I-41_18-06995-000_ProjRep-MicrobialSrcTrck_20210302

Project Report
FINAL
February 2021
Northeast Creek Microbial Source
Tracking Study
Durham, North Carolina
PREPARED FOR:
City of Durham
Public Works Department
Stormwater & GIS Services Division
101 City Hall Plaza, Third Floor
CITY OF
DURHAM Durham, North Carolina 27701

Note:
Some pages in this document have been purposely skipped or blank pages inserted
so that this document will print correctly when duplexed.

(NO-i
HERRERA
AOcom
Northeast Creek Microbial Source
Tracking Study
Project Report
City of Durham
Stormwater & GIS Services Division
Public Works Department
101 City Hall Plaza, Third Floor
CITY OF
DURHAM Durham, North Carolina 27701

AECOM—Herrera
Table of Contents
Acronymsand Abbreviations.................................................................................................................................... iv
1 Introduction..................................................................................................................................................... 1-1
2 Watershed Description................................................................................................................................... 2-1
2.1 Problem Statement....................................................................................................................................................................2-4
2.2 Project Objectives and Understanding..............................................................................................................................2-4
3 Study Methods................................................................................................................................................. 3-1
3.1 Sample Stations...........................................................................................................................................................................3-1
3.2 MST Method and Marker Selection....................................................................................................................................3-1
3.3 Sampling Schedule.....................................................................................................................................................................3-2
3.4 Sample Collection and Analysis............................................................................................................................................3-6
3.5 Data Quality Assessment.........................................................................................................................................................3-6
4 Results and Discussion.................................................................................................................................... 4-1
4.1 Data Quality Control..................................................................................................................................................................4-1
4.1.1 Fecal Coliform Bacteria............................................................................................................................................4-1
4.1.2 Biomarkers....................................................................................................................................................................4-1
4.2 Hydrology......................................................................................................................................................................................4-2
4.3 Fecal Coliform Bacteria.............................................................................................................................................................4-4
4.4 Human Biomarker.......................................................................................................................................................................4-6
4.5 Dog Biomarker.............................................................................................................................................................................4-7
4.6 Biomarker Comparison.............................................................................................................................................................4-8
4.7 Correlation Analysis.................................................................................................................................................................4-10
4.8 Management Implications....................................................................................................................................................4-10
5 Conclusions...................................................................................................................................................... 5-1
5.1 Hydrology......................................................................................................................................................................................5-1
5.2 Fecal Coliform Bacteria.............................................................................................................................................................5-1
5.3 Human Biomarker.......................................................................................................................................................................5-1
5.4 Dog Biomarker.............................................................................................................................................................................5-2
6 Recommendations...........................................................................................................................................6-1
6.1 Human Fecal Source Control.................................................................................................................................................6-1
6.2 Dog Fecal Source Control........................................................................................................................................................6-2
7 References........................................................................................................................................................7-1
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Appendices
Appendix A: Database
Appendix B: Field Notes and Photographs
Appendix C: Laboratory Reports and Quality Assurance Worksheets
Cover Photographs clockwise from upper left of stations MST-9, NE3.3NP, NE2.2NP, and MST-18 on March 9,
2020 by Rob Zisette, Herrera Environmental Consultants
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Figures
Figure 2-1. Northeast Creek Land Use (Source: Brown and Caldwell, 2012).............................................................................2-2
Figure 2-2. Northeast Creek Historical Sampling Stations and USGS Gages............................................................................2-3
Figure 3-1. Northeast Creek MST Study Sampling Stations . ........................................................................................................... 3-3
Figure 4-1. Hydrologic Data Summary for the Duration of the Northeast Creek MST Study............................................4-3
Figure 4-2. Precipitation in Relation to Sampling Times for Storm Sampling Events for the Northeast
CreekMST Study........................................................................................................................................................................4-4
Figure 4-3. Fecal Coliform Bacteria Box Plots for the Northeast Creek MST Study . .............................................................. 4-6
Figure 4-4. Human and Dog Biomarker Box Plots for the Northeast Creek MST Study . ..................................................... 4-8
Figure 4-7. Fecal Coliform Versus Human and Dog Biomarker Concentrations for the Northeast Creek
MSTStudy...................................................................................................................................................................................4-11
Tables
Table3-1.
Water Sample Locations . ........................................................................................................................................................
3-2
Table 3-2.
Northeast Creek MST Sampling Station Catchment Attributes . .............................................................................
3-5
Table 3-3.
Sampling Schedule and Precipitation Amounts . ...........................................................................................................
3-6
Table 4-1.
Fecal Source Sample Results for Dog Biomarker..........................................................................................................
4-2
Table4-2.
Precipitation Data......................................................................................................................................................................
4-3
Table 4-3.
Fecal Coliform Bacteria Statistics for Base and Storm Events..................................................................................
4-5
Table 4-4.
Biomarker Geomeans for Base and Storm Events........................................................................................................
4-7
Table 4-5.
Comparison of Host -Associated Biomarker and Fecal Coliform Bacteria Concentrations in
FecalSource Samples...............................................................................................................................................................4-9
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Acronyms and Abbreviations
ARA................. antibiotic resistance analysis
CAAE.............. North Carolina State University Center
for Applied Aquatic Ecology
cfs.................... cubic feet per second
CFU ................. colony forming unit
City .................. City of Durham Stormwater and GIS
Services
CV .................... coefficient of variation
DNQ............... detected not quantified
EPA .................. United States Environmental Protection
Agency
GIS................... geographic information systems
MEB................ method extraction blanks
mL................... milliliter
MPN ............... most probable number
MST ................ microbial source tracking
MQOs............ measurement quality objectives
NCSU.............. North Carolina State University
NCDWR.........
North Carolina Department of
Environmental Quality, Division of
Water Resources
ND ...................
not detected
NSW ...............
nutrient sensitive waters
NTC.................
no template controls
QA...................
quality assurance
QAPP..............
quality assurance project plan
QC ................... quality control
gPCR............... quantitative polymerase chain reaction
RPD................. relative percent difference
RTP.................. Research Triangle Park
SCM................
stormwater control measure
SSO.................
sanitary sewer overflow
TMDL..............
total maximum daily load
TNTC..............
too numerous to count
USGS..............
United States Geological Survey
WIP.................
watershed improvement plan
iv Northeast Creek Microbial Source Tracking Study —Project Report

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1 Introduction
The purpose of this document is to present the Project Report for the microbial source tracking (MST) study
conducted in Northeast Creek, Durham, North Carolina. This project was performed for the City of Durham
Stormwater and GIS Services staff (City). The Project Team consists of AECOM, Herrera Environmental Consultants
(Herrera), and Three Oaks Engineering (Three Oaks).
The City selected the Project Team to design and execute a quantitative MST study to improve water quality
management decision -making in the Northeast Creek Watershed. The goal of this study was to demonstrate the
ability and cost-effectiveness of quantitative DNA -based MST technology to characterize fecal bacteria pollution
sources in surface waters in the portion of the Northeast Creek Watershed located within the city.
The scope of work to meet this study goal included:
• Reviewing historical information about fecal coliform bacteria in the Northeast Creek Watershed to identify
potential fecal bacteria hot spots and sources, and recommending an MST study design to quantify key
sources of concern.
• Developing a Quality Assurance Project Plan (QAPP) that supplement ongoing monitoring activities with
molecular MST analysis using quantitative polymerase chain reaction (gPCR) and described sampling
locations and methods, analytical methods, quality assurance/quality control (QA/QC) requirements, and
other pertinent information.
• Conducting a site visit to train field staff on sampling protocols.
• Coordinating water sample collection and laboratory analysis to ensure high quality data are obtained.
• Conducting a series of meetings with the Project Team over the course of the study.
• Appling a holistic approach by integrating molecular MST results with fecal bacteria concentrations and
environmental factors using geospatial statistical analyses to evaluate how fecal sources vary spatially by
location and subwatershed characteristics, seasonally over time, and hydrologically over different types of
flow.
• Preparing a detailed report that presents the MST study methods, results, conclusions, and
recommendations for fecal source control actions and additional study as warranted by the results and in
consultation with the City.
This Project Report includes the following:
• Background Information —Watershed description, problem statement, and project objectives and
understanding.
• Study Methods —Field and laboratory methods for fecal coliform bacteria and gPCR MST analysis of water
samples collected at sample stations.
• Results and Discussion —Presentation and analysis of study data and discussion of management
implications.
• Conclusions —Summary of key study findings.
• Recommendations —Recommendations for fecal source control actions and additional study.
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2 Watershed Description
The entire Northeast Creek Watershed covers 50.2 square miles that includes large industrial and commercial
areas, undeveloped land, low density to high density residential areas, and rural residential areas (Figure 2-1). The
majority of the watershed is in Durham County, with almost 40 percent of the watershed within the City of
Durham limits (Brown and Caldwell, 2012). Research Triangle Park (RTP) encompasses nearly one-third of the
watershed along the eastern side, with the remaining portions in Wake and Chatham counties.
Northeast Creek flows north to south with Kit Creek and Burdens Creek as its two major tributaries, both flowing
from east to west. Interstate-40 passes through the middle of the watershed and NC-55 runs north to south
through the watershed within the city limits. The eastern border of the watershed is defined closely by Miami
Boulevard and Barbee Road outlines the western boundary. Highway NC-540 and O'Kelly Chapel Road border the
southern boundary of the watershed, and the northern tip of the watershed is the intersection of Ellis Road and
NC-147.
Northeast Creek drains to B. Everette Jordan Lake (Jordan Lake). Both Northeast Creek and Jordan Lake water
bodies have been identified as nutrient sensitive waters (NSW) by the North Carolina Department of
Environmental Quality. A fecal coliform bacteria total maximum daily load (TMDL) for Northeast Creek was
approved in 2003. The TMDL addresses an 8.4-mile stream reach from NC Highway 55 in the City of Durham to a
point 0.5 miles downstream of Panther Creek that flows through Chatham, Durham, and Wake counties.
The Northeast Creek Watershed is located within the Piedmont physiographic region of North Carolina that
typically receives a steady and moderate amount of annual precipitation (Brown and Caldwell, 2012). The fall
months of October, November and December are usually slightly drier months, although April is also often drier
than the annual average. Historically, one additional factor affecting precipitation in the watershed has been
Atlantic Ocean tropical storms. The Atlantic Hurricane Season extends from June 1 through November 30.
Northeast Creek is a United States Geological Survey (USGS) gauged stream with one gauging station located on
the lower main stem of the creek outside the City of Durham and the second located on a main tributary,
Northeast Creek North Prong, within the city. The upper USGS station 0209741387 is located on Northeast Creek
North Prong approximately 0.5 miles upstream of the confluence with Northeast Creek and approximately
9.5 miles upstream of Jordan Lake. Specifically, the gauge is located just off Carpenter Fletcher Road at the
intersection of NC-55. At this location, 3.25 square miles or 9 percent of the watershed area drains to the gauging
station. The USGS station 0209741955 is located approximately five miles upstream of Jordan Lake just south of
Scott King Road on Grandale Drive over Northeast Creek. At this location, 21.1 square miles or 60 percent of the
watershed area drains to the gauging station. Figure 2-2 shows the locations of both USGS stations.
Northeast Creek has a moderately high percentage of impervious area within its watershed (approximately
17 percent), which makes it prone to localized urban flooding problems, especially within the highly developed
areas (Brown and Caldwell, 2012). Soils in the Northeast Creek Watershed are highly erodible and poorly drained
with very low permeability (Brown and Caldwell, 2012). Low permeability soils increase surface runoff during
storms and cause low base flow in streams due to reduced groundwater available to feed streams in the intervals
between storm events.
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kn Ser5ia 9P
Hignway
Major Road
County Boundary
QSubwatersheds
Northeast and Crooked Creek Study Lmits
Existing Land Use
Very Low Density Residental
Low Den s+ty Reskden t a I
— Mod -urn Density Residenyal
— high Density Residental
— Agncultural
Parks and Open Space
— Commerual
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— k+sonjoonai
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Roadways
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Figure 2-1. Northeast Creek Land Use (Source: Brown and Caldwell, 2012).
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0209725960
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DURHAM
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Historical
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Figure 2-2. Northeast Creek Historical Sampling Stations and USGS Gages.
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The project QAPP (Herrera and AECOM 2020) includes additional background information on historical fecal
bacteria data, fecal sources identified by a previous MST study, and fecal bacteria control measures implemented
by the City in the Northeast Creek watershed.
2.1 Problem Statement
Fecal coliform bacteria concentrations are high in the upper reaches (North Prong) of Northeast Creek located
within the City of Durham. Effective control of fecal bacteria requires knowing how the amount of human and
animal fecal sources vary along the length of the stream under wet and dry conditions. Since this is an urban
watershed, human and dog are the primary fecal sources of concern for the city that can be controlled by SCMs
and illicit discharge detection.
2.2 Project Objectives and Understanding
The primary objective of this MST study was to quantify the abundance and contribution of human and dog
sources at locations throughout the Northeast Creek Watershed within the City of Durham to generate
investigative leads providing actionable outcomes. Project objectives included:
• Using an advanced MST method (single host marker quantification by gPCR) to quantify the abundance and
contribution of human and dog sources at locations throughout the Northeast Creek Watershed within the
City of Durham.
• Evaluating relationships between MST results and fecal coliform bacteria concentrations, hydrologic
conditions, land use conditions, and source control actions using geospatial statistical analysis.
• Prioritizing locations of the identified sources and identifying appropriate methods for controlling high
priority sources based on past experience.
This study design was based on the following understanding and assumptions:
• Fecal coliform bacteria concentrations in Northeast Creek are primarily affected by loadings in surface water
discharges to the stream from tributaries and drainage outfalls during storm and base flow events.
• Fecal coliform bacteria concentrations in Northeast Creek are higher during storm events than base flow
events.
• Sources of fecal coliform bacteria present in Northeast Creek will vary spatially, temporally, and
hydrologically.
• Human sources of fecal coliform bacteria present in Northeast Creek may primarily include sewer cross -
connections and overflows, but minor sources are possible from open defecation in the environment and
onsite septic systems located in subwatersheds draining to Northeast Creek upstream of Northeast Creek
North Prong.
• Domestic animal sources of fecal coliform bacteria present in Northeast Creek may primarily include dogs
from pet owners that do not properly dispose of pet waste because livestock are not expected to be
abundant in the watershed.
• The gPCR MST methods will accurately identify the relative abundance of human and dog fecal sources
present in the collected stream samples.
• Characterization of fecal sources in the collected samples will increase the ability to identify effective
corrective actions for meeting TMDL requirements.
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3 Study Methods
3.1 Sample Stations
Sample stations were selected based on the following objectives:
Include historical sample stations that have been monitored in the past to compare historical data to data
collected under this project.
• Include new sample stations draining specific subwatersheds of interest.
• Select stations with majority of drainage from the City of Durham.
The following S stations, from upstream to downstream, were selected in the Northeast Creek Watershed:
MST-9—new sample station located at East Cornwallis Road on upper Northeast Creek North Prong that
drains Subwatershed 9
NE3.3NP—historical sample station located at Odyssey Drive on middle Northeast Creek North Prong that
drains Subwatersheds 9 and 10.
• NE2.2NP—historical sample station located at Meridian Parkway near the mouth of Northeast Creek North
Prong that drains Subwatersheds 9, 10, and 11.
• MST-1 2—new sample station on a small tributary at Interstate 40 that drains to Northeast Creek from
Subwatershed 12.
• MST-18—new sampling station on a small tributary draining at Clermont Road that drains to Parkwood Lake
in the upper portion of Northeast Creek Subwatershed 18.
Sample stations are described in Table 3-1 and shown on Figure 3-1. Attributes of the catchment areas draining to
each station are presented in Table 3-2.
3.2 MST Method and Marker Selection
A state-of-the-art MST methodology was selected for this study: single host -associated markers by gPCR. The
gPCR method used was performed by Source Molecular, which is located in Miami, Florida and is the nation's only
A2LA-accredited gPCR MST analysis lab.
The gPCR method was selected because this technology is currently the most mature and effective method for
MST. Comprehensive method evaluation has been published to confirm its high performance (Griffith et al. 2013).
Standardized protocols have been developed and are available for many fecal sources. The gPCR method
protocols were recommended from the largest MST method evaluation study and by the California MST manual
(Griffith et al. 2013). Rigorous gPCR data acceptance criteria have also been published and are available to ensure
highest MST data quality (EPA, 2019 and Boehm et al. 2013). Method diagnostic sensitivity and specificity have
been recently reported for a comprehensive MST study in Washington (Herrera, 2019), along with a range of
measured marker concentrations and limits of quantification. The gPCR method is the most commonly used
molecular MST method, and is currently being used by the EPA. Source Molecular's involvement in this study
provided key advantages given they are a commercial laboratory offering high quality analysis and rapid data
turnaround, and they would be available for use in future studies without having to obtain an agreement with a
government or university laboratory.
Northeast Creek Microbial Source Tracking Study —Project Report 3-1

