environ-
mental health communities will be essential to
meeting One Health goals of the system, both in
the implementation of OHHABS and the opti-
mization of data collected in the system.
Biosketch
Ms. Virginia Roberts, MSPH is an epide-
miologist in the Waterborne Disease Prevention
Branch of the Centers for Disease Control and
Prevention (CDC). Ms. Roberts manages the
waterborne disease outbreak component of the
National Outbreak Reporting System (NORS)
and coordinates a CDC Great Lakes Restoration
Initiative project building waterborne disease
prevention capacity in Great Lakes states.
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U.S. EPA's 2016 Recreational Waters Conference
Importance of Staying Informed
Presenter: Gerald Ruiz
ATC Associates
Author: Gerald Ruiz
Abstract
As more than five cities in Texas each has a
population of almost 240,000 people (including
Houston, one of the largest cities in the coun-
try), programs such as Texas Beach Watch are
indispensable to the way of life that many enjoy
on the gulf coast. With the influx of recreational
activities such as fishing, surfing, and boating,
it has become essential for the public to be fully
aware of any hazards that could potentially
endanger such pastimes. Texas Beach Watch
and similar programs are designed to survey
and determine the quality of water in these
regions and to discover the dangers they could
pose to the public. However, more compelling
and accessible communication with the public
environment is necessary if those programs are
to effectively accomplish their goal of helping
the communities they serve. Possible meth-
ods of effectively informing the public include
(1) holding seminars in proximity to the water-
surveying locations, (2) promoting the pro-
grams through popular social media platforms,
and (3) reaching out to local businesses around
the area that would have an interest in the
quality of the water from which some of their
products might originate. To sum up, communi-
cation is key in helping promote environmental
programs such as Texas Beach Watch to better
serve the public.
Biosketch
Mr. Gerald Ruiz is an environmental
technician for ATC Group Services LLC, an
environmental engineering consulting firm
located in Houston, Texas. Mr. Riuz is a gradu-
ate of the University of Houston, where he
graduated with a bachelor of science degree in
environmental science. Prior to working with
ATC, Mr. Riuz interned at Joyce, McFarland,
and McFarland LLP, where he assisted with
trial work. He also is a member of the American
Meteorological Society and has volunteered at
the organization's environmental awareness
events, such as Earth Day festivals and weather
balloon launches aimed at informing young
adults and children about the environment they
live in. Currently, Mr. Ruiz is in charge of the
Texas Beach Watch, a program aimed at inform-
ing the public of Texas Gulf coasts beaches by
acquiring samples and determining the level of
bacteria that is sometimes present and can be
harmful to people.
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Day Three: Session X
J
iL2a
Washington State Marine Beach Bacteria
Trends, 2003-2014
Presenter: Debby Sargeant
Washington State Department of Ecology and Health
Authors: Deborah Sargeant, Julianne Ruffner
Abstract
The Beach Environmental Assessment,
Communication, and Health (BEACH) Program
is an ongoing monitoring and notification
program implemented throughout Puget
Sound and along the coast of Washington
State. Washington marine beaches are tested
for enterococci, a fecal bacteria, to determine
possible health risks to the public from water
contact recreation. The program began with a
pilot project in 2003.
The BEACH program consistently moni-
tors 51 core beaches, which were selected as
the highest use and highest risk beaches in the
state. Data from the core beaches were analyzed
to identify any long-term trends in marine
water quality based on summer bacteria data
from 2003 through 2014.
Results indicate that the core beaches
overall show no significant bacteria trends in
water quality for the period tested. Individual
core beaches were tested for bacteria trends over
time and showed mixed results.
Increasing trends in bacterial levels were
detected at the following beaches:
• Bay view State Park Beach in Skagit
County
• Dash Point Metro Park Beach in Pierce
County
• Freeland County Park Beach in Island
County
Decreasing trends in bacteria levels were
detected at Richey Viewpoint Beach in King
County and Howarth Park Beach in Snohomish
County.
Biosketch
Ms. Debby Sargeant is the program man-
ager of Washington State's Beach Environmental
Assessment, Communication, and Health
(BEACH) Program, which is administered by
the Washington departments of Ecology and
Health. Ms. Sargeant received her bachelor
of science degree in environmental studies
from The Evergreen State College, home of the
Geoducks. She has 25 years of experience work-
ing on bacteria-related issues in Washington,
including designing bacterial water quality
studies to track sources of pollution, bacterial
total maximum daily load studies, tracking
sources of bacteria to marine areas for shellfish
bed and beach water quality restoration, and
microbial source tracking.
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How to Achieve a Major Reduction in Shore
Pollution Using a Business Quality Approach
Presenter: Phillip Scanlan
Past AT^T Corporation Quality, Vice President
Past Ocean Conservancy Board Member
Author: Phillip Scanlan
U.S. EPA's 2016 Recreational Waters Conference
Abstract
In 1988,1 learned that half of the North
Atlantic bottlenose dolphins had been killed
due to pollution. I offered my help to the New
Jersey Department of Environmental Protection
to apply AT&T business quality methods to help
reduce the pollution problem.
The AT&T network of fiber and switches
is similar to the New Jersey network of sew-
age pipes and plants. However, the New Jersey
network is managed by over 100 different gov-
ernment municipalities and counties. We used
AT&T quality methods to set goals, establish
clear responsibilities for processes, share best
practices, and recognize improvement.
