Report on the Health of Hodges Creek:

Follow-up research on the fish kill that occurred July 2010 in Boone, NC
Erin Abernethy
Environmental Toxicology BIO3542-101/5542-101
April 27, 2011
In Collaboration with Anthony Marino, Matthew Cox, and Donna Lisenby

I have not violated the Appalachian State University Academic Integrity Code.

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ABSTRACT
Following the contamination of Hodges Creek with coal-tar based asphalt sealant, a fish kill
occurred in July of 2010 in Boone, NC. It is believed that all life was killed within the stream from the
PAHs released during the contamination event. We sampled sediments for PAHs and surveyed fish and
invertebrates in March of 2011 to assess the current ecological status of Hodges Creek. Although fish and
invertebrates have recolonized the stream in reduced numbers, it is believed that the high levels of PAHs
detected in the sediment samples will continue to negatively impact the wildlife of Hodges Creek by
causing low abundances and poor health. Monitoring should be continued to more fully map the extent of
contamination and determine the level of PAHs in the sediments in following years.
INTRODUCTION
On July 17, 2010, a fish kill occurred on Hodges Creek in Watauga County, Boone, NC. It was
believed that all life within the 1.5 mile section of Hodges Creek between the BB&T bank along Highway
105 and the Boone Mall was killed. A total of 97 dead fish, mostly trout, were collected. The cause of the
fish kill was a coal-tar based asphalt sealant that had been applied to the BB&T parking lot earlier that
day in the rain (Lisenby 2010). The rain caused this pollutant to wash off the parking lot and into Hodges
Creek. The asphalt sealant contained a chemical called benzo[a]pyrene, a polycyclic aromatic
hydrocarbon (PAH), that is listed by the EPA as a suspected carcinogen and a highly persistent toxin
(EPA 2010). Considering the potential negative health effects of this asphalt sealant on wildlife and
humans, the Town of Boone banned this product. Although the ban is not a full ban, the opportunities to
apply the sealant are so minimal that from now on other products will most likely be chosen in its stead
(Oakes 2011).
PAHs are released naturally into the environment from processes such as volcanic eruptions and
forest fires, but humans have drastically increased the amount of PAHs that are present through extensive
fossil fuel combustion (Menzie et al. 1992). While PAHs can be detected in the air, water, and soil, within
aquatic environments PAHs tend to accumulate in the sediments (Den Besten et al. 2003). Depending on
the substrate size composing contaminated sediments, PAHs are bioavailable at varying rates and

3
durations to different species of aquatic organisms (Landrum & Scavia 1983; Eadie et al. 1984; Landrum
et al. 1984). Certain species have the ability to metabolize PAHs at significantly different rates (Lu et al
1977; Southworth 1979; Dobroski & Epifanio 1980; Neff 1982; Maccubbin et al. 1985). Although PAHs
do not appear to biomagnify as they move up trophic levels, they do accumulate in the tissues of lower
trophic level aquatic invertebrates, possibly causing adverse health effects (Neff 1979; Varanasi et al.
1980; Baumann et al. 1982; Stegeman et al. 1982; Couch & Harshbarger 1985; McCarthy & Jimenez
1985; McCarthy et al. 1985). It is suggested that within aquatic systems PAHs have the potential to
biominimize when moving up trophic levels, but they can still have prolonged negative effects on the
health, abundance, and diversity of aquatic invertebrates (Eisler 1987).
We are interested as to whether PAHs from the BB&T spill in the summer of 2010 are still at
detectable levels within the sediments of Hodges Creek. We also want to know if fish and invertebrates
have recolonized the stream and if so in what abundance and diversity. We predict that PAH levels are
still high within sediments and that fish and invertebrates will have not recolonized the stream due to this
contamination.
METHODS
Six sites on Hodges Creek were sampled for PAHs. Two reference sites above the BB&T parking
lot were sampled and four sites downstream before the confluence with Boone Creek (Figure 1). To
collect the sediment samples for PAH analysis, we used a stainless steel pot and spoon to scoop sediment
from Hodges Creek into an amber colored wide mouth jar with a teflon lid. Before going into the field
and between each collection materials were washed with 1% Alconox solution and rinsed with acetone.
Jars were filled to the top with superfine mud-like sediment. Once labeled the jars were placed in sealable
plastic bags and stored in a cooler with ice. The samples were given to Dr. John Weinstein from the
Citadel for analysis.