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Table 3-1. Water Sample Locations.
MST-9 New Upper Northeast Creek North Prong on west bank at 100 feet 35.934027-78.890682
north (upstream) of East Cornwallis Road and 500 feet west of
Highway 55
NE3.3NP Existing
Middle Northeast Creek North Prong on west bank at 20 feet 35.928272-78.893501
north (upstream) of Odyssey Drive and 1,200 feet west of
Highway 55
NE2.2NP Existing
Lower Northeast Creek North Prong on west bank at 60 feet 35.910441-78.895253
north (upstream) of Meridian Parkway and 500 feet west of
Highway 55
MST-12 New
Northeast Creek Tributary on west bank at 100 feet north 35.907774-78.901568
(upstream) of its confluence, 500 feet south of Interstate 40,
and 1,600 feet west of Highway 55 (accessed via Hanmaum
Church parking lot)
MST-18 New
Northeast Creek Tributary on west bank at 200 feet north 35.890208-78.911527
(upstream) of Clermont Road and 50 feet upstream of pond
draining under Clermont Road to Parkwood Lake
The following two markers were selected for analysis:
Human marker HF183 EPA (Green et al. 2014a and EPA, 2019). Other human markers are available, but the
HF183 EPA marker was selected because it has high sensitivity (low false negatives) and high selectivity (low
false positives) (Griffith et al. 2013 and Herrera, 2019).
Dog marker DG3 (Green et al. 2014b). Other dog gPCR markers are available, but the DG3 marker was
selected because it is the most commonly used marker that has been shown to have relatively high
sensitivity and selectivity (Griffith et al. 2013 and Herrera, 2019).
3.3 Sampling Schedule
Water samples were collected from May through October 2020 during four base flow events and three targeted
storm events. Table 3-3 present the sampling schedule in comparison to that planned in the QAPP (Herrera and
AECOM, 2020). The four base flow sampling events were planned to occur in April, June, August, and October
2020. The Base 1 event was delayed from April to June due to COVID-19 work restrictions and the Base 3 event
was delayed from August to early September due to wet conditions. The three targeted storm events were
conducted as planned in April -May, June -August, and September -October.
Base flow events occurred after at least 72 hours of less than 0.04 inches of rain, which was assumed to be the
minimum amount of rain to generate runoff. Storm events occurred for storms with at least 0.20 inches of rain
within the previous 24-hour period. Larger storm events (greater than 0.50 inches) were targeted for sampling
based on local rainfall predictions to ensure the minimum rain amount is met. The previous 24-hour rainfall
amounts for storm events ranged from 0.77 to 2.08 inches (see Table 3-3). Precipitation data from USGS station
0209725960 was used to document storm event size and for data analysis. This rain gauge is a real-time station
located just west of the Northeast Creek Watershed at M.L. King Parkway in in the Third Creek Watershed (see
Figure 3-1).
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Table 3-2.
Northeast Creek MST Sampling Station Catchment Attributes.
Area (acres)
606
1,352
2,215
474
321
Sewer Density (feet/acre)
93
95
94
132
103
Sewer Line (feet)
56,523
128,438
207,744
62,660
32,993
Agricultural Parcels (number)
5
8
12
0
2
Land Use (Percent Cover)
Agricultural
15%
12%
9%
0%
14%
Parks and Open Space
0%
0%
4%
9%
13%
Very Low Density Residential
8%
6%
7%
3%
3%
Low Density Residential
17%
16%
22%
14%
27%
Medium Density Residential
6%
8%
7%
12%
10%
High Density Residential
17%
16%
11%
21%
10%
Institutional
3%
6%
6%
0%
5%
Commercial
9%
13%
13%
14%
3%
Industrial
12%
9%
7%
0%
1%
Roadways
14%
15%
15%
27%
14%
Land Cover (Percent Cover)
Open Water/Wetlands
0%
0%
1%
0%
0%
Barren Land (Rock/Sand/Clay)
0%
0%
0%
0%
0%
Cultivated Crops
0%
0%
0%
0%
0%
Pasture/Hay
2%
1%
1%
0%
0%
Shrub/Scrub
0%
0%
0%
0%
0%
Grassland/Herbaceous
2%
1%
1%
0%
1%
Mixed Forest
19%
13%
11%
4%
3%
Evergreen Forest
13%
8%
8%
3%
9%
Deciduous Forest
8%
10%
9%
2%
1%
Developed, Open Space
24%
29%
38%
39%
65%
Developed, Low Intensity
20%
23%
21%
29%
17%
Developed, Medium Intensity
10%
12%
10%
22%
3%
Developed, High Intensity
3%
2%
2%
1%
0%
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Base 1
Base 2
Base 3
Base 4
Storm 1
Storm 2
Storm 3
Dog Feces
Table 3-3. Sampling Schedule and Precipitation Amounts.
April 2020
6/3/20: 0930-1105
MST-12
0.00 (72-hr)
June 2020
6/23/20: 0925-1100
None
0.00 (72-hr)
August 2020
9/8/20: 1100-1240
None
0.00 (72-hr)
October 2020
10/15/20: 1025-1150
None
0.01 (72-hr)
April -May 2020
5/28/20: 0927-1121
None
0.65 (event)
June -August 2020
8/4/20: 0945-1100
MST-9
2.07 (event)
Sept. -Oct. 2020
9/30/20: 1130-1245
NE3.3
0.85 (event)
June 2020
6/23/20: 0603-1050
None
0.00 (72-hr)
3.4 Sample Collection and Analysis
Water sampling was conducted for all stations on the same day for each event by the Three Oaks field crew
according to procedures specified in the QAPP and following COVID-19 protocols. All samples were collected
using sterile technique as per the QAPP and in flowing water without incident. Compiled field notes are presented
in Appendix B and include weather observations, stream gauge data, sample locations and times, bottles filled,
photographs taken, flow conditions, water clarity, and other observations of interest.
Water samples were delivered within 6 hours of collection to Pace Analytical in Raleigh, NC for analysis of fecal
coliform bacteria by membrane filtration (Method SM 9222D-2006). The QAPP specified that this analysis would
be performed by North Carolina State University Center for Applied Aquatic Ecology, but this laboratory was
closed due to COVID-19 so Pace Analytical was selected as an alternate laboratory.
Water samples were shipped overnight to Source Molecular for sample filtration, DNA extraction, and gPCR
marker analysis of the following Bacteroidetes fecal biomarkers:
1. Human biomarker HF183 (Green et al. 2014a and EPA, 2019).
2. Dog marker DG3 (Green et al. 2014b).
The accuracy of these two markers has been well documented in terms of high sensitivity (less than 10 percent
false negatives) and high selectivity (less than 10 percent false positives) by Griffith et al. (2013) and Herrera
(2019).
To supplement accuracy results from the literature for the dog biomarker, 10 dog feces samples were collected
from a City maintained dog park (Northgate Dog Park located on Ellerbe Creek north of Northeast Creek) and
analyzed for the dog biomarker to measure the marker sensitivity to dog sources in the watershed. Fresh dog
feces samples were collected by the City of Durham in one day and shipped overnight to Source Molecular for
gPCR analysis according to the method described in the QAPP (Herrera and AECOM, 2020).
3.5 Data Quality Assessment
All data obtained from field and laboratory measurements were reviewed and verified for conformance to project
requirements, and then validated against the data quality objectives that are listed in the QAPP (Herrera and
AECOM, 2020). Only those data that are supported by appropriate quality control data and meet the
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measurement performance specification defined for this project were considered acceptable and used in the
project.
Laboratory data were verified and validated within 7 days of receiving the results from the laboratory. This review
was performed to ensure that all data are consistent, correct, and complete, and that all required quality control
information has been provided. Quality control reviews, including any problems or corrective actions, are
summarized in a Quality Assurance Worksheet for each sampling event (Appendix Q. Values associated with
minor quality control problems are considered estimates and assigned "J" qualifiers. Values associated with major
quality control problems are rejected and qualified "R". Estimated values are used for evaluation purposes,
whereas rejected values are not used. The data quality assessment was performed for the following quality control
elements:
• Completeness
• Methodology
• Holding times
• Negative controls (blanks)
• Positive controls
• Laboratory duplicates
• Fecal coliform bacteria enumeration
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4 Results and Discussion
4.1 Data Quality Control
4.1.1 Fecal Coliform Bacteria
Based on the data validation, all reported results were considered acceptable for use as reported and met
data quality objectives with the following exceptions:
Four fecal coliform bacteria results had plate colonies that were too numerous to count (TNTC) and
actual results may be higher than those reported by the laboratory. Results are reported as greater
than (>) the laboratory result.
• Two fecal coliform bacteria results were qualified as estimated (J) due to field duplicate criterion
exceedances (i.e., relative percent difference greater than 50 percent).
• One fecal coliform bacteria result was qualified as estimated (J) due to laboratory duplicate criterion
exceedances (i.e., relative percent difference greater than 35 percent).
4.1.2 Biomarkers
Based on the data validation, all reported results were considered acceptable for use as reported.
For quality control purposes, a positive control and a negative control are run alongside the samples to
ensure a properly functioning reaction and reveal any false negatives or false positives. Three method
extraction blanks (MEB) were included in each extraction batch. None of the extraction blanks generated
positive signals, meeting the measurement quality objective. For all host -associated markers, three No
Template Controls (NTC) were analyzed per gPCR plate as negative controls. None of the NTC generated
positive signals, meeting the measurement quality objective.
Marker sensitivity results for the fecal source sample analyses are presented in Table 4-1. Based on the
results, the dog marker (DG3) did not meet the 90 percent sensitivity objective identified in the QAPP as
50 percent (5 out of 10 samples) were not detected (ND). The low sensitivity of the dog marker indicates
that the presence of dog sources was underestimated in the water samples.
For comparison, dog feces samples recently analyzed for an MST Demonstration Project in Washington
State showed a sensitivity of 91 percent for the dog DG3 biomarker where the biomarker was detected in
10 of 11 samples (Herrera, 2019). In addition, septic tank samples analyzed for this MST Demonstration
Project also showed a sensitivity of 91 percent for the human HF183 biomarker where the biomarker was
detected in 10 of 11 samples (Herrera, 2019). These same source samples were also analyzed for seven
different source biomarkers (human, cow, horse, ruminant, pig, dog, and bird) and showed a specificity of
100 percent where none of the non -target biomarkers were detected in any of the dog and septic tank
samples (Herrera, 2019).
Northeast Creek Microbial Source Tracking Study —Project Report 4-1

AECOM—Herrera
Table 4-1. Fecal Source Sample Results for Dog Biomarker.
Dog-1
6/23/2020/06:03
ND
Dog-2
6/23/2020/07:45
ND
Dog-3
6/23/2020/8:20
ND
Dog-4
6/23/2020/8:50
DNQ
Dog-5
6/23/2020/8:58
4.75E+07
Dog-6
6/23/2020/9:24
ND
Dog-7
6/23/2020/9:30
8.99E+05
Dog-8
6/23/2020/9:45
4.59E+04
Dog-9
6/23/2020/10:40
3.36E+06
Dog-10
6/23/2020/10:50
ND
Geomeana
1.60E+06
Geomean calculated from quantified positive results.
DNQ = "Detected not quantified". The fecal biomarker was detected in both test replicates but in quantities below the limit of
quantification. This result indicates that fecal indicators associated with the respective host was present in the sample but in low
concentrations.
ND = "Not detected". The fecal biomarker was either not detected in test replicates, one replicate was detected at a cycle threshold
greater than 35 and the other was not, or one replicate was detected at a cycle threshold less than 35 and the other was not after
repeated analysis.
4.2 Hydrology
Hydrologic data compiled from other sources are summarized in Figure 4-1 showing the average daily
discharge of Northeast Creek (at USGS station 0209741955), daily precipitation totals (USGS station
0209725960), and base and storm sampling event dates. Also shown in Figure 4-1 is the average
discharge of Northeast Creek for the duration of the project (36.4 cubic feet per second [cfs]), which is
similar to the mean annual flow between October 1982 and October 2011 for Northeast Creek (30 cfs;
Brown and Caldwell, 2012).
The City compiled records of sanitary sewer overflows (SSOs) in the Northeast Creek Watershed
from March 1997 through November 2020 (L. Fontaine, City of Durham, personal communication).
No SSOs were recorded within any of the MST station catchments (subwatersheds) during the MST
sampling period. Only three SSOs were recorded in any of the MST station catchments in the past
23 years. These three SSO locations are shown in Figure 3-1 and include 3,000 gallons in catchment
MST-9 in February 2017, 90 gallons in catchment MST-18 in November 2017, and 2,060 gallons in
catchment MST-12 in November 2019. In addition to infrequent occurrence, these SSO volumes are
small in comparison to stream flow. For example, a 3,000 gallon spill represents 0.015 percent of the
average daily discharge of Northeast Creek (based on an annual average flow of 30 cfs or 20 million
gallons per day at USGS station 0209741955).
Precipitation data are presented for each of the three storm sampling events in Figure 4-2 showing rainfall
amounts (30-minute intervals and cumulative), stream discharge rates (at USGS station 0209741955), and
sample collection times for each of the five stations. The QAPP specified at least 0.20 inches of rain within
4-2 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

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the previous 24-hour period for storm events. As shown in Figure 4-2 and Table 3-3, the criteria were met
for all three storm events sampled. Sampling was initiated about 6 hours after rain stopped for Storms 1
and 2 and about 12 hours after rain stopped for Storm 3. Due to the travel time for stormwater to flow
into and along the creek, sampling was initiated on the rising limb or near the peak of the hydrograph for
lower Northeast Creek but likely after the hydrograph peak upstream in Northeast Creek North Prong.
s1 B1 B2 S2 B3 S3 64
Tao
Cao
E50D
rn
z40D
N
0
r
30D
2,
q
❑ zao
— Ds&arge
- Average ➢ischarge i �
Rainfall
I 1
1 i �
I l 1
l I 1
1 i 1
I I t
I I i
I 1 I
1 I I
1 1
1 1 ,
1 I 1
1 1 1
1 I I
I I 1
I 1 1
I 1
1 1
1 I
1 I
-- T — —r -- --- - -- - - -- - - -
35
3C
os
oa
2a20-05 M26-06 202a-07 2020-0S 2a20-09 2a20-10 2020-11
Month
Figure 4-1. Hydrologic Data Summary for the Duration of the Northeast Creek MST Study.
Table 4-2. Precipitation Data.
January
4.80
4.45
February
4.67
3.70
March
1.67
4.69
April
3.83
3.43
May
6.04
4.61
June
1.70
4.02
July
5.72
3.94
August
8.99
4.37
September
4.01
4.37
October
2.63
3.70
November
4.42
3.39
December
6.12
3.43
Total
54.6
48.1
a USGS Station 0209725960
US Climate Data, Durham, North Carolina (2021)
Northeast Creek Microbial Source Tracking Study —Project Report 4-3

AECOM-Herrera
0.7
0.6
.. 0.5
e
0.4
m
0.3
0.2
0.1
20
- 15
R
e 1.0
tY
0.5
Storm 1: 5/27/20 - 5/28/20
------------
lip
Ili
-- - - - -- - --
�.- ��' III•
II1
IIII
Storm 2: 8/03/20 - 8104/20
_- ---- ----44H
rl,' I I I
f III
� III
." IIII
IIII
-" IIII
� III
175
150
125 4
u
100 Iv
to
75 r
u
50
25
500
--- Cumulative Rainfall (in)
400 ,
- - MST-9
-- NE3.3NP
30D C7
-- NE22NP
R
200 N
MST-12
MST-18
C
M 30-Minute Rainfall (in)
100
— Discharge (cfs)
0.0 0
1.0
D.8
e
R
0.4
0.2
Storm 3: 9/29120 - 9130/20
rll
--- - - - - - - - - - - - - -
- - - - - - - - - - - - -T11
250
200 _
150 y
er
100 X
50
GA---- ...... - -I - I I I I I I I I - I I I I . • 0
1�0°Igo°rya°o°�o°q;'gti��c��'6o°��o�c�'�o°Rtia°�o°o°
Time EDT (HH:mm)
Figure 4-2. Precipitation in Relation to Sampling Times for Storm Sampling Events for the
Northeast Creek MST Study.
Monthly precipitation data for 2020 at USGS station 0209725960 is presented in Table 4-2. This rain
gauge is located just west of the Northeast Creek Watershed at M.L. King Parkway in in the Third Creek
Watershed (see Figure 3-1). Table 4-2 also presents average monthly precipitation amounts for Durham
for comparison. During the MST study sampling period, precipitation amounts were notably higher in May
(6.04 inches), July (5.72 inches), and August (8.99 inches) in 2020 than average monthly amounts
(4.61, 3.94, and 4.37 inches, respectively), while the June 2020 amount (1.7 inches) was notably lower than
the average amount (4.02 inches).
4.3 Fecal Coliform Bacteria
All fecal coliform bacteria results are presented in the project database in Appendix A. Geomeans and
80th percentiles of fecal coliform bacteria concentrations at each station for base and storm events are
4-4 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

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presented in Table 4-3. Box and whisker plots of fecal coliform bacteria concentrations at each station for
base and storm events are presented in Figure 4-3, which also includes geomeans, 80th percentiles, and
bold values exceeding 400 CFU/100 mL used for compliance with State criteria for freshwaters.
MST Study
MST-9
NE3.3NP
N E2.2 N P
MST-12
Table 4-3. Fecal Coliform Bacteria Statistics for Base and Storm Events.
Event Type
Fecal Coliform Geomean
8Uth Percei
Dn (number)
(CFU/100mL)
(CFU/100r
01
000
111
)I
0
Base (n=4)
Storm (n=3)
Base (n=4)
Storm (n=3)
MST-18 Base (n=4)
Storm (n=3)
Historical Data (12/2008 through 12/2018)
940
9,800
600
21,000
1,500
15,000
1,400
46,000
1,700
48,000
1,900
27,000
NE3.3NP Base (n=19)
485
11,200
Storm (n=1)
315
315
NE2.2NP Base (n=64)
290
870
Storm (n=35)
430
3,100
NE1.2NE Base (n=49)
170
440
Storm (n=27)
710
5,200
NEO.ONE Base (n=51) 170 440
Storm (n=24) 470 1,200
Bold values exceed 400 colonies/100 mL used for compliance with State standards.
North Carolina Surface Water Quality Standards include fecal coliform bacteria criteria for recreational
activities in freshwater streams. Northeast Creek is designated as Class C Secondary Recreation that
applies to all activities in surface waters that result in incidental body contact (which does not include
swimming). Fecal coliform bacteria concentrations shall not exceed a geometric mean of 200 colonies per
100 milliliters (ml-) based upon at least five samples taken over a 30-day period, nor exceed 400/100 mL in
more than 20 percent of the samples (i.e., 80th percentile shall not exceed 400/100 mL) examined during
such period. Violations of these criteria are expected during storm events and may be caused by
uncontrollable nonpoint source pollution. All fecal coliform bacteria concentrations shall be analyzed
using the membrane filter technique reporting units in colony forming units (CFU)/100 mL. The 80th
percentile criterion of 400 CFU/100 mL is commonly used for comparison to State standards for datasets
not meeting this required sampling frequency and for samples collected during storm events.
Northeast Creek Microbial Source Tracking Study —Project Report 4-5

