2014-2015 Final ReportNEW HANOVER COUNTY WATER QUALITY MONITORING PROGRAM
2014-2015
FINAL REPORT
Prepared by:
Coastal Planning & Engineering of North Carolina, Inc.
Marine Scientist: Brad Rosov, M.Sc.
Prepared For:
New Hanover County, North Carolina
Recommended Citation: Rosov, B., 2015. New Hanover County Water Quality Monitoring Program: 2014-2015 Final Report. New Hanover County, North Carolina: Coastal Planning & Engineering of North Carolina, Inc. 59p.
July 2015
COASTAL PLANNING & ENGINEERING OF NORTH CAROLINA, INC.
EXECUTIVE SUMMARY
This report represents the results of the New Hanover County Water Quality Monitoring
Program between July 2014 and June 2015. Nineteen (19) monitoring stations within seven (7) creeks in New Hanover County were monitored on a monthly basis for physical, chemical, and
biological parameters of water quality. The results presented in this report are described from a
watershed perspective.
In order to provide a quick-glance assessment of the water quality within a particular sampling station and watershed, a rating system has been established for a number of parameters. This
quantitative system assigns a rating of “GOOD”, “FAIR”, or “POOR” to a sampling station
depending on the percentage of samples exceeding the State standard for dissolved oxygen,
turbidity, chlorophyll-a, Enterococci, and fecal coliform bacteria. If the recorded value of a
parameter exceeds the State standard less than 10% of the times sampled, the station will receive a “GOOD” rating for the parameter. A “FAIR” rating is assigned when a parameter exceeds the
State standard 11-25% of the times sampled. Parameters measured that exceed the State
standard more than 25% of the sampling times are given a “POOR” rating.
As displayed in the table below, turbidity and chlorophyll-a were determined to be “good” within all watersheds throughout the study period. Dissolved oxygen was deemed to be “good”
in Motts Creek, Pages Creek, and Smith Creek while Barnards Creek, Futch Creek, and Lords
Creek were all deemed to be “fair”. Prince Georges Creek demonstrated “poor” levels of
dissolved oxygen during the study period. Enterococci was problematic within three of these
watersheds- Motts Creek, Pages Creek, and Prince Georges Creek. On the other hand, Futch Creek, Barnards Creek, and Lords Creek were deemed “good” for enterococci.
Ratings by Watershed
Parameter Barnards
Creek
Futch
Creek
Lords
Creek
Motts
Creek
Pages
Creek
Prince
Georges
Creek
Smith
Creek
Turbidity GOOD GOOD GOOD GOOD GOOD GOOD GOOD
Dissolved Oxygen FAIR FAIR FAIR GOOD GOOD POOR GOOD
Chlorophyll-a GOOD GOOD GOOD GOOD GOOD GOOD GOOD
Enterococci GOOD GOOD GOOD POOR POOR POOR FAIR
Long Term Trends
Using data collected on a monthly basis since at least November 2007, the long term trends of
select water quality monitoring parameters were assessed in this report as well. In general, dissolved oxygen, turbidity, and chlorophyll-a levels oscillate on a seasonal basis. Water quality, as it relates to these parameters, generally decreases during the warmer months when the
water temperatures increase. However, during the cooler months, when the water temperature
drops, these parameters improve.
Since 2007, dissolved oxygen levels exceeded the State standard within surface samples 36%, 23%, 19%, and 10% of the time within Prince Georges Creek, Pages Creek, Futch Creek, and
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COASTAL PLANNING & ENGINEERING OF NORTH CAROLINA, INC.
Smith Creek, respectively. Dissolved oxygen levels were better within Motts Creek where
samples exceeded the dissolved oxygen standard nine (9%) of the time. Barnards Creek and
Lords Creek each breached the standard only five (5%) of the times sampled.
Enterococci bacteria has been a chronic problem within several of the creeks monitored in this
study. Since November 2007, samples collected within Motts Creek exceeded the State standard
for Enterococci 53% of the time while Pages Creek, Barnards Creek, and Smith Creek exceeded
standard 36% of the time. Prince Georges Creek exceeded the standard 31% of the time while
Lords Creek and Futch Creek contained the least amount of bacteria with exceedances only 8% and 3% of the time, respectively.
Turbidity and chlorophyll-a were not problematic in any creeks. Since sampling began, only 20
exceedences of the chlorophyll-a standard were observed of the 1,772 samples collected. The
turbidity standard was only breached five times in total; two from within Smith Creek and three within Pages Creek.
Source Tracking
Separate from the monthly monitoring program, a source tracking study was conducted over the past year to help determine the origins of bacterial contamination within two sites that have demonstrated chronically high levels of bacterial contamination over previous monitoring years.
These sites, located within the Motts Creek and Pages Creek watersheds, were sampled during
rain events on four (4) occasions. High concentrations of Enterococcus bacteria were observed
over the four storms sampled and the array of storm events captured indicating a continued level of elevated concern about the microbial water quality of the waters in question. The source of these bacteria within many of these samples was determined to stem in part from human origins.
Results also indicate that rising groundwater during storm events could be playing a role in the
delivery of fecal contamination to this system possibly via connections among septic leachfields,
or sewage/stormwater conveyance systems. In order to hone in on the source of this anthropogenic contamination, additional studies may be warranted. One such study could include additional source tracking work up-stream from the known areas of contamination. An
alternative approach could use a tracer in the system of the household and follow the transport to
close by receiving waters. This approach would require a volunteer household close to either
Pages or Motts Creek.
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NEW HANOVER COUNTY WATER QUALITY MONITORING PROGRAM
FINAL REPORT
Table of Contents
1.0 Introduction .................................................................................................................................1 1.1 Parameters .............................................................................................................................4
1.2 Standards ...............................................................................................................................6
2.0 Methods.......................................................................................................................................8
2.1 Physical Parameters ..............................................................................................................8
2.2 Biological Parameters ...........................................................................................................8 2.3 Source Tracking ....................................................................................................................9
3.0 Results .........................................................................................................................................10
3.1 Rating System .................................................................................................................................... 10
3.2 Barnards Creek .......................................................................................................................10 3.3 Futch Creek ...........................................................................................................................12 3.4 Lords Creek ...........................................................................................................................17
3.5 Motts Creek ...........................................................................................................................20
3.6 Pages Creek ...........................................................................................................................23
3.7 Prince Georges ......................................................................................................................27 3.8 Smith Creek ..........................................................................................................................31 3.9 Comprehensive Rating by Watershed ...................................................................................36
3.10 Long Term Trends ..............................................................................................................37
3.10.1 Dissolved Oxygen ...................................................................................................37
3.10.2 Turbidity .................................................................................................................41 3.10.3 Chlorophyll-a ..........................................................................................................44 3.10.4 Enterococci .............................................................................................................48
3.11 Source Tracking ..................................................................................................................51
4.0 Discussion and Recommendations .............................................................................................53
5.0 Literature Cited ...........................................................................................................................58
List of Figures
Figure No.
1 Map of New Hanover County and watersheds included in this study ...................................3 2 Water Quality Sites within the Barnards Creek Watershed ...................................................11
3 Dissolved Oxygen at BC-CBR ..............................................................................................12
4 Enterococci at BC-CBR ........................................................................................................12
5 Water Quality Sites with the Futch Creek Watershed ..........................................................14 6 Dissolved Oxygen at FC-4 .....................................................................................................15 7 Dissolved Oxygen at FC-6 .....................................................................................................15
8 Dissolved Oxygen at FC-13 .................................................................................................. 15
9 Dissolved Oxygen at FC-FOY .............................................................................................. 16
10 Enterococci at FC-4 ............................................................................................................... 16
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NEW HANOVER COUNTY WATER QUALITY MONITORING PROGRAM
FINAL REPORT
Table of Contents (Cont’d)
11 Enterococci at FC-6 ...........................................................................................................16 12 Enterococci at FC-13 .........................................................................................................17
13 Enterococci at FC-FOY .....................................................................................................17
14 Water Quality Site within the Lords Creek Watershed .....................................................18
15 Dissolved Oxygen at LC-RR .............................................................................................19 16 Enterococci Levels at LC-RR ............................................................................................19 17 Water Quality Sites within the Motts Creek Watershed ....................................................21
18 Dissolved Oxygen at MOT-CBR .......................................................................................22
19 Dissolved Oxygen at MOT-ND .........................................................................................22
20 Enterococci at MOT-CBR .................................................................................................22 21 Enterococci at MOT-ND ...................................................................................................23 22 Water Quality Sites within the Pages Creek Watershed ....................................................24
23 Dissolved Oxygen at PC-BDDS ........................................................................................25
24 Dissolved Oxygen at PC-BDUS ........................................................................................25
25 Dissolved Oxygen at PC-M ...............................................................................................26 26 Enterococci at PC-BDDS ..................................................................................................26 27 Enterococci at PC-BDUS ..................................................................................................26
28 Enterococci at PC-M..........................................................................................................27
29 Water Quality Sites within the Prince Georges Creek Watershed .....................................28
30 Dissolved Oxygen at PG-CH .............................................................................................29 31 Dissolved Oxygen at PG-ML.............................................................................................29 32 Dissolved Oxygen at PG-NC .............................................................................................30
33 Enterococci at PG-CH .......................................................................................................30
34 Enterococci at PG-ML .......................................................................................................30
35 Enterococci at PG-NC .......................................................................................................31 36 Water Quality Sites within the Smith Creek Watershed ....................................................32 37 Dissolved Oxygen at SC-23 ...............................................................................................33
38 Dissolved Oxygen at SC-CD .............................................................................................33
39 Dissolved Oxygen at SC-CH .............................................................................................34
40 Dissolved Oxygen at SC-GR .............................................................................................34 41 Dissolved Oxygen at SC-NK .............................................................................................34 42 Enterococci at SC-23 .........................................................................................................35
43 Enterococci at SC-CD........................................................................................................35
44 Enterococci at SC-CH........................................................................................................35
45 Enterococci at SC-GR........................................................................................................36 46 Enterococci at SC-NK .......................................................................................................36 47 Long term surface dissolved oxygen data within Barnards Creek ....................................38
48 Long term surface dissolved oxygen data within Futch Creek ..........................................38
49 Long term surface dissolved oxygen data within Lords Creek..........................................39
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FINAL REPORT
Table of Contents (Cont’d)
50 Long term surface dissolved oxygen data within Motts Creek..........................................39
51 Long term surface dissolved oxygen data within Pages Creek..........................................40
52 Long term surface dissolved oxygen data within Prince Georges Creek ..........................40
53 Long term surface dissolved oxygen data within Smith Creek .........................................41
54 Long term surface turbidity data within Barnards Creek...................................................41 55 Long term surface turbidity data within Futch Creek ........................................................42
56 Long term surface turbidity data within Lords Creek ........................................................42
57 Long term surface turbidity data within Motts Creek ........................................................43
58 Long term surface turbidity data within Pages Creek ........................................................43
59 Long term surface turbidity data within Prince Georges Creek ........................................44 60 Long term surface turbidity data within Smith Creek .......................................................44
61 Long term chlorophyll-a data within Barnards Creek .......................................................45
62 Long term chlorophyll-a data within Futch Creek ............................................................45
63 Long term chlorophyll-a data within Lords Creek ............................................................46
64 Long term chlorophyll-a data within Motts Creek ............................................................46 65 Long term chlorophyll-a data within Pages Creek ............................................................47
66 Long term chlorophyll-a data within Prince Georges Creek .............................................47
67 Long term chlorophyll-a data within Smith Creek ............................................................48
68 Long term Enterococci data within Barnards Creek..........................................................49
69 Long term Enterococci data within Futch Creek ...............................................................49 70 Long term Enterococci data within Lords Creek ...............................................................50
71 Long term Enterococci data within Motts Creek ...............................................................50
72 Long term Enterococci data within Pages Creek ...............................................................51
73 Long term Enterococci data within Prince Georges Creek ...............................................51
74 Long term Enterococci data within Smith Creek ..............................................................52 75 Concentrations of molecular microbial source tracking markers in Motts Creek .............53 76 Concentrations of molecular microbial source tracking markers in Pages Creek .............54
77 Long term dissolved oxygen ratings ..................................................................................56
76 Long-term Enterococci ratings ..........................................................................................57
List of Tables
Table No.
1 List of Sampling Sites ............................................................................................................2 2 North Carolina Water Quality Standards ...............................................................................7 3 Single sample standards for Enterococci as determined by the US EPA ..............................7
4 Single sample standards for Enterococci as determined by the NC DENR Recreational
Water Quality Program .........................................................................................................7
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FINAL REPORT
Table of Contents (Cont’d)
5 Tier Classification for New Hanover County Water Quality Monitoring Sites ....................7
6 Mean values of select parameters from Barnards Creek .......................................................11
7 Ratings of parameters within sampling stations within Barnards Creek ...............................12
8 Mean values of select parameters from Futch Creek .............................................................14 9 Ratings of parameters within sampling stations within Futch Creek ....................................17
10 Mean values of select parameters from Lords Creek .............................................................19
11 Ratings of parameters within sampling stations within Lords Creek ....................................19
12 Mean values of select parameters from Motts Creek.............................................................21
13 Ratings of parameters within sampling stations within Motts Creek ....................................23 14 Mean values of select parameters from Pages Creek .............................................................25
15 Ratings of parameters within sampling stations within Pages Creek ....................................27
16 Mean values of select parameters from Prince Georges Creek .............................................29
17 Ratings of parameters within sampling stations within Prince Georges Creek .....................31
18 Mean values of select parameters from Smith Creek ............................................................33 19 Ratings of parameters within sampling stations within Smith Creek ....................................36
20 Ratings of parameters within each watershed ........................................................................37
List of Appendices Appendix No.
A Photographs of Sampling Sites
B Raw Data C Source Tracking Report
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1.0 INTRODUCTION
The creeks in New Hanover County, North Carolina provide a wide range of recreational
activities for thousands of local citizens and visiting tourists each year. Tidal creeks are rich areas in terms of aquatic, terrestrial and avian wildlife and can support complex food webs
(Odum et al, 1984; Kwak and Zedle, 1997). Protection of the water quality within these creeks is
a high priority for New Hanover County. As growth and development continue within the City
of Wilmington and the County, water quality has been increasingly threatened due to many
factors including aging infrastructure, increased impervious surface area and subsequent stormwater runoff. Furthermore, the County’s population in 2014 was estimated to be 209,635
and is expected to grow at a rate of 1.3% over the next 5 years (NC Division of Commerce,
Labor, and Economic Data and Site Information, 2015). To address these issues that impact
water quality, the County, since 1993, has administered a long-standing water quality monitoring
program designed to assess the water quality within the creeks located within the County.
Coastal Planning & Engineering of North Carolina, Inc. began monitoring seven (7) tidal creeks
within New Hanover County on a monthly basis in November 2007. The information presented
in this report represents the results of this monitoring between the months of July 2014 and June
2015. The creeks included in this study are Pages and Futch Creek, which drain into the Atlantic Intracoastal Waterway (ICW) and Lords, Motts, Barnards, Smith, and Prince Georges Creek,
which drain into the Cape Fear River (Figure 1) (Table 1). Thirteen (13) of the nineteen (19)
sampling sites were previously monitored by the University of North Carolina at Wilmington. In
order to assess any changes to historical trends within individual sites and entire watersheds, data
provided by UNCW has been analyzed and incorporated into the results and discussion section of this report. Photographs of each sampling site are found in Appendix A and all raw data is
found in Appendix B.
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Table 1. List of Sampling Sites
Creek Name Site Name Site Code Latitude Longitude
Motts Creek Carolina Beach Road MOT-CBR 34° 08.610 77° 53.830
Motts Creek Normandy Drive MOT-ND 34° 08.373 77° 54.580
Lords Creek River Road LC-RR 34° 05.185 77° 55.275
Barnards Creek Carolina Beach Road BC-CBR 34° 09.522 77° 54.712
Smith Creek Castle Hayne Road SC-CH 34° 15.541 77° 56.325
Smith Creek 23rd Street SC-23 34° 15.472 77° 55.178
Smith Creek Candlewood Drive SC-CD 34° 17.438 77° 51.332
Smith Creek North Kerr SC-NK 34° 15.744 77° 53.256
Smith Creek Gordon Road SC-GR 34° 16.639 77° 52.037
Prince Georges Creek Marathon Landing PG-ML 34° 21.088 77° 55.349
Prince Georges Creek Castle Hayne Road PG-CH 34° 20.675 77° 54.217
Prince Georges Creek North College PG-NC 34° 20.331 77° 53.607
Futch Creek 4 FC-4 34° 18.068 77° 44.760
Futch Creek 6 FC-6 34° 18.178 77° 45.038
Futch Creek 13 FC-13 34° 18.214 77° 45.451
Futch Creek Foy Branch FC-FOY 34° 18.405 77° 45.358
Pages Creek Mouth PC-M 34° 16.209 77° 46.270
Pages Creek Bayshore Drive Down Stream PC-BDDS 34° 16.685 77° 47.673
Pages Creek Bayshore Drive Up Stream PC-BDUS 34° 16.623 77° 48.104
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Figure 1. Map of New Hanover County and watersheds included in this study
The State of North Carolina has employed a series of classifications that apply to all waters in
the State including streams, rivers, and lakes (NC Administrative Code, section 15A NCAC 2B .0200). These classifications are meant to protect the specified uses within waterbodies. These include aquatic life survival and reproduction, secondary recreation, primary recreation,
shellfishing, and water supply. The classifications that apply to the creeks examined in this
study are:
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C Sw: Freshwater that is protected for aquatic life and secondary recreation uses. The
“Sw” supplemental classification indicates that these are swamp waters, and so are likely
to have lower dissolved oxygen and pH than non-swamp streams due to natural
conditions. However, a majority of the sites, including Lords Creek, Motts Creek, Barnards Creek, Smith Creek, and Prince Georges Creek, designated as C Sw by the
State, are tidally influenced and have a brackish salinity range.