In 1988, New Jersey had 803 beach block
days of closings due to pollution, the worst in
the United States. In 1991, that number had been
reduced to 10 and, in 1995, to 4. In the latest
National Resources Defense Council report for
2014, New Jersey is still one of the top U.S. states
in quality of beach water.
This presentation reviews the business
quality approach that was used to help the New
Jersey team achieve the greatest improvement in
water quality in the United States. The full story
is in my book The Dolphins are Back.
Biosketch
Mr. Phil Scanlan holds bachelor and
master of science degrees in engineering
from Northeastern University. He has spent
his 33-year business career at AT&T, starting
as an engineer and advancing to become the
company's quality vice president. Phil also has
been working on environmental projects for
more than 30 years, with the goal of making
the "quality of life" better wherever he lives by
using a quality approach to get results. As a
volunteer, he has helped apply that approach to
achieving major environmental improvements.
He spent 10 years supporting a team effort to
clean up the New Jersey shore after 2,500 North
Atlantic bottlenose dolphins died from pollu-
tion-related diseases, helping to turn the Jersey
shore from the worst in the nation to the best in
the nation. Phil was recognized for his leader-
ship efforts by the New Jersey Department of
Environmnetal Protection and the New Jersey
Secretary of Commerce. He also wrote a book
titled The Dolphins are Back to share how other
states could improve the health of their shores.
Because of his work on the Jersey shore, Phil
was invited to join the national board of the
Ocean Conservancy—the largest nonprofit
organization focused on protecting our ocean
wildlife—where he served for 10 years.
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* ^
Day Three: Session X
J
iL2a
Bacterial Source Tracking of Fecal
Contamination Using Digital PCR
Presenter: Samendra Sherchan, PhD
Tulane University
Author: Samendra Sherchan
Abstract
Identifying sources of fecal contamina-
tion—a serious problem in coastal and fresh
waters—is important in preventing disease
and ensuring water quality Using molecular
techniques to indicate the presence of human
fecal pollution and to determine water quality
has become increasingly popular. Detecting
Enterococci and Escherichia coli is a traditional
method of indicating fecal pollution. Both are
naturally found in the intestinal tracts and
feces of different types of animals and have
a high survivability rate in the environment.
The U. S. Environmental Protection Agency
(EPA) water quality criterion using membrane
filtration for Enterococci is 33 colony-forming
units (CFU)/100 milliliters (ml) and for E. coli
is 126 CFU/100 ml in freshwater systems.
However, determination of the amount of
Enterococci and E. coli in a water system indi-
cates only the presence of fecal contamina-
tion, is time-consuming, and will take days
to be concluded. Several molecular methods
have been developed to distinguish whether
the contamination is human or nonhuman.
Bifidobacteria and Bacteriodes are two examples
of bacterial indicators for host-specific identifica-
tion of human fecal pollution. Bifidobacteria are
gram-positive rods widely found in the intes-
tinal microflora of humans and some species
like Bifidobacteria adolescentis can be used as a
genetic marker in the Quantitative Polymerase
Chain Reaction (qPCR) technique, allowing
source tracking of human fecal contamination.
This review poster presentation will discuss
a recent advanced molecular method, Digital
PCR, a microbial source tracking technique that
can help to determine whether the source of
fecal contamination in estuaries and rivers is
human or nonhuman.
Biosketch
Dr. Samend Sherchan is an assis-
tant professor in the Department of Global
Environmental Health Sciences at Tulane
University. His areas of expertise include
environmental bioinformatics, environmental
microbiology, water quality, and water reuse.
He received his doctorate from the University of
Arizona in 2013. Dr. Sherchan's research inter-
ests include environmental monitoring, health-
related water microbiology, water quality, food
safety, microbial source tracking, environmental
metagenomics, environmental microbiology,
emerging contaminants, water reuse, water
treatment, waterborne diseases, and public
health.
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U.S. EPA's 2016 Recreational Waters Conference
Evaluation of Enterococcus and £. coli
Measurements at Recreational Beaches in Door
County by Quantitative Polymerase Chain Reaction
and Define Substrate Culture Analysis
Presenter: Nilay Sheth
University of Wisconsin Oshkosh, ERIC Lab
Author: Nilay Sheth
Abstract
Recently the U.S. Environmental Protection
Agency (EPA) has approved the use of the
qPCR monitoring method 1611 for surface water
regulatory decisions. This method allows for
results in as little as 4 hours, rather than the 24+
hours necessary for culture-based tests. Method
1611 is an enterococci-based test method, while
all Great Lake states have adopted E. coli as the
indicator organism of choice for recreational
water monitoring. While EPA qPCR method
C is an E. coli specific method, it has not been
approved as a method for regulatory decisions.
The change in the basis for the test (molecular
versus culture-based), and the heterogeneous
nature of Great Lake beach communities
presents significant challenges to implementa-
tion of the rapid method. The objective of this
study was to determine the feasibility of using
both qPCR method 1611 and C for beach water
monitoring and management in northern Lake
Michigan communities. Currently, EPA has
utilized 1,000 CCE/100 mL as a target value for
enterococci-based qPCR method. However, no
such target value has been determined for E.
coli qPCR method C. The study will also aim to
find a target value for qPCR E. coli method C for
northern Lake Michigan beaches. This presen-
tation will present results from multiple years
with samples collected from northern Lake
Michigan beaches (Door County, Wisconsin),
comparing both qPCR EPA method 1611 and
C with culture based enterococci and E. coli
method.