AECOM—Herrera
100,000
10,000
E
G
0
7
1,000
100
Fecal Coliform
MST-9 NE3.3NP NE2 2NP MST-12 MST-18
Station ID
Base Flow
Storm Flow
O Geometric Mean
80th Percentile
--- 200 CFU/10 OmL
- - 400 CFU/100 mL
f Maximum
F 7511 Percentile
— Median
— 25a' Percentile
r Minimum
Figure 4-3. Fecal Coliform Bacteria Box Plots for the Northeast Creek MST Study.
Geometric means and 80th percentiles exceeded 400 CFU/100 mL at all stations for both base and storm
events. Fecal coliform concentrations were generally an order of magnitude higher during storm events
than base events at all stations. Maximum concentrations were observed in the first storm event in May
2020 for station NE3.3NP, in the second storm event in August 2020 for stations NE2.2NP and MST-12,
and in the third storm event in September 2020 for stations MST -9 and MST-18. Thus, fecal coliform
concentrations were high during both base and storm flow conditions, and storm flow concentrations
were much higher than base flow concentrations.
Historical fecal coliform bacteria data were compiled for four stations in the Northeast Creek Watershed
from December 2008 through December 2018 (City of Durham, 2020). The stations include two MST
stations on North Prong (NE3.3NP and NE2.2NP) and two stations downstream on Northeast Creek
(NE1.2NE and NEO.ONE). Each value was identified as whether rain occurred within 24 hours of sample
collection. For comparison to the MST study results, values identified as no rain were assigned to base
events and values identified as positive rain were assigned to storm events. Geomeans and 80th
percentiles of the historical data are also presented in Table 4-3. For the two MST stations with historical
data (NE3.3NP and NE2.2NP), geomeans exceeded State criteria and are somewhat (2-3 times) higher for
base events and an order of magnitude higher for storm events for the MST study compared to historical
data. The much higher geomeans observed for the MST study is likely due to the large storm events
sampled compared to historical data assigned as a storm event regardless of the amount of recent rain.
4.4 Human Biomarker
All human biomarker HF183 results are presented in Appendix A. Geomeans of human biomarker
concentrations at each station for base and storm events are presented in Table 4-4. Box and whisker
plots of human biomarker concentrations at each station for base and storm events are presented in
Figure 4-4. Some results were reported as ND at a detection limit of 50 copies/100 mL and some results
were reported as DNQ at a quantitation limit of 500 copies/100 mL. The ND results were assigned a value
of one-half the detection limit and the DNQ results were assigned a value of one-half the quantitation
limit as best estimates of the actual concentrations. A red line for the ND value of 25 copies/100 mL is
included in Figure 4-4 for reference.
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Table 4-4. Biomarker Geomeans for Base and Storm Events.
MST-9
Base
25
25
Storm
1,310
250
NE3.3NP
Base
25
25
Storm
1,210
634
NE2.2NP
Base
299
25
Storm
1,120
654
MST-12
Base
590
25
Storm
650
1,150
MST-18
Base
560
25
Storm
2,460
1,140
Human biomarker HF183 exhibited a wide range of concentrations in the collected samples. Results for
HF183 ranged from ND to a maximum of 7,730 copies/100ml- at station MTS-12 (base event). Samples
collected during the storm events from station MST-18 had the highest HF183 geomean of
2,460 copies/100 mL. HF183 was not detected during any base flow events at the two uppermost stations
MST-9 and NE3.3NP.
Comparison of human biomarker geomeans for storm and base events show that human sources were
substantially more abundant during storm events than base events at the three stations on Northeast
Creek North Prong (MST-9, NE2.2NP, and NE3.3NP), but not at the Northeast Creek tributary stations
(MST-12 and MST-18) where human sources were present in similar amounts during both base and storm
events. Thus, human biomarker concentrations followed the same hydrologic pattern as fecal coliform
bacteria with much higher concentrations during storm flow than base flow for all North Prong stations
but not for tributary stations MST-12 and MST-18 where human biomarker concentrations were similar in
base and storm flow.
4.5 Dog Biomarker
All dog biomarker DG3 results are presented in Appendix A. Geomeans of dog biomarker concentrations
at each station for base and storm events are presented with human biomarker results in Table 4-4. Box
and whisker plots of dog biomarker concentrations at each station for base and storm events are
presented with human biomarker results in Figure 4-4.
Dog biomarker DG3 was not detected (detection limit of 50 copies/100mQ in any samples collected
during base flow events. Dog biomarker DG3 was detected in all samples collected during storm events
ranging from DNQ at less than 500 copies/100mL to a maximum of 1,870 copies/100 mL at station
NE3.3NP. Samples collected during the storm events from station MST-12 had the highest geometric
mean value for DG3 of 1,150 copies/100 mL. Thus, dog biomarker concentrations followed the same
hydrologic pattern as fecal coliform bacteria with much higher concentrations during storm flow than
base flow at all stations except the uppermost station MST-9 where low concentrations were observed
during storm and base flow.
Northeast Creek Microbial Source Tracking Study —Project Report 4-7

AECOM-Herrera
MOW
E 11,0130
0
li
V
u 1O0
X
MOW
14
Human Biomarker (HF183)
L J?
MST-9 NE3.3NP NE2.2NP MST-12 MST-18
Station ID
Dog Biomarker (DG3)
® Base Flow
Stour~ Flow
O Geometric Mean
- - DetWivn Limit
# Mabmum
751h Fercen#ile
Median
+ 2Y' Percentile
+ minimum
E:::::::) Base Flow
r___1 Storm Flow
O Geometric Mean
Detection Limit
— Maximum
— 75�' Percentile
Median
.- 251h Percfmlilr
4- Minimum
VAST-9 NUAP NE2 2NP V45T-12 MST-18
Station ID
Figure 4-4. Human and Dog Biomarker Box Plots for the Northeast Creek MST Study.
4.6 Biomarker Comparison
Host -associated biomarker concentrations are not directly comparable because the amount of fecal DNA
present in fecal sources varies considerable among types of hosts (Griffith et al. 2013). Table 4-5 presents
fecal coliform bacteria concentrations in municipal wastewater (human) and dog feces reported by EPA
(2001, 2002), compared to human and dog biomarker DNA concentrations in fecal source samples
recently reported for an MST Demonstration Project in Washington (Herrera 2019) and dog biomarker
DNA concentrations in fecal source samples collected for this MST study.
Fecal bacteria concentrations are much higher in human wastewater at 400 million CFU/gram than dog
feces at 18 million CFU/gram (EPA, 2001 and EPA, 2002). The Herrera (2019) MST study in Washington
State found a much higher concentration for dog biomarker DG3 in dog feces (640 million copies/gram)
than human biomarker HF183 in septic tank samples (20 million copies/gram). Thus, human source
4-8 New Hope Creek and Little Creek Watershed Improvement Plan -Data Control Plan

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samples had higher fecal coliform and lower biomarker amounts than dog, indicating that human
biomarker results may represent higher fecal coliform bacteria concentrations than an equivalent dog
biomarker concentration.
Table 4-5. Comparison of Host -Associated Biomarker and Fecal Coliform
Bacteria Concentrations in Fecal Source Samples.
Human 400 20 (Herrera, 2019)b 20
Dog 18 640 (Herrera, 2019)c 0.03
Dog 18 1.6 (Current MST Study)d 11
a Average dog value reported from various sources and used as default variables in the BASINS model (EPA, 2001). Average human
value based on mid -point of ranges reported for fecal coliform bacteria (10' MPN/100 mL) and total suspended solids (250 mg/L)
in residential wastewater (EPA, 2002).
Human biomarker HF183 from Washington MDT Demonstration Project (Herrera, 2019), calculated average from 10 septage
source samples.
Dog biomarker DG3 from Herrera (2019), calculated average from 10 dog fecal source samples.
a Dog biomarker DG3 from the Northeast Creek MST study, calculated geomean from quantified dog fecal source samples (ND and
DNQ results not included in calculation).
The dog source samples collected for the Northeast Creek MST study had much lower dog biomarker
concentrations at 1.6 million copies/gram, which is an overestimate because it excludes the lower ND and
DNQ results. Using this lower dog biomarker concentration for dog feces, the fecal coliform to biomarker
concentration for dog (11 CFU/copies) is similar to that reported by Herrera (2019) for human
(20 CFU/copies). This suggests that human and dog biomarker concentrations may represent similar fecal
coliform bacteria concentrations in water samples, but actual fecal coliform bacteria and human
biomarker concentrations in Durham wastewater is not known and may be much different than that
reported for fecal coliform bacteria by EPA (2001, 2002) and for human biomarkers in septic tanks samples
by Herrera (2019).
Human biomarker HF183 exhibited a wide range of concentrations in the collected samples. Results for
HF183 ranged from ND to a maximum of 7,730 copies/100mL at station MTS-12 (base event). Samples
collected during the storm events from station MST-18 had the highest HF183 geomean of
2,460 copies/100 mL. HF183 was not detected during any base flow events at the two uppermost stations
MST-9 and NE3.3NP.
Comparison of human biomarker geomeans for storm and base events show that human sources were
substantially more abundant during storm events than base events at the three stations on Northeast
Creek North Prong (MST-9, NE2.2NP, and NE3.3NP), and at station MST-18 but not at station MST-12.
However, at the Northeast Creek tributary stations (MST-12 and MST-18) human sources were present
during both base and storm events. Thus, human biomarker concentrations followed the same hydrologic
pattern of much higher concentrations during storm flow as shown by fecal coliform bacteria at all
stations except station MST-12.
The dog biomarker was detected in only 50 percent of dog fecal source samples collected for the
Northeast Creek MST study (see Section 4.1. Data Quality Control). The low sensitivity of the DG3
biomarker in these samples indicates that dog biomarker concentrations were underestimated and actual
amounts of dog fecal DNA present at the sample were higher than reported. Low sensitivity is a Type II
Northeast Creek Microbial Source Tracking Study —Project Report 4-9

AECOM—Herrera
error causing false negatives where dog sources actually may have been present in samples with an
undetected dog biomarker.
4.7 Correlation Analysis
Scatter plots were prepared for fecal coliform bacteria versus biomarker concentrations on log scales and
are presented separately for human and dog biomarkers in Figure 4-7. Geomean values were plotted
separately for base and storm events at each MST station. These plots show a positive relationship
between fecal coliform bacteria and both biomarkers when base and storm events are combined, but not
within either hydrologic group of data. A lack of correlation between fecal coliform bacteria and DNA
concentrations has been typically observed elsewhere and is not surprising given the differences in the
populations tested and methods used for quantification (Herrera, 2019).
4.8 Management Implications
Fecal coliform bacteria are comprised primarily of E. coli at typically over 90 percent. However, this
percentage can be lower due to Klebsiella species that are also present in the gut but can grow in a warm,
nutrient -enriched environment such as a pile of decaying vegetation or wood chips. E. coli also has been
shown to grow in some extremely enriched conditions. There is some evidence that they can grow in
enriched lake or catch basin sediments, but this is rare and very slow in comparison to typical sediment
input rates. Freshwater mesocosm experiments have shown that fecal coliform bacteria die -off rates are
10 times higher in water than sediment inoculated with sanitary wastewater and dog feces, and no net
growth of fecal coliform bacteria occurred in any mesocosm over the 28-day experiment (Anderson,
et al. 2005). Investigations by the City of Durham indicate that sediment suspension during storm flow
increased fecal coliform bacteria concentrations in the monitored streams, but actual growth of fecal
coliform bacteria in stream sediments was not determined (City of Durham, 2012 and City of Durham,
2021). The fine clay content, highly erodible, and low permeable soils in the watershed result in a high
suspended solids load and throughput in the stream during storm events that far exceeds the amount of
suspended stream sediment. Therefore, suspension of stream sediment bacteria is likely small relative to
the input of watershed derived bacteria.
Fecal coliform bacteria die in surface waters from ultraviolet light exposure and predation by other
microbes, which likely makes up for some propagation in most cases. Fecal coliform bacteria die -off
(inactivation) rates in the literature are highly variable, ranging from 0.01 to 0.90 per hour (1 to 90 present
reduction in an hour), and generally increase with light radiation, water temperature, salinity, and bacteria
concentration (Selvakumar et al. 2007). Microcosm experiments of lake water and sediment showed that
suspended sediment reduced indicator bacteria decay rates, which ranged from 0.02 to 0.04 per hour for
E. coli (Kinnaman et al. 2012). Although Northeast Creek watershed waters are very turbid with low light
penetration during storm flow, there likely is not enough organic matter to yield a net growth of fecal
coliform bacteria. Modeling of fecal coliform bacteria in Northeast Creek showed that bacterial
associations with suspended sediment particles was relatively small (< 10%) (Characklis et al. 2009). This
same study of Northeast Creek reported increases of indicator bacteria one to four orders of magnitude
from base to storm flow and concluded that urban nonpoint sources of contamination (i.e., stormwater
contaminated by sanitary wastewater and animal feces) are likely primarily responsible for the observed
increase and resulting impairment. Thus, it is expected that principally all fecal bacteria enumerated in the
MST study samples originated from human or animal fecal sources.
4-10 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

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100,000
• • •
10,000
0LL
: • •
u 1,000
M •
v
U
f6
m .
E 100
0
4-
0
U
U
LL 10
1
1 10 100 1,000
Human Biomarker (HF183) (copies/100mL)
100,000
3 10,000
E
0
0
ZD
U_
u 1,000
v
+1
ra
CO
E 100
0
0
U
ro
U
ii 10
Figure 4-7
•
10,000
♦ MST-12 Base Events
• MST-12 Storm Events
♦ MST-18 Base Events
• MST-18 Storm Events
♦ MST-9 Base Events
• MST-9 Storm Events
♦ NE2.2NP Base Events
• NE2.2NP Storm Events
♦ NE3.3NP Base Events
• NE3.3NP Storm Events
♦ MST-12 Base Events
• MST-12 Storm Events
♦ MST-18 Base Events
• MST-18 Storm Events
♦ MST-9 Base Events
• MST-9 Storm Events
♦ NE2.2NP Base Events
• NE2.2NP Storm Events
♦ NE3.3NP Base Events
• NE3.3NP Storm Events
10 100 1000 10000
Dog Biomarker (DG3) (copies/100mQ
Fecal Coliform Versus Human and Dog Biomarker Concentrations for the Northeast
Creek MST Study.
Northeast Creek Microbial Source Tracking Study —Project Report 4-11

AECOM—Herrera
All Bacteroidetes species are obligate anaerobes which means that oxygen is toxic to them and they do
not grow outside the gut in aerobic conditions (Hagedorn et al. 2011). This fact, along with their higher
abundance than E. coli in the gut, is why they have been selected as the preferred MST method by EPA.
Although anaerobes die within hours to days in the presence of oxygen outside the gut, biomarker
analysis measures DNA from both live and dead cells and DNA may take days to months to degrade to
undetectable concentrations in the environment (Hagedorn et al. 2011). However, a die -off study of
manure -amended soils showed that host associated gPCR biomarkers decayed much faster than indicator
bacteria (Rogers et al. 2011). Regardless of their relative decay rates, both fecal coliform bacteria and
Bacteroidetes DNA were transported from the source to the sampling point in Northeast Creek within
hours when relatively little inactivation likely occurred for both. Thus, Bacteroidetes DNA detected in
water samples was most likely from fecal sources discharged into the stream within a day of sample
collection. The significant differences in base and storm flow concentrations further suggest that the DNA
quantified in the water samples were recently discharged to the stream.
DNA of only two of likely many fecal sources present in Northeast Creek were evaluated for this MST
study. Fecal bacteria from birds, rodents, and other wildlife may be more abundant in the stream than
human or dog. Thus, it is not known if fecal coliform bacteria concentrations would significantly decline if
both human and dog sources were controlled. However, the hydrologic and spatial patterns in human and
dog biomarker results from this MST study provide evidence of where control of these sources is needed
to address the chronically high concentrations of fecal coliform bacteria observed in Northeast Creek.
Human sources of fecal bacteria may include SSOs, illicit connections of sanitary sewers to storm drains,
exfiltration or leakage from sanitary sewers, or unsanitary practices in the watershed where human waste
is deposited on land, such as from homeless encampments. SSOs would be expected to primarily impact
storm flow conditions due to insufficient sewer conveyance capacity under those conditions. Illicit
connections and leaky sewers would be expected to primarily impact base flow conditions when
proportions of sewer to stream flow are greatest. Unsanitary practices would be expected to primarily
impact storm flow conditions when land deposits wash into the stream via runoff.
Human biomarkers were not detected in base flow at the two uppermost MST stations (MST-9 and
NE3.3NP), suggesting that illicit connections are not a significant source of fecal coliform bacteria in these
subwatersheds. The consistently detected and moderately high human biomarker concentrations
observed during base flow at the three lowermost MST stations suggest that illicit connections may be a
significant source of fecal coliform bacteria in these subwatersheds.
Based on the evaluation of SSO data for the past 10 years, the recorded occurrence of only three SSOs in
the MST study subwatersheds is much too infrequent to be a major source of the human biomarkers
observed in the stream samples. The relatively high human biomarker concentrations observed at all five
MST stations during storm flow suggests that SSOs occur much more frequently than records show. While
it is possible that unsanitary practices contribute to the observed human biomarker concentrations, it
seems unlikely that they would be significant source of the moderately high human biomarker
concentrations observe at all five MST stations. Illicit connections or leaky sewers may be significant
sources at tributary station MST-12 and less so in tributary station MST-18 where human biomarker
concentrations were elevated during base flow compared to the North Prong stations.
Dogs do not appear to be a significant source of fecal bacteria during base flow, but likely are a
substantial source during storm flow at all stations except the uppermost station MST-9. Dog feces were
apparently washed into the stream from runoff during storm events, indicating that fecal bacteria
concentrations could be reduced during storm events through better compliance with pet waste best
management practices.
4-12 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

AECOM—Herrera
5 Conclusions
Water quality monitoring was conducted from May through October 2020 during four base flow events and three
targeted storm events. Base flow sampling occurred after at least 72 hours of less than 0.04 inches of rain, and
storm event sampling occurred for storms with 0.65 to 2.07 inches of rain. Monitoring was conducted at five
stations of interest located in the Northeast Creek Watershed and within the City of Durham. Three stations were
located on the Northeast Creek North Prong (MST-9, NE3.3NP, and NE2.2NP) and two stations were located on
small tributaries that drain to Northeast Creek downstream of North Prong (MST-12 and MST-18). The samples
were analyzed for fecal coliform bacteria, human biomarker HF183, and dog biomarker DG3. The results generally
met data quality objectives with some data qualifications due to low precision of laboratory or field duplicate
analyses, or bacteria reported as too numerous to count.
Key study findings are listed below for hydrology, fecal coliform bacteria, human biomarker, and dog biomarker.
5.1 Hydrology
• The average discharge of Northeast Creek for the duration of the project (36 cfs) was similar to the mean
annual flow between October 1982 to October 2011 for Northeast Creek (30 cfs).
• Precipitation amounts were notably higher in May (6.04 inches), July (5.72 inches), and August (8.99 inches)
in 2020 than average monthly amounts (4.61, 3.94, and 4.37 inches, respectively), while the June 2020
amount (1.7 inches) was notably lower than the average amount (4.02 inches).
5.2 Fecal Coliform Bacteria
• The geometric mean criterion (200 CFU/100 mL) and 80th percentile criterion (400 CFU/100 mL) were
exceeded at all stations for both base and storm events.
Fecal coliform concentrations were generally an order of magnitude higher during storm events than base
events at all stations.
• Fecal coliform concentrations were high during both base and storm flow conditions, and storm flow
concentrations were much higher than base flow concentrations.
• Fecal coliform bacteria concentrations were higher for the MST study than historical fecal coliform bacteria
data compiled for the two North Prong stations from 2008 through 2018.
5.3 Human Biomarker
Human biomarker HF183 exhibited a wide range of concentrations in samples ranging from not detected to
a maximum of 7,730 copies/100mL.
• Human biomarker concentrations followed the same hydrologic pattern as fecal coliform bacteria with much
higher concentrations during storm flow than base flow for all three North Prong stations but not tributary
station MST-12 where human biomarker concentrations were similar in base and storm flow.
Samples collected during the storm events from station MST-18 had the highest geometric mean value for
HF183 of 2,460 copies/100 mL and this tributary station also exhibited elevated human biomarker
concentration during base flow.
Northeast Creek Microbial Source Tracking Study —Project Report 5-1