SA: Saline water bodies that are protected for shellfishing uses. This use requires a more
stringent standard for fecal coliform. Areas protected for shellfishing are also subject to the protection requirements for the less stringent classifications of SC and SB, which
include aquatic life, secondary recreation, and primary recreation. This designation
applies to Futch Creek and Pages Creek.
1.1 Parameters Physical, chemical, and biological water quality monitoring data are currently being collected for
this study. Physical parameters include temperature, salinity, conductivity, pH, turbidity, and
dissolved oxygen. Chemical parameters monitored in this study include orthophosphate and
nitrate/nitrite. Biological parameters include Chlorophyll-a and two suites of fecal indicator
bacteria: Enterococci and fecal coliform bacteria. Due to limited funding, fecal coliform samples were only collected from sampling sites located within Futch Creek and Pages Creek.
Temperature:
Thermal pollution can result in significant changes to the aquatic environment. Most aquatic
organisms are adapted to survive within a specific temperature range. Thermal pollution may also increase the extent to which fish are vulnerable to toxic compounds, parasites, and disease.
If temperatures reach extremes of heat or cold, few organisms will survive.
Thermal pollution may be caused by stormwater runoff from warm surfaces such as streets and
parking lots. Soil erosion is another cause, since it can cause cloudy conditions in a water body. Cloudy water absorbs the sun's rays, resulting in a rise in water temperature. Thermal pollution
may even be caused by the removal of trees and vegetation which normally shade the water
body. In addition to the direct effects of thermal pollution on aquatic life, there are numerous
indirect effects. Thermal pollution results in lowered levels of dissolved oxygen, since cooler
water can hold more oxygen than warmer water.
Salinity:
Salinity is a measure of the amount of sodium chloride ions dissolved in water. This is important
to monitor since changes in the levels of salt concentration can impact the ability of salt sensitive
species to survive. An estuary, such as the lower Cape Fear River, usually exhibits a gradual change in salinity throughout its length, as freshwater entering the estuary from tributaries mixes with seawater moving in from the ocean. Salinity levels control, to a large degree, the types of
plants and animals that can live in different zones of the estuary. Freshwater species may be
restricted to the upper reaches of the estuary, while marine species inhabit the estuarine mouth.
Some species tolerate only intermediate levels of salinity while broadly adapted species can acclimate to any salinity ranging from freshwater to seawater.
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Conductivity:
Specific conductance is a measure of the ability of water to conduct an electrical current.
Similar to salinity, it measures the amount of dissolved ions (including sodium chloride) in the
water.
pH:
The pH of water is a measurement of the concentration of H+ ions, using a scale that ranges from
0 to 14. Natural water usually has a pH between 6.5 and 8.5. While there are natural variations
in pH, many pH variations are due to human influences. Unanticipated decreases in pH could be indications of acid rain, runoff from acidic soils, or contamination by agricultural chemicals.
Turbidity:
Turbidity is the amount of particulate matter that is suspended in water. Turbidity measures the
scattering effect that suspended solids have on light: the higher the intensity of scattered light, the higher the turbidity. During a rainstorm, particles from the surrounding land are washed into the river making the water a muddy brown color, indicating higher turbidity.
Dissolved Oxygen:
Dissolved oxygen (DO) refers to the volume of oxygen that is contained in water. Oxygen enters the water as rooted aquatic plants and algae undergo photosynthesis and as oxygen is transferred across the air-water interface. The amount of oxygen that can be held by the water depends on
the water temperature, salinity, and pressure.
Rapidly moving water, such as in a flowing stream, tends to contain a lot of dissolved oxygen, while stagnant water contains little. Oxygen levels are also affected by the diurnal (daily) cycle. Plants, such as rooted aquatic plants and algae produce excess oxygen during the daylight hours
when they are photosynthesizing. During the dark hours they must use oxygen for life processes.
Bacteria in water can consume oxygen as organic matter decays. Thus, excess organic material
in waterbodies can cause oxygen deficits. Aquatic life can become stressed or die in stagnant water containing high levels of rotting, organic material in it, especially in summer, when dissolved-oxygen levels are at a seasonal low.
Chlorophyll-a:
Chlorophyll-a is a green pigment found in plants. It absorbs sunlight and converts it to sugar during photosynthesis. Chlorophyll-a concentrations are an indicator of phytoplankton
abundance and biomass in coastal and estuarine waters. High levels often indicate an algal
bloom which can induce the depletion of oxygen in the water column due to the microbial
degradation of plant cells. Chlorophyll-a concentrations are often higher after rainfall,
particularly if the rain has flushed nutrients into the water. Higher chlorophyll-a levels are also common during the summer months when water temperatures and light levels are high because
these conditions lead to greater phytoplankton numbers.
Enterococci: Enterococci are distinguished from fecal coliform bacteria by their ability to survive in saltwater, and in this respect they more closely mimic many pathogens than do the other indicators.
Enterococci are typically more human-specific than the larger fecal streptococcus group. EPA
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recommends Enterococci as the best indicator of health risk in saltwater used for recreation and
as a useful indicator in freshwater as well. In 2004, Enterococci took the place of fecal coliform
as the new federal standard for water quality at public beaches. It is believed to provide a higher
correlation than fecal coliform with many of the human pathogens often found in sewage (Jeng, et al., 2004). Results indicated that Enterococci might be a more stable indicator than fecal
coliform and, consequently, a more conservative indicator under brackish water conditions.
1.2 Standards
Water quality standards have been established legislatively for a number of these parameters (Table 2). Many of the water quality standards are described in the NC Administrative Code,
section 15A NCAC 2H .0100. The water quality standards for Enterococci bacteria are
described by the US EPA (US EPA, 1986) and in the NC Administrative Code, section 15A
NCAC 18A .3402. The US EPA standards for Enterococci bacteria are based on incidents of
gastrointestinal illness following contact with bathing waters. Bacterial contamination is quantified by “colony forming units” or CFU. Single sample maximum allowable Enterococci
density is 104 CFU/100ml, 158 CFU/100ml, 276 CFU/100ml, and 501 CFU/100ml for
designated beach areas, swimming areas with moderate to full body contact, lightly used full
body contact swimming areas, and infrequently used full body contact swimming areas,
respectively (Table 3). When at least five samples are collected within a 30 day period, the US EPA recommends utilizing a geometric mean standard of 35 CFU/100ml. Geometric means are
often useful summaries for highly skewed data, as are often found with bacteriological datasets.
The North Carolina Recreational Water Quality Program (RWQ) adopted similar standards for
Enterococci bacteria, also determined by the frequency of swimming activity. As defined by
RWQ, Tier I swimming areas are used daily during the swimming season, Tier II swimming areas are used three days a week during the swimming season, and Tier III swimming areas are
used on average four days a month during the swimming season. Single sample standards for
Tiers I, II, and III are 104 CFU/100ml, 276 CFU/100ml, and 500 CFU/100ml, respectively
(Table 4). A geometric mean of 35 CFU/100ml within Tier I swimming areas may also be
utilized if at least five samples are collected within 30 days. The creeks included in this study have not been classified within the RWQ tier system; however an analysis of accessibility as an
indicator of swimming and boating usage has been performed (Table 5). Based on this analysis,
of the nineteen (19) sampling sites, two (2) could be considered Tier II and seventeen (17) could
be considered Tier III.
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Table 2. North Carolina Water Quality Standards
Parameter Standard for SA Waters Standard for C Sw Waters
Dissolved Oxygen 5.0 mg/l 4.0 mg/la
Turbidity 25 NTU 50 NTU
pH 6.8-8.5 6.0-9.0b
Chlorophyll-a 40.0 ug/l 40.0 ug/l
Fecal Coliform Geometric Mean (5 samples within
30 days) <14 CFU/100ml; or 10%
of samples <43 CFU/100ml
Geometric Mean (5 samples within
30 days) <200 CFU/100ml; or single
sample <400 CFU/100ml
Enterococci c Geometric Mean (5 samples within
30 days) <35 CFU/100ml
Geometric Mean (5 samples within
30 days) <35 CFU/100ml (a) Swamp waters may have lower values if caused by natural conditions (b) For swamp streams, pH may be as low as 4.3 if caused by natural conditions (c) See Table 4 for single sample standards based off the tiered system employed by NC DENR Recreational Water
Quality Program
Table 3. Single sample standards for Enterococci as determined by the US EPA
Single sample maximum
Designated beach areas < 104 CFU/100ml
Swimming areas with moderate full body contact < 158 CFU/100ml
Lightly used full body contact swimming areas < 276 CFU/100ml
Infrequently used full body contact swimming areas < 501 CFU/100ml
Table 4. Single sample standards for Enterococci as determined by the NC DENR Recreational Water Quality Program
Single sample maximum
Tier I, swimming areas used daily during the
swimming season <104 CFU/100ml
Tier II, swimming areas used three days a week
during the swimming season <276 CFU/100ml
Tier III, swimming areas used on average four days
a month during the swimming season <500 CFU/100ml
Table 5. Tier Classification for New Hanover County Water Quality Monitoring Sites
Site Name
Proposed
Tier
Classification
Accessible for
Boating or
Swimming Comments
MOT-CBR Tier III No Adjacent to culvert off Carolina Beach Road
MOT-ND Tier III No Adjacent to small bridge on Normandy Drive
LC-RR Tier III No Adjacent to bridge on River Road
BC-CBR Tier III No Adjacent to culvert off Carolina Beach Road
SC-CH Tier III No Adjacent to bridge on Castle Hayne Road
SC-23 Tier III No Adjacent to bridge on 23rd Street
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SC-CD Tier III No Narrow, shallow. Adjacent to Candlewood Drive
SC-NK Tier III No Adjacent to bridge on North Kerr
SC-GR Tier III No Adjacent to culvert on Gordon Road
PG-ML Tier III No Small boat launch site on private property
PG-CH Tier III No Adjacent to culvert on Castle Hayne Road
PG-NC Tier III No Adjacent to culvert on North College Road
FC-4 Tier III No Private docks are the only means of direct access
FC-6 Tier III No Private docks are the only means of direct access
FC-13 Tier III No Private docks are the only means of direct access
FC-FOY Tier III No No clear access points (no docks on Foy branch)
PC-M Tier II Yes Direct access via docks and boat ramp at Pages Creek Marina
PC-BDDS Tier III No Private docks are the only means of direct access
PC-BDUS Tier II Yes Public boat ramp off Bayshore Drive
2.0 METHODS
The seven creeks included in this study were selected by County staff and individual sampling
sites were selected by County staff in consultation with Coastal Planning & Engineering of North
Carolina, Inc. These seven creeks are primarily located in the unincorporated portion of New
Hanover County. Sampling sites were accessed from land, generally near a bridge or culvert
crossing, or by boat. Each site was sampled one time per month during a high ebb tide. Tides were determined utilizing the National Oceanic and Atmospheric Administration’s (NOAA)
Tides and Currents website (http://tidesandcurrents.noaa.gov/).
Due to time constraints, monthly sampling events were conducted on three subsequent days each month. Lords Creek, Motts Creek, and Barnards Creek were visited on the first sampling day while Smith Creek and Prince Georges Creek were visited the second day. Futch Creek and
Pages Creek were visited on the third day. Rainfall totals for the 24 hours prior to each sampling
event were obtained from observations recorded at Wilmington International Airport as reported
by NOAA’s National Weather Service web site (http://www.srh.noaa.gov/data/RAH/RTPRAH). 2.1 Physical Parameters All physical measurements (temperature, salinity, conductivity, turbidity, dissolved oxygen, and
pH) were taken in situ utilizing a 6820 YSI Multiparameter Water Quality Probe linked to a YSI
650 MDS display unit. The YSI Probe was calibrated each day prior to use. Physical measurements were taken from the surface at all sites (depth = 0.1m) and near the creek bottom at sites with depths greater than 0.5m. Following each sampling trip, the YSI Probe was post-
calibrated following each sampling date to ensure that the physical parameters measured were
within an acceptable range.
2.2 Biological Parameters Water samples were obtained for the laboratory analysis of biological (Enterococci and
Chlorophyll-a) parameters. These grab samples were collected in sterile bottles during a high
ebb tide from the surface at each site (depth = 0.1m). Water samples were placed on ice
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immediately following collection and were delivered in coolers to Environmental Chemists, Inc.
of Wilmington, North Carolina for analysis. All analyses performed by Environmental
Chemists, Inc. were conducted utilizing the following standard EPA approved methods:
Chlorophyll-a: SM 10200H
Enterococci: EnterolertE
2.3 Source Tracking
Water samples were collected at each of the four sites during or directly following six storm events. Subsamples were collected for the quantification of Enterococci bacteria and for batch
analysis using well described molecular methods to determine the source of the bacteria.
Samples of 100ml were filtered in triplicate through 0.45 μm polycarbonate filters and frozen at -
20 C and delivered to UNC Chapel Hill IMS within two weeks of sample collection. The filters
were subjected to the following molecular analyses:
1. Fecal Bacteroides qPCR (Converse et al. 2009)
2. BacHum analyses (Kildare et al. 2007)
3. Human specific marker for Bacteroides (HF183, Layton et al. 2013)
Quantitative polymerase chain reaction (qPCR) methods were used to conduct the Bacteroides
assays:
• Fecal Bacteroides qPCR assay (Converse et al. 2009) relies on Taqman chemistry and all
the reagents are in a liquid formulation, except the OmniMix. The assay quantifies a
cohort of bacteria found in high concentrations in the human gut, including Bacteroides thetaiotaomicron, Bacteroides distastonis, and Bacteroides fragilis. However, the method is not human specific. The assay has been tested against a range of different fecal samples types, and has been shown to be capable of quantifying over a wide range of
concentrations, and to be sensitive at concentrations relevant to water quality source
tracking studies. When using the qPCR approach for fecal Bacteroides, strong
relationships have been observed with a wide array of human sewage collected from areas on both east and west US coastlines. The assay is highly sensitive and the target bacteria that are enumerated have been shown to be a predictor of human health in both
sand and recreational waters (Wade et. al. 2011, Heaney et al. 2011) during large-scale
EPA-run epidemiology studies. This is a fully quantitative qPCR-based assay that is
being used in an array of studies in stormwater contaminated areas and that, with the use of other additional confirmatory methods, can be used to both identify potential hot spots of human fecal contamination (Converse et al, 2009).
• BacHum Human Marker: A separate qPCR assay was utilized to quantify the BacHum molecular markers reported by Kildare et al., 2007. The assay has been widely tested for
specificity against a range of fecal sample types and has shown high capacity for
discrimination against human and animal fecal types (Ahmed et al., 2009).
• HF (human fecal) 183: Human specific marker by qPCR has been conducted previously by
Bernhard and Field (2000) and updated by Seurinck et al., 2006. This assay is specific to a
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region of ribosomal rDNA within the Bacteroides spp. that is found almost exclusively in
human feces. The assay has been tested repeatedly in a range of different environments for
cross reactivity with other types of fecal material, and various researchers have found either a
90 to 100% ability to discriminate between human and animal feces when using this assay. The assay, however, can be problematic when used alone, because the target copy concentration in fecal material contributed to receiving water environments can be quite low
due to dilution and the assay has a relatively low sensitivity.
3.0 RESULTS
The results described in this report represent the physical, biological, and chemical data collected
from all sampling sites on a monthly basis between July 2014 and June 2015. These results are
primarily organized by watershed. All raw data, including parameters not summarized in this
section, are included in Appendix B.
3.1 Rating System
In order to provide a quick-glance assessment of the water quality within a particular sampling
station or watershed, a rating system for a number of parameters has been employed. This
quantitative system assigns a rating of “GOOD”, “FAIR”, or “POOR” to a sampling station depending on the percentage of samples exceeding the State standard for dissolved oxygen,
turbidity, Chlorophyll-a, Enterococci, and fecal coliform bacteria. If the recorded value of a
parameter exceeds the State standard less than 10% of the times sampled, the station will receive
a “good” rating for the parameter. A “fair” rating is assigned when a parameter exceeds the State
standard 11-25% of the times sampled. Parameters measured that exceed the State standard more than 25% of the sampling times are given a “poor” rating.
3.2 Barnards Creek
The Barnards Creek watershed includes 4,953 acres and is located in the southwestern portion of
the County, just along the City line. The watershed drains portions of Carolina Beach Road at its headwaters and flows towards River Road before entering into the Cape Fear River. Zoning
within the watershed is comprised of a mix of residential and commercial uses. The land is
classified as a mix of transition, urban, and conservation according to the CAMA land use plan.
This watershed contains approximately 16.9% impervious surface coverage (Hume, 2009).
Sampling was conducted at one site (BC-CBR) within the Barnards Creek watershed (Figure 2).
Dissolved oxygen within BC-CBR ranged between 1.6 mg/l and 9.8 mg/l with a mean value of
5.1 mg/l (Table 6). Samples collected during July and August 2014 and June 2015 contained
dissolved oxygen levels below the State standard of 4.0 mg/l for C Sw waters at both the surface
and near the bottom of the water column (Figure 3).
Chlorophyll-a ranged between 1.0 ug/l and 7.0 ug/l with a mean value of 3.0 ug/l at BC-CBR
(Table 6). These values did not approach the 40ug/l standard.
Enterococci ranged between 5 CFU/100ml and 2282 CFU/100ml with a geometric mean value of 42 CFU/100ml, which is below the NCDENR standard of 500 CFU/100ml for Tier III waters
(Figure 4, Table 6). Only one (1) of the twelve (12) samples collected during this period
exceeded this standard.
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Turbidity values were generally good ranging between 1 and 26 NTU with a mean value of 9
NTU (Table 6). No observations exceeded the State standard of 50 NTU for C SW waters.
Table 7 depicts the ratings for these parameters for the watershed.
Figure 2. Water Quality Sites within the Barnards Creek Watershed
Table 6. Mean values of select parameters from Barnards Creek. Range in parentheses.