Biosketch
Mr. Nilay Sheth is a research scientist at
the Environmental Research and Innovation
Center at the University of Wisconsin- (UW-)
Oshkosh. He received his bachelor of science
degree in cell biology from UW-Whitewater and
his master of science degree in microbiology
from UW-Oshkosh. Mr. Sheth currently super-
vises the Door County, Wisconsin, laboratory,
where they adhere to regulatory body methods
to detect coliform and E. coli in surface water
using defined substrate methods and rapid
molecular assay (qPCR) to quantify Enterococci
and E. coli in surface water. For the past 5 years,
Mr. Sheth has worked as a research scientist at
UW-Oshkosh, where he manages various proj-
ects in the fields of drinking water and surface
water.
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Day Three: Session X
J
iL2a
Assessing Potential Sources and Influential
Parameters of Fecal Contamination at F.W. Kent Park
Lake, Oxford, Iowa
Presenter: Reid Simmer
University of Iowa, Department of Geographic and Sustainability Sciences
Authors: Reid Simmer, Mary Skopec, David Bennett
Abstract
Fecal contamination of recreational water
bodies in Iowa poses a threat to water quality
as well as human health. Concern for the health
effects of waterborne pathogens resulted in 149
beach advisories across 39 state-owned beaches
during the 2015 beach season. While the pres-
ence of pollution is often clear, its cause and
source are difficult to identify. Furthermore, the
current practice in Iowa of sampling once per
week allows for high error and does little to pre-
vent swimmer exposure. The objectives of this
study were to identify the environmental factors
causing spikes in fecal contamination as well as
the sources of pollution at F.W. Kent Park Lake
in Oxford, Iowa. Water samples were collected
twice per week at the swimming beach and
throughout the contributing watershed from
May to October 2015. All samples were analyzed
for Escherichia coli using the IDEXX Colilert enu-
meration method. The resulting data as well as
numerous environmental parameters are being
analyzed using Virtual Beach. The two expected
sources of fecal contamination are agricultural
runoff and the wild geese that frequent the
beach. We will consider the spatiotemporal pat-
tern of rainfall and E. coli concentrations in our
analysis to determine the relative impact the
two alternative sources have on water quality.
The ultimate goal of this research is to develop
a predictive model of water conditions to better
inform beachgoers of the potential threat of
exposure to fecal contamination.
Biosketch
Mr. Reid Simmer is a graduate student
in geography at the University of Iowa, where
he received his bachelor of science degree in
environmental science. Mr. Simmer has worked
as a summer field and lab technician for the
Iowa Department of Natural Resources Beach
Monitoring Program for the past 4 years. His
main research interests include water quality
monitoring and modeling as well as pollution
source tracking.
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U.S. EPA's 2016 Recreational Waters Conference
Evaluation of Raw Sewage, Highly Treated Reclaimed
Water, and Sewage-Impacted Surface Waters for
Indicator and Pathogenic Microorganisms: Implications
for Recreational, Agricultural, and Drinking Water Use
Presenter: Mark Sobsey, PhD
University of North Carolina, Chapel Hill
Authors: Emily Bailey, Mark Sobsey
Abstract
The microbial quality of reclaimed
wastewaters and wastewater-impacted sur-
face waters is a health concern for beneficial
uses in recreation or agriculture, or as drink-
ing water sources. In this study indicator and
pathogenic bacteria, viruses, and parasites were
quantified in periodic samples of (1) raw sew-
age, (2) tertiary-treated, dual-disinfected (free
chlorine and UV radiation) reclaimed water,
and (3) wastewater-impacted surface waters
used for recreation or as water supply sources
collected from representative facilities in the
North Carolina (NC) Research Triangle region.
Specified treatment processes for high-quality
reclaimed water under NC regulations effec-
tively reduced indicator bacteria, virus, and
protozoan parasite surrogate concentrations
(to less than or equal to 3 E. coll! 100 milliliter
(mL), less than or equal to coliphages/100 mL,
and less than or equal to Clostridium perfrin-
gens/100 mL). However, human enteric viruses
(adenoviruses and noroviruses) and protozoan
parasites (Cryptosporidium and Giardia) were
still detectable in 10-liter samples of reclaimed
water by nucleic acid (RT-PCR/PCR) and immu-
nofluorescent microscopy methods, respectively.
Indicators met NC state loglO reduction targets
for high-quality reclaimed water, but loglO
pathogen reductions were lower and highly
variable, and did not meet loglO reduction per-
formance targets. Pathogen concentration data,
however, are not based on infectivity or cultur-
ability and may not predict human health risks.
Microbial analysis to date of sewage-impacted
surface water samples used recreationally and
near drinking water supply intakes shows
that they do not meet bacteriological quality
requirements for recreational use and that dual-
disinfected reclaimed water is of higher quality
based on indicator microorganism concentra-
tions. Developing better microbial water quality
criteria for wastewater, recreation, and reuse
requires further investigation.