Figure 1. Map of sampling sites on Hodges and Winklers Creeks in Boone, NC. The location of sampling
sites and BB&T are shown by the pushpin icons. Site 1 & 2 are the upstream reference sites for PAH soil
sampling along Poplar Grove Rd. The site labeled Upstream Reference is where fish and invertebrates
were sampled upstream of BB&T. Sites 3, 4, 5, & 6 were where soil, invertebrate, and fish sampling was
done. Fish were also sampled on Winklers Creek at the site marked by the appropriately labeled pushpin.
In Dr. Weinsteins lab the samples will be subjected to ultrasonication in acetonitrile three times
for 10 minutes each time. After being centrifuged, the supernatant will be filtered through a 0.5 micron
filter and run through a high-performance liquid chromatography (HPLC). The HPLC will detect the
presence and concentrations of the 16 EPA priority PAHs. QA/QC will be run with standards from NIST.
Fish surveys were performed on Hodges Creek by electro-shocking. Five sites were sampled, one
upstream of the BB&T parking lot and four downstream (Figure 1). Winklers Creek was also sampled as
a separate reference site. Shocking was carried out for approximately 150 seconds throughout all habitat
types available, riffles, runs, and pools. Fish were captured in hand nets and seines. The species, lengths,
and weights of each fish were recorded. Condition, diversity, and abundance indexes were calculated for
each site from this data. For the condition index, Fultons Condition Factor (K) was calculated (Carlander
1950).
K = (W/L3) x 100,000

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W represents the weight in grams. Length (L) was measured in millimeters. 100,000 is a constant value
that brings the condition value close to a single digit that was then rounded to one decimal place. The
condition indexes for each individual fish were averaged to calculate a final condition index for each site.
The Shannon-Wiener Diversity Index (H) was calculated using the standard equation.
H = -Pi(lnPi)
Pi is the proportion of each species within the sample collected at each site. The abundance value for each
site was calculated by dividing the total number of fish captured by the number of seconds spent
searching. Crayfish and salamander species and numbers were recorded. The presence of snails, Elimia,
was also noted.
Invertebrate surveys were performed in the same reaches as the fish surveys the following day. A
fine mesh seine was set up downstream, and two scientists kicked over rocks upstream while walking
towards the scientist holding the seine for approximately 30 seconds. This technique was performed five
times at each site, and invertebrates were preserved in 70% ethanol for identification in the lab. The
Shannon-Wiener Diversity Index and abundance was calculated from this data using the same methods as
with the fish data. A biotic index (BI) was calculated using tolerance values from the North Carolina
Department of Environment and Natural Resources Standard Operating Procedures (2006) and the Digital
Key to Aquatic Insects of North Dakota from Valley City State University Macro-Invertebrate Lab.
BI = (TVi)(ni) / N
TVi represents the ith taxas tolerance value. ni represents the ith taxas abundance value of 1, 3, or 10,
where 1 is rare, 3 is common, and 10 is abundant. N is the sum of all abundance values. Based on the
biotic index each site was rated on a scale of excellent to poor.
RESULTS
The PAH results from Dr. Weinstein are preliminary due to machine failure and time constraints.
Qualitative results were obtained from the two reference sites upstream of BB&T and two sites
downstream of BB&T. The reference sites showed very low levels of PAH content. The sites downstream
of BB&T showed some of the highest levels of PAH that Dr. Weinstein had seen in his work, which is