Parameter BC-CBR
Turbidity (NTU) 9 (1-26)
Dissolved Oxygen (mg/l) 5.1 (1.6-9.8)
Chlorophyll-a (ug/l) 3.0 (1.0-7.0)
Enterococci (#CFU/100ml) 42 (5-2282)1
(1)Enterococci values expressed as geometric mean
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Figure 3. Dissolved Oxygen at BC-CBR
Figure 4. Enterococci at BC-CBR
Table 7. Ratings of parameters within sampling stations within Barnards Creek
Parameter BC-CBR
Turbidity GOOD
Dissolved Oxygen FAIR
Chlorophyll-a GOOD
Enterococci GOOD
3.3 Futch Creek Futch Creek is located on the New Hanover-Pender County line and drains into the Intracoastal
Waterway. The Futch Creek watershed encompasses approximately 3,136 acres extending from
Scotts Hill Loop Road and Highway 17 on the north and east, to Porters Neck Road on the south.
Zoning within the Futch Creek watershed is predominately residential with a small business district along Highway 17. The land within the Futch Creek watershed is classified as watershed resource protection or transition in the CAMA land use plan. This watershed contains
approximately 11.0% impervious surface coverage (Hume, 2009). Sampling was conducted at
four (4) sites (FC-4, FC-6, FC-13, and FC-FOY) within the Futch Creek watershed (Figure 5) on
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eleven (11) occasions over the twelve (12) month study. No samples were collected during
March, 2014 due to mechanical issues with the boat.
Dissolved oxygen within Futch Creek ranged between 4.2 mg/l and 8.9 mg/l with a mean value of 6.5 mg/l (Figures 6-9, Table 8). Samples collected during July and September 2014 dissolved
oxygen levels below the State standard of 5.0 mg/l for SA waters at both the surface and near the
bottom of the water column in FC-FOY and FC-13 (Figures 8 and 9). In addition, the standard
was breached September at FC-6 (Figure 7).
Chlorophyll-a ranged between 1.0 ug/l and 14.0 ug/l with a mean value of 4.0 ug/l (Table 8).
None of these values approached the 40ug/l Chlorophyll-a standard.
Enterococci ranged between 5 CFU/100ml and 243 CFU/100ml with a geometric mean value of
14 CFU/100ml. None of the samples collected within Futch Creek exceeded the NCDENR Enterococci standard of 500 CFU/100ml for Tier III waters (Figures 10-13, Table 8).
Turbidity values were generally low ranging between 0 and 51 NTU with a mean value of 6
NTU (Table 8). One observation exceeded the State standard of 25 NTU for SA waters.
Table 9 depicts the ratings for these parameters for the watershed.
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Figure 5. Water Quality Sites within the Futch Creek Watershed
Table 8. Mean values of select parameters from Futch Creek. Range in parentheses.
Parameter FC-4 FC-6 FC-13 FC-FOY
Turbidity (NTU) 5 (1-19) 4 (0-19) 8 (0-51) 7 (0-31)
Dissolved Oxygen (mg/l) 6.7 (5.4-8.3) 6.7 (4.8-8.3) 6.4 (4.2-8.9) 6.2 (4.2-8.3)
Chlorophyll-a
(ug/l) 3.0 (1.0-6.0) 3.0 (1.0-5.0) 5.0 (1.0-14.0) 4.0 (1.0-14.0)
Enterococci
(#CFU/100ml) 9 (5-52)1 12 (5-161)1 25 (5-243)1 12 (5-109)1
(1)Enterococci values expressed as geometric mean
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Figure 6. Dissolved Oxygen at FC-4
Figure 7. Dissolved Oxygen at FC-6
Figure 8. Dissolved Oxygen at FC-13
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Figure 9. Dissolved Oxygen at FC-FOY
Figure 10. Enterococci at FC-4
Figure 11. Enterococci at FC-6
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Figure 12. Enterococci at FC-13
Figure 13. Enterococci at FC-FOY Table 9. Ratings of parameters within sampling stations within Futch Creek
Parameter FC-4 FC-6 FC-13 FC-FOY
Turbidity GOOD GOOD GOOD GOOD
Dissolved Oxygen GOOD GOOD FAIR FAIR
Chlorophyll-a GOOD GOOD GOOD GOOD
Enterococci GOOD GOOD GOOD GOOD
3.4 Lords Creek
The Lords Creek Watershed is located in the southwestern portion of the County and encompasses approximately 3,047 acres. Zoning within the watershed is completely residential. This watershed contains approximately 12.6% impervious surface coverage (Hume, 2009).
According to the CAMA land use plan, the land in the watershed is classified as a mix of
conservation, transition, watershed resource protection and a small natural heritage resource
protection designation. Sampling was conducted at one (1) site (LC-RR) within the Lords Creek watershed (Figure 14).
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Dissolved oxygen LC-RR ranged between 1.6 mg/l and 11.2 mg/l with a mean value of 6.9 mg/l
(Table 10). All surface samples were within an acceptable level above the State standard of 4.0
mg/l for C Sw waters during both the surface and near the bottom of the water column with the
exception of data collected in August, 2014 and June, 2015 (Figure 15).
Chlorophyll-a ranged between 2.0 ug/l and 21.0 ug/l with a mean value of 6.0 ug/l (Table 10).
No samples exceeded the State standard of 40ug/l for Chlorophyll-a.
Enterococci ranged between 5 CFU/100ml and 275 CFU/100ml with a geometric mean value of 47 CFU/100ml (Table 10). No samples contained high levels of Enterococci beyond the
NCDENR standard of 500 CFU/100ml for Tier III waters.
Turbidity values were generally moderate ranging between 2 and 27 NTU with a mean value of
12 NTU (Table 10). No observations exceeded the State standard of 50 NTU for C Sw waters in Lords Creek during the study period.
Table 11 depicts the ratings for these parameters for the watershed.
Figure 14. Water Quality Site within the Lords Creek Watershed
Table 10. Mean values of select parameters from Lords Creek. Range in parentheses.
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Parameter LC-RR
Turbidity (NTU) 12 (12-27)
Dissolved Oxygen (mg/l) 6.9 (1.6-11.2)
Chlorophyll-a (ug/l) 6 (2.0-21.0)
Enterococci (#CFU/100ml) 47 (5-275)1
(1)Enterococci values expressed as geometric mean
Figure 15. Dissolved Oxygen at LC-RR
Figure 16. Enterococci Levels at LC-RR
Table 11. Ratings of parameters within sampling stations within Lords Creek
Parameter LC-RR
Turbidity GOOD
Dissolved Oxygen FAIR
Chlorophyll-a GOOD
Enterococci GOOD
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3.5 Motts Creek Motts Creek watershed encompasses approximately 2,389 acres and is located in the
southwestern portion of the County, just below Sanders Road. The Creek drains portions of
Carolina Beach Road at its headwaters and then drains toward River Road before entering into the Cape Fear River. Zoning in the watershed is predominately residential with commercial
business districts along Carolina Beach Road. Land in the watershed is classified as transition,
conservation or wetland resource protection according to the CAMA land use plan. This
watershed contains approximately 12.6% impervious surface coverage (Hume, 2009). Sampling
was conducted at two (2) sites (MOT-CBR, MOT-ND) within the Motts Creek watershed (Figure 17).
Dissolved oxygen within Motts Creek ranged between 2.3 mg/l and 12.0 mg/l with a mean value
of 6.9 mg/l (Figures 18 and 19, Table 12). Two samples collected at MOT-CBR contained
dissolved oxygen levels below the standard (May and June 2015) (Figure 18).
Chlorophyll-a ranged between 0.0 ug/l and 7.0 ug/l with a mean value of 3.0 ug/l (Table 12). No
samples exceeded the 40ug/l standard.
Enterococci ranged between 20 CFU/100ml and 19,863 CFU/100ml with a geometric mean value of 419 CFU/100ml (Table 12). MOT-CBR exceeded the NCDENR standard of 500
CFU/100ml for Tier III waters during six (6) of the twelve (12) times it was sampled. MOT-ND
exceeded this standard four (4) of the twelve (12) sample events (Figures 20 and 21).
Turbidity values were generally good ranging between 2 and 31 NTU with a mean value of 10 NTU (Table 12). No turbidity observations exceeded the State standard of 50 NTU for C Sw
waters.
Table 13 depicts the ratings for these parameters for the watershed.
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Figure 17. Water Quality Sites within the Motts Creek Watershed
Table 12. Mean values of select parameters from Motts Creek. Range in parentheses.
Parameter MOT-CBR MOT-ND
Turbidity (NTU) 10 (2-30) 11 (2-31)
Dissolved Oxygen (mg/l) 6.6 (2.3-10.1) 7.2 (4.8-12.0)
Chlorophyll-a (ug/l) 3.0 (0.0-7.0) 3.0 (0.0-6.0)
Enterococci (#CFU/100ml) 399 (20-19863)1 440 (31-7270)1
(1)Enterococci values expressed as geometric mean
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Figure 18. Dissolved Oxygen at MOT-CBR
Figure 19. Dissolved Oxygen at MOT-ND
Figure 20. Enterococci at MOT-CBR
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Figure 21. Enterococci at MOT-ND
Table 13. Ratings of parameters within sampling stations within Motts Creek
Parameter MOT-CBR MOT-ND
Turbidity GOOD GOOD
Dissolved Oxygen GOOD GOOD
Chlorophyll-a GOOD GOOD
Enterococci POOR POOR
3.6 Pages Creek
Located in northeastern New Hanover County and encompassing 2,044 acres, Pages Creek
watershed drains into the Intracoastal Waterway, north of Middle Sound Loop Road. Zoning
within the Pages Creek watershed is predominately residential, with commercial zoning along Highway 17. The land within the Pages Creek watershed is predominately classified as
watershed resource protection and conservation, with a small portion classified as transitional
according to the CAMA land use plan. This watershed contains approximately 23.2%
impervious surface coverage (Hume, 2009). Sampling was conducted at three (3) sites (PC-
BDDS, PC-BDUS, and PC-M) within the Motts Creek watershed (Figure 22).
Dissolved oxygen within Pages Creek ranged between 4.4 mg/l and 8.8 mg/l with a mean value
of 6.6 mg/ (Table 14) (Figures 23 through 25). Of the three (3) sites monitored over the twelve
(12) month study, the dissolved oxygen levels exceeded the standard only twice; once in May
2015 at PC-BDDS and once in June 2015 at PC-BDUS (Figures 23 and 24).
Chlorophyll-a ranged between 1.0 ug/l and 56.0 ug/l with a mean value of 11.0 ug/l (Table 14).
Three (3) sample exceeded the State standard of 40 ug/l for chlorophyll-a.
Enterococci ranged between 5 CFU/100ml and 9,804 CFU/100ml with a geometric mean value of 120 CFU/100ml (Figures 26-28, Table 14). While samples collected from PC-M did not
contain high levels of Enterococci, five (5) and nine (9) samples from PC-BDDS and PC-BDUS,
respectively, contained levels higher than the NCDENR standards.
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Turbidity values were generally good ranging between 0 and 28 NTU with a mean value of 7
NTU (Table 14). Two (2) of the observed turbidity values exceeded the State standard of 25
NTU for class SA waters.
Table 15 depicts the ratings for these parameters for the watershed.
Figure 22. Water Quality Sites within the Pages Creek Watershed
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Table 14. Mean values of select parameters from Pages Creek. Range in parentheses.
Parameter PC-BDUS PC-BDDS PC-M
Turbidity (NTU) 10 (2-26) 5 (0-28) 6 (0-21)
Dissolved Oxygen (mg/l) 5.9 (2.9-8.4) 6.5 (4.3-9.4) 7.3 (4.6-10.0)
Chlorophyll-a (ug/l) 15 (1.0-52.0) 15 (1.0-56.0) 3.0 (1.0-5.0)
Enterococci (#CFU/100ml) 602 (41-9804)1 188 (20-2613)1 15 (5-3968)1
(1)Enterococci values expressed as geometric mean
Figure 23. Dissolved Oxygen at PC-BDDS
Figure 24. Dissolved Oxygen at PC-BDUS
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Figure 25. Dissolved Oxygen at PC-M
Figure 26. Enterococci at PC-BDDS
Figure 27. Enterococci at PC-BDUS
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Figure 28. Enterococci at PC-M
Table 15. Ratings of parameters within sampling stations within Pages Creek
Parameter PC-BDDS PC-BDUS PC-M
Turbidity GOOD GOOD GOOD
Dissolved Oxygen GOOD GOOD GOOD
Chlorophyll-a GOOD GOOD GOOD
Enterococci POOR POOR GOOD
3.7 Prince Georges
Prince Georges Creek drains into the Cape Fear River. The Prince Georges Creek watershed is
approximately 14,589 acres and drains most of Castle Hayne, extending eastward across I-40 into the Blue Clay Road area. Zoning within the Prince Georges Creek watershed is predominately residential with some business and light industrial districts within Castle Hayne.
Most of the land within the Prince Georges Creek watershed is classified as aquifer resource
protection, conservation or transition according to the CAMA land use plan. This watershed
contains approximately 10.1% impervious surface coverage (Hume, 2009). Sampling was conducted at three (3) sites (PG-CH, PG-ML, and PG-NC) within the Prince Georges Creek watershed (Figure 29).
Dissolved oxygen within Prince Georges Creek ranged between 0.0 mg/l and 9.8 mg/l with a
mean value of 4.9 mg/l (Table 16). Surface dissolved oxygen values at PG-NC and PG-CH were below the State standard of 4.0 mg/l for C Sw during seven (7) and three (3) sampling events, respectively. PG-ML was below the standard on two (2) occasions (Figures 30 through 32).
Chlorophyll-a ranged between 0.0 ug/l and 19.0 ug/l with a mean value of 3.0 ug/l (Table 16).
No samples from Prince Georges Creek exceeded the 40ug/l standard. Enterococci ranged between 5 CFU/100ml and 8,164 CFU/100ml with a geometric mean value
of 92 CFU/100ml (Table 16). During this study, four (4) samples from PG-CH and PG-ML,
respectively, contained Enterococci levels above the NCDENR standard of 500 CFU/100ml for
Tier III waters. Two (2) samples from PG-NC exceeded this value during the same time period (Figures 33 through 35).
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Turbidity values were generally good ranging between 1 and 57 NTU with a mean value of 10
NTU (Table 16). One (1) sample exceeded the State standard of 50 NTU for C Sw waters.
Table 17 depicts the ratings for these parameters for the watershed.
Figure 29. Water Quality Sites within the Prince Georges Creek Watershed
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Table 16. Mean values of select parameters from Prince Georges Creek. Range in parentheses.
Parameter PG-CH PG-ML PG-NC
Turbidity (NTU) 13 (1-57) 6 (1-18) 11 (1-25)
Dissolved Oxygen (mg/l) 5.2 (1.2-9.8) 5.8 (2.4-8.8) 3.9 (0.0-8.4)
Chlorophyll-a (ug/l) 1.0 (1.0-4.0) 3.0 (1.0-8.0) 4.0 (1.0-19.0)
Enterococci (#CFU/100ml) 139 (10-1725)1 146 (10-3255)1 38 (5-8164)1
(1)Enterococci values expressed as geometric mean
Figure 30. Dissolved Oxygen at PG-CH
Figure 31. Dissolved Oxygen at PG-ML
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Figure 32. Dissolved Oxygen at PG-NC
Figure 33. Enterococci at PG-CH
Figure 34. Enterococci at PG-ML
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Figure 35. Enterococci at PG-NC
Table 17. Ratings of parameters within sampling stations within Prince Georges Creek
Parameter PG-CH PG-ML PG-NC
Turbidity GOOD GOOD GOOD
Dissolved Oxygen POOR FAIR POOR
Chlorophyll-a GOOD GOOD GOOD
Enterococci POOR POOR FAIR
3.8 Smith Creek
Located in north-central New Hanover County and containing approximately 14,665 acres, the
Smith Creek watershed drains into the lower northeast Cape Fear River, just north of the Isabelle
Holmes Bridge. The watershed drains land within the City limits and the unincorporated County, including the Wilmington International Airport. Zoning within the Smith Creek
watershed is a mix of industrial, residential, and commercial. The land within the watershed is
predominately classified as urban and transition, with a small portion classified as conservation.
This watershed contains approximately 21.9% impervious surface coverage (Hume, 2009).
Along with increased development and impervious surfaces, water quality in Smith Creek has declined in recent years. High bacteria levels have been reported, as well as low dissolved
oxygen levels. As a result, Smith Creek has been listed on the 303(d) list for impaired waters
due to impaired biological integrity. Sampling was conducted at five (5) sites (SC-CH, SC-23,
SC-NK, SC-GR, SC-CD) within the Smith Creek watershed (Figure 36).
Dissolved oxygen within the creek ranged between 3.6 mg/l and 11.5 mg/l with a mean value of
7.2 mg/l (Table 18). Dissolved oxygen levels within SC-23 and SC-CH fell below State standard
of 4.0 mg/l for C Sw waters on one (1) and two (2) occasions, respectively (Figures 37 and 39)).
Chlorophyll-a ranged between 0.0 ug/l and 30.0 ug/l with a mean value of 4.0 ug/l (Table 18).
No samples exceeded the State Standard for chlorophyll-a.
Enterococci ranged between 5 CFU/100ml and 11,199 CFU/100ml with a geometric mean value
of 119 CFU/100ml (Table 18). A number of samples exceeded the NCDENR standard of 500
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CFU/100ml for Tier III waters including five (5) from SC-GR, four (4) from SC-CD, three (3)
from SC-NK, and two (2) from SC-23 (Figures 42 through 46).
Turbidity values were generally good ranging between 1 and 31 NTU with a mean value of 11 NTU (Table 18). No observations exceeded the State standard of 50 NTU for SW class C
waters.
Table 19 depicts the ratings for these parameters for the watershed.
Figure 36. Water Quality Sites within the Smith Creek Watershed
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Table 18. Mean values of select parameters from Smith Creek. Range in parentheses.