Biosketch
Dr. Mark Sobsey is a Kenan Distinguished
Professor of Environmental Sciences and
Engineering specializing in environmental
health microbiology and water, sanitation, and
hygiene in the Department of Environmental
Sciences and Engineering, Gillings School
of Global Public Health, University of North
Carolina at Chapel Hill (UNC-CH). He received
his bachelor of science degree in biology (1965)
and master of science degree in hygiene (1967)
from the University of Pittsburgh. Dr. Sobsey
received his doctorate in environmental health
sciences from the School of Public Health,
University of California, Berkeley (1971). He
held a postdoctoral position (1971), instructor-
ship (1972), and assistant professorship (1973) in
the Department of Virology and Epidemiology,
Baylor College of Medicine, Houston, Texas, and
joined the UNC-CH faculty in 1974 as an assis-
tant professor. Dr. Sobsey's research, teaching,
and service encompass the detection, char-
acterization, occurrence, environmental sur-
vival/transport/fate, treatment, human health
effects characterization, and risk assessment of
viruses, bacteria, and parasites of public health
concern in water, wastewater, biosolids, soil,
air, and food for the prevention and control of
244
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water-, food-, and excreta-borne disease. His
most recent research focuses on household
water treatment for improved water quality
and health; new, improved, and rapid microbial
detection technologies for water and wastewa-
ter; wastewater reclamation and reuse; virus
survival in fecal wastes and sewage; and anti-
microbial resistance of bacteria associated with
fecal waste sources in the environment.
Day Three: Session X
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U.S. EPA's 2016 Recreational Waters Conference
Pathogenic Viruses and Bacteria in Stormwater
Discharging to Beaches with Year-Round Surfer
Populations in San Diego, California
Presenter: Joshua Steele, PhD
Southern California Coastal Water Research Project
Authors: Joshua Steele, Kenneth Schiff, A. Denene Blackwood, Laila Othman,
Rachel Noble, John Griffith
Abstract
Microbial water quality, usually measured
by fecal indicator bacteria (FIB), at California
beaches near storm drains worsens follow-
ing storms, leading to well-known 72-hour
rain advisories. Until recently, however, the
pathogenic bacteria and viruses in stormwater
that likely cause illness could not be reliably
measured, primarily because of the difficulty
in quantifying dilute pathogens in complex
water matrices. Digital PCR assays now have
the sensitivity to enable direct quantification of
pathogenic viruses and bacteria. Using digi-
tal PCR assays, we measured microbial water
quality in stormwater discharges that drained
into two beaches with large, year-round surfer
populations during wet-weather seasons in San
Diego, California. Tourmaline Creek drained
a small urban watershed and the San Diego
River drained a large mixed urban/undeveloped
watershed. Stormwater was collected during six
events with precipitation ranging from 0.19 to
2.5" from January to March 2014 and December
2014 to March 2015. Microbial water quality was
determined by quantifying FIB, Campylobacter,
Salmonella, human Norovirus, adenovirus, and
enterovirus. We found high FIB and pathogen
concentrations in stormwater from both the
large and small watersheds during and in the
3 days following rainfall. Norovirus type Gil
and Campylobacter spp. were detected most
frequently, while Salmonella and adenovirus
were rarely detected, and enterovirus was
not detected at all. We found no relationship
between FIB and pathogen concentration. The
ability to quantify pathogens in stormwater
provides the ability to more precisely determine
the microbial contamination. This direct quan-
tification also will enable better risk estimates
and improve management at beaches affected
by nonpoint source pollution.
Biosketch
Dr. Joshua Steele is a scientist at the
Southern California Coastal Water Research
Project who specializes in environmental
microbiology and microbial ecology, including
cultivation—independent tracking of patho-
gens and natural microorganisms in the coastal
ocean and watersheds. He received his bachelor
of science degree in molecular biology from
the University of California, San Diego and his
doctorate in biological oceanography from the
University of Southern California. Dr. Steele's
current research focuses on developing and
applying molecular techniques to detect and
track pathogenic bacteria and viruses, deter-
mine their viability and infectivity, and link
them to beachgoer risk using statistical mod-
els; detecting microbes of emerging concern
in coastal water; employing next generation
sequencing to connect the environmental micro-
biome to water quality and ecosystem health;
and applying bioinformatics tools in environ-
mental assessment.
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Day Three: Session X
J
iL2a
Using Canine Scent Tracking To Determine Bacteria
Source Contamination: A Case Study from the Grand
Traverse Bay Watershed, Michigan
Presenter: Sarah U'Ren
The Watershed Center Grand Traverse Bay
Author: Sarah U'Ren
Abstract
The Watershed Center Grand Traverse
Bay (TWC) is a nonprofit organization based in
Traverse City Michigan, whose mission is to
advocate for clean water in Grand Traverse Bay
and protect and preserve the bay's watershed.
During the past 8 years, we have implemented
our Healthy Beaches Program, which includes
bacteria monitoring, source tracking work at
local beaches, education via advertising and
social media, and large-scale best management
practices at beaches to reduce bacterial contami-
nation. One of the first things TWC did in reac-
tion to high bacteria counts at local beaches was
to begin a source tracking monitoring program
to determine if the bacteria was from human or
animal sources, which would in turn dictate our
management approach. In addition to partner-
ships with research agencies like Michigan State
University and the U.S. Geological Survey, our
source tracking program included the use of
a canine unit trained to detect the presence of
human sewage in water. Using the canine team
from Environmental Canine Services proved to
be a cost-effective and timely way to pinpoint
and eliminate areas in which human sources
may or may not have been contributing to
bacteria counts. This poster summarizes TWC's
source tracking efforts with the canine team to
help protect local beaches from bacterial con-
tamination and explains how those results were
used to implement management efforts.