AECOM—Herrera
• Human biomarker HF183 was not detected during any base flow event at the two uppermost stations MST-9
and NE3.3NP.
• The presence of human biomarkers at similar concentrations for both base and storm events at station
MST-12 indicates the potential presence of illicit connections or leaky sewer pipes in this subwatershed
where sewer density and medium intensity development cover is highest among the five subwatersheds
studied.
The relatively high human biomarker concentrations at all five stations during storm flow and the low
frequency of SSOs recorded in their subwatersheds suggests that SSOs occur much more frequently than
records show.
• Unsanitary practices may be an additional human source from runoff during storm flow but seems unlikely
to be as important as SSOs based on the widespread occurrence of human biomarkers during storm flow
conditions.
5.4 Dog Biomarker
• Dog biomarker concentrations followed the same hydrologic pattern as fecal coliform bacteria with much
higher concentrations during storm flow than base flow at all stations except the uppermost station MST-9
where low concentrations were observed during storm and base flow.
• Dogs do not appear to be a significant source of fecal bacteria during base flow, but likely are a substantial
source during storm flow at all stations except the uppermost station MST-9.
• Dog biomarker DG3 was not detected above the quantitation limit in any samples collected during base
flow events.
• Dog biomarker DG3 was detected in all samples collected during storm events ranging from DNQ to a
maximum of 1,870 copies/100mL.
• Samples collected during the storm events from station MST-12 had the highest geometric mean value for
DG3 of 1,150 copies/100 mL.
• The low sensitivity of the DG3 biomarker in dog feces samples analyzed for this MST study indicate that dog
biomarker concentrations were underestimated and actual amounts of dog fecal DNA present at the sample
were higher than reported.
5-2 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

AECOM—Herrera
6 Recommendations
It is recommended that the sources of high concentrations of fecal coliform bacteria throughout the study
area be located and controlled, if possible. Human and dog fecal sources should be targeted for control in
the upper subwatersheds of the Northeast Creek watershed based on the MST study findings.
6.1 Human Fecal Source Control
Human source control should include SSO reduction in all sampled subwatersheds based on the higher
human biomarker concentrations observed during storm events than base events. Priority subwatersheds
should include subwatersheds 9 and 10, the two uppermost subwatersheds draining to North Prong,
where storm event increases in human biomarkers were greatest. SSOs occasionally occur in almost every
sewer system, even though systems are intended to collect and contain all the sewage that flows into
them (EPA 2021). When SSOs happen frequently, it means something is wrong with the system. Problems
that can cause SSOs include:
• Inappropriate materials sent to the sewers — materials such as fats, oils, and grease (FOG), and some
household products (including some marked 'flushable') such as baby wipes, facial wipes, facial
masks, sanitary pads, and tampons. All of these may create blockages. Tree roots entering through
defects or openings in a sewer line also may cause blockages.
• Leaky sewers — stormwater, ground water, and snowmelt entering the sanitary sewer from cracks
and faults in the sewer or leaky sewerjoints can overload a sanitary sewer.
• Inappropriate connections — Connections of sources of water such as sump pumps, roof drains,
foundation drains, and area drains can overload a sanitary sewer.
• Improper or inadequate maintenance and cleaning of sewers.
• Inadequate pump maintenance and lack of backup power.
• Undersized sewers and/or pumps.
• Equipment failures and breaks.
Many avoidable SSOs are caused by inadequate or improper operation or maintenance, inadequate
system capacity, and improper system design and construction (EPA 2021). SSOs can be reduced by:
• Sewer system cleaning and maintenance.
• Reducing infiltration and inflow through system rehabilitation and repairing broken or leaking
service lines.
• Enlarging or upgrading sewer, pump station, or sewage treatment plant capacity and/or reliability.
• Controls limiting FOG into the sewer systems.
• Educating the public on how FOG and certain household products can clog sewers.
• Construction of wet weather storage facilities.
• Expanding the capacity of the treatment works.
Sewer pipe condition, clogging, and capacity should be inspected with closed-circuit television (CCTV). A
plan should be developed to address the observed issues. Pipe clogging can be addressed by hydro -
Northeast Creek Microbial Source Tracking Study —Project Report 6-1

AECOM—Herrera
jetting with a combination jet -vacuum truck. Pipe cracks or failed joints causing infiltration and inflow can
be addressed through system rehabilitation and repairing broken or leaking service lines. Inappropriate
inputs of stormwater sources can be redirected to the storm drain system.
Human biomarkers were observed during base flow at three MST stations (NE2.2NP, MST-12, and
MST-18) but not the two uppermost MST stations (MST-9 and NE3.3NP). The presence of human sources
during base flow at these lower stations suggests there may be leaky sanitary sewer pipes or illicit
connections from sanitary sewers to storm drains. If the CCTV inspection shows no sanitary sewer leaks in
the associated three lower subwatersheds (11, 12, and upper 18), then illicit connections to storm drains
also should be investigated in the associated lower subwatersheds as part of the Illicit Discharge
Detection and Elimination (IDDE) Program.
6.2 Dog Fecal Source Control
Dog fecal source control should include improved pet waste management in all subwatersheds except the
uppermost subwatershed 9 draining to station MST-9 where the dog biomarker was not detected above
the quantitation limit. It is recommended that the City continue to educate citizens about pet waste
management and include notification and enforcement of the Durham County Canine Waste Ordinance,
which includes fines of $50 to $150 for ordinance violations. Additional dog waste stations should be
installed and maintained for use by homeowner associations and multifamily housing units to better
support proper disposal of dog waste by residents.
6-2 Northeast Creek Microbial Source Tracking Study —Project Report

AECOM—Herrera
7 References
Anderson, K.L., J.E. Whitlock, and V.J. Harwood. 2005. Persistence and Differential Survival of Fecal Indicator
Bacteria in Subtropical Waters and Sediments. Appl. Environ. Microbiol. 71(6): 3041-3048. June.
Boehm, A.B., L.C. Van De Werfhorst, J.F. Griffith, P. A. Holden, J. A. Jay, O. C. Shanks, D. Wang, and S. B. Weisberg.
2013 Performance of Forty -One Microbial Source Tracking Methods: A Twenty -Seven Laboratory Evaluation
Study. Southern California Coastal Water Research Project. Technical Report804. December.
<httl2s://pdfs.semanticscholar.org/62f3/dO44c981 fe9cbdOe6dded799c6b2b6l e8O4f.pdf>.
Brown and Caldwell. 2012. Characterization and Assessment Report for the Northeast Creek and Crooked Creek
Watersheds. Prepared for the City of Durham by Brown and Caldwell, Raleigh, NC. April 2, 2012.
Characklis, G.W., L.H. Krometis, M.D. Sobsey, and O.D. Simmons. 2009. Watershed Restoration to Address Fecal
Coliform Impairment of Northeast Creek in Durham, NC. Prepared for North Carolina Department of
Environment and Natural Resources, Division of Water Quality by University of North Carolina at Chapel Hill,
Chapel Hill, North Carolina. September.
City of Durham. 2012. Attenuation of Bacteria Following Remediation of a Sanitary Sewer Discharge (08-003a).
Technical Report by the City of Durham Department of Public Works, Durham, North Carolina. May 22.
City of Durham. 2020. Water Quality Data Web Portal. City of Durham, Public Works Department.
<http://durhamwaterquality.orgZ>.
City of Durham. 2021. Characterization and Correlation of Bacteria Levels in Durham Stream Sediments, Project
Report 08-002a. Technical Report by the City of Durham Department of Public Works, Durham, North Carolina.
Draft, February 11.
EPA. 2001. Bacterial Indicator Tool. Spreadsheet obtained from BASINS User Information and Guidance Website by
the US Environmental Protection Agency, Office of Water, Washington, DC. December.
<https://www.epa.gov/ceam/basins-user-information-and-guidance>.
EPA. 2002. Onsite Wastewater Treatment Systems Manual. EPA/625/R-00/008. US Environmental Protection
Agency, Office of Water, Washington, DC. February.
EPA. 2019. Method 1696: Characterization of Human Fecal Pollution in Water by HF183/BacR287 TagMans
Quantitative Polymerase Chain Reaction (gPCR) Assay. EPA821 R19002. US Environmental Protection Agency,
Office of Water, Washington, DC. March. <https://www.epa.gov/sites/production/files/2019-
03/documents/method 1696 draft 2019.pdf>.
EPA. 2021. National Pollutant Discharge Elimination System (NPDES), Sanitary Sewer Overflow (SSO) Frequent
Questions. Website by the US Environmental Protection Agency, Office of Water, Washington, DC. Accessed
February 24. <https://www.epa.gov/npdes/sanitary-sewer-overflow-sso-frequent-questions#why>.
Green, H.C., R. Haugland, M. Varma, H.T. Millen, M.A. Borchardt, K.G. Field, C.A. Kelty, M. Sivaganesan, and
O.C. Shanks. 2014a. Improved HF183 Quantitative Real -Time PCR Assay for Characterization of Human Fecal
Pollution in Ambient Surface Water Samples. Applied and Environmental Microbiology 80(10):3086-3094. May.
Green, H.C., K.M. White, C.A. Kelty, and O.C. Shanks. 2014b. Development of Rapid Canine Fecal Source
Identification PCR-Based Assays. Environmental Science and Technology 48(19):11453-11461. September.
Northeast Creek Microbial Source Tracking Study —Project Report 7-1

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Griffith, J.F., B.A. Layton, A.B. Boehm, P.A. Holder, J.A. Jay, C. Hagedorn, C.D. McGee, S.B. Weisberg. 2013. The
California Microbial Source Identification Manual: A Tiered Approach to Identifying Fecal Pollution Sources to
Beaches. Southern California Coastal Water Research Project. Technical Report 804. December 2013.
<https://www.waterboards.ca.gov/water issues/programs/beaches/cbi projects/docs/sipp manual.pdf
Hagedorn, C., A.R. Blanch, and V.J. Harwood, ed. 2011. Microbial Source Tracking: Methods, Applications, and Case
Studies. Springer, New York, New York.
Herrera. 2019. Data Analysis Report, Quantitative Microbial Source Tracking Demonstration Project. Prepared for
Washington State Conservation Commission by Herrera Environmental Consultants, Inc., Seattle, WA. July 11,
Herrera and AECOM. 2020. Quality Assurance Project Plan, Northeast Creek Microbial Source Tracking Study.
Prepared for the City of Durham, Durham, North Carolina by Herrera Environmental Consultants, Seattle,
Washington and AECOM, Morrisville, North Carolina. April.
Kinnaman, A.R., C.Q. Surbeck, and D.C. Usner. 2012. Coliform Bacteria: The Effect of Sediments on Decay Rates and
on REquried Detention Times in Stormwater BMPs. Journal of Environmental Protection. 3:787-797. August.
Rogers, S.W., M. Donnelly, L. Peed, C.A. Kelty, S. Mondal, Z. Zhong, and O.C. Shanks. 2011. Decay of Bacterial
Pathogens, Fecal Indicators, and Real -Time Quantitative PCR Genetic Markers in Manure -Amended Soils. Appl.
Environ. Microbiol. 77(14): 4839-4848. July.
US Climate Data. 2021. Climate Data for Durham, North Carolina. Monthly Averages, 1981 through 2010 Normals.
<https://www.usclimatedata.com/climate/durham/north-carolina/united-states/usnc0192>.
7-2 New Hope Creek and Little Creek Watershed Improvement Plan —Data Control Plan

AECOM
Appendix A:
Database

AECOM—Herrera

Table A-1. Northeast Creek MST Study Results.
Event
Number
Sample Date
Sample
Time
Station ID
Sample ID
Human HF183
co ies 100 mL
Dog DG3
co ies 100 mL
Fecal Coliform
CFU 100 mL
Storm 1
5/28/2020
9:27 AM
MST-9
MST-9-51
4,270
DNQ
21,000
Storm 1
5/28/2020
9:46 AM
NE3.3NP
NE3.3NP-52
2,680
749
32,000
Storm 1
5/28/2020
10:10 AM
NE2.2NP
NE2.2NP-53
1,440
DNQ
26,000
Storm 1
5/28/2020
11:21 AM
MST-12
MST-12-55
837
1,260
5,500
Storm 1
5/28/2020
10:36 AM
MST-18
MST-18-54
2,730
1,610
19,000
Base 1
6/3/2020
11:05 AM
MST-9
MST-9-51
ND
ND
3,800
Base 1
6/3/2020
10:52 AM
NE3.3NP
NE3.3NP-52
ND
ND
3,000
Base 1
6/3/2020
10:30 AM
NE2.2NP
NE2.2NP-53
512
ND
2,400
Base 1
6/3/2020
10:05 AM
MST-12
MST-12-55
7,730
ND
3000 J
Base 1
6/3/2020
9:30 AM
MST-12
MST-X-56
DNQ
ND
1090 J
Base 1
6/3/2020
9:30 AM
MST-18
MST-18-54
ND
ND
1,360
Base 2
6/23/2020
10:00 AM
MST-9
MST-9-51
ND
ND
460
Base 2
6/23/2020
10:15 AM
NE3.3NP
NE3.3NP-52
ND
ND
1,180
Base 2
6/23/2020
10:30 AM
NE2.2NP
NE2.2NP-53
DNQ
ND
727
Base 2
6/23/2020
9:25 AM
MST-12
MST-12-55
DNQ
ND
162
Base 2
6/23/2020
11:00 AM
MST-18
MST-18-54
5,180
ND
1,270
Storm 2
8/4/2020
11:10 AM
MST-9
MST-9-51
DNQ
DNQ
16000 J
Storm 2
8/4/2020
10:10 AM
MST-9
MST-X-56
DNQ
DNQ
>60000
Storm 2
8/4/2020
10:50 AM
NE3.3NP
NE3.3NP-52
DNQ
DNQ
24,000
Storm 2
8/4/2020
10:30 AM
NE2.2NP
NE2.2NP-53
431
812
>60000
Storm 2
8/4/2020
10:00 AM
MST-12
MST-12-55
1,330
1,310
>60000
Storm 2
8/4/2020
9:45 AM
MST-18
MST-18-54
1,870
1,120
5,460
Base 3
9/8/2020
11:10
MST-9
MST-9-51
ND
ND
1,800
Base 3
9/8/2020
11:30
NE3.3NP
NE3.3NP-52
ND
ND
520
Base 3
9/8/2020
11:45
NE2.2NP
NE2.2NP-53
DNQ
ND
800
Base 3
9/8/2020
12:40
MST-12
MST-12-55
DNQ
ND
800
Base 3
9/8/2020
12:10
MST-18
MST-18-54
DNQ
ND
1,200
Storm 3
9/30/2020
11:30
MST-9
MST-9-51
2,100
DNQ
28,000
Storm 3
9/30/2020
11:35
NE3.3NP
NE3.3NP-52
2,640
1,360
18,000
Storm 3
9/30/2020
11:35
NE3.3NP
NEX-S3
1,640
1,870
22,000
Storm 3
9/30/2020
11:55
NE2.2NP
NE2.2NP-53
2,260
1,380
>600
Storm 3
9/30/2020
12:20
MST-12
MST-12-55
DNQ
927
29,000
Storm 3
9/30/2020
12:45
MST-18
MST-18-54
2,920
817
32,000
Base 4
10/15/2020
10:25
MST-9
MST-9-51
ND
ND
153
Base 4
10/15/2020
10:40
NE3.3NP
NE3.3NP-52
ND
ND
290
Base 4
10/15/2020
11:00
NE2.2NP
NE2.2NP-53
DNQ
ND
560
Base 4
10/15/2020
11:25
MST-12
MST-12-55
DNQ
ND
330
Base 4
10/15/2020
11:50
MST-18
MST-18-54
2,970
ND
2,600
Base 4
10/15/2020
11:50
MST-18
MST-X
3,650
ND
2,100
J = Sample results is estimated
ND = Not Detectable at less than the method detection limit of 50 copies/100 mL
DNQ = Detected Not Quantified at less than the limit of quantification of 500 copies/100 mL
> = Plate count was too numerous to count, result is likely higher

AECOM
Appendix B:
Field Notes and Photographs

AECOM—Herrera

New Hope Creek: Microbial Source Tracking: 19-320 Task 9.3
Storm Sample May 28, 2020; weather overcast and 75 degrees Fahrenheit
3 Oaks personnel: Tom Dickinson, Lizzy Stokes -Cawley
5/28/2020 Storm Sample (5 Samples): Significant recent rain, moderately turbid streams at sample sites
USGS rain gauge 40209725960: 24-hour precipitation total: 0.77 inches
USGS station #0209741387: Gage height 0.91ft to 2.06 ft between 00:00 May 28 and 00:00 May 29
USGS station #0209741955: Gage height 5.70 ft to 7.59 ft between 00:00 May 28 and 00:00 May 29
Discharge: 59.6 to 163 cubic feet/ second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
5/28 at 9:27 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water levels moderate and flowing, water turbid
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
5/28 at 9:46 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken Digital photos taken, water levels moderate and flowing, water turbid
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
5/28 at 10:10 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water levels moderate and flowing, water turbid
MST-18 Sample site 4 Lat/Long: 35.907774,-78.901568
5/28 at 10:36 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, Parkwood lake evidence of blue-green algae with dead freshwater mussels floating
in water. Water at site flowing and clear.
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
5/28 at 11:21 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, flood plain inundated with water, water levels high and flowing, moderately turbid

New Hope Creek: Microbial Source Tracking: 19-320 Task 9.3
Baseflow sampling June 3, 2020; weather sunny and 80 degrees Fahrenheit
3 Oaks personnel: Tom Dickinson, Lizzy Stokes -Cawley
6/3/2020 Base Flow Sample (6 Samples, inc. duplicate): No recent rain, three upstream sites significantly
turbid
USGS rain gauge #0209725960: 72-hour precipitation total: 0.00 inches
USGS station #0209741387: Gage height 0.47 ft to 0.54 ft between 00:00 June 2 and 00:00 June 3
USGS station #0209741955: Gage height 5.70 ft to 7.59 ft between 00:00 June 2 and 00:00 June 3
Discharge: 3.76 to 4.05 cubic feet/ second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
6/03 at 11:05 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed at site, photos in folder
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
6/03 at 10:52 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed, photos of turbid water in folder
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
6/03 at 10:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed, photos of turbid water in folder
MST-18 Sample site 4 Lat/Long: 35.907774,-78.901568
6/03 at 9:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, Parkwood lake evidence of blue-green algae with dead freshwater mussels floating
in water, photos of blue-green algae in folder. Water at site low and clear, box turtle in water.
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
6/03 at 10:05 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water levels low but flowing, moderately turbid
DUPLICATE MST-X Sample site 6 taken at MST-12 Sample site 5
6/03 at 9:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water levels low but flowing, moderately turbid (duplicate of above)