Parameter SC-23 SC-CD SC-CH SC-GR SC-NK
Turbidity
(NTU) 11 (4-19) 10 (1-25) 14 (4-31) 10 (2-20) 8 (1-23)
Dissolved
Oxygen (mg/l) 6.6 (3.6-10.2) 8.0 (5.4-9.9) 6.7 (3.6-11.5) 8.0 (5.9-10.7) 6.7 (4.3-11.4)
Chlorophyll-a
(ug/l) 7.0 (2.0-14.0) 2.0 (0.0-5.0) 3.0 (1.0-16.0) 2.0 (1.0-6.0) 9.0 (1.0-30.0)
Enterococci
(#CFU/100ml) 53 (5-2481)1 272 (5-15531)1 50 (5-488)1 251 (10-8164)1 142 (5-11199)1
(1)Enterococci values expressed as geometric mean
Figure 37. Dissolved Oxygen at SC-23
Figure 38. Dissolved Oxygen at SC-CD
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Figure 39. Dissolved Oxygen at SC-CH
Figure 40. Dissolved Oxygen at SC-GR
Figure 41. Dissolved Oxygen at SC-NK
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Figure 42. Enterococci at SC-23
Figure 43. Enterococci at SC-CD
Figure 44. Enterococci at SC-CH
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Figure 45. Enterococci at SC-GR
Figure 46. Enterococci at SC-NK
Table 19. Ratings of parameters within sampling stations within Smith Creek
Parameter SC-23 SC-CD SC-CH SC-GR SC-NK
Turbidity GOOD GOOD GOOD GOOD GOOD
Dissolved Oxygen GOOD GOOD FAIR GOOD GOOD
Chlorophyll-a GOOD GOOD GOOD GOOD GOOD
Enterococci FAIR POOR GOOD POOR POOR
3.9 Comprehensive Rating by Watershed
When combining all results from each site within individual watersheds, it is possible to obtain a
“snapshot” of water quality within each watershed (Table 20). As displayed in the table below,
turbidity and chlorophyll-a were determined to be “good” within all watersheds throughout the study period. Dissolved oxygen was deemed to be “good” in Motts Creek, Pages Creek, and
Smith Creek while Barnards Creek, Futch Creek, and Lords Creek were all deemed to be “fair”.
Prince Georges Creek demonstrated “poor” levels of dissolved oxygen during the study period.
Enterococci was problematic within three of these watersheds- Motts Creek, Pages Creek, and
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Prince Georges Creek. On the other hand, Futch Creek, Barnards Creek, and Lords Creek were
deemed “good” for enterococci.
Table 20. Ratings of parameters within each watershed
Parameter Barnards
Creek
Futch
Creek
Lords
Creek
Motts
Creek
Pages
Creek
Prince
Georges
Creek
Smith
Creek
Turbidity GOOD GOOD GOOD GOOD GOOD GOOD GOOD
Dissolved Oxygen FAIR FAIR FAIR GOOD GOOD POOR GOOD
Chlorophyll-a GOOD GOOD GOOD GOOD GOOD GOOD GOOD
Enterococci GOOD GOOD GOOD POOR POOR POOR FAIR
3.10 Long Term Trends Water quality data has been collected within New Hanover County since the mid 1990’s.
Several of the historical monitoring sites continue to be utilized for the ongoing monitoring
effort. In order to assess the long term trends in water quality, a database has been created to
include the data collected within the seven (7) tidal creeks under current investigation. Prior to 2007, UNCW collected data within three (3) of the tidal creeks included in the present study. These include Pages Creek, Futch Creek, and Smith Creek. Accordingly, the data from these
three creeks dating to 2004 has been incorporated in the analysis of long term trends. The long
term trends from the remaining creeks (Motts Creek, Lords Creek, Prince Georges Creek, and
Barnards Creek) have been derived from data obtained between November 2007 and June 2015. For each parameter examined, data was plotted on a line graph over time and a trend line was
created. Trend lines, also known as regression lines, can be used as a way of visually depicting
the relationship between the independent (x) and dependent (y) variables in the graph. In this
case the independent variable is time and the dependent variable is the water quality parameter. A trend in water quality is defined as an increase or decrease in a particular constituent concentration over time. Statistical analysis was not performed; therefore the significance of
these long term trends should be interpreted with caution.
3.10.1 Dissolved Oxygen Figures 47-53 depicts the long term trends in dissolved oxygen within the seven (7) creeks examined within this study. The figures illustrate a distinct seasonal pattern including higher
dissolved oxygen during the cooler winter months and lower dissolved oxygen during the
warmer summer months. Generally speaking, the dissolved oxygen levels within each creek
have not changed drastically from year to year. The apparent increasing trend line associated with Smith Creek is not necessarily representative of an actual improvement in dissolved oxygen levels due to the fact that sampling was only conducted seasonally between 2004 and 2006
thereby skewing the data. Since 2007, dissolved oxygen levels exceeded the State standard
within surface samples 36%, 23%, 19%, and 10% of the time within Prince Georges Creek,
Pages Creek, Futch Creek, and Smith Creek, respectively. Dissolved oxygen levels were better within Motts Creek where samples exceeded the dissolved oxygen standard nine (9%) of the
time. Barnards Creek and Lords Creek each breached the standard only five (5%) of the times
sampled.
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Figure 47. Long term surface dissolved oxygen data within Barnards Creek
Figure 48. Long term surface dissolved oxygen data within Futch Creek
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Figure 49. Long term surface dissolved oxygen data within Lords Creek
Figure 50. Long term surface dissolved oxygen data within Motts Creek
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Figure 51. Long term surface dissolved oxygen data within Pages Creek
Figure 52. Long term surface dissolved oxygen data within Prince Georges Creek
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Figure 53. Long term surface dissolved oxygen data within Smith Creek
3.11.2 Turbidity
Figures 54-60 depict the long term trends in turbidity within the seven (7) creeks examined
within this study. In general, the long term trend of turbidity has remained fairly constant within each creek on an annual basis, however seasonal patterns emerge. This includes higher turbidity
observations during the warmer months and lower turbidity during the cooler months. The
turbidity within Smith Creek has demonstrated a slight decrease over time while turbidity in
Prince Georges Creek has increased slightly. Turbidity within all other creeks included within
the study have maintained roughly the same level since 2007, however, these long term turbidity trends have not been verified to be statistically significant.
Figure 54. Long term surface turbidity data within Barnards Creek
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Figure 55. Long term surface turbidity data within Futch Creek
Figure 56. Long term surface turbidity data within Lords Creek
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Figure 57. Long term surface turbidity data within Motts Creek
Figure 58. Long term surface turbidity data within Pages Creek
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Figure 59. Long term surface turbidity data within Prince Georges Creek
Figure 60. Long term surface turbidity data within Smith Creek
3.11.3 Chlorophyll-a Figures 61-67 depict the long term trends in chlorophyll-a within the seven (7) creeks examined
within this study. In general, the long term trend of turbidity has remained fairly constant within
each creek. Similar to the trend observed with dissolved oxygen, chlorophyll-a levels appear to
increase during the warmer months and decrease during the cooler months. Since sampling began, only 20 exceedences of the chlorophyll-a standard were observed of the 1,772 samples collected.
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Figure 61. Long term chlorophyll-a data within Barnards Creek
Figure 62. Long term chlorophyll-a data within Futch Creek
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Figure 63. Long term chlorophyll-a data within Lords Creek
Figure 64. Long term chlorophyll-a data within Motts Creek
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Figure 65. Long term chlorophyll-a data within Pages Creek
Figure 66. Long term chlorophyll-a data within Prince Georges Creek
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Figure 67. Long term chlorophyll-a data within Smith Creek
3.10.4 Enterococci
Figures 68-74 depict the long term trends in Enterococci within the seven (7) creeks examined
within this study. Motts Creek, Smith Creek, Barnards Creek, and Prince Georges Creek have all maintained a relatively high level of bacteria over time. Pages Creek and Futch Creek contain
levels of bacteria which have apparently increased within recent years while the levels of
bacteria within Smith Creek and Barnards Creek have decreased slightly. Lords Creek and Futch
Creek which, on average, have contained relatively lower bacteria levels compared to the other
creeks included within this study. Since November 2007, samples collected within Motts Creek exceeded the State standard for Enterococci 53% of the time while Pages Creek, Barnards Creek,
and Smith Creek exceeded standard 36% of the time. Prince Georges Creek exceeded the
standard 31% of the time while Lords Creek and Futch Creek contained the least amount of
bacteria with exceedances only 8% and 3% of the time, respectively.
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Figure 68. Long term Enterococci data within Barnards Creek
Figure 69. Long term Enterococci data within Futch Creek
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Figure 70. Long term Enterococci data within Lords Creek
Figure 71. Long term Enterococci data within Motts Creek
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Figure 72. Long term Enterococci data within Pages Creek
Figure 73. Long term Enterococci data within Prince Georges Creek
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Figure 74. Long term Enterococci data within Smith Creek 3.11 Source Tracking
High concentrations of Enterococcus bacteria were observed over the four storms sampled
indicating a continued level of elevated concern about the microbial water quality in the sites included within this source tracking study (MOT-ND and PC-BDDS). Enterococcus
concentrations ranged from 54.6 to >24196 MPN/100 ml, with 93% of the samples exhibiting
Enterococcus concentrations that exceeded EPA single sample standards for recreational waters.
This compares with the 96% observed exceedance in the first study, and demonstrates that
patterns were roughly the same. Rainfall amounts for the storms sampled ranged from 0.5-1.6 inches over the course of the study. These are rainfall amounts that are typical of storms in the
Wilmington, NC area, and would not be expected to be categorized as an extreme event. In
Motts Creek, the mean Enterococcus sp. concentration over all samples collected was 848
MPN/100 ml, while at Pages Creek, it was over 5-fold higher at 5204 MPN/100 ml.
A trio of Bacteroides-based molecular markers were quantified in order to assess the potential for
the presence of human fecal contamination at the four sites studied. A notable result was from
the smallest of the four storms, where the 3rd sample collected (2/17/2015) at both Motts Creek
(Figure 75) and Pages Creek (Figure 76) resulted in contamination for all four markers
measured. While this was a small storm, and only a single storm, these results indicate that the tail of the storm event is important. This also indicates that rising groundwater, and subsequent connections among septic leachfields, or sewage/stormwater conveyance systems could be
playing a role in the delivery of fecal contamination to this system. All three human markers
were positive, and the gull fecal contamination marker was also positive. The HF183 marker,
along with the Fecal Bacteroides spp. marker were also both strongly positive at the beginning (Sample 1) for the storm from 1/12/2015, a rainfall event of our 0.6 inches total rainfall. For the storm event of March 17-18, 2015 (see Figure 75 and 76), the HF183 marker was positive in
both Sample 1 and Sample 2, occurring with a steady spring-like rainfall event of a total of 1.3
inches. The other human markers during this event were non-detectable for Pages Creek, but in
Motts Creek for this single storm the BacHum results corroborated the HF183 results.
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It is interesting to note that several of the other samples collected over the course of this study
did not demonstrate any indication of a human fecal contamination signal. The human fecal
contamination signal is indeed ephemeral and appears to be driven specifically by rainfall patterns. Methodological limitations prevented accurate quantification of the gull marker in a
few instances, because the water sample collected and purified had to be diluted in order to gain
a proper qPCR signal from the sample. This is called “qPCR inhibition”. Inhibition is typically
caused by large high molecular weight molecules such as humic and fulvic acids, which are
present in high quantities in the Creeks studied here.
Over the duration of all four storms, samples obtained from both sites contained very high
concentrations of the Fecal Bacteroides spp. concentrations with concentrations well in excess of
100,000 CE/100 ml (Figures 75 and 76). Most of the samples that demonstrated very high Fecal
Bacteroides spp. concentrations also had high concentrations of the BacHum marker, ranging from 4,120 to 15,378 gene copies/100 ml (Figures75 and 76). There was no statistical
relationship between Bacteroides-based molecular markers and the Enterococcus concentrations
or total rainfall, but this study includes a small sample size, upon which it is often difficult to
conduct statistical analyses appropriately (p > 0.05, correlation analysis). This is a small study,
and the sites were selected based upon previous historical data showing contamination. All of the sites indicate that human fecal contamination has the potential to pose a serious problem to
these receiving waters.
Figure 75. Concentrations of molecular microbial source tracking markers Fecal Bacteroides spp.,
BacHum, HF183, and Gull-2 gene copies/100 ml for four storm events in Motts Creek, NC. Note axis is on logarithmic scale.
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Figure 76. Concentrations of molecular microbial source tracking markers Fecal Bacteroides spp., BacHum, HF183, and Gull-2 gene copies/100 ml for four storm events in Pages Creek, NC. Note axis is on logarithmic scale.
4.0 DISCUSSION AND RECOMMENDATIONS
Water quality is an important issue in the region due to the fact that there are many economic and recreational opportunities that are supported by the aquatic resources in and around these
waterways. One of the greatest threats to water quality in this area is stormwater runoff created
by increased impervious surface coverage (Mallin et al., 2000). Due to many of the
contaminants found in stormwater runoff, adverse effects can be imposed upon plants, fish, animals and people. Excess nutrients can cause algal blooms while bacteria and other pathogens can wash into swimming areas and create health hazards. New Hanover County has experienced
rapid growth and development over the past several decades. In 1990, the population within the
County was 120,284. By 2006, the population grew over 50% to 182,591 (U.S. Census Bureau,
2006). The County’s population in 2012 was estimated to be 209,635 and is expected to grow at a rate of 1.3% over the next 5 years (NC Division of Commerce, Labor, and Economic Data and Site Information, 2015). Along with this population growth came increased stormwater runoff,
increase in septic tanks, aging wastewater infrastructure, and other issues that potentially
impacted the water quality within the County’s creeks. Since this time, New Hanover County’s
water quality within its tidal creeks has become altered. This has led to a strong community desire for greater protection and enhancement of surface and ground water resources. The County is working towards providing the Marquis Hill subdivision within the Motts Creek
watershed with sewer service which will replace a number of failing septic systems within the
neighborhood. Furthermore, the County continues to work toward preventing further
deterioration and loss of public uses in surface water through initiatives such as riparian buffer land acquisition projects and promoting low impact development. With this in mind, it is important to monitor the water quality of these local systems to determine potential impacts to
both human health and ecosystem function.
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Typically, water quality degrades as the water temperature increases and oxygen is not as readily
dissolved in the water column. This was observed while investigating the long term trends of
water quality in this study. The dissolved oxygen along with chlorophyll-a and turbidity levels
increased during the warmer summer months. Furthermore, longer days allow for increased photosynthetic activity allowing for an increase in phytoplankton blooms. While often more
problematic in the summer months, algal blooms are less common in the fall and winter when
water temperature decreases. High levels of chlorophyll-a and nutrients along with increases in
pH and turbidity may indicate the presence of an algal bloom. Throughout the course of this
study, pH values were generally found to be within acceptable ranges as were turbidity values. However, two sites in Pages Creek (PC-BDUS and PC-BDDS) contained high levels of
chlorophyll-a above the State standard. The turbidity at these sites was relatively high, although
within the State standard. Accordingly, it is possible that an algal bloom was occurring within
the creek during that sampling period. This would mark the first algal bloom identified within
the network of creeks included within this monitoring program since 2007.
Fourteen percent (14%) of all samples collected during the 12 month study contained dissolved
oxygen levels below the State standard. Of the 31 samples that fell below this standard, two-
thirds (67%) were observed during June, July, and August when water temperatures were the
highest. The lowest dissolved oxygen, on average, was observed at PG-NC where the standard was breached seven (7) of twelve (12) sampling events. This portion of the creek is
characterized by a broad shallow bank in a swamp-like setting. It is typical of swamps to contain
low levels of dissolved oxygen and higher levels of pH, as observed. Therefore, the low
dissolved oxygen observed in Prince Georges Creek, particularly at PG-NC, could be regarded as
a natural phenomenon. Compared to last year, the DO levels within Barnards Creek and Lords Creek declined from a “Good” rating to a “Fair” rating while all other creeks remained the same.
Lords Creek was the only creek that did not include a sample below this level.
High levels of Enterococci bacteria persisted within three (3) of the seven (7) watersheds
throughout the study period. Enterococci levels exceeded the State standard in individual sampling sites within Barnards Creek, Smith Creek, Prince Georges Creek, Motts Creek, and
Pages Creek 8%, 23%, 28%, 42%, and 42% of the time, respectively. The sites with the most
frequent high concentrations of Enterococci bacteria were located within Pages Creek at BDUS
where nine (9) of the twelve (12) samples obtained at each sample exceeded the State standard.
The other site located within the Bayshore community at pages Creek is at the neighborhood boat ramp (PC-BDUS). Five (5) of the twelve (12) samples collected at this site exceeded the
State standard. Collectively, these two sampling sites in the Bayshore neighborhood exceeded
the standard 58% of the time. Samples collected at MOT-CBR also contained high levels of
Enterococci on a consistent basis as six (6) of the twelve (12) sampling events exceeded the
standard. Over the past several years, two sites have continued to improve their water quality in terms of bacterial contamination: SC-CD and MOT-ND. SC-CD exceeded the standard ten (10)
times during 2012-2013 and only four (4) times the past two years. MOT-ND also exceeded the
standard ten (10) times in 2012-2013 but improved to only six (6) exceedences this during 2013-
2014 and four (4) times this past year. Samples collected from Futch Creek, Barnards Creek,
and Lords Creek demonstrated the best water quality in terms of bacteria. No samples containing high levels of enterococcus bacteria were collected from Futch Creek and Lord Creek
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while with only one (1) sample from Barnards Creek contained high levels of Enterococci above
the State standard.
In general, the water quality ratings ascribed for each creek was the same between the 2013-2014 study period and this period were similar with only two exceptions. The dissolved oxygen rating
declined from “Good” to “Fair” at both Barnards and Lords Creek. Aside from these two
changes, all other ratings within the creeks remained unchanged.