Biosketch
Ms. Sarah U'Ren has served as the pro-
gram director for The Watershed Center Grand
Traverse Bay (TWC) for the past 14 years. She is
responsible for overseeing and coordinating all
watershed projects and grant activities at TWC
and has 16 years of experience in watershed
project management, research, and fieldwork.
Ms. U'Ren authored the Grand Traverse Bay
Watershed Protection Plan, specializes in beach
and stormwater management and stormwater-
related restoration activities, and has overseen
more than 60 grant-funded projects in her ten-
ure at TWC. She earned her bachelor's degree
in biology from Alma College and her master's
degree in environmental science from the
University of Maryland.
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Utilizing Green Infrastructure to Protect Public
Health at Beaches: A Case Study in Suttons Bay, MI
Presenter: Sarah U'Ren
The Watershed Center Grand Traverse Bay
Author: Sarah U'Ren
U.S. EPA's 2016 Recreational Waters Conference
Abstract
The Watershed Center Grand Traverse
Bay (TWC) is a nonprofit organization based
in Traverse City Michigan, whose mission is
to advocate for clean water in Grand Traverse
Bay and protect and preserve the bay's water-
shed. In 2013, TWC and the Village of Suttons
Bay installed 18 rain gardens and 3,600 feet of
underground infiltration trenches throughout
the village in an effort to use green infrastruc-
ture to protect public health and reduce harmful
stormwater runoff from reaching local beaches.
Funded by the U.S. Environmental Protection
Agency's Great Lakes Restoration Initiative, the
rain gardens and trenches allow infiltration
of most of the stormwater originating in the
village and prevent it from reaching the bay.
Additionally, the village formed a Friends of
the Rain Gardens group in 2015 that consists of
volunteers who will take care of the rain gar-
dens as they become established over the next
few years. This poster will display the types
and locations of installed green infrastructure,
lessons learned, and a look at the status of the
project 2 years after implementation.
Biosketch
Ms. Sarah U'Ren has served as the pro-
gram director for The Watershed Center Grand
Traverse Bay (TWC) for the past 14 years. She is
responsible for overseeing and coordinating all
watershed projects and grant activities at TWC
and has 16 years of experience in watershed
project management, research, and fieldwork.
Ms. U'Ren authored the Grand Traverse Bay
Watershed Protection Plan, specializes in beach
and stormwater management and stormwater-
related restoration activities, and has overseen
more than 60 grant-funded projects in her ten-
ure at TWC. She earned her bachelor's degree
in biology from Alma College and her master's
degree in environmental science from the
University of Maryland.
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Day Three: Session X
J
iL2a
Methods to Sample E. coli in Foreshore Sand
and Pore Water
Presenter: Laura Vogel
University of Western Ontario
Author: Laura Vogel
Abstract
In recent years, a number of studies have
shown that microbial contaminants, includ-
ing fecal indicator bacteria (FIB) such as E.
coli, accumulate in sand and pore water close
to the shoreline at beaches. This reservoir of
bacteria can act as a nonpoint source, resulting
in elevated microbial levels in adjacent surface
waters. While health units following recom-
mended sampling guidelines routinely monitor
surface waters at many recreational beaches,
there is no widely accepted method for collect-
ing sand/sediment or pore water samples for
FIB enumeration. The efficiency with which FIB
attaches to different types of sand grains and
the different moisture conditions under which
FIB attachment can occur add to the uncertainty
of characterizing the abundance of E. coli in the
foreshore reservoir. Some studies have quanti-
fied the abundance of E.coli in the foreshore
reservoir by collecting unsaturated surface
sand samples, while other studies test the pore
water and disturbed saturated sand samples.
The utility of the different sampling strategies
in providing quantifiable information related to
the abundance of FIB in the foreshore reservoir
and associated risk is not well understood. Field
sampling was conducted to evaluate techniques
for characterizing the abundance of E. coli in the
foreshore reservoir. Sampling was performed
at a range of beaches with different sand types
including fine grain (0.125
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U.S. EPA's 2016 Recreational Waters Conference
Differential Decay of Bacterial and Viral Fecal
Indicators in Common Human Pollution Sources
Presenter: Pauline Wanjugi, PhD
U.S. Environmental Protection Agency
Author: Pauline Wanjugi
Abstract
Understanding the decomposition of
microorganisms associated with different
human fecal pollution sources is necessary for
proper implementation of many water quality
management practices, as well as predicting
associated public health risks. Here, the decay
of select cultivated and molecular indicators of
fecal pollution originating from fresh human
feces, septage, and untreated sewage in a sub-
tropical marine environment was assessed over
a 6-day period with an emphasis on the influ-
ence of ambient sunlight and indigenous micro-
biota. Ambient water mixed with each fecal
source (1:10 dilution) was placed in dialysis bags
and incubated in situ in a submersible aquatic
mesocosm. Genetic and cultivated fecal indica-
tors, including fecal indicator bacteria (entero-
cocci, E. coli, and Bacteroidales), coliphage
(somatic and F+), and human-associated genetic
indicators (HF183/BacR287 and HumM2), were
measured in each sample. Simple linear regres-
sion assessing treatment trends in each fecal
type over time showed significant decay trends
(p < 0.05) in most treatments of feces and sew-
age sources (27/28 and 32/40, respectively), com-
pared to the septage source (6/26). A two-way
analysis of variance comparison of loglO reduc-
tion values for sewage and fecal experiments
indicated that treatment effects differentially
impact survival of cultivated bacteria, cultivated
coliphage, and genetic indicators. Findings sug-
gest that sunlight is critical for coliphage decay,
while indigenous microbiota play a greater role
in decay of genetic indicators in sewage than in
fecal source. This study offers new insights into
the decay of common human fecal pollution
sources in subtropical marine waters with
important implications for future water quality
management applications.