6
mainly with stormwater pond sediments. Dr. Weinstein will be re-running these samples in May 2011 and
will be able to give us more specific results.
The fish survey on Hodges Creek yielded 7 species of fish for a total of 31 individuals caught in
818 seconds of electroshocking. Winklers Creek yielded 27 individuals of 6 species within 150 seconds
of shocking. The abundance of fish on Hodges Creek was found to be 0.0379, compared to 0.1800 at
Winklers Creek, a value 4.7 times higher. The abundance per site on Hodges Creek was lowest at the
Upstream Reference, 0.0054, and highest at Site 6, 0.0859. The abundance was higher directly
downstream of BB&T at Site 3 than at Sites 4 or 5 (Table 1 and Figure 2). The number of species caught
and the Shannon-Wiener Diversity Index increase as you move downstream within Hodges Creek (Table
1 and Figure 3). The same number of species was caught at Winklers Creek as Site 6, but the diversity
index is lower for Winklers Creek. The average condition index is similar across sites, ranging from 1.0 to
1.2 on Hodges Creek and 1.1 at Winklers Creek (Table 1 and Figure 3).
Table 1. Fish survey data collected and calculated for Hodges and Winklers Creeks.
Site

Number of
Species

Upstream Reference
Site 3
Site 4
Site 5
Site 6
Winklers Creek

1
1
3
4
6
6

ShannonWiener
Diversity Index
0
0
1.01
1.33
1.73
1.55

Number of
Individuals

Abundance

Average
Condition

1
4
6
6
14
27

0.0054
0.0370
0.0313
0.0353
0.0859
0.1800

1.0
1.1
1.1
1.2
1.1
1.1

Fish Abundance in Hodges and Winklers Creeks

0.2
0.18
0.16
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
Upstream
Reference

Site 3

Site 4

Site 5

Site 6

Winklers Creek

Figure 2. Fish abundance in Hodges and Winklers Creeks. Abundances for each site sampled are shown.
The y-axis represents abundance values calculated by dividing the number of individuals caught by the
amount of time spent electroshocking.

Fish Survey Data for Hodges and Winklers Creeks

7
6
5

Number of Species

4
Shannon-Wiener
Diversity Index

3
2

Average Condition

1
0
Upstream
Reference

Site 3

Site 4

Site 5

Site 6

Winklers
Creek

Figure 3. Fish survey data for Hodges and Winklers Creeks. Fish survey results showing how many
species were caught along with the diversity index and average condition index that was calculated for
each site on Hodges and Winklers Creeks.
The invertebrate survey yielded 6 Orders and 16 Families with a total of 508 individuals collected
in 12.5 minutes of search time within Hodges Creek. The greatest numbers of taxon groups were collected
at Site 4 and the Upstream Reference, and Site 3 had more invertebrates than Site 5 or 6 (Table 2 and

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Figure 4). The Shannon-Wiener Diversity Index was greatest at Site 4 and the Upstream Reference and
slightly higher at Site 5 compared to Site 3. Site 6 had the lowest diversity. The abundance was highest at
the Upstream Reference and lowest at Site 3. The Biotic Index was best at Site 3 and worst at Site 6.
Table 2. Invertebrate survey data collected and calculated for Hodges Creek.
Site

Number
of Taxa

Shannon-Wiener
Diversity Index

Number of
Individuals

Abundance

Biotic Index

10

1.81

164

1.09

4.22 good

7
12
6
3

1.41
1.83
1.57
0.52

44
120
50
130

0.29
0.80
0.33
0.87

3.77 excellent
5.02 good/fair
5.87 fair
7.24 poor

Upstream
Reference
Site 3
Site 4
Site 5
Site 6

Invertebrate Survey Data for Hodges Creek

14
12

Number of Taxa

10

Shannon-Wiener
Diversity Index

Abundance
6
Biotic Index

4
2
0
Upstream
Reference

Site 3

Site 4

Site 5

Site 6

Figure 4. Invertebrate survey data for Hodges Creek. Invertebrate survey results showing the number of
families collected and the calculated diversity index, abundance, and biotic index for each site along
Hodges Creek.
Elimia were present at all sites (Table 3). Two taxon of salamanders were collected,
Desmognathus and Eurycea wilderae. The highest number of salamanders, 6, were collected at Site 3.
Two species of crayfish were collected Cambarus bartonii and Cambarus chasmodactylus. The greatest
number of crayfish, 30, were found at Site 4. No crayfish or salamanders were collected from Sites 5 or 6.