An assessment of the past seven years of water quality monitoring has revealed some long term trends regarding the ratings for dissolved oxygen and Enterococci bacteria within each creek. In
general, the dissolved oxygen within Barnards Creek, Lords Creek, and Smith Creek has been
rated “Good” over time, with few exceptions (Figure 77). Both Barnards Creek and Lords Creek
demonstrated a “Fair” rating this past year, however. Futch Creek has maintained a “Fair” rating
for six (6) of the eight (8) years with one year rated as “Poor” and one as “Good”. Motts Creek has shown improvement over the past three (3) years while Pages Creek seems to have improved
to “Good” after several years of “Poor” dissolved oxygen. Prince Georges Creek has maintained
"Poor" dissolved oxygen for this long term period with the exception of one “Fair” rating in
2007-2008 (Figure 77).
Figure 77. Long-term dissolved oxygen ratings
The long term trends for enterococci ratings over the past seven years have shown that a number
of creeks have basically maintained “Poor” ratings. These include Motts Creek, Pages Creek, Prince Georges Creek, and Smith Creek. Smith Creek, however, has improved over the past two
(2) years with a “Fair” rating. Barnards Creek and Lords Creek have demonstrated various
conditions over the past seven years while Futch Creek has maintained a “Good” rating
consistently (Figure 78).
Good
Fair
Poor
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Figure 78. Long-term Enterococci ratings
Results of the source tracking study have suggested that the impairment of water quality at the
sites examined (PC-BDDS and MOT-ND) is complex, and likely driven by the presence of
sources of 1) human fecal contamination, 2) bird fecal contamination, and 3) potential regrowth
of fecal indicator bacteria in the organic matter rich estuarine and creek systems. While human
fecal contamination has been determined to be present, the signal is ephemeral, meaning that it is driven by a series of complex hydrological factors. It is difficult to pinpoint these factors with
small scale study efforts such as what has been performed. Further targeted upstream sampling
efforts would need to follow one of two main approaches to be fruitful. The first potential
approach would be to begin to conduct adaptive sampling for storms in 5-10 sampling locations
to hone in on upstream “hot spots”. This is a tried and true water quality sampling approach. The same sites currently being sampled would be tested, and in addition, sites upstream that represent
possible areas where illicit stormwater/sewage conduit connections might exist would also be
targeted with high spatial frequency (i.e. sampling at all potential locations of source input). In
Motts Creek, where a septic system to sewerage system transformation is in the process of taking
place, it would be most useful to conduct this assessment after the installation of sewage treatment. In Pages Creek, there may be sewage lines that are exfiltrating during wet weather.
During those periods that contamination can become connected with stormwater outflows,
causing impairment of coastal waters.
A second recommendation would be to conduct a limited set of tracer studies using tracer viruses in the local septic systems. This approach would require a volunteer household close to either
Pages or Motts Creek. The approach would be to use a tracer in the system of the household and
follow the transport to close by receiving waters. This would be particularly applicable in areas
where a mixture of septic and sewage systems exist, and where it is desired to hone in on specific
sources of human fecal contamination to the estuary.
Good
Fair
Poor
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5.0 LITERATURE CITED
Ahmed, W., A. Goonetilleke, D. Powell and T. Gardner. 2009. Evaluation of multiple sewage-
associated Bacteroides PCR markers for sewage pollution tracking. Water Research 43(19):4872-4877.
Bernhard, A.E. and K. G. Field. 2000. A PCR assay to discriminate human and ruminant feces on
the basis of host differences in Bacteroides-Prevotella genes encoding 16S rRNA. Applied and
environmental microbiology. 66(10):4571-4574.
Converse, R.R., J.F. Griffith, and R.T. Noble (2009) Rapid QPCR-based assays for fecal
Bacteroides and Enterococcus speciation as tools for assessing fecal contamination in
recreational waters. J Water Research. 43:4828-4837.
Grizzard, T.J., Randall, C.W., Helsel, D.R., and Hartigan, J.P. 1980. Analysis of non-point
pollution export from small catchments. Journal of Water Pollution Control Federation, 52:
780-790.
Howarth, R.W. and Marino, R. 2006. Nitrogen as the limiting nutrient for eutrophication in coastal marine ecosystems: Evolving views over three decades. Limnology and Oceanography,
51: 364-376.
Hume, A. 2008. Determination of Impervious Surface in New Hanover County, North Carolina.
Report submitted to New Hanover County. Wilmington, North Carolina.
Jeng, J.G., Bradford, H, and Englande, A.J. 2004. "Comparison of E. coli, enterococci, and fecal
coliform as indicators for brackish water quality assessment". Water Environmental Research.
76: 245–55.
Kelsey, H., Porter, D.E, Scott, G., Neet, M., and White, D. 2004. Using geographic information systems and regression analysis to evaluate relationships between land use and fecal coliform
bacterial pollution. Journal of Experimental Marine Biology and Ecology. 298:197-209.
Kildare, B.J., C.M. Leutenegger, B.S. McSwain, D.G. Bambic, V.B. Rajal and S. Wuertz. 2007.
16S rRNA-based assays for quantitative detection of universal, human-, cow- and dog-specific fecal Bacteroidales: a Bayesian approach. Water Research. 41(16):3701–3715.
Kwak, T.J. and Zedler, J.B. 1997. Food web analysis of southern California coastal wetlands
using multiple stable isotopes. Oecologia 110: 262–277.
Mallin, M.A.; Williams, K.E.; Esham, C.E.; and Lowe, P.R., 2000. Effect of human
development on bacteriological water quality in coastal watersheds. Ecological Applications
10:1047-1056.
Mallin, M.A., Ensign, S.H., McIver, M.R., Shank, G.C., and Fowler, P.K. 2001. Demographic, landscape, and meteorological factors controlling the microbial pollution of coastal waters.
Hydrobiologia. 460: 185-193.
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Mallin, M.A., 2010. University of North Carolina at Wilmington, Aquatic Ecologist. Personal
communication regarding findings of water samples obtained within PG-NC.
NC Division of Commerce, Labor, Economic Data and Site Information. 2015. Thrive in North
Carolina, County Demographics Report.
http://accessnc.commerce.state.nc.us/docs/countyProfile/NC/37129.pdf. Last visited July 8,
2015.
Odum, W.E., Smith, T.J., Hoover, J.K., and McIvor, C.C. 1984. The Ecology of Tidal Freshwater Marshes of the United States East Coast: A Community Profile. U.S. Fish and
Wildlife Service FWS/OBS-83/17, 177 pp.
Ricks, C., 2011. Cape Fear Public Utility Authority. Personal communication regarding sewage spills in New Hanover County.
Schueler, T., 1994. The importance of imperviousness. Water Protection Technology. 1: 100-
111.
Spivey, 2008. The use of PCR and T-RFLP as a means of identifying sources of fecal bacteria pollution in the tidal creeks of New Hanover County, North Carolina. Masters Thesis. University of North Carolina at Wilmington. 54pp.
U.S. Census Bureau, 2006 Population Estimates, Census 2000, 1990 Census.
U.S. Environmental Protection Agency. 1984. Health effects criteria for fresh recreational waters. EPA-600/1-84-004, U.S. Environmental Protection Agency, Washington, D.C.
U.S. Environmental Protection Agency. 1986. Ambient Water Quality Criteria for Bacteria-
1986. EPA-440/5/84-002, U.S. Environmental Protection Agency, Washington, D.C. Wade, T. J., Sams, E., Brenner, K. P., Haugland, R., Chern, E. Beach, M., Wymer, L., Rankin,
C. C., Love, D., Li, Q., Noble, R., and A.P. Dufour. 2010. Rapidly measured indicators of
recreational water quality and swimming-associated illness at marine beaches. Journal of
Environmental Health Perspectives. 9:66-80.
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APPENDIX A
Photographs of Sampling Sites
Barnards Creek at Carolina Beach Road (BC-CBR)
Futch Creek 4 (FC-4)
Futch Creek 6 (FC-6)
Futch Creek 13 (FC-13)
Futch Creek at Foy Branch (FC-FOY)
Lords Creek at River Road (LC-RR)
Motts Creek at Carolina Beach Road (MOTT-CBR)
Motts Creek at Normandy Drive (MOT-ND)
Pages Creek at Bayshore Drive Upstream (PC-BDUS)
Pages Creek at Bayshore Drive (PC-BDDS)
Pages Creek Mouth (PC-M)
Prince Georges Creek at Castle Hayne Road (PG-CH)
Prince Georges Creek at Marathon Landing (PG-ML)
Prince Georges Creek at North College Road (PG-NC)
Smith Creek at Candlewood Drive (SC-CD)
Smith Creek at Castle Hayne Road (SC-CH)
Smith Creek at 23rd Street (SC-23)
Smith Creek at North Kerr Ave. (SC-NK)
Smith Creek at Gordon Road (SC-GR)
APPENDIX B
Raw Data
APPENDIX B
2014-2015 Raw Data
Date Site Depth Temp. Cond. Salinity DO mg/L DO% pH Turb. Entero. Chl-a
7/10/14 BC-CBR 0.1 25.9 240 0.1 2.1 26% 8 6 108 2
7/10/14 BC-CBR 1.5 25.7 225 0.1 1.8 26% 8 6 N/A N/A
7/10/14 LC-RR 0.1 27.8 28328 16.4 4.8 68% 6.5 27 5 21
7/10/14 LC-RR 1.8 27.9 28468 16.5 4.8 68% 6.6 27 N/A N/A
7/10/14 MOT-CBR 0.1 25.7 182 0.1 5.4 70% 7.5 29 2495 7
7/10/14 MOT-ND 0.1 25.4 248 0.1 5.3 65% 7.5 31 7270 6
7/11/14 PG-CH 0.1 24.3 254 0.1 3.4 41% 7.9 42 1169 4
7/11/14 PG-CH 1.8 24.3 253 0.1 3.4 41% 7.7 57 N/A N/A
7/11/14 PG-ML 0.1 26.4 242 0.1 3.7 47% 8.4 3 420 3
7/11/14 PG-NC 0.1 23.6 148 0.1 1.9 22% 7.3 14 663 6
7/11/14 PG-NC 3.6 20.7 335 0.2 0.5 5% 7.9 11 N/A N/A
7/11/14 SC-23 0.1 27.9 1975 0.9 4.3 55% 8.2 12 373 13
7/11/14 SC-23 2.1 27.9 2018 1 4.3 55% 8.1 12 N/A N/A
7/11/14 SC-CD 0.1 24.7 150 0.1 6.5 78% 7.7 12 441 2
7/11/14 SC-CH 0.1 28.1 12447 6.7 4.3 57% 7.8 10 359 6
7/11/14 SC-CH 2.4 28.1 13034 7 4.1 54% 7.8 23 N/A N/A
7/11/14 SC-GR 0.1 24.4 129 0.1 6.2 73% 7.7 15 563 2
7/11/14 SC-NK 0.1 25.6 287 0.1 4.6 56% 7.7 10 1169 30
7/11/14 SC-NK 1.8 25.6 283 0.1 4.5 55% 7.7 11 N/A N/A
7/12/14 FC-13 0.1 28.7 56878 34.6 4.9 76% 8.2 12 10 7
7/12/14 FC-13 0.9 28.7 56350 34.6 4.9 76% 8.3 12 N/A N/A
7/12/14 FC-4 0.1 28.3 56310 34.7 6 95% 8.7 6 5 4
7/12/14 FC-4 1.5 28.1 56110 34.7 6.2 96% 8.7 5 N/A N/A
7/12/14 FC-6 0.1 28.6 56317 34.7 5.6 87% 8.6 8 5 5
7/12/14 FC-6 1.3 28.4 56296 34.7 5.7 89% 8.6 9 N/A N/A
7/12/14 FC-FOY 0.1 28.7 54706 33.5 4.7 73% 8.4 10 5 9
7/12/14 FC-FOY 1.2 28.7 55314 33.9 4.8 74% 8.4 31 N/A N/A
7/12/14 PC-BDDS 0.1 28.5 52760 32 5.3 82% 8.4 8 2613 24
7/12/14 PC-BDUS 0.1 28.1 43006 25.8 5.4 80% 8.4 13 689 30
7/12/14 PC-M 0.1 28.4 56071 34.6 6.3 86% 8.5 2 20 5
7/12/14 PC-M 1.7 28.4 56161 34.7 6.3 86% 8.6 7 N/A N/A
8/8/14 BC-CBR 0.1 24.3 177 0.1 3.8 45% 7.3 5 223 2
8/8/14 BC-CBR 2.2 24.3 175 0.1 3.6 43% 7.1 26 N/A N/A
8/8/14 LC-RR 0.1 26.8 11666 6.4 3.5 46% 6 19 163 5
8/8/14 LC-RR 1.8 26.8 11628 6.4 3.5 46% 6 19 N/A N/A
8/8/14 MOT-CBR 0.1 25.1 208 0.1 5.6 68% 6.8 12 717 4
8/8/14 MOT-ND 0.1 24.8 208 0.1 6.9 83% 6.8 14 1305 3
8/11/14 FC-13 0.1 26 32632 20.7 5.1 70% 7.1 10 110 14
8/11/14 FC-13 1.1 26 33216 20.4 5 70% 7.2 14 N/A N/A
8/11/14 FC-4 0.1 26.3 39459 24.6 6.4 90% 7.5 6 10 5
8/11/14 FC-4 1.5 36.3 41633 26 6.3 89% 7.5 6 N/A N/A
8/11/14 FC-6 0.1 26.1 36722 22.6 6.2 86% 7.3 8 30 5