Biosketch
Dr. Pauline Wanjugi is a National Research
Council postdoctoral research fellow working
at the U.S. Environmental Protection Agency
(EPA) Office of Research and Development in
Cincinnati, Ohio. Dr. Wanjugi received her
undergraduate degree from the University of
Idaho, her master's degree from Pennsylvania
State University, and her doctorate from the
University of South Florida. Her current
research pertains to the identification and quan-
tification of sources of fecal pollution in recre-
ational waters using traditional and molecular
microbiology approaches. Prior to her current
tenure at EPA, Dr. Wanjugi worked in a similar
capacity at Texas A&M University, where she
conducted research on assessment of sources
of fecal pollution in Texas watersheds and
expansion of the Texas bacterial source tracking
library.
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Day Three: Session X
J
iL2a
Trash Free Beaches via Trash Free Waters
Presenter: Kelsey Watts-FitzGerald
U.S. Environmental Protection Agency, Office of Water
Authors: Margaret Murphy, Laura Johnson, Robert Benson, Noemi Mercado,
Rahul Madhusudanan
Abstract
Aquatic trash pollution is a global prob-
lem. Approximately 80% of trash in the world's
oceans comes from inland sources. The U.S.
Environmental Protection Agency's (EPA's)
Trash Free Waters (TFW) Program pursues
innovative strategies to prevent and reduce
the amount of trash entering U.S. rivers, lakes,
and coastal waters with the aim of moving
towards a zero loading of trash (particularly
plastic trash) entering our watersheds and
ultimately the ocean. The four major objectives
of the program are to 1) support stakeholder-
driven initiatives and projects in cities and
states; 2) support research on the ecological,
economic, and potential human health impacts
of trash in aquatic environments; 3) support
U.S. Government engagement on marine trash/
debris issues internationally; and 4) foster pub-
lic/private partnerships to generate innovative
ideas for technology, litter prevention, and mate-
rial reuse. With these as the tenets of TFW, our
program's progress includes having developed
regional strategies with projects in the Mid-
Atlantic, California and Pacific Islands, Gulf
of Mexico, New York/New Jersey, and Puerto
Rico; an assessment of the state of research
pertaining to aquatic plastic impacts in order to
explore the issue and inform EPA and non-EPA
research; Our Oceans commitments to follow
on the success of our Puerto Rico strategy by
expanding the TFW approach into the wider
Caribbean, as well as a partnership with China
to share information and best practices on
reducing aquatic trash; and partnerships with
industry stakeholders by convening an execu-
tive dialogue with our Administrator to explore
breakthrough ideas to achieve trash-free waters
in the U.S.
Biosketch
Ms. Kelsey Watts-FitzGerald is an Oak
Ridge Institute for Science and Education par-
ticipant in the U.S. Environmental Protection
Agency's Office of Water. She received her
bachelor of arts degree in biology and environ-
mental studies from Oberlin College, and her
master of public health degree in environmental
science and policy from Columbia University.
Ms. Watts-FitzGerald's fellowship placement is
in the Marine Pollution Control Branch, which
is focused on programs related to trash-free
waters; ocean acidification; corals; ocean dump-
ing; and vessels, marinas, and ports.
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U.S. EPA's 2016 Recreational Waters Conference
Vancomycin-resistant Enterococci in a
Domestic Sewage Spill in South Florida
Co-Presenters: Suzanne Young and Valerie ). Harwood, PhD
Department of Intergrative Biology, University of South Florida
Authors: Suzanne Young, Valerie ). Harwood
Abstract
Culturable Enterococcus faecium harbor-
ing the van A gene were detected in water and
sediment after a sewage spill in south Florida.
The site was sampled for 7 weeks after over 1
million gallons of domestic sewage was released
through a broken pipe. This is only the second
instance in which vancomycin-resistant entero-
cocci (VRE) harboring van A genes that confer
high-level resistance have been reported outside
of clinical settings in the United States. Putative
VRE were isolated on mEI agar amended with
32 |jg-ml-l vancomycin and could be cultured
from water and sediment as much as 5 days
after the event. Fecal indicator bacteria also
were monitored at the site and in receiving
waters and sediment. Culturable enterococci
levels in water at the site exceeded recreational
water quality guidelines for 4 weeks follow-
ing the spill. Analysis of bacterial species via
16S rRNA gene sequencing showed changes in
community structure through time following
the sewage spill in both sediment and water.
Communities in sediment were distinct from
those in water, with different dominant fami-
lies. Enteric pathogen families Enterococcaceae
and Enterobacteriaceae were present at low levels
in both water and sediment over the course of
the study. Six sewage-associated families identi-
fied by sequencing results represented a small
proportion of total reads, but declined over time
and on a similar timescale to culturable entero-
cocci and VRE in both water and sediment.