New Hope Creek: Microbial Source Tracking: 19-320 Task 9.3
Baseflow sampling June 23, 2020; weather overcast and 85 degrees Fahrenheit, high humidity
3 Oaks personnel: Tom Dickinson, Lizzy Stokes -Cawley
6/23/2020 Base Flow Sample (5 Samples): No recent rain, sites only slightly turbid.
USGS rain gauge #0209725960: 72-hour precipitation total: 0.00 inches
USGS station #0209741387: Gage height 0.38 ft to 0.42 ft between 00:00 June 22 and 00:00 June 23
USGS station #0209741955: Gage height 3.56 ft to 3.99 ft between 00:00 June 22 and 00:00 June 23
Discharge: 6.83 to 13.1 cubic feet / second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
6/23 at 10:00 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, low flow and low turbidity in water at site
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
6/23 at 10:15 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, low flow and low turbidity in water at site
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
6/23 at 10:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, moderate turbidity observed, moderate flow
MST-18 Sample site 4 Lat/Long: 35.907774,-78.901568
6/23 at 9:25 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, Parkwood lake numerous dead freshwater mussels floating in water, however blue -
algae not visible. Water at site low and clear, low flow.
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
6/23 at 11:00 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water flow slow to moderate, beaver dam is present, slightly turbid water

New Hope Creek: Microbial Source Tracking:19-320 Task 9.3
Stormwater sampling August 4, 2020; weather partly sunny and 78 degrees Fahrenheit, morning
following tropical storm Isaias.
3 Oaks personnel: Tom Dickinson, Lizzy Stokes -Cawley
8/4/2020 Storm Samples (6 Samples, including duplicate): Significant recent rain associated with
Hurricane Isaias, turbid waters at all five sample locations
USGS rain gauge #0209725960: 24-hour precipitation total: 2.08 inches
USGS station #0209741387: Gage height 0.42 ft to 4.12 ft between 00:00 August 3 and 00:00 August 4
USGS station #0209741955: Gage height 3.54 ft to 9.51 ft between 00:00 August 3 and 00:00 August 4
Discharge: 6.80 to 617 cubic feet/second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
8/04 at 11:00 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed at site, water starting to recede, high levels but water not in
the floodplain
DUPLICATE MST-X Sample site 6 of MST-9 Sample site 1
8/04 at 11:00 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Duplicate conditions of MST-9
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
8/04 at 10:50 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed, water levels high but not into the floodplain.
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
8/04 at 10:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, floodplain inundated with floodwaters, moderate to high turbidity observed
MST-18 Sample site 4 Lat/Long: 35.907774,-78.901568
8/04 at 9:45 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water flowing, slightly turbid, elevated water levels
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
8/04 at 10:05 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, surrounding floodplain completely inundated, water connection between stream of
floodplain, water turbid at the site.

New Hope Creek: Microbial Source Tracking: 19-320 Task 9.3
Baseflow sampling September 8, 2020; weather mostly cloudy, —80 degrees Fahrenheit
3 Oaks personnel: Tom Dickinson, Nancy Oberle
9/8/2020 Base Flow Sample (5 Samples): No recent rain, sites relatively clear to turbid.
USGS rain gauge #0209725960: 72-hour precipitation total: 0.00 inches
USGS station #0209741387: Gage height approximately 0.37 ft between 00:00 September 7 and 00:00
September 8
USGS station #0209741955: Gage height approximately 3.85 ft and 10.5 cubic feet / second between
00:00 September 7 and 00:00 September 8
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
9/8 at 11:10 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, low flow, but marked turbidity (nearby construction?)
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
9/8 at 11:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, very low flow, slight turbidity, algae growth noted on rocky substrate
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
9/8 at 11:45 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, discernable flow in rip -rap, light turbidity
MST-18 Sample site 4 Lat/Long: 35.907774,-78.901568
9/8 at 12:10 pm Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, very low flow, clear
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
9/8 at 12:40 pm Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, low flow, relatively clear, beaver dam is present

New Hope Creek: Microbial Source Tracking:19-320 Task 9.3
Stormwater sampling September 30, 2020; weather partly sunny and 70 degrees Fahrenheit, morning
following a heavy rainstorm.
3 Oaks personnel: Tom Dickinson, Kate Montieth Sevick
9/30/2020 Storm Samples (6 Samples, including duplicate): Significant recent rain,turbid waters at all
five sample locations
USGS rain gauge #0209725960: 24-hour precipitation total: 0.86 inches
USGS station #0209741387: Gage height 3.7ft to 1.0 ft between 00:00 September 30 and 00:00 Oct 1
USGS station #0209741955: Gage height 8 ft to 5 ft between 00:00 September 30 and 00:00 Oct 1
Discharge: 200 to 45 cubic feet/second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
9/30 at 11:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed at site, water starting to recede, high levels but water not in
the floodplain
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
9/30 at 11:35 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, high turbidity observed, water levels high but not into the floodplain.
DUPLICATE NEX Sample site 3 of NE3.3 Sample site 2
9/30 at 11:35 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Duplicate conditions of NE3.3
NE2.2 Sample site 4 Lat/Long: 35.910441,-78.895253
9/30 at 10:30 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, floodplain partially inundated with floodwaters, moderate to high turbidity observed
MST-12 Sample site 5 Lat/Long: 35.890208,-78.911527
9/30 at 12:20 pm Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF 183 EPA, DG3) samples taken
Digital photos taken, surrounding floodplain partially inundated, moderate turbidity at the site.
MST-18 Sample site 6 Lat/Long: 35.907774,-78.901568
9/30 at 12:45 pm Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water flowing, slightly turbid, elevated water levels

New Hope Creek: Microbial Source Tracking:19-320 Task 9.3
Baseflow sampling October 15, 2020 weather clear and sunny
30aks personnel: Tom Dickinson, Nancy Scott
10/15/2020 Baseflow Samples (6 Samples, including duplicate):
USGS rain gauge #0209725960: 72-hour precipitation total: 0.01 inches
USGS station #0209741387: Gage height 0.56 ft to 0.52 ft between 00:00 October 15 and 00:00 October
16
USGS station #0209741955: Gage height 4 ft to 4.25 ft between 00:00 October 15 and 00:00 Oct 16
Discharge: 13 to 18 cubic feet/second
MST-9 Sample site 1 Lat/Long: 35.934027,-78.890682
10115 at 10:25 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, very light staining, low but discernable flow at culvert
NE3.3 Sample site 2 Lat/Long: 35.928272,-78.893501
10115 at 10:40 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, very light staining, good flow at culvert
NE2.2 Sample site 3 Lat/Long: 35.910441,-78.895253
10115 at 11:00 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, very light turbidity, good flow
MST-12 Sample site 4 Lat/Long: 35.890208,-78.911527
10115 at 11:25 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, good flow, very light turbidity, beaver dam intact
MST-18 Sample site 5 Lat/Long: 35.907774,-78.901568
10115 at 11:50 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Digital photos taken, water flowing, clear
DUPLICATE MST-X Sample 6 at Sample site MST-18
10115 at 11:50 am Fecal Coliform (1 grab sample 250 mL bottle) and MST (1 grab sample 250 mL bottle)
(HF183 EPA, DG3) samples taken
Duplicate conditions of site 5

Field Notes for Dog Feces Sampling- MST Dog marker verification
Location: Northgate Dog Park
400 W Lavender Avenue, Durham NC 27704
Date: 6/23/20
General Conditions: Low 72°, high of 90° Partly Cloudy, wind <10 mph.
Staff: J. Smith/ C. Cailleret
Staff arrived at Northgate dog park at 7:40 am, dog park is a converted baseball field with adjacent
greenway and parking lot. Samples stored on wet ice in field cooler following processing.
Dog-1 sample collected at 06:03, staff pet (CC) prior to arrival, process at dog park.
Dog-2 Collected at 07:45, dog passing on greenway, resident donated prepackaged fecal material.
Dog- 3 collected at 8:20 after observed bowel movement (BM), first dog located inside of dog park.
Dog- 4 collected at 08:50after observed BM, dog currently being treated for hookworms.
Dog-5 collected at 08:58, passing on greenway collected pre bagged sample, no comment.
Dog-6 collected 09:24 after observed BM in dog park, no comment.
Dog-7 collected at 09:30, no comment.
JS left to meet with Three Oaks staff at Durham Rescue Mission for recommended shipping cooler and
label. Dog fecal samples shipped separate from MST project samples to avoid potential cross
contamination.
Dog-8 collected at 09:45, no comment.
JS returns to dog park, CC has not been able to collect a fecal sample for roughly an hour, staff agreed to
walk dog park for last 2 samples.
Dog-9 collected at 10:40 from prebagged sample donated by resident. (picture)
Dog-10 observed resident not bagging feces and confirmed recent BM at location. Feces noted as very
red in color, most likely due to food dye used in dog food.
Samples packaged at 11:15. shipped from Fed Ex location at 610 9th St, Durham, NC 27705, Shipping #
394137918330.

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06/23/20: Basef low
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AECOM
Appendix C:
Laboratory Reports and Quality Assurance
Worksheets

AECOM—Herrera

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
June 02, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on May 28, 2020. The results relate only to the
samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project:
Pace Project No.:
Durham MST Study (5/28/20)
92479340
Analytes
Lab ID
Sample ID
Method
Analysts
Reported
Laboratory
92479340001
MST-9-S1-05282020
SM 9222D-2006
WT
1
PASI-R
92479340002
NE3.3.NP-S2-05282020
SM 9222D-2006
WT
1
PASI-R
92479340003
NE2.2.NP-S3-05282020
SM 9222D-2006
WT
1
PASI-R
92479340004
MST-18-S4-05282020
SM 9222D-2006
WT
1
PASI-R
92479340005
MST-12-S5-05282020
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical
Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Sample: MST-9-S1-05282020 Lab ID: 92479340001 Collected: 05/28/20 09:27 Received: 05/28/20 12:34 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 21000 CFU/100 mL 1.0 1 05/28/20 14:22 05/29/20 14:18
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 4 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Sample: NE3.3.NP-S2-05282020 Lab ID: 92479340002 Collected: 05/28/20 09:46 Received: 05/28/20 12:34 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 32000 CFU/100 mL 1.0 1 05/28/20 14:22 05/29/20 14:18
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 5 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Sample: NE2.2.NP-S3-05282020 Lab ID: 92479340003 Collected: 05/28/20 10:10 Received: 05/28/20 12:34 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 26000 CFU/100 mL 1.0 1 05/28/20 14:22 05/29/20 14:18
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 6 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Sample: MST-18-S4-05282020 Lab ID: 92479340004 Collected: 05/28/20 10:36 Received: 05/28/20 12:34 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 19000 CFU/100 mL 1.0 1 05/28/20 14:22 05/29/20 14:18
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 7 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Sample: MST-12-S5-05282020 Lab ID: 92479340005 Collected: 05/28/20 11:21 Received: 05/28/20 12:34 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 5500 CFU/100 mL 1.0 1 05/28/20 14:22 05/29/20 14:18
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 8 of 12

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
QC Batch: 544618 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92479340001, 92479340002, 92479340003, 92479340004, 92479340005
METHOD BLANK: 2900048 Matrix: Water
Associated Lab Samples: 92479340001, 92479340002, 92479340003, 92479340004, 92479340005
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 05/29/20 14:18
SAMPLE DUPLICATE: 2900049
92479340001 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 21000 23000 9
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC.
Page 9 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/02/2020 04:13 PM without the written consent of Pace Analytical Services, LLC. Page 10 of 12

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Durham MST Study (5/28/20)
Pace Project No.: 92479340
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92479340001
MST-9-S1-05282020
SM
9222D-2006
544618
SM
9222D-2006
544621
92479340002
NE3.3.NP-S2-05282020
SM
9222D-2006
544618
SM
9222D-2006
544621
92479340003
NE2.2.NP-S3-05282020
SM
9222D-2006
544618
SM
9222D-2006
544621
92479340004
MST-18-S4-05282020
SM
9222D-2006
544618
SM
9222D-2006
544621
92479340005
MST-12-S5-05282020
SM
9222D-2006
544618
SM
9222D-2006
544621
Date: 06/02/2020 04:13 PM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 11 of 12

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Page 12 of 12

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: May 29, 2020
Report Generated: July 9, 2020
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Sample ID Date Time Analysis Requested Marker Quantified Result Unit
Collected Collected
SM20F25055 MST-9-S1-05282020 5/28/2020 9:27 AM Human HF183 4.27E+03 copies er 100m1
SM20F25056
NE 3.3 NP-S2-05282020
5/28/2020
9:46 AM
Human HF183
2.68E+03
copies per 100ml
SM20F25057
NE 2.2 NP-S3-05282020
5/28/2020
10:10 AM
Human HF183
1.44E+03
copies per 100ml
SM20F25058
MST-18-S4-05282020
5/28/2020
10:36 AM
Human HF183
2.73E+03
copies per 100ml
SM20F25059
MST-12-S5-05282020
5/28/2020
11:21 AM
Human HF183
8.37E+02
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: May 29, 2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
�: ACCR ED
"\�� Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20F25055
MST-9-S1-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25056
NE 3.3 NP-S2-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25057
NE 2.2 NP-S3-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25058
MST-18-S4-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25059
MST-12-S5-05282020
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: May 29, 2020
Report Generated: July 9, 2020 DNQ: Detected Not Quantified
SM20E29012
Sample ID Date Time Analysis Requested Marker Quantified Result Unit
Collected Collected IL 7
MST-9-S1-05282020 5/28/2020 9:27 AM Dog DG3 EPA DNQ copies per 100ml
SM20E29013
NE 3.3 NP-S2-05282020
5/28/2020
9:46 AM
Do DG3 EPA
7.49E+02
copies per 100ml
SM20E29014
NE 2.2 NP-S3-05282020
5/28/2020
10:10 AM
Do DG3 EPA
DNQ
copies per 100ml
SM20E29015
MST-18-S4-05282020
5/28/2020
10:36 AM
Do DG3 EPA
1.61 E+03
copies per 100ml
SM20E29016
MST-12-S5-05282020
5/28/2020
11:21 AM
Do DG3 EPA
1.26E+03
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: May 29, 2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
�: ACCR ED
"\�� Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20E29012
MST-9-S1-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM20E29013
NE 3.3 NP-S2-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM20E29014
NE 2.2 NP-S3-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM20E29015
MST-18-S4-05282020
Water
7/7/2020
7/8/2020
100
ml
2
SM20E29016
MST-12-S5-05282020
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date W0I2018
PSource
Molecular
Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Sample ID
Analysis Company Name
Re ueste
("a pg. 2) Contact Name(s)
Mark
t Send Results To
Phone
Address
City/State/Zip
Billing Info
amments
d.a. s schial r{e-
�� GS
Tc-; vt'` ; GUCti v1 Ci t�
(email) O � - � t C,Li VO v 9-1��� ((�
c
ff��Z�
�j�V� �o-c, *ti/e{( 5 �� -e- "L t�
,,A\n v✓1 IN TTi
-c
PO#:
Will call with credit card
of
Collection
Collection
Time
?
_ -Coniners
V "'L
-t 5 Cow j-•
$ f
c�c
1 ` i a '--1 40- C)
5 � . , - I
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Lt r - -�S GL-2-.. Zv
5 T-
Completed by Client:
Relinquished By ( 0y' f��
Signature+� J- -- --
Date/Time / �3 CZUZO f'2;(50 �2
To protect confidentiality, confirmation and results will only be sent to email address provided
or authorized by contact provided. Signed form indicates agreement with the test limitations on
the back of this form and the oempanys terms of use found here: seurcemelecular.comfabout-
Temperature a:e_1
Thermometer go!
Cooler Number 1211, 18
Received/Flitered 1/i Us
Signature -Ai A t
DateMme
i ;c
sourcemolecular,privacy staterrtentl.