8/11/14 FC-6 1.5 26.2 37477 23.2 5.7 81% 7.3 10 N/A N/A
8/11/14 FC-FOY 0.1 26 33337 20 5.1 71% 7.2 10 86 14
8/11/14 FC-FOY 1.5 26 34498 21.2 5 70% 7.2 11 N/A N/A
8/11/14 PC-BDDS 0.1 25.8 35091 21.7 5.5 77% 7.3 10 697 38
8/11/14 PC-BDUS 0.1 25 11899 6.9 5.4 70% 7.7 20 1259 5
8/11/14 PC-M 0.1 26.4 45860 28.7 6.6 96% 7.4 7 63 4
8/11/14 PC-M 1.8 26.5 45909 28.8 6.3 92% 7.5 8 N/A N/A
8/12/14 PG-CH 0.1 24.7 105 0.1 4.5 54% 7 10 728 3
8/12/14 PG-CH 1.7 24.5 105 0.1 4.5 54% 7 10 N/A N/A
8/12/14 PG-ML 0.1 25.5 98 0 5.2 63% 7.2 9 1063 8
8/12/14 PG-NC 0.1 24.3 94 0 3.9 46% 6.4 9 369 5
8/12/14 PG-NC 3.7 24.3 96 0 3.9 46% 6.3 16 N/A N/A
8/12/14 SC-23 0.1 25 112 0.1 4.1 51% 7.3 13 73 5
8/12/14 SC-23 2.2 25 114 0.1 4 50% 7.4 15 N/A N/A
8/12/14 SC-CD 0.1 25.5 98 0 5.4 76% 6.4 19 52 4
8/12/14 SC-CH 0.1 25.2 104 0.1 4 49% 7.4 12 63 1
8/12/14 SC-CH 2.8 25.2 103 0.1 3.9 48% 7.3 12 N/A N/A
8/12/14 SC-GR 0.1 24.1 104 0.1 6.5 77% 6.5 11 N/A 5
8/12/14 SC-NK 0.1 25.2 110 0.1 4.7 57% 6.8 11 189 5
8/12/14 SC-NK 2.8 25.2 103 0.1 4.6 55% 6.8 11 N/A N/A
9/8/14 BC-CBR 0.1 24.8 111 0.1 5.5 66% 8.8 3 2282 3
9/8/14 BC-CBR 1.9 24.9 116 0.1 5.2 64% 8.6 12 N/A N/A
9/8/14 LC-RR 0.1 28.1 32964 19.3 4.9 70% 7.6 4 62 5
9/8/14 LC-RR 1.9 28 32988 19.3 4.9 71% 7.6 5 N/A N/A
9/8/14 MOT-CBR 0.1 25 107 0.1 6.8 82% 8 15 5172 5
9/8/14 MOT-ND 0.1 24.8 128 0.1 6.1 73% 7.7 10 5475 6
9/9/14 PG-CH 0.1 24.1 147 0.1 4.1 49 7.5 6 291 4
9/9/14 PG-CH 1.9 24.1 147 0.1 4.1 48 7.5 6 N/A N/A
9/9/14 PG-ML 0.1 24.3 206 0.1 4.7 56 8 10 651 2
9/9/14 PG-NC 0.1 24 130 0.1 3.5 41 7.2 7 131 5
9/9/14 PG-NC 3.5 24 129 0.1 3.4 40 7.1 8 N/A N/A
9/9/14 SC-23 0.1 24.9 517 0.3 4.7 57 8.3 15 2481 4
9/9/14 SC-23 2.1 24.9 518 0.3 4.7 57 8.2 18 N/A N/A
9/9/14 SC-CD 0.1 23.9 114 0.1 5.9 70 7 16 957 5
9/9/14 SC-CH 0.1 27.5 12824 7 3.7 4.9 7.5 16 488 4
9/9/14 SC-CH 2.8 27.5 12930 7 3.6 48 7.4 26 N/A N/A
9/9/14 SC-GR 0.1 23.8 114 0.1 5.9 70 6.7 15 959 4
9/9/14 SC-NK 0.1 24.3 119 0.1 5.1 60 7.1 9 3255 3
9/9/14 SC-NK 2.8 24.3 118 0.1 4.9 59 7 10 N/A N/A
9/10/14 FC-13 0.1 26.1 533.1 34.3 4.2 63 7.8 4 73 6
9/10/14 FC-13 1.1 26.1 53730 34.7 4.2 63 7.9 6 N/A N/A
9/10/14 FC-4 0.1 26.7 58613 37.6 5.4 82 8.2 1 20 2
9/10/14 FC-4 2.3 26.9 59808 38.4 5.4 85 8.2 2 N/A N/A
9/10/14 FC-6 0.1 26.4 56908 36.7 4.8 73 8.1 3 10 4
9/10/14 FC-6 1.7 26.5 56997 36.8 4.8 73 8.1 3 N/A N/A
9/10/14 FC-FOY 0.1 26 53331 34.4 4.1 62% 8 5 10 1
9/10/14 FC-FOY 1.2 26.1 54250 35.1 4.2 63 8 5 N/A N/A
9/10/14 PC-BDDS 0.1 25.6 45567 29 5.2 7.4 8 3 1281 4
9/10/14 PC-BDUS 0.1 24.7 6467 3.5 7.1 86 89 9 788 1
9/10/14 PC-M 0.1 26.7 58531 37.6 5.4 84 8.1 3 20 2
9/10/14 PC-M 2.3 26.7 58058 37.2 5.3 81 9.1 5 N/A N/A
10/21/14 BC-CBR 0.1 17.5 244 0.1 6.2 65% 9.6 3 5 1
10/21/14 BC-CBR 2 16.6 210 0.1 4.7 48% 9.5 17 5 1
10/21/14 LC-RR 0.1 20.4 23494 15.8 6.9 84% 7.8 5 41 3
10/21/14 LC-RR 1.6 20.4 23501 15.8 6.9 84% 7.8 5 41 3
10/21/14 MOT-CBR 0.1 18.8 324 0.2 6 65% 8.4 5 173 2
10/21/14 MOT-ND 0.1 17.3 294 0.2 7.2 74% 8.5 6 173 0
10/22/14 PG-CH 0.1 16.6 285 0.2 4 42% 10 5 612 1
10/22/14 PG-CH 1.5 16.6 285 0.2 3.5 34% 9.8 12 612 1
10/22/14 PG-ML 0.1 17.6 287 0.2 4.6 48% 10.3 1 3076 1
10/22/14 PG-NC 0.1 15.4 153 0.1 2.6 25% 9.6 14 5 0
10/22/14 PG-NC 3.3 15.2 159 0.1 1.3 13% 9.6 14 5 0
10/22/14 SC-23 0.1 24.9 517 0.3 4.7 57% 8.3 15 41 5
10/22/14 SC-23 2.3 24.9 518 0.3 4.7 57% 8.2 18 41 5
10/22/14 SC-CD 0.1 16.6 186 0.1 9.1 93% 9.4 1 3255 0
10/22/14 SC-CH 0.1 21.1 11679 7.3 5.7 67% 8.9 4 109 1
10/22/14 SC-CH 2.5 21.1 11920 7.4 5.6 65% 8.9 12 109 1
10/22/14 SC-GR 0.1 16.7 154 0.1 8.3 85% 9.3 2 6131 6
10/22/14 SC-NK 0.1 18 397 0.2 5.4 57% 9.1 2 63 1
10/22/14 SC-NK 1.7 18 399 0.2 5.1 54% 9.1 2 63 1
10/23/14 FC-13 0.1 17.9 44367 33.8 6.5 84% 9.6 4 10 2
10/23/14 FC-13 1.1 17.9 44583 33.9 6.5 84% 9.6 11 10 2
10/23/14 FC-4 0.1 19 46862 35 7.4 98% 9.6 1 5 4
10/23/14 FC-4 1.2 19 46859 35 7.2 95% 9.6 1 5 4
10/23/14 FC-6 0.1 19.2 46912 34.8 7.6 101% 9.6 1 5 2
10/23/14 FC-6 1.2 19.1 46723 34.7 7.1 94% 9.6 1 5 2
10/23/14 FC-FOY 0.1 17.9 44543 34 7.7 99% 9.6 2 5 1
10/23/14 FC-FOY 1.2 18 45124 34.2 7.1 92% 9.6 15 5 1
10/23/14 PC-BDDS 0.1 17.8 42640 32.4 7.6 97% 9.5 3 199 3
10/23/14 PC-BDUS 0.1 18.4 34601 25.3 6.8 84% 9.8 6 96 2
10/23/14 PC-M 0.1 19 46655 34.7 7.4 98% 8.8 2 5 2
10/23/14 PC-M 1.7 19.1 46704 34.7 7 93% 8.8 2 5 2
11/24/14 BC-CBR 1.9 17 157 0.1 5.1 53% 9.2 14 N/A N/A
11/24/14 PG-CH 0.1 13.3 145 0.1 4.5 43% 9.1 11 1725 1
11/24/14 PG-CH 1.8 13.3 145 0.1 4.2 40% 9.1 11 N/A N/A
11/24/14 PG-ML 0.1 13 318 0.2 6.3 59% 9.2 5 3255 1
11/24/14 PG-NC 0.1 14.1 127 0.1 5.8 56% 9 12 8164 2
11/24/14 PG-NC 3.4 13.7 129 0.1 4.6 44% 9 15 N/A N/A
11/24/14 SC-23 0.1 13.6 3912 2.7 8.1 79% 9.8 12 1439 4
11/24/14 SC-23 1.8 13.6 4032 2.8 8.1 78% 9.8 13 N/A N/A
11/24/14 SC-CD 0.1 16.6 107 0.1 8.3 84% 8.4 25 15531 2
11/24/14 SC-CH 0.1 13.6 18310 23.4 8.2 86% 8.7 9 318 3
11/24/14 SC-CH 3.3 13.6 18503 23.7 8.2 86% 8.8 23 N/A N/A
11/24/14 SC-GR 0.1 16.4 97 0.1 8.1 83% 8.1 18 8164 2
11/24/14 SC-NK 0.1 16.3 121 0.1 6.9 70% 8.4 22 11199 2
11/24/14 SC-NK 2.6 16.3 123 0.1 6.7 68% 8.8 23 N/A N/A
11/25/14 BC-CBR 0.1 17.2 196 0.1 5.5 56% 9.2 1 120 3
11/25/14 FC-13 0.1 15.9 42038 33.4 7.7 96% 9 2 243 5
11/25/14 FC-13 1.2 15.9 42060 33.4 7.7 96% 9 8 N/A N/A
11/25/14 FC-4 0.1 15.2 43129 34.9 8.3 101% 9.1 3 52 2
11/25/14 FC-4 1.8 14.8 43522 35 8.3 101% 9.1 5 N/A N/A
11/25/14 FC-6 0.1 15.7 42224 33.8 8.3 101% 9.1 1 161 3
11/25/14 FC-6 1.8 15.6 42575 34.2 8.3 101% 9.1 2 N/A N/A
11/25/14 FC-FOY 0.1 16 41303 33.2 8.1 100% 9 2 109 3
11/25/14 FC-FOY 1.2 15.8 41827 33.3 7.9 98% 9 3 N/A N/A
11/25/14 LC-RR 0.1 15.1 29238 22.8 8.6 98% 8.7 4 73 4
11/25/14 LC-RR 1.8 15.1 29358 22.9 8.6 98% 8.7 5 N/A N/A
11/25/14 MOT-CBR 0.1 16.9 192 0.1 8.1 84% 8.8 2 629 1
11/25/14 MOT-ND 0.1 17.1 201 0.1 6.5 67% 8.2 2 388 3
11/25/14 PC-BDDS 0.1 16.4 39030 30.4 7.1 88% 8.8 2 20 2
11/25/14 PC-BDUS 0.1 17.1 30176 22.4 6.3 75% 9 8 2098 2
11/25/14 PC-M 0.1 15.2 43122 35 8 98% 8.8 7 3968 3
11/25/14 PC-M 1.8 15.1 43158 35 8 98% 8.8 9 N/A N/A
12/8/14 BC-CBR 0.1 10.8 288 0.2 9 81% 8.1 2 10 1
12/8/14 BC-CBR 1.7 10.6 201 0.1 8.1 73% 8 3 N/A N/A
12/8/14 LC-RR 0.1 11.6 21641 18 9.9 102% 8.2 2 97 2
12/8/14 LC-RR 1.9 11.8 23583 19.6 9.8 101% 8.2 3 N/A N/A
12/8/14 MOT-CBR 0.1 12.8 280 0.2 7.8 72% 8 2 63 2
12/8/14 MOT-ND 0.1 10.7 244 0.2 7.2 65% 8 4 110 1
12/9/14 PG-CH 0.1 9.9 179 0.1 7 62% 8.1 2 52 3
12/9/14 PG-CH 2 9.9 179 0.1 6.7 59% 8 7 N/A N/A
12/9/14 PG-ML 0.1 10.4 208 0.1 7.3 65% 8 2 20 0
12/9/14 PG-NC 0.1 9.4 142 0.1 7.3 63% 8.2 1 <10 1
12/9/14 PG-NC 3.3 9.4 145 0.1 3.7 28% 8.2 17 N/A N/A
12/9/14 SC-23 0.1 11.6 1885 1.3 8.4 78% 7.8 8 10 2
12/9/14 SC-23 2.3 11.7 3407 2.4 8.3 77% 7.9 6 N/A N/A
12/9/14 SC-CD 0.1 11.7 169 0.1 9.3 86% 7.9 1 109 1
12/9/14 SC-CH 0.1 12.1 13162 10.5 8.4 84% 8 6 10 2
12/9/14 SC-CH 2.9 12.3 14780 11.7 8.2 82% 7.9 22 N/A N/A
12/9/14 SC-GR 0.1 12 141 0.1 8.3 74% 8.2 2 31 1
12/9/14 SC-NK 0.1 11.2 381 0.3 7.9 72% 7.8 1 <10 2
12/9/14 SC-NK 1.8 11.2 382 0.3 7.8 71% 7.8 3 N/A N/A
12/10/14 FC-13 0.1 10.8 42337 38.5 8.2 95% 7.9 0 <10 2
12/10/14 FC-13 1.1 10.8 42418 38.6 8.1 94% 7.9 0 N/A N/A
12/10/14 FC-4 0.1 12.3 45310 40 8 96% 8 3 <10 3
12/10/14 FC-4 1.4 12.4 45357 40 7.9 95% 8.1 3 N/A N/A
12/10/14 FC-6 0.1 12 44870 39.8 8.1 97% 8 2 <10 3
12/10/14 FC-6 1.4 12 44981 39.9 8.1 97% 8 2 N/A N/A
12/10/14 FC-FOY 0.1 10.8 42773 38.9 8.3 96% 7.9 0 <10 1
12/10/14 FC-FOY 1.6 10.9 42981 39 8.2 94% 8 0 N/A N/A
12/10/14 PC-BDDS 0.1 10.8 42170 38.4 7.9 91% 7.9 0 52 1
12/10/14 PC-BDUS 0.1 12.2 37468 32.3 7.9 91% 7.9 2 41 1
12/10/14 PC-M 0.1 11.7 44029 39.3 7.9 93% 8.1 0 <10 2
12/10/14 PC-M 1.6 11.7 44126 39.4 7.8 82% 8 1 N/A N/A
1/21/15 BC-CBR 0.1 10.2 163 0.1 7.4 66% 7 2 10 4
1/21/15 BC-CBR 1.8 9.9 182 0.1 7 61% 7 5 N/A N/A
1/21/15 LC-RR 0.1 8.4 4763 3.8 10 11.50% 7.2 16 275 2
1/21/15 LC-RR 1.5 8.3 4752 3.8 10 11.50% 7.2 16 N/A N/A
1/21/15 MOT-CBR 0.1 11.1 237 0.2 9.2 84% 7.1 5 20 2
1/21/15 MOT-ND 0.1 10.2 233 0.2 9.5 85% 7.2 7 31 2
1/21/15 PG-CH 0.1 9.9 129 0.1 9.1 80% 6.6 5 10 3
1/21/15 PG-CH 1.4 9.9 129 0.1 8.7 77% 6.5 6 N/A N/A
1/21/15 PG-ML 0.1 9.8 136 0.1 8.8 77% 6.8 5 20 2
1/21/15 PG-NC 0.1 9.6 112 0.1 8.4 72% 6 3 5 3
1/21/15 PG-NC 3.2 9.5 112 0.1 7.3 64% 5.9 8 N/A N/A
1/21/15 SC-23 0.1 8.9 131 0.1 10 87% 6.8 13 134 2
1/21/15 SC-23 2.1 8.9 131 0.1 9.9 85% 6.8 12 N/A N/A
1/21/15 SC-CD 0.1 12 131 0.1 9.9 92% 6.7 7 5 2
1/21/15 SC-CH 0.1 8.2 103 0.1 11.5 98% 6.7 13 30 1
1/21/15 SC-CH 2.5 8.2 103 0.1 11 93% 6.7 20 N/A N/A
1/21/15 SC-GR 0.1 11.7 117 0.1 9.8 90% 6.6 5 30 1
1/21/15 SC-NK 0.1 10.8 144 0.1 11.3 102% 7 9 197 2
1/21/15 SC-NK 2.1 10.8 144 0.1 10.6 96% 2 10 N/A N/A
1/22/15 FC-13 0.1 10.1 40880 37.8 8.9 100% 8.2 1 5 1
1/22/15 FC-13 1.1 10.1 40978 37.9 8 91% 8.2 1 N/A N/A
1/22/15 FC-4 0.1 10 43027 39.1 6.6 73% 8.2 1 10 1
1/22/15 FC-4 1.4 10.1 43030 39.1 6.6 73% 8.3 1 N/A N/A
1/22/15 FC-6 0.1 10.1 40899 37.8 7.2 81% 8.3 0 5 1
1/22/15 FC-6 1.4 10 40946 37.9 7.2 81% 8.3 1 N/A N/A
1/22/15 FC-FOY 0.1 10.1 40210 37.1 5.9 66% 8.2 1 5 1
1/22/15 FC-FOY 1 10.1 40754 37.2 6 67% 8.2 1 N/A N/A
1/22/15 PC-BDDS 0.1 10.3 38098 34.7 5.1 56% 8.1 0 72 1
1/22/15 PC-BDUS 0.1 13.3 34076 28.3 7.4 86% 7.9 3 650 3
1/22/15 PC-M 0.1 10.4 40916 37.6 5.2 57% 8.3 0 5 1
1/22/15 PC-M 1.7 10.4 40932 37.6 5.8 63% 8.3 0 N/A N/A
2/3/15 BC-CBR 0.1 7.4 125 0.1 9.8 81% 7.1 3 20 3
2/3/15 BC-CBR 1.8 7.4 126 0.1 9.3 78% 7.1 20 N/A N/A
2/3/15 LC-RR 0.1 7.1 3953 3.3 10.8 91% 7.3 16 20 3
2/3/15 LC-RR 1.9 7.1 3973 3.3 11.2 95% 7.3 17 N/A N/A
2/3/15 MOT-CBR 0.1 8.6 175 0.1 10.1 88% 6.9 5 30 2
2/3/15 MOT-ND 0.1 7.4 158 0.1 12 100% 7.1 11 107 2
2/4/15 PG-CH 0.1 6 116 0.1 7.8 62% 6.7 6 179 1
2/4/15 PG-CH 1.6 6 116 0.1 7.7 61% 6.7 10 N/A N/A
2/4/15 PG-ML 0.1 7.4 134 0.1 7.5 63% 6.9 7 31 1
2/4/15 PG-NC 0.1 5.6 102 0.1 7 55% 6.2 6 <10 1
2/4/15 PG-NC 3.5 5.6 103 0.1 6.8 54% 6.2 6 N/A N/A
2/4/15 SC-23 0.1 8.7 168 0.1 10.2 88% 7.4 8 <10 2
2/4/15 SC-23 2.6 8.7 167 0.1 10.1 87% 7.3 8 N/A N/A
2/4/15 SC-CD 0.1 9.5 119 0.1 7.9 69% 6.7 8 31 1
2/4/15 SC-CH 0.1 7.8 87 0.1 10.6 89% 6.7 6 20 1
2/4/15 SC-CH 2.3 7.5 87 0.1 10.5 88% 6.6 7 N/A N/A
2/4/15 SC-GR 0.1 9.1 106 0.1 10.7 93% 6.8 7 10 1
2/4/15 SC-NK 0.1 8.1 124 0.1 11.4 96% 7 6 41 1
2/4/15 SC-NK 2.3 8.1 124 0.1 11 93% 7 5 N/A N/A
2/5/15 FC-13 0.1 10.2 33034 29.7 7.8 84% 8.1 2 97 3
2/5/15 FC-13 0.8 10.2 33288 30.4 7.1 77% 8.1 5 N/A N/A
2/5/15 FC-4 0.1 10.3 36765 33.4 5.8 63% 8.2 1 <10 2
2/5/15 FC-4 1.2 10.4 36894 33.5 5.8 63% 8.2 1 N/A N/A
2/5/15 FC-6 0.1 10.2 36128 33.4 6.5 72% 8.2 1 20 1
2/5/15 FC-6 1 10.2 36130 33.4 6.6 73% 8.2 0 N/A N/A
2/5/15 FC-FOY 0.1 10.2 32621 29.2 6.2 67% 8.1 2 75 2
2/5/15 FC-FOY 0.9 10.2 34932 31.6 5.8 62% 8.2 1 N/A N/A
2/5/15 PC-BDDS 0.1 10.3 33340 30.2 8.3 90% 8.1 1 504 2
2/5/15 PC-BDUS 0.1 11.5 23680 19.9 8.2 85% 7.7 7 771 2
2/5/15 PC-M 0.1 10.4 37215 33.6 7.5 83% 8.2 1 <10 2
2/5/15 PC-M 1.2 10.6 37502 33.8 7 78% 8.3 5 N/A N/A
3/18/15 BC-CBR 0.1 16 181 0.1 4.2 43% 7.3 3 20 2
3/18/15 BC-CBR 1.6 15.4 182 0.1 4 40% 7.2 4 N/A N/A
3/18/15 LC-RR 0.1 14.1 8567 6.2 7.1 72% 7.2 12 41 5
3/18/15 LC-RR 2.1 14.1 8568 6.2 6.7 67% 7.2 13 N/A N/A
3/18/15 MOT-CBR 0.1 15.3 250 0.2 7.2 73% 7 5 1989 2
3/18/15 MOT-ND 0.1 14.2 241 0.2 7.1 70% 7.2 9 369 2
3/19/15 PG-CH 0.1 13.8 182 0.1 9.8 94% 6.9 1 52 3
3/19/15 PG-CH 1.6 13.8 182 0.1 6 58% 6.9 11 N/A N/A
3/19/15 PG-ML 0.1 15.3 173 0.1 6.5 65% 7.2 3 160 2
3/19/15 PG-NC 0.1 13.4 134 0.1 3.8 36% 6.4 7 5 2