Biosketch
Ms. Suzanne Young is a fourth-year PhD
candidate in Dr. Valerie (Jody) Harwood's
water quality microbiology laboratory at the
University of South Florida in Tampa, Florida.
She completed her master's degree at Queens
College/City University of New York in New
York City, studying sewage contamination in
the Hudson River in collaboration with the
Riverkeeper water quality monitoring program.
Her publication topics include antibiotic resis-
tance in the environment, alternative habitats
of Vibrio vulnificus, microbial source tracking,
and disease ecology. Ms. Young received the
2013 EPA STAR Graduate Research Fellowship
to study the ecology of antibiotic resistant
bacteria in aquatic environments. The focus of
her doctoral dissertation is antibiotic resistance
gene transfer and the survival of vancomycin-
resistant enterococci under varying environ-
mental condition.
Dr. Valerie J. (Jody) Harwood is a profes-
sor and chair of the Department of Integrative
Biology at the University of South Florida,
where she has worked since 1998. She is an
environmental microbiologist whose research
focuses on microbial ecology, water quality, and
the use of molecular biology tools for assess-
ing the extent and sources of fecal contamina-
tion in water. She is a major contributor to the
U.S. Environmental Protection Agency (EPA)
guide document on microbial source tracking
(MST) and is a coeditor of the book, Microbial
Source Tracking: Methods, Applications & Case
Studies (Springer 2011). Dr. Harwood also has
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Day Three: Session X
J
La
conducted extensive research on the persistence
and ecology of enteric organisms in secondary
habitats such as water and sediments, and is at
the forefront of efforts to implement quantita-
tive PCR methods as a tool in environmental
microbiology research. She is the author of
more than 80 peer-reviewed publications on
subjects including MST, environmental per-
sistence and fate of fecal indicators and water-
borne pathogens, and Vibrio vulnificus ecol-
ogy Dr. Harwood collaborates with agencies
ranging from local to international (e.g., EPA,
WHO, UNESCO) on water quality issues. She
is a member of the Council Policy Committee
(executive board), a distinguished lecturer
for the American Society for Microbiology,
and a Fulbright Specialist Award recipi-
ent for work in Queensland, Australia, with
the Commonwealth Scientific and Industrial
Research Organisation.
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U.S. EPA's 2016 Recreational Waters Conference
Predictive Modeling of Microbial Indicators for
Timely Beach Notifications and Advisories at Marine
Beaches
Presenter: Richard Zepp, PhD
U.S. Environmental Protection Agency, National Exposure Research Laboratory/Office of Research and
Development, Ecosystems Research Division
Authors: Richard Zepp, Mike Cyterski, Marirosa Molina, Chris Fitzgerald, Gene Whelan,
Rajbir Parmar Kurt Wolfe, Mike Galvin
Abstract
Marine beaches are occasionally contami-
nated by levels of fecal indicator bacteria (FIB)
that exceed U.S. Environmental Protection
Agency water quality criteria. Here, we describe
application of a recent version of the software
package Virtual Beach tool, VB 3.0.5, to build
and evaluate multiple linear regression (MLR)
and generalized boosted models to predict
microbial water quality for selected marine
beaches in the eastern United States and Puerto
Rico. For comparison, we used the same model-
ing technique to evaluate selected freshwater
beaches of the Great Lakes. Culturable and
qPCR methods were used to measure entero-
cocci concentrations. Environmental variables
for the beach sites such as rainfall, wind and
current speed/direction, and solar radiation
were obtained concurrently. VB 3.0.5 can be
used to develop site-specific relationships
—"models"—that link the densities of FIB to
the environmental variables. VB was used to
evaluate the best models for these beaches from
an extensive list of choices. Comparisons of
MLR modeling results for predicted FIB densi-
ties at the marine beaches and results obtained
at selected Great Lakes beaches indicate that,
on the basis of adjusted R2 values for predicted
versus observed levels of FIB, model perfor-
mance was better for the freshwater Great Lakes
beaches than for the marine beaches (freshwa-
ter average adjusted R2 = 0.5, marine average
adjusted R2 = 0.39). Modeling results for the
culturable FIB data at the marine beaches also
were somewhat better than for qPCR data (cul-
turable average adjusted R2 = 0.46, qPCR aver-
age adjusted R2 = 0.42). Lower values for marine
beaches likely reflect the interplay of several
factors discussed in the presentation.
Biosketch
Dr. Richard G. Zepp is a Senior Research
Scientist at the U.S. Environmental Protection
Agency National Exposure Research Laboratory
in Athens, Georgia. He received his bachelor of
science degree Furman University and his doc-
torate from Florida State University. Dr. Zepp's
research interests include predictive model-
ing and fate and transport of pathogens and
fecal indicators in aquatic environments. He is
a member of the United Nations Environment
Programme Environmental Effects Assessment
Panel and is an adjunct professor at the
Rosenstiel School of Marine and Atmospheric
Sciences at the University of Miami, Florida,
and the Department of Chemistry at the State
University of New York in Syracuse, New York.
He is also a member of AGU, ACS, SETAC, ISES,
ASM, ASLO, Sigma Xi, and AAAS.