(LJ%A_ Data Quality Assurance Worksheet
HERRERA
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms ; Source Molecular / gPCR DNA
Sample Date/Sample ID: 5/28/2020 / MST-9, NE3.3NP, NE2.2NP, MST-18, MST-12 (Lab No. 92479340)
By G. Catarra
Date
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
NONE
Fecal
OK /
NA
NA
1
<
100mL
NA
NA
NA
NA
MST-9
< 35
NS
< 50
OK
10 cfu/
Coliform
SM 9222D
_1
9
100mL
<50
NONE
O 3
copies/100
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
<50
OK
<50
<50
NONE
OK / HF 183
copies/100
Human
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
< 50
OK
<50
1 If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-le necreek gawkst 05282020.doc
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
June 05, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on June 03, 2020. The results relate only to the
samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette. Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project:
Pace Project No.:
Durham MST Study (6/3/20)
92480137
Analytes
Lab ID
Sample ID
Method
Analysts
Reported
Laboratory
92480137001
MST-9-S1
SM 9222D-2006
WT
1
PASI-R
92480137002
NE3.3.ND-S2
SM 9222D-2006
WT
1
PASI-R
92480137003
NE2.2.ND-S3
SM 9222D-2006
WT
1
PASI-R
92480137004
MST-18-S4
SM 9222D-2006
WT
1
PASI-R
92480137005
MST-12-S5
SM 9222D-2006
WT
1
PASI-R
92480137006
MST-X-S6
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical
Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: MST-9-S1 Lab ID: 92480137001 Collected: 06/03/20 11:05 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 3800 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 4 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: NE3.3.ND-S2 Lab ID: 92480137002 Collected: 06/03/20 10:52 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 3000 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 5 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: NE2.2.ND-S3 Lab ID: 92480137003 Collected: 06/03/20 10:30 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 2400 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 6 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: MST-18-S4 Lab ID: 92480137004 Collected: 06/03/20 09:30 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 1360 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 7 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: MST-12-S5 Lab ID: 92480137005 Collected: 06/03/20 10:05 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 3000 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 8 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Sample: MST-X-S6 Lab ID: 92480137006 Collected: 06/03/20 09:30 Received: 06/03/20 12:05 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 1090 CFU/100 mL 1.0 1 06/03/20 15:48 06/04/20 14:38
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 9 of 13

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
QC Batch: 545444 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92480137001, 92480137002, 92480137003, 92480137004, 92480137005, 92480137006
METHOD BLANK: 2904259 Matrix: Water
Associated Lab Samples: 92480137001, 92480137002, 92480137003, 92480137004, 92480137005, 92480137006
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 06/04/20 14:38
SAMPLE DUPLICATE: 2904260
92480137006 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 1090 1640 40
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC.
Page 10 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/05/2020 04:12 PM without the written consent of Pace Analytical Services, LLC. Page 11 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Durham MST Study (6/3/20)
Pace Project No.: 92480137
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92480137001
MST-9-S1
SM
9222D-2006
545444
SM
9222D-2006
545446
92480137002
NE3.3.ND-S2
SM
9222D-2006
545444
SM
9222D-2006
545446
92480137003
NE2.2.ND-S3
SM
9222D-2006
545444
SM
9222D-2006
545446
92480137004
MST-18-S4
SM
9222D-2006
545444
SM
9222D-2006
545446
92480137005
MST-12-S5
SM
9222D-2006
545444
SM
9222D-2006
545446
92480137006
MST-X-S6
SM
9222D-2006
545444
SM
9222D-2006
545446
Date: 06/05/2020 04:12 PM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 12 of 13

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Page 13 of 13

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: June 4, 2020 ND: Not Detected
Report Generated: July 9, 2020 DNQ: Detected Not Quantified
Sample ID Date Time Analysis Requested Marker Quantified Result Unit
Collected Collected
SM20F25044 MST-18-54 6/3/2020 9:30 AM Human HF183 ND copies per 100ml
SM20F25047 MST-12-55
6/3/2020
10:05 AM
Human HF183
7.73E+03
copies per 100ml
SM20F25048 NE2.2NP-53
6/3/2020
10:30 AM
Human HF183
5.12E+02
copies per 100ml
SM20F25049 MST-X-56
6/3/2020
9:30 AM
Human HF183
DNQ
copies per 100ml
SM20F25050 NE3.3NP-52
6/3/2020
10:52 AM
Human HF183
ND
copies per 100ml
SM20F25051 MST-9-51
6/3/2020
11:05 AM
Human HF183
ND
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: June 4,2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM201`25044
MST-18-54
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25047
MST-12-55
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25048
NE2.2NP-53
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25049
MST-X-56
Water
7/7/2020
7/8/2020
100
ml
2
SM201`25050
NE3.3NP-52
Water
7/7/2020
7/8/2020
100
ml
2
SM20F25051
MST-9-51
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: June 4, 2020 ND: Not Detected
Report Generated: July 9, 2020
Sample ID Date Time Analysis Requested Marker Quantified Result Unit
Collected Collected
E ■
SM20FO4093 MST-18-54 6/3/2020 9:30 AM Do DG3 EPA ND copies per 100ml
SM20FO4094
MST-12-55
6/3/2020
10:05 AM
Do DG3 EPA
ND
copies per 100mil
SM20FO4095
NE2.2NP-53
6/3/2020
10:30 AM
Do DG3 EPA
ND
copies per 100mil
SM20FO4096
MST-X-56
6/3/2020
9:30 AM
Do DG3 EPA
ND
copies per 100ml
SM20FO4097
NE3.3NP-52
6/3/2020
10:52 AM
Do _DG3 EPA
ND
copies per 100ml
SM20FO4098
MST-9-51
6/3/2020
11:05 AM
Do DG3 EPA
ND
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: June 4,2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20FO4093
MST-18-54
Water
7/7/2020
7/8/2020
100
ml
2
SM20FO4094
MST-12-55
Water
7/7/2020
7/8/2020
100
ml
2
SM20FO4095
NE2.2NP-53
Water
7/7/2020
7/8/2020
100
ml
2
SM20FO4096
MST-X-56
Water
7/7/2020
7/8/2020
100
ml
2
SM20FO4097
NE3.3NP-52
Water
7/7/2020
7/8/2020
100
ml
2
SM20FO4098
MST-9-51
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

�* Source Molecular Corporation
*,
Chain 4f Custody Record ource 15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Revision 1.2 Molecular Tel: [phone] Fax: [phone]
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sourcemoleculatfpdvacy_stetement/.
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(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms ; Source Molecula / gPCR DNA
Sample Date/Sample ID: 6/03/2020 / MST-9-S1, NE3.3NP-S2, NE2.2NP-S3, MST- 18-S4, MST-12-S5, MST-X-
S6 (field duplicate of MST-12) (Lab No. 92480137)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
FLAG MST-12 AND MST-X
F
Fecal
Fecal
/
OK /
NA
NA
1
-1
100mL
NA
NA
NA
NA
MST-X
< 35
MST-12
< 50
OK
"J" DUE TO LAB AND FIELD
10
rm
SM 92
40
93
100mL
DUPLICATE EXCEEDANCES
<50
NONE
OK / DG3
copies/100
MST-18
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NC
5 50
OK
<50
<50
NONE
OK / HF 183
co p ies/100
MST -18
Human
(EPA)
NA
NA
NA
NA
mL,
NA
NA
NA
NA
NA
NA
NC
< 50
OK
<50
I If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-2e necreek gawkst 06032020.do"
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
June 25, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Bacteria
Pace Project No.: 92483054
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on June 23, 2020. The results relate only to the
samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette. Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Bacteria
Pace Project No.: 92483054
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project: Bacteria
Pace Project No.: 92483054
Analytes
Lab ID Sample ID
Method
Analysts
Reported
Laboratory
92483054001 MST-9-S1
SM 9222D-2006
WT
1
PASI-R
92483054002 NE2.2.NP-S3
SM 9222D-2006
WT
1
PASI-R
92483054003 NE3.3 NP-S2
SM 9222D-2006
WT
1
PASI-R
92483054004 MST-18-S4
SM 9222D-2006
WT
1
PASI-R
92483054005 MST-12-S5
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Bacteria
Pace Project No.: 92483054
Sample: MST-9-S1 Lab ID: 92483054001 Collected: 06/23/20 10:00 Received: 06/23/20 12:00 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 460 CFU/100 mL 1.0 1 06/23/20 12:45 06/24/20 14:35
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 4 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Bacteria
Pace Project No.: 92483054
Sample: NE2.2.NP-S3 Lab ID: 92483054002 Collected: 06/23/20 10:30 Received: 06/23/20 12:00 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 727 CFU/100 mL 1.0 1 06/23/20 12:45 06/24/20 14:35
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 5 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Bacteria
Pace Project No.: 92483054
Sample: NE3.3 NP-S2 Lab ID: 92483054003 Collected: 06/23/20 10:15 Received: 06/23/20 12:00 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 1180 CFU/100 mL 1.0 1 06/23/20 12:45 06/24/20 14:35
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 6 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Bacteria
Pace Project No.: 92483054
Sample: MST-18-S4 Lab ID: 92483054004 Collected: 06/23/20 11:00 Received: 06/23/20 12:00 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 1270 CFU/100 mL 1.0 1 06/23/20 12:45 06/24/20 14:35
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 7 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Bacteria
Pace Project No.: 92483054
Sample: MST-12-S5 Lab ID: 92483054005 Collected: 06/23/20 09:23 Received: 06/23/20 12:00 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 162 CFU/100 mL 1.0 1 06/23/20 12:45 06/24/20 14:35
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 8 of 13

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Bacteria
Pace Project No.: 92483054
QC Batch: 549576 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92483054001, 92483054002, 92483054003, 92483054004, 92483054005
METHOD BLANK: 2922547 Matrix: Water
Associated Lab Samples: 92483054001, 92483054002, 92483054003, 92483054004, 92483054005
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 06/24/20 14:35
SAMPLE DUPLICATE: 2922548
92483054005 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 162 126 25
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC.
Page 9 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Bacteria
Pace Project No.: 92483054
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 06/25/2020 10:28 AM without the written consent of Pace Analytical Services, LLC. Page 10 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Bacteria
Pace Project No.: 92483054
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92483054001
MST-9-S1
SM
9222D-2006
549576
SM
9222D-2006
549585
92483054002
NE2.2.NP-S3
SM
9222D-2006
549576
SM
9222D-2006
549585
92483054003
NE3.3 NP-S2
SM
9222D-2006
549576
SM
9222D-2006
549585
92483054004
MST-18-S4
SM
9222D-2006
549576
SM
9222D-2006
549585
92483054005
MST-12-S5
SM
9222D-2006
549576
SM
9222D-2006
549585
Date: 06/25/2020 10:28 AM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 11 of 13

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Page 13 of 13

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: June 24, 2020 ND: Not Detected
Report Generated: July 9, 2020 DNQ: Detected Not Quantified
Sample ID DaW Time Analysis Requested Marker Quantified Result Unit
Collected Collected
SM20F16447 MST-9 6/23/2020 10:00 AM Human HF183 ND copies per 100ml
SM20F16448
NE2.2NP
6/23/2020
10:30 AM
Human HF183
DNQ
copies per 100ml
SM20F16449
NE3.3NP
6/23/2020
10:15 AM
Human HF183
ND
copies per 100ml
SM20F16450
MST-18
6/23/2020
11:00 AM
Human HF183
5.18E+03
copies per 100ml
SM20F16451
MST-12
6/23/2020
9:25 AM
Human HF183
DNQ
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: June 24,2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
�: ACCR ED
"\�� Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20F16447
MST-9
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16448
NE2.2NP
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16449
NE3.3NP
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16450
MST-18
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16451
MST-12
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: June 24, 2020 ND: Not Detected
Report Generated: July 9, 2020
SM20F16452
Sample ID DaNK Time Analysis Requested Marker Quantified Result Unit
Collected Collected
MST-9 6/23/2020 10:00 AM Do DG3 EPA ND copies per 100ml
SM20F16453
NE2.2NP
6/23/2020
10:30 AM
Do DG3 EPA
ND
copies per 100ml
SM20F16454
NE3.3NP
6/23/2020
10:15 AM
Do DG3 EPA
ND
copies per 100ml
SM20F16455
MST-18
6/23/2020
11:00 AM
Do DG3 EPA
ND
copies per 100ml
SM20F16456
MST-12
6/23/2020
9:25 AM
Do DG3 EPA
ND
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: June 24,2020
Report Generated: July 9, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
�: ACCR ED
"\�� Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20F16452
MST-9
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16453
NE2.2NP
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16454
NE3.3NP
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16455
MST-18
Water
7/7/2020
7/8/2020
100
ml
2
SM201`16456
MST-12
Water
7/7/2020
7/8/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: July s, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody RecordilC)UrC' Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Revision 1.2 Tel: [phone] Fax: [phone]
Effective Date 8/2012018 Email: [e-mail]
Sample IDk
Analysis Company Marne
Rec�ueste�d .
[se pg.1 Contact Name(s)
Mark l3✓r� C :.rr��
W.MpX„ � Send Results TO (email)
Phone
rEr�
Address
City/State/Zipc�r'dn; (' 2 --7 Q
Billing Info PO#: Will call with credit card
:ta€greiu`%� Collection Collection
r_-, - ar- sr tp.. Date TIme
N is 2 .
Y<
23
• ?, 0
�'5, y N P
X
X
2112-3
Twoo
Completed by Client. y
Relinquished By T c,%,
Signature
Date/Time (,2-/; &L)
To protect confidentiality, confirmation and results will ohiy be sent to email address provided
or authorized by contact provided, Signed form indicates agreement with the test limitations on
the back of this form and the company's terms of use found here: sourcemolecular.com/about-
sourcemalecularlpfivac:y__§tlterrenU.
Completed by Source Q Molecular:
Temperature C— ReceivedlFlltered
Thermometer 0 0 ( Signature c}}knit
Cooler lumber K0 Date/Time ci�i

(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms ; Source Molecular / gPCR DNA
Sample Date/Sample ID: 6/23/2020 / MST-9-S1, NE2.2NP-S3, NE3.3NP-S2, MST- 18-S4, MST- 12-S5
(Lab No.92483054)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
NONE
Fecal
OK /
NA
NA
1
100mL
NA
NA
NA
NA
MST-12
< 35
NS
< 50
OK
10 cfa/
Coliform
SM 9222D
25
100mL
<50
NONE
OK / DG3
copies/100
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
< 50
OK
<50
<50
NONE
OK / HF 183
copies/100
Human
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
< 50
OK
<50
' If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-3e necreek gawkst 06232020.doc
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
August 06, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on August 04, 2020. The results relate only to
the samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette. Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project:
Pace Project No.:
Durham MST Study (8/4/20)
92489027
Analytes
Lab ID
Sample ID
Method
Analysts
Reported
Laboratory
92489027001
MST-18-54
SM 9222D-2006
WT
1
PASI-R
92489027002
MST-12-55
SM 9222D-2006
WT
1
PASI-R
92489027003
NE2.2 NP-93
SM 9222D-2006
WT
1
PASI-R
92489027004
NE3.3 NP-52
SM 9222D-2006
WT
1
PASI-R
92489027005
MST-9-51
SM 9222D-2006
WT
1
PASI-R
92489027006
MST-X-56
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical
Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: MST-18-54 Lab ID: 92489027001 Collected: 08/04/20 09:45 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 5460 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 4 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: MST-12-55 Lab ID: 92489027002 Collected: 08/04/20 10:00 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 60000 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01 u2
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 5 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: NE2.2 NP-93 Lab ID: 92489027003 Collected: 08/04/20 10:30 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 60000 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01 u2
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 6 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: NE3.3 NP-52 Lab ID: 92489027004 Collected: 08/04/20 10:50 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 24000 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 7 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: MST-9-51 Lab ID: 92489027005 Collected: 08/04/20 11:00 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 16000 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 8 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Sample: MST-X-56 Lab ID: 92489027006 Collected: 08/04/20 11:00 Received: 08/04/20 11:52 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 60000 CFU/100 mL 1.0 1 08/04/20 13:57 08/05/20 12:01 u2
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 9 of 13

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
QC Batch: 558319 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92489027001, 92489027002, 92489027003, 92489027004, 92489027005, 92489027006
METHOD BLANK: 2962694 Matrix: Water
Associated Lab Samples: 92489027001, 92489027002, 92489027003, 92489027004, 92489027005, 92489027006
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 08/05/20 12:01
SAMPLE DUPLICATE: 2962695
92489027001 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 5460 4910 11
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC.
Page 10 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
ANALYTE QUALIFIERS
u2 Colonies are too numerous to count. Actual result may be greater than reported.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 08/06/2020 01:39 PM without the written consent of Pace Analytical Services, LLC. Page 11 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Durham MST Study (8/4/20)
Pace Project No.: 92489027
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92489027001
MST-18-54
SM
9222D-2006
558319
SM
9222D-2006
558320
92489027002
MST-12-55
SM
9222D-2006
558319
SM
9222D-2006
558320
92489027003
NE2.2 NP-93
SM
9222D-2006
558319
SM
9222D-2006
558320
92489027004
NE3.3 NP-52
SM
9222D-2006
558319
SM
9222D-2006
558320
92489027005
MST-9-51
SM
9222D-2006
558319
SM
9222D-2006
558320
92489027006
MST-X-56
SM
9222D-2006
558319
SM
9222D-2006
558320
Date: 08/06/2020 01:39 PM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 12 of 13

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Page 13 of 13

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: August 5, 2020
Report Generated: August 18, 2020 DNQ: Detected Not Quantified
Collected Collected
SELM A Sample ID Date T Time Analysis Requested Marker Quantified Result Unit
SM20HO5025 MST-18 080420 8/4/2020 9:45 AM Human HF183 1.87E+03 copies per 100ml
SM20HO5024 MST-12 080420
8/4/2020
10:00 AM
Human HF183
1.33E+03
copies per 100ml
SM20HO5028 NE2.2NP 080420
8/4/2020
10:30 AM
Human HF183
4.31 E+02
copies per 100ml
SM20HO5029 NE3.3NP 080420
8/4/2020
10:50 AM
Human HF183
DNQ
copies per 100ml
SM20HO5030 MST-9 080420
8/4/2020
11:10 AM
Human HF183
DNQ
copies per 100ml
SM20HO5031 MST-X 080420
8/4/2020
11:10 AM
Human HF183
DNQ
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: August 5, 2020
Report Generated: August 18, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20HO5025
MST-18 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5024
MST-12 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5028
NE2.2NP 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5029
NE3.3NP 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5030
MST-9 080420
Water
8/17/2020
8/18/2020
100
ml
2
SM20HO5031
MST-X 080420
Water
8/17/2020
8/17/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: August 18, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: August 5, 2020
Report Generated: August 18, 2020 DNQ: Detected Not Quantified
M # Sample ID Date T Time Analysis Requested Marker Quantified Result Unit
AL Collected Collected IL E
SM20HO5034 MST-18 080420 8/4/2020 9:45 AM Dog DG3 EPA 1.12E+03 copies per 100ml
SM20HO5035
MST-12 080420
8/4/2020
10:00 AM
Do DG3 EPA
1.31 E+03
copies per 100ml
SM20HO5039
NE2.2NP 080420
8/4/2020
10:30 AM
Dog DG3 EPA
8.12E+02
copies per 100ml
SM20HO5040
NE3.3NP 080420
8/4/2020
10:50 AM
Do DG3 EPA
DNQ
copies per 100ml
SM20HO5041
MST-9 080420
8/4/2020
11:10 AM
Do _DG3 EPA
DNQ
copies per 100ml
SM20HO5042 MST-X 080420
8/4/2020
11:10 AM
Do DG3 EPA
DNQ
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: August 5, 2020
Report Generated: August 18, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20HO5034
MST-18 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5035
MST-12 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5039
NE2.2NP 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5040
NE3.3NP 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5041
MST-9 080420
Water
8/17/2020
8/17/2020
100
ml
2
SM20HO5042
MST-X 080420
Water
8/17/2020
8/17/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: August 18, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date 8120/2018
oMolecular
Source
Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Tel: [phone] Fax: [phone]
)email: [e-mail]
Sample ID
nalysis
e�ueste
e.
spg. 2.
Mark
"fj X.,
+"
vComments
Company Name
rq-Q� S A ti.neAj-i U 1
Contact Name(s)
TO �111 ad �
Send Results To
(email) t+r�
Phone
I j7
Address-
�4 C
V
City/State/Zip
�e- ka, L­ Lid r� l
Billing Info
PO#:
Q Will tali with credit card
i.e. 6M.cW reau s.
# of containers
Collection
Collection
Ti
2 Y-IS0
_t{tea
5T- I Z u42-
�� � Z-o
16
64"0
:,0
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642,76
Completed by Client:
Relinquished By Tit D( 4r'>'J✓�
Signature
Date/Time
To protect confidentiality, confirmation dndfesultswlil only be sent to email address provided
or authorized by contact provided. Signed form indicates agreement with the test limitations on
the back of this form and the company's terms of use found here: sourcemolecular.comlabout-
sourcemolecularlprivacy statement!.
Completed by Source MWecular: `
Temperature � Ret ived/Filtered
Thermometer Signature
Cooler Number M �.(Q Date[Time