3/19/15 PG-NC 3.3 13 174 0.1 3 29% 6.5 20 N/A N/A
3/19/15 SC-23 0.1 15.8 245 0.1 6.8 69% 7.2 10 20 4
3/19/15 SC-23 1.8 15.8 246 0.1 6.9 70% 7.1 12 N/A N/A
3/19/15 SC-CD 0.1 14.3 144 0.1 8.5 83% 7 5 158 1
3/19/15 SC-CH 0.1 `15.0 775 0.5 7.4 74% 7 16 31 6
3/19/15 SC-CH 2.1 15 788 0.5 7.4 74% 6.9 17 N/A N/A
3/19/15 SC-GR 0.1 13.9 145 0.1 8.9 86% 7.2 4 108 3
3/19/15 SC-NK 0.1 15.6 193 0.1 6.3 63% 7.2 3 84 7
3/19/15 SC-NK 2.3 15.7 193 0.1 6.3 63% 7.2 5 N/A N/A
3/20/15 FC-13 0.1 n/a n/a n/a n/a n/a n/a n/a N/A N/A
3/20/15 FC-4 0.1 n/a n/a n/a n/a n/a n/a n/a N/A N/A
3/20/15 FC-6 0.1 n/a n/a n/a n/a n/a n/a n/a N/A N/A
3/20/15 FC-FOY 0.1 n/a n/a n/a n/a n/a n/a n/a N/A N/A
3/20/15 PC-BDDS 0.1 12.4 38328 33.1 7.7 88% 8 1 355 1
3/20/15 PC-BDUS 0.1 13.4 28128 22.8 5.5 61% 7.4 4 9804 2
3/20/15 PC-M 0.1 11.8 39533 34.7 8.5 97% 8.2 1 10 2
3/20/15 PC-M 2 11.8 39535 34.7 8.4 97% 8.2 21 N/A N/A
4/1/15 LC-RR 0.1 15.4 6877 4.7 8.1 84% 7.4 13 30 4
4/1/15 LC-RR 1.6 15.5 7192 4.9 8.1 84% 7.3 12 N/A N/A
4/2/15 BC-CBR 0.1 15.3 176 0.1 4.2 42% 7.4 2 10 1
4/2/15 BC-CBR 1.9 13.3 181 0.1 3.8 37% 7 23 N/A N/A
4/2/15 MOT-CBR 0.1 15.1 251 0.2 7 69% 7 4 75 5
4/2/15 MOT-ND 0.1 14.5 245 0.2 7.3 72% 7.1 7 134 4
4/2/15 PG-CH 0.1 13.1 155 0.1 6.1 59% 7.1 6 10 2
4/2/15 PG-ML 0.1 14.6 170 0.1 7.3 72% 7.4 4 20 3
4/2/15 PG-NC 0.1 13 123 0.1 4.5 41% 6.6 6 5 2
4/2/15 PG-NC 3.1 12.7 143 0.1 3.6 37% 6.5 6 N/A N/A
4/2/15 SC-23 0.1 15.2 209 0.1 7.2 72% 7.3 4 5 9
4/2/15 SC-23 1.9 15.2 211 0.1 7.2 72% 7.3 4 N/A N/A
4/2/15 SC-CD 0.1 15 148 0.1 9.1 89% 7.1 5 97 1
4/2/15 SC-CH 0.1 15.1 133 0.1 7.1 71% 7.7 5 20 1
4/2/15 SC-CH 2.3 15.1 133 0.1 7.2 72% 7.4 7 N/A N/A
4/2/15 SC-GR 0.1 13.7 131 0.1 9 86% 7 9 74 2
4/2/15 SC-NK 0.1 14.6 172 0.1 6.8 67% 7.1 3 52 5
4/2/15 SC-NK 2.2 14.6 172 0.1 6.7 66% 7.1 3 N/A N/A
4/6/15 FC-13 0.1 16.2 38640 30.2 7.1 87% 8 1 31 3
4/6/15 FC-13 0.6 16.2 39186 30.7 7 85% 8 1 N/A N/A
4/6/15 FC-4 0.1 15.9 43726 35 8 99% 8.1 2 10 2
4/6/15 FC-4 1.1 15.8 43671 35.1 7.9 98% 8.1 3 N/A N/A
4/6/15 FC-6 0.1 16 43540 34.7 7.9 98% 8.1 1 5 2
4/6/15 FC-6 1 16 43669 34.9 7.9 98% 8.1 2 N/A N/A
4/6/15 FC-FOY 0.1 16.2 41450 32.7 7.6 95% 8.1 0 10 2
4/6/15 FC-FOY 0.5 16.2 41739 32.9 7.6 95% 8.1 1 N/A N/A
4/6/15 PC-BDDS 0.1 17 37281 28.5 6.1 73% 7.7 2 502 32
4/6/15 PC-BDUS 0.1 20 31095 21.7 8.8 101% 7.8 11 231 48
4/6/15 PC-M 0.1 16 43950 35.1 8.2 102% 8.1 1 5 2
4/6/15 PC-M 1.4 15.9 43923 35.1 8.2 102% 8.1 1 N/A N/A
5/4/15 BC-CBR 0.1 16.9 184 0.1 4.5 46% 7.6 3 5 2
5/4/15 BC-CBR 1.8 16.4 193 0.1 4.1 42% 7.4 21 N/A N/A
5/4/15 LC-RR 0.1 20 16666 10.9 6.8 80% 7.2 8 20 8
5/4/15 LC-RR 1.8 19.9 16687 10.9 6.7 78% 7.2 9 N/A N/A
5/4/15 MOT-CBR 0.1 18.2 306 0.2 3.3 35% 6.9 6 146 2
5/4/15 MOT-ND 0.1 17.7 269 0.2 6.7 71% 7.2 10 246 2
5/5/15 PG-CH 0.1 18.1 258 0.1 3.8 42% 7.2 5 20 1
5/5/15 PG-CH 1.6 17.2 255 0.1 3.8 42% 7.1 17 N/A N/A
5/5/15 PG-ML 0.1 19.5 158 0.1 5.2 56% 7.4 3 10 5
5/5/15 PG-NC 0.1 16.4 158 0.1 3.8 39% 6.8 5 41 3
5/5/15 PG-NC 3.1 14.5 421 0.3 1 10% 6.8 7 N/A N/A
5/5/15 SC-23 0.1 20.9 960 0.5 7.1 79% 7.9 4 10 14
5/5/15 SC-23 1.7 20.8 970 0.5 6.9 77% 7.6 5 N/A N/A
5/5/15 SC-CD 0.1 18.4 165 0.1 8.4 89% 7.4 5 226 1
5/5/15 SC-CH 0.1 20.8 1112 0.6 6 68% 7.3 4 5 1
5/5/15 SC-CH 2 20.3 1277 0.7 6 68% 7.2 15 N/A N/A
5/5/15 SC-GR 0.1 17.3 148 0.1 8.2 85% 7.4 6 231 1
5/5/15 SC-NK 0.1 19.5 283 0.2 6.8 74% 7.4 4 10 23
5/5/15 SC-NK 2.6 19.5 283 0.2 6.7 73% 7.3 4 N/A N/A
5/6/15 FC-13 0.1 21.4 45529 32 5.8 78% 8 4 5 5
5/6/15 FC-13 1 21.4 45716 32.1 5.7 77% 8 13 N/A N/A
5/6/15 FC-4 0.1 21.6 49186 34.7 6.4 89% 8.1 5 5 3
5/6/15 FC-4 1.2 21.6 49196 34.7 6.2 86% 8.1 6 N/A N/A
5/6/15 FC-6 0.1 21.6 49097 34.8 6 84% 8.1 4 5 2
5/6/15 FC-6 1.2 21.6 49033 34.8 6.1 85% 8.1 4 N/A N/A
5/6/15 FC-FOY 0.1 21.4 46244 32.5 5.7 78% 8 5 5 3
5/6/15 FC-FOY 1 21.6 47599 33.5 5.8 79% 8.1 8 N/A N/A
5/6/15 PC-BDDS 0.1 21.6 46071 32.3 4.4 60% 7.7 6 31 12
5/6/15 PC-BDUS 0.1 23.1 27721 17.8 5.8 75% 7.5 6 857 35
5/6/15 PC-M 0.1 21.8 49288 34.6 6.4 89% 8.1 4 5 2
5/6/15 PC-M 1.5 21.8 49298 34.7 6.2 88% 8.1 8 N/A N/A
6/15/15 BC-CBR 0.1 25.5 212 0.1 2.4 29% 7.3 18 397 7
6/15/15 BC-CBR 1.5 24.4 223 0.1 1.6 19% 7.2 24 N/A N/A
6/15/15 LC-RR 0.1 25.5 212 0.1 2.4 29% 7.3 18 63 9
6/15/15 LC-RR 1.5 24.4 223 0.1 1.6 19.00% 7.2 24 N/A N/A
6/15/15 MOT-CBR 0.1 24.5 355 0.2 2.3 28% 6.8 30 19863 7
6/15/15 MOT-ND 0.1 25.1 313 0.2 4.8 59% 7.3 23 3448 3
6/16/15 PG-CH 0.1 25.6 364 0.2 1.5 18% 7.1 17 199 1
6/16/15 PG-CH 1.5 24.7 363 0.2 1.2 15% 7.1 32 N/A N/A
6/16/15 PG-ML 0.1 28.2 199 0.1 2.4 30% 7.3 18 84 4
6/16/15 PG-NC 0.1 25.5 181 0.1 0.9 11% 6.7 25 61 19
6/16/15 PG-NC 3.1 17.8 580 0.3 0 0% 6.7 15 N/A N/A
6/16/15 SC-23 0.1 29.7 480 0.2 3.6 48% 7.5 18 20 14
6/16/15 SC-23 1.7 29.6 478 0.2 3.6 48% 7.4 19 N/A N/A
6/16/15 SC-23 1.7 29.6 478 0.2 3.6 48% 7.4 19 N/A N/A
6/16/15 SC-CD 0.1 26.1 200 0.1 7.4 91% 7.4 19 2481 2
6/16/15 SC-CH 0.1 28.4 3864 1.9 3.6 47% 7 20 31 3
6/16/15 SC-CH 2.2 28.2 4164 2.1 3.6 46% 6.9 31 N/A N/A
6/16/15 SC-GR 0.1 24.1 189 0.1 6.5 78% 7.3 20 538 1
6/16/15 SC-NK 0.1 29 287 0.1 5.1 67% 7.3 12 75 23
6/16/15 SC-NK 2.1 29 288 0.1 4.3 56% 7.2 18 N/A N/A
6/17/15 FC-13 0.1 29 57560 35.1 4.7 74% 7.9 21 31 7
6/17/15 FC-13 1.1 29 57479 35.1 4.7 74% 8 51 N/A N/A
6/17/15 FC-4 0.1 28.9 58600 36.1 5.6 88% 8.1 19 10 6
6/17/15 FC-4 1.6 28.6 58523 36.3 5.8 91% 8.2 19 N/A N/A
6/17/15 FC-6 0.1 28.8 58493 35.9 5.4 86% 8.1 18 63 5
6/17/15 FC-6 1.3 28.8 58302 35.9 5.4 86% 8.1 19 N/A N/A
6/17/15 FC-FOY 0.1 28.9 57540 35.2 4.9 78% 8 20 5 6
6/17/15 FC-FOY 1 28.9 57664 35.3 4.9 78% 8 28 N/A N/A
6/17/15 PC-BDDS 0.1 30.3 51989 30.6 5.1 80% 7.8 26 20 56
6/17/15 PC-BDUS 0.1 29.3 40872 24 4.8 73% 7.7 28 414 52
6/17/15 PC-M 0.1 28.3 57871 35.9 5.7 89% 8.1 18 10 5
6/17/15 PC-M 1.5 28.3 57669 35.8 5.7 89% 8.1 19 N/A N/A
APPENDIX C
Source Tracking Report
Final Report: Microbial Source Tracking Investigation in Wilmington, NC
Dr. Rachel T. Noble, University of North Carolina Chapel Hill
Contact Information:
UNC Chapel Hill – Dr. Rachel Noble, Professor
3431 Arendell Street, Morehead City, NC 28557
June 22, 2015
Introduction The concept for this small-scale fecal contamination study was to continue the use of a trio of
Bacteroidales based quantitative markers to confirm the presence of human fecal contamination
and to quantify that contamination in a framework of other available environmental parameter and fecal indicator bacteria (FIB) data. This information is being interpreted in the context of
available local knowledge to deduce possible mechanisms of fecal contamination delivery and
transport dynamics. Bacteroides species are non-sporing, obligate anaerobes that are the
numerically dominant bacteria in the human large intestine with up to 1011cells per gram of
human feces. Previous research conducted by PI Noble has demonstrated a strong correlation between Bacteroides thetaiotamicron concentrations to respective human sewage influent amounts (Converse et al. 2009). While this is true, it has also been noted that the Fecal
Bacteroides spp. qPCR assay does quantify bird and dog feces with high affinity. Therefore, it is
always necessary to pair the use of the Fecal Bacteroides spp. qPCR assay with other more
specific assays of human fecal contamination. To accomplish this, we used two additional Bacteroides based assays in order to determine the presence of human fecal contamination, the BacHum assay and the HF183 assay. For more methodological details see Converse et al. 2009,
Kildare et al. 2007, and Layton et al. 2013). In addition, in this supplemental work, we also
quantified gull or seabird contamination through the use of the Gull-2 assay which targets C. marimammalium. Study Areas Initially, there were four sampling locations that were identified by Coastal Planning &
Engineering of North Carolina, Inc. as chronically contaminated during stormwater events, and
as such were areas of concern. These four sites were: Motts Creek, Smith Creek, and an upstream and downstream location each in Pages Creek. During the first study, a limited amount of sampling was conducted, therefore a major goal of this supplemental effort was to hone in on
specific areas and continue to collect more information about the possibility of human fecal
contamination being present at two sites, Motts Creek at Normandy Drive, and Pages Creek
Downstream Site at Bayshore Drive (Table 1). The aim over this project was to sample fewer locations, but attend to sample collection over the course of storms, in hopes that it would illuminate patterns of contamination.
Table 1. List of Sampling Sites
Creek Name Site Name Site Code Latitude Longitude
Motts Creek Normandy Drive MOT-ND 34° 08.373 77° 54.580
Pages Creek Bayshore Drive Down Stream PC-BDDS 34° 16.685 77° 47.673
Figure 1: Map depicting general study area and the locations of Motts and Pages Creeks
Motts Creek Motts Creek watershed encompasses approximately 2,389 acres and is located in the
southwestern portion of the County, just below Sanders Road (Figure 1). The Creek drains
portions of Carolina Beach Road at its headwaters and then drains toward River Road before
entering into the Cape Fear River. Zoning in the watershed is predominately residential with
commercial business districts along Carolina Beach Road. Land in the watershed is classified as
transition, conservation or wetland resource protection according to the CAMA land use plan.
This watershed contains approximately 12.6% impervious surface coverage (Hume, 2009, Figure 2). Sampling was conducted at MOT-ND within the Motts Creek watershed (Figure 2)
Figure 2: Water Quality Sites within the Motts Creek Watershed
Pages Creek
Located in northeastern New Hanover County and encompassing 2,044 acres, Pages Creek
watershed drains into the Intracoastal Waterway, north of Middle Sound Loop Road. Zoning within the Pages Creek watershed is predominately residential, with commercial zoning along
Highway 17. The land within the Pages Creek watershed is predominately classified as
watershed resource protection and conservation, with a small portion classified as transitional
according to the CAMA land use plan. This watershed contains approximately 23.2%
impervious surface coverage (Hume, 2009). Sampling was conducted only at PC-BDDS in Pages Creek (Figure 3).
Figure 3: Water Quality Sites within the Pages Creek Watershed
Sampling and Storm Event Analysis Approach:
Sampling was conducted by Coastal Planning & Engineering of North Carolina, Inc. over the
course of four storm events at the two sites, PC-BDDS and MOT-ND, and sampling was
conducted multiple times (n=3) over the duration of each of the four storm events. The storm
events occurred on March 17-18 and September 8, 2014, January 12 and February 17, 2015.
Environmental data collected and rainfall data available electronically is presented in Table 1. Water samples were collected for enumeration via Enterolert™, and additional samples were
collected and filtered and processed aseptically (as guided by the Noble laboratory) by Coastal
Planning & Engineering of North Carolina, Inc. for later batch analysis using well described
molecular methods (see below). 100 ml sample volumes were filtered in triplicate through 0.45
µm polycarbonate filters and frozen at -20 C and delivered to UNC Chapel Hill IMS within four weeks of sample collection.