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Day Three: Session X
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iL2a
Process Relationships for Evaluating the Role of
Light-induced Inactivation of Coliphages at Selected
Beaches and Nearby Tributaries of the Great Lakes
Presenter: Richard Zepp, PhD
U.S. Environmental Protection Agency, National Exposure Research Laboratory/Office of Research and
Development, Ecosystems Research Division
Authors: Richard Zepp, Marirosa Molina, Kelvin Wong, Mike Cyterski, Gene Whelan,
Rajbir Parmar, Brad Acrey, Rania Georgacopoulos
Abstract
One approach to predictive modeling of
biological contamination of recreational waters
and drinking water sources involves applying
process-based models that consider microbial
sources, hydrodynamic transport, and microbial
fate. Coliphages are possible indicators of fecal
contamination of recreational waters. Past stud-
ies have indicated that sunlight plays an impor-
tant role in altering densities of coliphages,
other indicator microorganisms, and pathogens
in aquatic environments. Here, we report on
laboratory studies of light-induced inactivation
of two species of coliphage, an F-specific RNA
coliphage and a somatic coliphage, under vari-
ous conditions in phosphate-buffered solution
and natural waters. Inactivation rates of the coli-
phages were determined in a series of irradia-
tions that used simulated solar radiation passed
through light filters that blocked different parts
of the ultraviolet and visible spectral region.
Inactivation rates and spectral irradiance were
then analyzed by the Rundel technique to
develop biological weighting functions (BWFs)
for the light-induced inactivations. Direct expo-
sure to solar radiation at midday and shallow
depths in all waters resulted in rapid inactiva-
tion, with half-lives of hours or less. In the dark,
inactivation half-lives were much longer—on
the order of several days. BWFs were com-
bined with data for underwater solar spectral
irradiance to model light-induced inactivation
of phages in selected recreational waters. We
report on use of this approach to help evaluate
potential effects of sunlight-induced inactiva-
tion on phage densities at Great Lakes sites
located near Milwaukee, Wisconsin; Michigan
City, Indiana; Cleveland, Ohio; Duluth,
Minnesota; and the Manitowoc River basin.
Biosketch
Dr. Richard G. Zepp is a Senior Research
Scientist at the U.S. Environmental Protection
Agency National Exposure Research Laboratory
in Athens, Georgia. He received his bachelor of
science degree Furman University and his doc-
torate from Florida State University. Dr. Zepp's
research interests include predictive model-
ing and fate and transport of pathogens and
fecal indicators in aquatic environments. He is
a member of the United Nations Environment
Programme Environmental Effects Assessment
Panel and is an adjunct professor at the
Rosenstiel School of Marine and Atmospheric
Sciences at the University of Miami, Florida,
and the Department of Chemistry at the State
University of New York in Syracuse, New York.
He is also a member of AGU, ACS, SETAC, ISES,
ASM, ASLO, Sigma Xi, and AAAS.
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U.S. EPA's 2016 Recreational Waters Conference
Development of a Human-Specific B. thetaiotaomicron
IMS/ATP Assay for Measuring Viable Human
Contamination in Surface Waters in Baja California,
Mexico
Presenter: Amity Zimmer-Faust, PhD
U.S. Environmental Protection Agency, National Health and Environmental Effects Research Laboratory
Authors: Amity Zimmer-Faust, Vanessa Thulsiraj, Leopoldo Espinosa-Mendoza,
Jennifer Jay
Abstract
Immunomagnetic separation/ adenosine
triphosphate (IMS/ATP) assays utilize paramag-
netic beads and target-specific antibodies to iso-
late target organisms. Following isolation, ade-
nosine tri-phosphate (ATP) is extracted from the
target population and quantified. An inversely
coupled (Inv-IMS/ATP) assay for detection of
Bacteroides thetaiotaomicron was developed and
applied for rapid detection of human-associated
fecal contamination in surface waters in Baja
California. Specificity of the assay was tested
against challenge solutions of varying concen-
tration of dog, gull, horse, and chicken feces,
and a field validation survey of assay measure-
ments in coastal and wastewater treatment plant
effluent water quality in Rosarito and Ensenada,
Baja California, was conducted. Inv-IMS/ATP
measurements were shown to be specific and
sensitive to human fecal contamination. At test
concentrations of challenge feces less than 1000
MPN ENT/100 mL, sensitivity and specificity
of the assay both exceeded 80%. Moreover, the
Inv-MS/ATP assay yielded measurements of
viable B. thetaiotaomicron that were comparable
to the HF183 human marker in complex surface
waters impacted with both wastewater and
stormwater runoff, and the Inv-IMS/ATP assay
was able to effectively differentiate between sur-
face waters impacted with adequately and inad-
equately treated wastewater. The Inv-IMS/ATP
assay shows promise for rapid evaluation of
recreational water quality in areas where access
to more expensive methods is limited and in
areas where water quality is unpredictable.
Biosketch
Dr. Amy Zimmer-Faust is a postdoc-
toral researcher for the U.S. Environmental
Protection Agency at the National Health and
Environmental Exposure Research Lab, Pacific
Coastal Ecology Branch located in Newport,
Oregon. She received her bachelor of science
degree in aquatic biology from the University
of California at Santa Barbara and her doctorate
in environmental science and engineering from
the University of California at Los Angeles.
Dr. Zimmer-Faust's PhD focused on the applica-
tion of rapid methods (MST markers and IMS/
ATP assays) for evaluation of coastal water qual-
ity. Her main research interests include evalua-
tion of the sources and fate of fecal pollution in
coastal systems and the environmental param-
eters that control fecal inputs.
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