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Table 3. Snail, Salamander, and Crayfish data collected for Hodges and Winklers Creeks.
Site

Upstream
Reference
3
4
5
6
Winklers Creek

Elimia
Present

Salamanders
Number of Taxa

Salamanders
Number of Individuals

Yes

Yes
Yes
Yes
Yes
Yes

1
2
0
0
1

6
4
0
0
1

Crayfish
Number of
Species
1
1
1
0
0
1

Crayfish
Number of
Individuals
1
10
30
0
0
9

Water chemistry data was collected for all sites (Table 4). pH ranged from 7.2 to 7.68. Percent
dissolved oxygen ranged from 82.4 to 86.9, and dissolved oxygen measured in miligrams per liter ranged
from 9 to 10.06. Both measures of DO were highest at Winklers Creek. Salinity was 0.1 for the Hodges
Creek sites and zero for Winklers Creek. Conductivity ranged from 221.2 to 252.8 on Hodges Creek but
was only 33.2 on Winklers Creek. The temperature ranged from 9.1 to 10.4 on Hodges Creek, being
highest at Site 4, and was 9C on Winklers Creek.
Table 4. Water chemistry and temperature is shown for Hodges and Winklers Creeks.
Site
Upstream
Reference
Site 3
Site 4
Site 5
Site 6
Winklers Creek

pH
7.43

DO%
82.4

DO (mg/L)
9.49

Salinity
0.1

Conductivity
221.2

Temperature (C)
9.1

7.54
7.68
7.3
7.2
7.54

84.8
85.6
84
83.1
86.9

9.67
9.59
9.63
9
10.06

0.1
0.1
0.1
0

234.2
233.9
230.8
252.8
33.2

9.9
10.4
9.5
9.8
9

DISCUSSION
Based on the preliminary PAH analyses from Dr. Weinstein, concentrations of PAHs are still at
extremely high levels within the sediments of Hodges Creek. We suggest that this could have a negative
effect on the health of the aquatic invertebrates and possibly the fishes within Hodges Creek. Although
recolonization has taken place, the health of the invertebrate organisms for cancer or other potential health
problems was not examined. The fish that were found appeared to have a similar condition index as fish

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collected from the reference stream, Winklers Creek. The abundances of fish within Hodges Creek are
drastically lower than that of Winklers Creek, but the diversity does not appear to differ. We suggest that
the differences seen in abundance and diversity for fishes and invertebrates within Hodges Creek is more
a result of stream habitat. The stream is highly urbanized and subject to significant runoff and
sedimentation from roads, parking lots, and construction sites along its banks. Once the results come back
from Dr. Weinstein, we will be able to tell if the concentrations of PAHs in the sediments decreased as
distance increased from the spill site. If concentrations are reasonably constant, then this will confirm our
hypothesis that the fluctuations in abundance and diversity are a function of habitat and not a result of the
spill.
We believe that over time the invertebrates in Hodges Creek will suffer long term health effects,
and fish will continue to show reduced abundances due to diminished food resources, such as
invertebrates, due to the PAH contamination from the coal-tar based asphalt sealant that washed off the
BB&T parking lot in July 2010. Monitoring should take place every 6 months to determine whether
PAHs are still present in the sediments. Bioremediation techniques could be explored to purify the
sediments within Hodges Creek, but due to the lack of evidence supporting the effectiveness of these
tactics, residents may simply have to demand that testing be done every couple months in order to
determine when the PAHs have desorbed from the sediments. More testing should occur downstream and
within the New River to determine the extent of PAH contamination within this system.
With respect to human health, there are high levels of possible carcinogens within the sediments
of Hodges Creek, but due to the lack of long term exposure to humans playing in the creek, PAHs may
pose little risk compared to our regular daily intake. It is estimated that the average daily exposure to
carcinogenic PAHs is between 3 and 15 g for adult American males. The majority of this exposure
comes from food and air (Menzie et al. 1992). Although dermal exposure to PAHs from the sediments is
not a very effective route of uptake, it might be prudent to not allow barefoot children to play in the creek
for the foreseeable future, and the consumption of any organisms from Hodges Creek is not advised
(Weinstein, personal communication).

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While invertebrates and fish have recolonized Hodges Creek, PAHs and poor stream management
practices have condemned this ecosystem to be highly impacted for a significant period of time. More
research is needed to show the extent, with respect to stream distance and time, that this stream will be
impacted by PAH contamination.

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