The filters were subjected to the following molecular analyses:
1. Fecal Bacteroides qPCR (Converse et al. 2009)
2. BacHum analyses (Kildare et al. 2007) 3. Human specific marker for Bacteroides (HF183, Layton et al. 2013)
4. Gull-2 qPCR (Sinigalliano et al. 2013)
Methodology: Bacteroides spp. comprise approximately one-third of the human fecal microflora, considerably outnumbering Enterococcus and E. coli. The Bacteroides group belongs to a group of non-spore-forming, gram-negative, obligate anaerobes, so there is little concern over regrowth
in the environment. Quantitative polymerase chain reaction (qPCR) methods are used to conduct
the Bacteroides assays:
• Fecal Bacteroides qPCR assay (Converse et al. 2009) relies on Taqman chemistry and all
the reagents are in a liquid formulation, except the OmniMix. The assay quantifies a
cohort of bacteria found in high concentrations in the human gut, including Bacteroides thetaiotaomicron, Bacteroides distastonis, and Bacteroides fragilis. However, the method is not human specific. The assay has been tested against a range of different fecal samples types, and has been shown to be capable of quantifying over a wide range of
concentrations, and to be sensitive at concentrations relevant to water quality source
tracking studies. When using the qPCR approach for fecal Bacteroides, strong
relationships have been observed with a wide array of human sewage collected from areas on both east and west US coastlines. The assay is highly sensitive and the target bacteria that are enumerated have been shown to be a predictor of human health in both
sand and recreational waters (Wade et. al. 2010, Heaney et al. 2012) during large-scale
EPA-run epidemiology studies. This is a fully quantitative qPCR-based assay that is
being used in an array of studies in stormwater contaminated areas and that, with the use of other additional confirmatory methods, can be used to both identify potential hot spots of human fecal contamination (Converse et al, 2009).
• BacHum Human Marker: A separate qPCR assay was utilized to quantify the BacHum
molecular markers reported by Kildare et al., 2007. The assay has been widely tested for
specificity against a range of fecal sample types and has shown high capacity for discrimination against human and animal fecal types (Ahmed et al., 2009).
• HF (human fecal) 183: Human specific marker by qPCR has been conducted previously
by Bernhard and Field (2000) and updated by Seurinck et al., 2006. This assay is specific
to a region of ribosomal rDNA within the Bacteroides spp. that is found almost exclusively in human feces. The assay has been tested repeatedly in a range of different environments for cross reactivity with other types of fecal material, and various
researchers have found either a 90- to 100-percent ability to discriminate between human
and animal feces when using this assay. The assay, however, can be problematic when
used alone, because the target copy concentration in fecal material contributed to receiving water environments can be quite low due to dilution and the assay has a relatively low sensitivity.
• Gull-2: This is an assay that has demonstrated specificity to gull and seabird fecal
contamination. The assay is specific to a 16S rDNA gene sequence within Catellicoccus marimammalium. While, this assay has been demonstrated to cross react with some other types of fecal contamination, reported specificities are roughly 89-95%.
Results:
The data generated over the course of this project is presented in Table 2. High
concentrations of Enterococcus spp. were observed over the four storms sampled and the
array of storm events captured, indicating a continued level of elevated concern about the
microbial water quality of the waters in question. Enterococcus concentrations ranged from 54.6 to >24196 MPN/100 ml, with 93% of the samples exhibiting Enterococcus
concentrations that exceeded EPA single sample standards for recreational waters. This
compares with the 96% observed exceedance in the first study, and demonstrates that
patterns were roughly the same. Rainfall amounts for the storms sampled ranged from
0.5-1.6 inches over the course of the study. These are rainfall amounts that are typical of storms in the Wilmington, NC area, and would not be expected to be categorized as an
extreme event. In Motts Creek, the mean Enterococcus sp. concentration over all
samples collected was 848 MPN/100 ml, while at Pages Creek, it was over 5-fold higher
at 5204 MPN/100 ml (Figure 4 a and b).
A trio of Bacteroides-based molecular markers was quantified in order to assess the
potential for the presence of human fecal contamination at the four sites studied. To
interpret molecular microbial source tracking data, it is useful to cover a few terms used
in the field as related to sensitivity and specificity. These Bacteroides-based markers
have been selected based upon previous successfully conducted blind studies of human and animal fecal contamination quantification and discrimination, and for their coverage
of a range of specificities and sensitivities. The Bacteroides-based methods chosen for
this project have been recently included in a range of publications (Layton et al. 2013,
Boehm et al. 2013, Stewart et al. 2013, Converse et al. 2009, Kildare et al. 2007, to name
a few). These methods are well represented in the peer reviewed literature. The three assays cover a range of specificities for human fecal contamination. One way to look at
specificity is to use a percentage attribution. That is, if 100 water samples containing
human or animal fecal contamination were tested using each of the three assays, what is
the number of samples upon which the correct discrimination between the two types of
samples would have been made. For the Fecal Bacteroides spp., BacHum, and HF183 assays, the specificities increase across the trio from 85%, 92% and 96% respectively.
Conversely, sensitivity is the chance that in a water sample containing human fecal
contamination that the marker will be detectable or present. Sensitivity is the lowest for
the HF183 assay, and increases strongly from the BacHum to the Fecal Bacteroides spp.
assay. This means that while HF183 is the most specific to human fecal contamination, that there is danger in using it alone as a source tracking marker, because in some human
fecal contamination, it will occur at such low concentrations that a negative result may
occur simply due to the amount of sample analyzed. Alternatively, it would not be useful
to base an entire source tracking study upon Fecal Bacteroides spp. quantification,
because while it occurs in the highest concentrations in human fecal contamination of the three markers, it is also present in dogs and seagulls. To date, we have observed that the
Gull-2 molecular marker of gull fecal contamination has excellent specificity and
sensitivity, i.e. in a recent study of Wilmington-based fecal material from dogs, gulls, and
humans, the assay was found to be over 90% specific to bird fecal contamination.
The importance of sensitivity and sensitivity cannot be understated because of
implications of false negatives during real world fecal contamination studies such as the
one conducted here. False negatives are essentially a non-detect result or low
concentration result that occurs on a sample due to methodological limitations. In a
human fecal contamination source tracking study, a non-detect, or negative result can
lead someone to believe that human fecal contamination was not present in water tested, when it the outcome could be a result of heterogeneity of the original water sample, an
inappropriate sampling and concentration approach, or poor methodological constraints.
Every effort has been made with this work to generate quantitative results on water
samples collected, even those that pose methodological limitations such as qPCR. While
a full discussion of false negatives and their implications is beyond the scope of this document, the reader can refer to a series of well written articles resulting from the
Source Identification Pilot Project (SIPP) in the Journal of Water Research in the coming
months in which PI Noble was a major participant (Layton et al. 2013, Stewart et al.
2013, Boehm et al. 2013).
None of the assays utilized in this study for human fecal contamination are 100 %
specific to human fecal contamination. However, when combined, they become more
powerful. Previous studies conducted blindly on human and animal fecal contamination
(HRSD AH Environmental Report 2012), all three markers measured in this study were
positive and correctly identified human fecal contamination 100% of the time when applied in a similar manner. Based upon these results, and other results of blind studies
conducted in the past five years, samples for which all three assays yield positive results
are confirmed to contain at least some level of human fecal contamination. If two of the
assays are positive, it is likely that there is the potential for human fecal contamination to
have existed in the water sample. Finally, the concentrations of the markers can be used to examine the levels of human fecal contamination in the water tested and to prioritize
remediation strategies.
A notable result was from the smallest of the four storms, where the 3rd sample collected
(2/17/2015) at both Motts Creek (Figure 5 a) and Pages Creek (Figure 5b) resulted in contamination for all four markers measured. While this was a small storm and only a
single storm, this result indicates that the tail of the storm event is important. This also
indicates that rising groundwater, and subsequent connections among septic leachfields,
or sewage/stormwater conveyance systems could be playing a role in the delivery of fecal
contamination to this system. All three human markers were positive, and the gull fecal contamination marker was also positive. The HF183 marker, along with the Fecal
Bacteroides spp. marker were also both strongly positive at the beginning (Sample 1) for
the storm from 1/12/2015, a rainfall event of our 0.6 inches total rainfall. For the storm
event of March 17-18, 2015 (see Figure 5 a and b), the HF183 marker was positive in
both Sample 1 and Sample 2, occurring with a steady spring-like rainfall event of a total of 1.3 inches. The other human markers during this event were non-detectable for Pages
Creek, but in Motts Creek for this single storm the BacHum results corroborated the
HF183 results.
It is interesting to note that several of the other samples collected over the course of this study did not demonstrate any indication of a human fecal contamination signal. The
human fecal contamination signal is indeed ephemeral and appears to be driven
specifically by rainfall patterns. Methodological limitations prevented accurate
quantification of the gull marker in a few instances, because the water sample collected
and purified had to be diluted in order to gain a proper qPCR signal from the sample.
This is called “qPCR inhibition”. Inhibition is typically caused by large high molecular weight molecules such as humic and fulvic acids, which are present in high quantities in
the Creeks studied here.
Motts and Pages Creek over the duration of all four storms had very high concentrations
of the Fecal Bacteroides spp. concentrations at both sites with concentrations well in excess of 100,000 CE/100 ml (Figure 5 a and b). Most of the samples that demonstrated
very high Fecal Bacteroides spp. concentrations also had high concentrations of the
BacHum marker, ranging from 4,120 to 15,378 gene copies/100 ml (Figure 5 and b).
There was no statistical relationship between Bacteroides-based molecular markers and
the Enterococcus concentrations or total rainfall, but this study includes a small sample size, upon which it is often difficult to conduct statistical analyses appropriately (p >
0.05, correlation analysis). This is a small study, and the sites were selected based upon
previous historical data showing contamination. All of the sites indicate that human fecal
contamination has the potential to pose a serious problem to these receiving waters.
Recommendation: At this time, it has been shown over multiple years that rainfall and
storm events translate to poor water quality in Motts and Pages Creek. During this study,
a limited amount of sampling was conducted during dry weather, but these samples were
not analyzed using the molecular markers. In one of the dry weather samples, the
Enterococcus concentration was 631 MPN per 100 ml, but in the other samples from Pages Creek, the concentration of Enterococcus was <10 MPN/100 ml.
The impairment of water quality at these sites is complex and likely driven by the
presence of sources of 1) human fecal contamination, 2) bird fecal contamination, and 3)
potential regrowth of fecal indicator bacteria in the organic matter rich estuarine and creek systems. While human fecal contamination has been determined to be present, the
signal is ephemeral, meaning that it is driven by a series of complex hydrological factors.
It is difficult to pinpoint these factors with small scale study efforts. It is clear that the
effort to increase sampling frequency during storm events has been useful, but further
targeted upstream sampling efforts would need to follow one of two main approaches to be fruitful.
1) The first potential approach would be to begin to conduct adaptive sampling for storms
in 5-10 sampling locations to hone in on upstream “hot spots”. This is a tried and true
water quality sampling approach. The same sites as currently being sampled would be tested, and in addition, sites upstream that represent possible areas where illicit
stormwater/sewage conduit connections might exist would also be targeted with high
spatial frequency (i.e. sampling at all potential locations of source input). In Motts
Creek, where a septic system to sewerage system transformation will take place, it would
be most useful to conduct this assessment after the installation of sewage treatment. In Pages Creek, there may be sewage lines that are exfiltrating during wet weather, and
during those periods, that contamination can become connected with stormwater
outflows, causing impairment of coastal waters.
2) A second recommendation would be to conduct a limited set of tracer studies using tracer viruses, in the local septic systems. This approach would require a volunteer
household close to either Pages or Motts Creek. The approach would be to use a tracer in
the system of the household and follow the transport to closeby receiving waters. This
would be particularly applicable in areas where a mixture of septic and sewage systems
exist, and where it is desired to hone in on specific sources of human fecal contamination to the estuary.
Figure 4A: Enterococcus sp. concentrations as measured via Defined Substrate Technology (Enterolert™) reported in Most probable number/100 ml for all four sampling events for Motts Creek.
Figure 4B: Enterococcus sp. concentrations as measured via Defined Substrate
Technology (Enterolert™) reported in Most probable number/100 ml for all four
sampling events for Pages Creek.
Figure 5A: Concentrations of molecular microbial source tracking markers Fecal Bacteroides spp., BacHum, HF183, and Gull-2 gene copies/100 ml for four storm events in Motts Creek, NC. Note axis is on logarithmic scale.
Figure 5B: Concentrations of molecular microbial source tracking markers Fecal Bacteroides spp., BacHum, HF183, and Gull-2 gene copies/100 ml for four storm
events in Pages Creek, NC. Note axis is on logarithmic scale.
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Table 2: Summary of site, date, environmental parameters, and molecular marker data at the two study sites, Motts and Pages
Creek.
1 Station
Sample
#Date
Rain
(inches) Time
Depth
m
Temp
oC
Cond
mS
Salinity
psu
DO
mg/L DO%pH
Turbidit
y NTU Entero.
Fitler
Volume
Filter
time .CD /E/100ml .CD /E/100ml
.CD /E/100ml
Average
HC183
/E/100ml
HC183
/E/100ml
HC183
/E/100ml
Average
.acHum
/E/100ml
.acHum
/E/100ml
.acHum
/E/100ml
Average
Dull-2
/E/100ml
Dull-2
/E/100ml
Dull-2
/E/100ml
Average
PC-BDDS N/A 1/27/2014 0.0 14:39 0.1 18.3 5592 3.5 5.2 56 8 10 631 100ml 5 min.MOT-ND N/A 1/27/2014 0.0 14:03 0.1 10.7 296 0.2 9 87 8 9 10 100ml 5 min.
Dry weather baseline data
2 PC-BDDS 1 3/17/2014 1.3 10:08 0.1 11.3 36989 32.8 8.1 91 7.5 5 >24196 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T bhb DETE 8186 8186 bhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETE/T
PC-BDDS 2 3/17/2014 1.3 15:43 0.1 9.4 798 0.6 10.6 93 7.8 4 8297 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T 8509 9653 9081 bhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETE/T
PC-BDDS 3 3/18/2014 1.3 10:45 0.1 8.9 34785 32.6 8.7 93 7.7 1 457 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T bhb DETEbhb DETEbhb DETEbhb DETE 4120 4120 141481 bhb DETE 141481
MOT-ND 1 3/17/2014 1.3 9:37 0.1 11.4 178 0.1 7.7 71 7.5 7 263 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T bhb DETE 38770 38770 65791 bhb DETE 65791 292798 481308 387053
MOT-ND 2 3/17/2014 1.3 15:11 0.1 11.1 177 0.1 8.7 79 7.8 7 419 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T bhb DETE 9672 9672 11788 12124 11956 40106 bhb DETE 40106
MOT-ND 3 3/18/2014 1.3 10:07 0.1 8.8 169 0.1 8.5 73 8 6 110 100ml 5 min.bhb DETE/T bhb DETE/T bhb DETE/T bhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETEbhb DETE/T
steady spring-like rain lasting more than 24 hrs.
Samples were inhibited
Single replicate positive
3 PC-BDDS 1 9/8/2014 1.5 9:05 0.1 24.5 6068 5.9 7.3 89 7.4 36 788 100ml 5 min.893970bhb DETE/T 893970 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0PC-BDDS 2 9/8/2014 1.5 12:44 0.1 24.8 151 0.1 8.3 88 7.9 24 54.6 100ml 5 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETELnhibitedPC-BDDS 3 9/8/2014 1.5 15:15 0.1 25.1 225 0.1 7.4 89 7.8 15 866.4 100ml 5 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETEinhibited
MOT-ND 1 9/8/2014 1.5 10:10 0.1 24.8 128 0.1 6.1 73 7.7 10 5475 100ml 5 min.bhb DETE/T 691114 691114 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
MOT-ND 2 9/8/2014 1.5 12:18 0.1 24.8 136 0.1 6.4 77 8.1 11 136.4 100ml 5 min.bhb DETE/T 478163 478163 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
MOT-ND 3 9/8/2014 1.5 14:47 0.1 25 132 0.1 5.7 69 7.8 9 104.1 100ml 5 min.bhb DETE/T 1948083 1948083 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
heavy rains starting the evening before- rained hard during first sampling and then lessened throughout the day.
4 PC-BDDS 1 1/12/2015 0.6 8:21 0.1 1207 100ml 2 min.951591 635586 793589 bhb DETEbhb DETE 0 bhb DETE 48840 48840 bhb DETEbhb DETE 0PC-BDDS 2 1/12/2015 0.6 11:42 0.1 24196 100ml 2 min. 90579 214412 152495 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 265287 94631 179959
PC-BDDS 3 1/12/2015 0.6 15:28 0.1 907 100ml 2 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
MOT-ND 1 1/12/2015 0.6 8:50 0.1 309 100ml 2 min.784696 882863 833780 bhb DETEbhb DETE 0 28848 bhb DETE 28848 bhb DETEbhb DETE 0
MOT-ND 2 1/12/2015 0.6 11:08 0.1 638 100ml 2 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
MOT-ND 3 1/12/2015 0.6 14:51 0.1 277 100ml 2 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0
rains starting in the early morning, 0.3" before sampled. - rain lessened between 1st and 2nd sample, but increased heavily before 3rd sample..
NO PHYSICAL DATA RECORDED (bad YSI)
5 PC-BDDS 1 2/17/2015 0.5 9:01 0.1 6.5 32371 32.2 8.1 82 7.9 4 3873 100ml 2 min.bhb DETE/T bhb DETE/T 0 25649 bhb DETE 25649 6641 17076 11859 102390 46721 74555PC-BDDS 2 2/17/2015 0.5 10:48 0.1 5.3 246 0.2 12.7 100 7.8 8 1951 100ml 5 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0PC-BDDS 3 2/17/2015 0.5 15:34 0.1 5.2 210 0.2 13.3 105 7.8 3 233 100ml 5 min.bhb DETE/T 12561 12561 21864 42857 32360 6846 3599 5223 bhb DETEbhb DETE 0
MOT-ND 1 2/17/2015 0.5 8:29 0.1 8.1 185 0.1 11.7 99 7.7 17 1376 100ml 2 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 bhb DETEbhb DETE 0 442490 bhb DETE 442490
MOT-ND 2 2/17/2015 0.5 10:21 0.1 7.6 138 0.1 9.6 80 7.9 25 1664 100ml 5 min.bhb DETE/T bhb DETE/T 0 bhb DETEbhb DETE 0 15378 bhb DETE 15378 2006178 1771530 1888854
MOT-ND 3 2/17/2015 0.5 15:01 0.1 7.1 147 0.1 10.2 84 7.4 18 243 100ml 5 min.573177bhb DETE/T 573177 bhb DETE 61699 61699 26929 47307 37118 bhb DETE 685089 685089