(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms ; Source Molecular / gPCR DNA
Sample Date/Sample ID: 8/04/2020 / MST-9, NE3.3NP, NE2.2NP, MST-18, MST-12, MSTA (field duplicate of
MST-9) (Lab No. 92489027)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate Recovery
Samples
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
(%)
Recovery (%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goal
Reporte
Goal
Reported
Goal'
Report
Goal'
Calibration/
Parameter
Methodology
Limit
d
ed
Performance
ACTION
<_1.0 cfa/
FLAG MST-12, NE2.2NP,
100mL
MST-X AS <60,000 DUE TO
Fecal
OK/
MST-18
MST-9
TNTC PLATES
Coliform
SM 9222D
NA
NA
1
:91
10 cfu/
NA
NA
NA
NA
11
S 35
NC
S 50
OK
FLAG MST-9 J DUE TO
100ml-
FIELD DUPLICATE
EXCEEDANCE DUE TO
TNTC
<50
NONE
OK / DG3
copies/100
MST-9
Dog
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
S 50
OK
(EPA)
NC
<50
<50
NONE
OK
co ies/100
p
MST -9
Human
NA
NA
NA
NA
ML
NA
NA
NA
NA
NA
NA
550
OK
(EPA)
A)
NC
<50
I If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc4e necreek gawkst 08042020.do"
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
September 16, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (9/8)
Pace Project No.: 92494324
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on September 08, 2020. The results relate only
to the samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
Some analyses were subcontracted outside of the Pace Network. The test report from the external subcontractor is
attached to this report in its entirety.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette, Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 3

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Page 2 of 3

EMSL Analytical, Inc. EMSL Order: 292009305
2500 Gateway Centre Blvd., Suite 600, Morrisville, NC 27560 CUStomerlD: pace54
Phone/Fax: [phone] CustomerPO: NMG4324
htto://www.EMSL.com raleiahlab(Wemsl.com ProjectlD:
Attn: Nicole Gasiorowski
Phone:
[phone]
Pace Analytical Services, LLC
Fax:
[phone]
9800 Kincey Avenue, Suite 100
Received:
9/8/2020 02:55 PM
Huntersville, NC 28078
Analysis Date:
9/11/2020
Collected:
9/8/2020
Durham MST Stu
Test Report: Fecal Coliform (Membrane Filtration Technique)
Method:
SM-9222 D for Water Samples
Sample
Sampling Location
Amount Received Amount Sampled
Fecal Coliform
Date/Time Collected
mL mL
CFU/100ml
292009305-0001
250 1
1800
MST-9-S 1
9/8/2020
292009305-0002
250 10
520
NE3.3-NP-S2
9/8/2020
292009305-0003
250 1
800
NE2.2-NP-S3
9/8/2020
292009305-0004
250 10
800
MST-18-S4
9/8/2020
292009305-0005 250 1 1200
MST-12-S5 9/8/2020
Analyst(s)
Virginia Causey (5)
Gs ct 'Irk q -'] (ck � —� t t' -�—
Alan Goldstein, Ph.D., Laboratory Manager
or other approved signatory
EMSL maintains liability limited to cost of analysis. Interpretation and use of test results are the responsibility of the client. This report relates only to the samples reported above, and may not be
reproduced, except in full, without written approval by EMSL. EMSL bears no responsibility for sample collection activities or analytical method limitations. The report reflects the samples as received.
Results are generated from the field sampling data (sampling volumes and areas, locations, etc.) provided by the client on the Chain of Custody. Samples are within quality control criteria and met method
specifications unless otherwise noted.
NA = not applicable. The detection limit is equal to 1 colony forming unit (CFU) per 100 mL of sample.
Samples analyzed by EMSL Analytical, Inc. Morrisville, NC
Initial report from 09/14/2020 16:55:29
Test Report EMSL MR19-7.26.0 Printed: 9/14/2020 4:55:29 PM Page 1 of 1
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
�1 ACCREDITED
�lln�`� Cartl/lcwte•72e.a7
ISO 17025 Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: September 9, 2020 ND: Not Detected
Report Generated: September 23, 2020 DNQ: Detected Not Quantified
Sample ID Date I Time Analysis Requested Marker Quantified Result Unit
Collected Collected
SM20109097 MST-9 090820 9/8/2020 11:10 AM Human HF183 ND copies per 100ml
SM20109098
NE3.3NP 090820
9/8/2020
11:30 AM
Human HF183
ND
copies per 100ml
SM20109099
NE2.2NP 090820
9/8/2020
11:45 AM
Human HF183
DNQ
copies per 100ml
SM20109100
MST-18 090820
9/8/2020
12:10 PM
Human HF183
DNQ
copies per 100ml
SM20109101
MST-12 090820
9/8/2020
12:40 PM
Human HF183
DNQ
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: September 9, 2020
Report Generated: September 23, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20109097
MST-9 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109098
NE3.3NP 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109099
NE2.2NP 090820
Water
9/22/2020
9/23/2020
100
ml
2
SM20109100
MST-18 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109101
MST-12 090820
Water
9/22/2020
9/22/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: September 23, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: September 9, 2020 ND: Not Detected
Report Generated: September 23, 2020
Sample ID Date I Time Analysis Requested Marker Quantified Result Unit
Collected Collected IL E
SM20109101 MST-9 090820 9/8/2020 11:10 AM Do DG3 EPA ND copies per 100ml
SM20109103
NE3.3NP 090820
9/8/2020
11:30 AM
Do DG3 EPA
ND
copies per 100ml
SM20109104
NE2.2NP 090820
9/8/2020
11:45 AM
Do DG3 EPA
ND
copies per 100ml
SM20109105
MST-18 090820
9/8/2020
12:10 PM
Do DG3 EPA
ND
copies per 100ml
SM20109106
MST-12 090820
9/8/2020
12:40 PM
Do DG3 EPA
ND
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: September 9, 2020
Report Generated: September 23, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20109101
MST-9 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109103
NE3.3NP 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109104
NE2.2NP 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109105
MST-18 090820
Water
9/22/2020
9/22/2020
100
ml
2
SM20109106
MST-12 090820
Water
9/22/2020
9/22/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: September 23, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date 8t2012018
oSource
Molecular
Source Molecular Corporations
15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Sam le 0
Analysis
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Se$ 21
Mark �'
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Company Name
4VrefA,
Contact Name(s)U
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Send Results To
(email) ton,,atdkk'." � �� @N a 1pQri.n oA, Z(his
Phone
q 15 4 17
Address
�2°,�r
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CitylStatelZip
�€nl�hC✓w. N C—
Billing Info
PO#:
= Will call with credit card
co
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ants
# of ontainers
Collection
Collection
Tim/
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/,
� :�
1 J 1• / V
2 v
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Z.
Completed by Client: —
Relinquished By I G (L Y
Signature
Date/Time 2. v l-
To protect confidentiality, confirmation and results will only be sent to email address provided
or authorized by contact provided. Sigred form indicates agreement with the test limitations on
the back of this form and the company's terms of use found here: sourcemoiecular.comlabout-
seurcemolecularFprivacy_statement!.
Completed by Source Molecula
Temperature % 1 Rec(ed/Filtered .SL
Thermometer _) Signature
Cooler Numbe-r� �" Date/Time L s

(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms; Source Molecular / gPCR DNA
Sample Date/Sample ID: 9/08/2020 / MST-9-S1, NE3.3NP-S2, NE2.2NP-S3, MST- 18-S4, MST- 12-S5
(Lab No.92494324)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
NONE
Fecal
OK /
NA
NA
1
100mL
NA
NA
NA
NA
NS
< 35
NS
< 50
OK
10 cfu/
Coliform
SM 9222D
100mL
<50
NONE
OK / DG3
copies/100
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
< 50
OK
<50
<50
NONE
OK / HF 183
copies/100
Human
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NS
< 50
OK
<50
' If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-5e necreek gawkst 09082020.do"
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
October 05, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on September 30, 2020. The results relate only
to the samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette. Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project:
Pace Project No.:
Durham MST Study (9/30)
92498060
Analytes
Lab ID
Sample ID
Method
Analysts
Reported
Laboratory
92498060001
MST-9-S1
SM 9222D-2006
WT
1
PASI-R
92498060002
NE3.3 NP-S2
SM 9222D-2006
WT
1
PASI-R
92498060003
NEX-S3
SM 9222D-2006
WT
1
PASI-R
92498060004
NE2.2 NP-S4
SM 9222D-2006
WT
1
PASI-R
92498060005
MST-12-S5
SM 9222D-2006
WT
1
PASI-R
92498060006
MST-18-S6
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical
Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: MST-9-S1 Lab ID: 92498060001 Collected: 09/30/20 11:30 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 28000 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47 D6
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 4 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: NE3.3 NP-S2 Lab ID: 92498060002 Collected: 09/30/20 11:35 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 18000 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 5 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: NEX-S3 Lab ID: 92498060003 Collected: 09/30/20 11:35 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 22000 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 6 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: NE2.2 NP-S4 Lab ID: 92498060004 Collected: 09/30/20 11:55 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 600 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47 u2
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 7 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: MST-12-S5 Lab ID: 92498060005 Collected: 09/30/20 12:20 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 29000 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 8 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Sample: MST-18-S6 Lab ID: 92498060006 Collected: 09/30/20 12:45 Received: 09/30/20 13:45 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 32000 CFU/100 mL 1.0 1 09/30/20 14:50 10/01/20 13:47
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 9 of 13

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
QC Batch: 570501 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92498060001, 92498060002, 92498060003, 92498060004, 92498060005, 92498060006
METHOD BLANK: 3022114 Matrix: Water
Associated Lab Samples: 92498060001, 92498060002, 92498060003, 92498060004, 92498060005, 92498060006
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 10/01/20 13:47
SAMPLE DUPLICATE: 3022115
92498060001 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 28000 21000 29 D6
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC.
Page 10 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
ANALYTE QUALIFIERS
D6 The precision between the sample and sample duplicate exceeded laboratory control limits.
u2 Colonies are too numerous to count. Actual result may be greater than reported.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/05/2020 09:55 AM without the written consent of Pace Analytical Services, LLC. Page 11 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Durham MST Study (9/30)
Pace Project No.: 92498060
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92498060001
MST-9-S1
SM
9222D-2006
570501
SM
9222D-2006
570507
92498060002
NE3.3 NP-S2
SM
9222D-2006
570501
SM
9222D-2006
570507
92498060003
NEX-S3
SM
9222D-2006
570501
SM
9222D-2006
570507
92498060004
NE2.2 NP-S4
SM
9222D-2006
570501
SM
9222D-2006
570507
92498060005
MST-12-S5
SM
9222D-2006
570501
SM
9222D-2006
570507
92498060006
MST-18-S6
SM
9222D-2006
570501
SM
9222D-2006
570507
Date: 10/05/2020 09:55 AM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 12 of 13

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Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: October 1, 2020
Report Generated: October 14, 2020 DNQ: Detected Not Quantified
SM20JO1045
Sample ID Date I Time Analysis Requested Marker Quantified Result Unit
Collected Collected
MST-9 093020 9/30/2020 11:30 AM Human HF183 2.10E+03 copies er 100ml
SM20JO1057
NE3.3 NP 093020
9/30/2020
11:35 AM
Human HF183
2.64E+03
copies per 100ml
SM20JO1058
NEX 093020
9/30/2020
Human HF183
1.64E+03
copies per 100ml
SM20J01058
NE2.2NP 093020
9/30/2020
11:55 AM
Human HF183
2.26E+03
copies per 100m1
SM20JO1060
MST-12
9/30/2020
12:20 PM
Human HF183
DNQ
copies per 100ml
SM20J01061 MST-18
9/30/2020
12:45 PM
Human HF183
2.92E+03
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
)Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 1, 2020
Report Generated: October 14, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
�: ACCR ED
"\�� Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20JO1045
MST-9 093020
Water
10/12/2020
10/12/2020
100
ml
2
SM20JO1057
NE3.3 NP 093020
Water
10/12/2020
10/12/2020
100
ml
2
SM20JO1058
NEX 093020
Water
10/12/2020
10/12/2020
100
ml
2
SM20JO1058
NE2.2NP 093020
Water
10/12/2020
10/12/2020
100
ml
2
SM20JO1060
MST-12
Water
10/12/2020
10/12/2020
100
ml
2
SM20JO1061
MST-18
Water
10/12/2020
10/12/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: October 14, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
�1 ACCREDITED
�lln�`� Cartl/lcwte•72e.a7
ISO 17025 Accredited
Testing Laboratory
Submitter: Three Oaks Engineering
Date Received/Processed: October 1, 2020
Report Generated: October 14, 2020 DNQ: Detected Not Quantified
M #7 Sample ID Coll C w d Analysis Requested Marker Quantified Result Unit
SM20JO1062 MST-9 093020 9/30/2020 11:30 AM Do DG3 EPA DNQ copies per 100ml
SM20JO1063 NE3.3 NP 093020
9/30/2020
11:35 AM
Do DG3 EPA
1.36E+03
copies per 100ml
SM20J01065 NEX 093020
9/30/2020
Do DG3 EPA
1.87E+03
copies per 100ml
SM20JO1066 NE2.2NP 093020
9/30/2020
11:55 AM
Do DG3 EPA
1.38E+03
copies per 100ml
SM20JO1067 MST-12
9/30/2020
12:20 PM
Do _DG3 EPA
9.27E+02
copies per 100ml
SM20JO1068 MST-18
9/30/2020
12:45 PM
Do DG3 EPA
8.17E+02
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 1, 2020
Report Generated: October 14, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
�.NE3.3
NP 093020
1111100 •
�r
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Three Oaks Engineering
Report Generated: October 14, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date 812012018
0 Source
Molecular
Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami lakes, FL 33016
Tel: [phone] Fax: [phone]
Email: lnfo@sourcemolecular-com
Sample ID
Analysis Company Name tftC a ak 5
Reec�uest�d
p9• 1 . f� Contact Names) •T�;,,.� i � f k �.,y,,,, t� � i•� � � �{�..e
Mark e
b?ggX„ `�'� Send Results To (email) m� q ;� rss, (���M40,1h1?1, ;111A�A , E 0 oN -L
^� Phone VZ— 2— 69 9 1
Address 3 2-+ e fa t J' -t,41 1 S �a i k z-
CitylStaterzip D zx 4
Billing Info P4#: i a wit] call with credit card
gmments Collection Collection
r
0`.1 0
'0 2r)
1*
0 y a A
-
FY D )302-v
tti
[52 ,2 P O'3 3
XX
tc
1z.2- 0
01c��
ll,'
Completed by Client;
Relinquished By 16^-1 D 1 d 1^5
Signature
17
OatelTime
To protect eonffdentiality, confirmation and results will only be sent to email address provided
or authorized by contact provided. Signed form indicates agreement with the test limitations on
the back of this form and the company's terms of use fouhd here: sourcemolecufar.comlabout-
sourcemolecularlpdvacy_statemmtl.
Completed by Sourge Mg
)ecular,
Temperature Re d/Filtered
Thermometer b h —.—Signature
Cooler Number a Q� Date/Time 0 f ' t
2)
At,Con�

(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms; Source Molecular / gPCR DNA
Sample Date/Sample ID: 9/30/2020 / MST-9, NE3.3NP, NE2.2NP, MST-18, MST-12, MSTA (field duplicate of
NE3.3NP) (Lab No. 92498060)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
FLAG NE2.2NP < DUE To
Fecal
OK /
NA
NA
1
100mL
NA
NA
NA
NA
MST
< 35
NE3.3NP
< 50
OK
TNTC PLATE COUNT
10 cfu/
Coliform
SM 9222D
29
20
100mL
<50
NONE
OK / DG3
co p ies/100
NE3.3NP
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
32
< 50
OK
<50
<50
NONE
OK / HF 183
copies/100
NE3.3NP
Human
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
48
< 50
OK
<50
' If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-6e necreek gawkst 09302020.do"
Herrera Environmental Consultants

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
October 19, 2020
Tom Dickinson
Three Oaks Engineering
324 Blackwell St
Durham, NC 27701
RE: Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Dear Tom Dickinson:
Enclosed are the analytical results for sample(s) received by the laboratory on October 15, 2020. The results relate only to
the samples included in this report. Results reported herein conform to the applicable TNI/NELAC Standards and the
laboratory's Quality Manual, where applicable, unless otherwise noted in the body of the report.
The test results provided in this final report were generated by each of the following laboratories within the Pace Network:
• Pace Analytical Services - Raleigh
If you have any questions concerning this report, please feel free to contact me.
Sincerely,
Nicole Gasiorowski
[e-mail]
[phone]
Project Manager
Enclosures
cc: Rob Zisette. Herrera Environmental
REPORT OF LABORATORY ANALYSIS
r
e
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 1 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
CERTIFICATIONS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Pace Analytical Services Raleigh
6701 Conference Drive, Raleigh, NC 27607 North Carolina Bioassay Certification #: 16
North Carolina Wastewater Certification #: 67 North Carolina Drinking Water Certification #: 37731
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 2 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
SAMPLE ANALYTE COUNT
Project:
Pace Project No.:
Durham MST Study (10/15)
92500686
Analytes
Lab ID
Sample ID
Method
Analysts
Reported
Laboratory
92500686001
MST-9-S1
SM 9222D-2006
WT
1
PASI-R
92500686002
NE 3.3 NP-S2
SM 9222D-2006
WT
1
PASI-R
92500686003
NE 2.2 NP-S3
SM 9222D-2006
WT
1
PASI-R
92500686004
MST-12-S4
SM 9222D-2006
WT
1
PASI-R
92500686005
MST-18-S5
SM 9222D-2006
WT
1
PASI-R
92500686006
MST-X-S6
SM 9222D-2006
WT
1
PASI-R
PASI-R = Pace Analytical
Services - Raleigh
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC. Page 3 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: MST-9-S1 Lab ID: 92500686001 Collected: 10/15/20 10:25 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 153 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 4 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: NE 3.3 NP-S2 Lab ID: 92500686002 Collected: 10/15/20 10:40 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 290 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 5 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: NE 2.2 NP-S3 Lab ID: 92500686003 Collected: 10/15/20 11:00 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 560 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 6 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: MST-12-S4 Lab ID: 92500686004 Collected: 10/15/20 11:25 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 330 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 7 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: MST-18-S5 Lab ID: 92500686005 Collected: 10/15/20 11:50 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 2600 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 8 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
ANALYTICAL RESULTS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Sample: MST-X-S6 Lab ID: 92500686006 Collected: 10/15/20 11:50 Received: 10/15/20 12:43 Matrix: Water
Parameters Results Units Report Limit DF Prepared Analyzed CAS No. Qual
MBIO 9222D Fecal Coliform RAL Analytical Method: SM 9222D-2006 Preparation Method: SM 9222D-2006
Pace Analytical Services - Raleigh
Fecal Coliforms 2100 CFU/100 mL 1.0 1 10/15/20 14:07 10/16/20 13:10
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 9 of 13

lV11.1 Analytical
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
QC Batch: 573878 Analysis Method: SM 9222D-2006
QC Batch Method: SM 9222D-2006 Analysis Description: 9222D Fecal Coliform(MF) - RAL
Laboratory: Pace Analytical Services - Raleigh
Associated Lab Samples: 92500686001, 92500686002, 92500686003, 92500686004, 92500686005, 92500686006
METHOD BLANK: 3038429 Matrix: Water
Associated Lab Samples: 92500686001, 92500686002, 92500686003, 92500686004, 92500686005, 92500686006
Blank Reporting
Parameter Units Result Limit Analyzed Qualifiers
Fecal Coliforms CFU/100 mL ND 1.0 10/16/20 13:10
SAMPLE DUPLICATE: 3038430
92500686001 Dup
Parameter Units Result Result RPD Qualifiers
Fecal Coliforms CFU/100 mL 153 153 0
Results presented on this page are in the units indicated by the "Units" column except where an alternate unit is presented to the right of the result.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC.
Page 10 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALIFIERS
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
DEFINITIONS
DF - Dilution Factor, if reported, represents the factor applied to the reported data due to dilution of the sample aliquot.
ND - Not Detected at or above adjusted reporting limit.
TNTC - Too Numerous To Count
J - Estimated concentration above the adjusted method detection limit and below the adjusted reporting limit.
MDL -Adjusted Method Detection Limit.
PQL - Practical Quantitation Limit.
RL - Reporting Limit - The lowest concentration value that meets project requirements for quantitative data with known precision and
bias for a specific analyte in a specific matrix.
S - Surrogate
1,2-Diphenylhydrazine decomposes to and cannot be separated from Azobenzene using Method 8270. The result for each analyte is
a combined concentration.
Consistent with EPA guidelines, unrounded data are displayed and have been used to calculate % recovery and RPD values.
LCS(D) - Laboratory Control Sample (Duplicate)
MS(D) - Matrix Spike (Duplicate)
DUP - Sample Duplicate
RPD - Relative Percent Difference
NC - Not Calculable.
SG - Silica Gel - Clean -Up
U - Indicates the compound was analyzed for, but not detected.
Acid preservation may not be appropriate for 2 Chloroethylvinyl ether.
A separate vial preserved to a pH of 4-5 is recommended in SW846 Chapter 4 for the analysis of Acrolein and Acrylonitrile by EPA
Method 8260.
N-Nitrosodiphenylamine decomposes and cannot be separated from Diphenylamine using Method 8270. The result reported for
each analyte is a combined concentration.
Pace Analytical is TNI accredited. Contact your Pace PM for the current list of accredited analytes.
TNI -The NELAC Institute.
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
Date: 10/19/2020 11:17 AM without the written consent of Pace Analytical Services, LLC. Page 11 of 13

lceAnalyircal
www.pacelabs.com
Pace Analytical Services, LLC
9800 Kincey Ave. Suite 100
Huntersville, NC 28078
[phone]
QUALITY CONTROL DATA CROSS REFERENCE TABLE
Project: Durham MST Study (10/15)
Pace Project No.: 92500686
Analytical
Lab ID
Sample ID
QC
Batch Method
QC Batch
Analytical Method
Batch
92500686001
MST-9-S1
SM
9222D-2006
573878
SM
9222D-2006
573879
92500686002
NE 3.3 NP-S2
SM
9222D-2006
573878
SM
9222D-2006
573879
92500686003
NE 2.2 NP-S3
SM
9222D-2006
573878
SM
9222D-2006
573879
92500686004
MST-12-S4
SM
9222D-2006
573878
SM
9222D-2006
573879
92500686005
MST-18-S5
SM
9222D-2006
573878
SM
9222D-2006
573879
92500686006
MST-X-S6
SM
9222D-2006
573878
SM
9222D-2006
573879
Date: 10/19/2020 11:17 AM
REPORT OF LABORATORY ANALYSIS
This report shall not be reproduced, except in full,
without the written consent of Pace Analytical Services, LLC.
Page 12 of 13

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Page 13 of 13

0 Source
%Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 16, 2020
Report Generated: October 28, 2020
%� /^� � ACCRRDITEPI
�hh,\�° cerencateat2s.ot
ISO 17025 Accredited
Testing Laboratory
ND: Not Detected
DNQ: Detected Not Quantified
Sample ID Mro,,.Ct.d C—olle'�cted Analysis Requested Marker Quantified Result Unit
MST-9 101520 10/15/2020 10:25 AM Human_HF183 ND copies per 100ml
NE3.3NP 101520 10/15/2020 10:40 AM Human_HF183 ND copies per 100ml
SM20J16100
SM20J16101
SM20J16102
NE2.2NP 101520
10/15/2020
11:00 AM
Human_HF183
DNQ
copies per 100ml
SM20J16103
MST-12 101520
10/15/2020
11:25 AM
Human_HF183
DNQ
copies per 100ml
SM20J16104
MST-18 101520
10/15/2020
11:50 AM
Human_HF183
2.97E+03
copies per 100ml
SM20J16105
MST-X 101520
10/15/2020
11:50 AM
Human_HF183
3.65E+03
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 16, 2020
Report Generated: October 28, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCREDITED
Certificate 4120.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20J16100
MST-9 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16101
NE3.3NP 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16102
NE2.2NP 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16103
MST-12101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16104
MST- 18101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16105
MST-X 101520
Water
10/27/2020
10/27/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submitter: Three Oaks Engineering
Report Generated: October 28, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate
was detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents
or representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid
by him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be
maintained by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research
purposes. The client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted
sample(s) to be used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed
in a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is
homogenized for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per
manufacturer's protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory
extraction procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating
target gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

0 Source
%Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 16, 2020
Report Generated: October 28, 2020
%� /^� � ACCRRDITEPI
�hh,\�° cerencateat2s.ot
ISO 17025 Accredited
Testing Laboratory
ND: Not Detected
Sample IDJS We Analysis Requested �rMarker
"011ected Collected
SM20J16106 MST-9 101520 10/15/2020 10:25 AM Dog_DG3 (EPA)
SM20J16107 NE3.3NP 101520 10/15/2020 10:40 AM Dog_DG3 (EPA)
Quantified Result Unit
ND copies per 100ml
ND copies per 100ml
SM20J16108
NE2.2NP 101520
10/15/2020
11:00 AM
Dog_DG3 (EPA)
ND
copies per 100ml
SM20J16109
MST-12 101520
10/15/2020
11:25 AM
Dog_DG3 (EPA)
ND
copies per 100ml
SM20J16110
MST-18 101520
10/15/2020
11:50 AM
Dog_DG3 (EPA)
ND
copies per 100ml
SM20J16111
MST-X 101520
10/15/2020
11:50 AM
Dog_DG3 (EPA)
ND
copies per 100ml
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Three Oaks Engineering
Date Received/Processed: October 16, 2020
Report Generated: October 28, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCREDITED
Certificate 4120.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20J16106
MST-9 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16107
NE3.3NP 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16108
NE2.2NP 101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16109
MST-12101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16110
MST- 18101520
Water
10/27/2020
10/27/2020
100
ml
2
SM20J16111
MST-X 101520
Water
10/27/2020
10/27/2020
100
ml
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submitter: Three Oaks Engineering
Report Generated: October 28, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate
was detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents
or representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid
by him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be
maintained by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research
purposes. The client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted
sample(s) to be used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed
in a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is
homogenized for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per
manufacturer's protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory
extraction procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating
target gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date 8/20/2018
14 Source
Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami Lakes, FL 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Sam le ID
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Mark �� l ll1 t "LsScyit ;
Send Results To (email) -�,Qnl .d't;•[.i+v�•�}�["CC�AI*�l1• .+5 1r-,r f, F�, �� xn�'. C':,*
Phone
Address �a :Z S-U i f C j 2 CJ ()
City/State/Zip
Billing Info FO#: F-1 Will call with credit card
Comments Collection Collection
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Completed by Client; --r..
Relinquished By I G l <' 1
Signature
Date/Time
To protect confidentiality, confirmation and results will only be sent to email address provided
or authorized by contact provided. Signed form indicates agreement with the test llmilalions on
the back of this form and the companVs terms of use found here: sourcemolecular.comlabout-
sourcemotecularlprivacy_statetnentl.
Completed by Source Molecular:
Temperature 7 __ Received/Filtered
Thermometer CK!,I Signature_
Cooler Number Date/Time
,Z
am

(LJ%A_ Data Quality Assurance Worksheet
HERRERA By
Project Name/No./Client: Northeast Creek MST Study / 18-06995-000 / City of Durham Date
Laboratory/Parameters: Pace Analytical Services / Fecal coliforms
Sample Date/Sample ID: 10/15/2020 / MST-9, NE3.3NP, NE2.2NP, MST-18, MST-12, MSTA (field duplicate
of MST-18) (Lab No. 92500686)
G. Catarra
Checked: initials
date
Page 1 of 1
Pre -preservation
Matrix Spikes/
Lab Control
Holding Times
Total Holding
Surrogate
Samples Recovery
Lab Duplicates
Field Duplicates
(minutes)
Times (days)
Method
Recovery (%)
(%)
RPD (%)
RPD (%)
Blanks
Instrument
Completeness/
Reported
Goal
Reported
Goal
Reporting
Reported
Goa
Reported
Goa
Reported
Goal'
Reported
Goal'
Calibration/
Parameter
Methodology
Limit
1
1
Performance
ACTION
<_1.0 cfu/
NONE
Fecal
OK /
NA
NA
1
100mL
NA
NA
NA
NA
MST
< 35
MST-18
< 50
OK
10 cfa/
Coliform
SM 9222D
0
21
100mL
<50
NONE
OK / DG3
co p ies/100
MST -18
Dog
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
NC
< 50
OK
<50
<50
NONE
OK / HF 183
copies/100
MST-18
Human
(EPA)
NA
NA
NA
NA
mL
NA
NA
NA
NA
NA
NA
21
< 50
OK
<50
' If the sample or duplicate value is less than five times the reporting limit, the difference is calculated rather than the relative percent difference (RPD). The QA goal is a difference <2 times the
detection limit instead of the number indicated in the goal column.
NA — not applicable or not available; NC — not calculable due to one or more values below the detection limit; NS — field duplicate not sampled.
MDF https:llherreraine.sharepoint.con✓18-06995-0001shareddocuments/data report/apx c/apxc-7e necreek gawkst 10152020.do"
Herrera Environmental Consultants

Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by quantitative
Polymerase Chain Reaction (gPCR) DNA analytical technology
� 1/aGGREpITEp�
�/eili+\`�CertlOcate 4128.0'1
ISO 1702! Accredited
Testing Laboratory
Submitter: Herrera Environmental
Date Received/Processed: June 24, 2020 ND: Not Detected
Report Generated: July 10, 2020 DNQ: Detected Not Quantified
Sample ID Date I Time Analysis Requested Marker Quantified Result Unit
Collected Collected
SM20F16459 Dog-1 6/23/2020 6:03 AM Do DG3 EPA ND copies per ram
SM20F16460
Dog-2
6/23/2020
7:45 AM
Do DG3 EPA
ND
copies per ram
SM20F16461
Dog-3
6/23/2020
8:20 AM
Do DG3 EPA
ND
copies per ram
SM2
Dog-4
6/23/2020
8:50 AM
Do DG3 EPA
DNQ
copies per gram
SM20F16463
Dog-5
6/23/2020
8:58 AM
Do DG3 EPA
4.75E+07
copies per gram
SM20F16464
Dog-6
6/23/2020
9:24 AM
Do DG3 EPA
ND
copies per gram
SM20F16465
Dog-7
6/23/2020
9:30 AM
Do DG3 EPA
8.99E+05
copies per gram
SM20F16466
Dog-8
6/23/2020
9:45 AM
Do DG3 EPA
4.59E+04
copies per gram
SM20F16467
Dog-9
6/23/2020
10:40 AM
Dog DG3 EPA
3.36E+06
copies per gram
SM20F16468
Do -10
6/23/2020
10:50 AM
Do DG3 EPA
ND
copies per gram
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and included in the analysis.
Revision 1.4
Effective Date 12/12/19
Page 1 of 3

0 Source
Molecular
15280 NW 79th Court, Suite 107 Miami Lakes, Florida 33016
Tel: [phone] Fax: [phone]
Email: [e-mail]
Fecal Host Quantification ID Test Results Report
Detection and quantification of the fecal host associated gene biomarker by
quantitative Polymerase Chain Reaction (gPCR) DNA analytical technology
Submitter: Herrera Environmental
Date Received/Processed: June 24, 2020
Report Generated: July 10, 2020
SM # Sample ID Sample Extraction Analysis
Type Date Date
ACCR ED
Certificate 4126.01
ISO 17025 Accredited
Testing Laboratory
Amount Amount Analytical
Processed Processed Unit Volume (ul)
SM20F16459
Dog-1
Fecal Material
6/26/2020
7/1/2020
0.57
gram
2
SM20F16460
Dog-2
Fecal Material
6/26/2020
7/1/2020
0.51
gram
2
SM20F16461
Dog-3
Fecal Material
6/26/2020
7/1/2020
0.49
gram
2
SM20F16462
Dog-4
Fecal Material
6/26/2020
7/1/2020
0.63
gram
2
SM20F16463
Dog-5
Fecal Material
6/26/2020
7/1/2020
0.58
gram
2
SM201`16464
Dog-6
Fecal Material
6/26/2020
7/1/2020
0.52
gram
2
SM20F16465
Dog-7
Fecal Material
6/26/2020
7/1/2020
0.57
gram
2
SM20F16466
Dog-8
Fecal Material
6/26/2020
7/1/2020
0.48
gram
2
SM20F16467
Dog-9
Fecal Material
6/26/2020
7/1/2020
0.46
gram
2
SM20F16468
Dog-1 0
Fecal Material
6/26/2020
7/1/2020
0.45
gram
2
Reported Results Authorized By: Anda Quintero, Quality Manager
Results reported herein apply only to the sample matrices as received.
Results reported herein relate to the genetic material extracted from the sample matrix processed and
included in the analysis.
Revision 1.4
Effective Date
Page 2 of 3

Laboratory Comments
Submltter: Herrera Environmental
Report Generated: July 10, 2020
Non -Detect Results
In sample(s) classified as non -detect, the host -associated fecal gene biomarker(s) was either not detected in test
replicates, one replicate was detected at a cycle threshold greater than 35 and the other was not, or one replicate was
detected at a cycle threshold less than 35 and the other was not after repeated analysis.
Detected Results
In sample(s) classified as detected, the host -associated fecal gene biomarker(s) was detected in both test replicates
suggesting that the host's fecal contamination is present in the sample(s). Copy number measurements reported
are relative, not absolute, quantification.
Detected Not Quantified (DNQ) Results
In sample(s) classified as Detected Not Quantified (DNQ), the host -associated fecal biomarker was detected in both
test replicates but in quantities below the limit of quantification. This result indicates that fecal indicators associated
with the respective host was present in the sample(s) but in low concentrations.
Fecal Reference Samples
The client is encouraged to submit fecal samples from suspected sources in the surrounding area in order to gain a
better understanding of the concentration of the host -associated biomarker with the regional population. A more
precise interpretation would be available to the client with the submittal of such baseline samples.
Result Interpretations
The presence of the biomarker does not signify the presence or absence of that form of fecal pollution conclusively.
The most reliable way to accurately test for contamination is to combine genetic testing with scientifically sound and
adequate study design appropriate for the water quality questions to be answered or issues to be resolved.
Additional Testing
A portion of all samples has been frozen and will be archived for 3 months. The client is encouraged to perform
additional tests on the sample(s) for other hosts suspected of contributing to the fecal contamination.
Limitation of Damages — Repayment of Service Price
It is agreed that in the event of breach of any warranty or breach of contract, or negligence of Source Molecular Corporation, as well as its agents or
representatives, the liability of the company shall be limited to the repayment, to the purchaser (submitter), of the individual analysis price paid by
him/her to Source Molecular Corp. The company shall not be liable for any damages, either direct or consequential. Source Molecular Corp.
provides analytical services on a PRIME CONTRACT BASIS ONLY. Terms are available upon request. The sample(s) cited in this report may be
used for research purposes after an archiving period of 3 months from the date of this report. Research includes, but is not limited to internal
validation studies and peer -reviewed research publications. Anonymity of the sample(s), including the exact geographic location will be maintained
by assigning an arbitrary internal reference. These anonymous samples will only be grouped by state / province of origin for research purposes. The
client must contact Source Molecular in writing within 10 days from the date of this report if he/she does not wish for their submitted sample(s) to be
used for any type of future research.
DNA Analytical Method Explanation
Water Samples: Each submitted water sample is filtered through 0.45 micron membrane filter(s). Each filter is placed in
a separate, sterile 2ml disposable tube containing a unique mix of beads and lysis buffer. The sample is homogenized
for 1 min and the DNA extracted using the Generite DNA-EZ ST1 extraction kit (GeneRite, NJ), as per manufacturer's
protocol. Devitations to these procedures may occur at the client's request.
Non -Water Samples: Each non -water sample submitted by the client is processed as per internal laboratory extraction
procedures. An extracted DNA sample is proceed directly to PCR analysis. Details available upon request.
Amplifications to detect the target gene biomarker were run on an Applied Biosystems StepOnePlus real-time thermal
cycler (Applied Biosystems, Foster City, CA) in a final reaction volume of 20ul sample extract, forward primer, reverse
primer, probe and an optimized buffer. All assays are run in duplicate. Quantification is achieved by extrapolating target
gene copy numbers from a standard curve generated from serial dilutions of known gene copy numbers.
For quality control purposes, a positive control and a negative control, were run alongside the sample(s) to ensure a
properly functioning reaction and reveal any false negatives or false positives.
Page 3 of 3

Chain Of Custody Record
Revision 1.2
Effective Date 8/2012018
e�MolecularSource
Source Molecular Corporation
15280 NW 79th CT Suite 107 Miami Lakes, Ft 33016
Tel: [phone] Fax: [phone]
Email. [e-mail]
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Completed by Client: ti
Relinquished By
Signature n.
Date/Time 2 7 T I Ij
To protect confidentiality, confirmation and results will only be sent to email address provided
or authorized by contact provided. Signed form Indicates agreement vrith the test limitations on
the back of this form and the companys terms of use found here: scurcemolecular.comlabout-
soufcemolecularlprivacy_statemenV.
Completed by Source Molecular,
Temperature I'.3_G Received/Filtered
Thermometer�Z_ Signature
Cooler Number. �_ Date/Time