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Effects of stream acidification on loticsalamander assemblages in a coal-mined watershed in the CumberlandPlateauMark S. Schorr a , Melissa C. Dyson b , Charles H. Nelson a , GeneS. Van Horn a , David E. Collins c & Sean M. Richards aa Department of Biological and Environmental Sciences ,University of Tennessee at Chattanooga , Chattanooga ,Tennessee , USAb Unit for Laboratory Animal Medicine , University of Michigan ,Ann Arbor , Michigan , USAc Tennessee Aquarium , Chattanooga , Tennessee , USAPublished online: 02 May 2013.

To cite this article: Mark S. Schorr , Melissa C. Dyson , Charles H. Nelson , Gene S. Van Horn ,David E. Collins & Sean M. Richards (2013): Effects of stream acidification on lotic salamanderassemblages in a coal-mined watershed in the Cumberland Plateau, Journal of Freshwater Ecology,DOI:10.1080/02705060.2013.778219

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Effects of stream acidification on lotic salamander assemblages in

a coal-mined watershed in the Cumberland Plateau

Mark S. Schorra�, Melissa C. Dysonb, Charles H. Nelsona, Gene S. Van Horna,

David E. Collinsc and Sean M. Richardsa

aDepartment of Biological and Environmental Sciences, University of Tennessee at Chattanooga,Chattanooga, Tennessee 37403 USA; bUnit for Laboratory Animal Medicine, University ofMichigan, Ann Arbor, Michigan 48109 USA; cTennessee Aquarium, Chattanooga,Tennessee 37402 USA

(Received 3 September 2012; final version received 23 January 2013)

We studied the effects of acid mine drainage (AMD) from abandoned coal mines onlotic salamanders and environmental conditions in the upper watershed (CumberlandPlateau) of North Chickamauga Creek (NCC; Tennessee River drainage) in southeas-tern Tennessee, USA, from 1996–97. Study sites (2nd- or 3rd-order reaches) were sam-pled in an AMD-influenced section (five sites) and in two reference streams (twominimally disturbed sites). A total of 212 plethodontids (premetamorphic larvae) rep-resenting four species were collected by kicknetting in riffles (n ¼ 99) and electrofish-ing in mixed habitats (n ¼ 113). The dusky salamander (Desmognathus fuscus) wasthe most abundant species in both AMD and reference reaches (> 80 – 90% of totalcatches), successively followed by the southern two-lined salamander (Euryceacirrigera), spring salamander (Gyrinophilus porphyriticus), and red salamander(Pseudotriton ruber). Mining-influenced reaches were characterized by acidic flows(mean pH ¼ 3.8–5.6), zero to low alkalinity, and elevated conductivity, sulfate, hard-ness, aluminum, and manganese, as well as very low abundances of salamanders.Reference reaches were slightly acidic to circumneutral (mean pH ¼ 6.0–6.9) withlow to moderate alkalinity, low levels of conductivity, hardness, sulfate, and metals,and high salamander abundances. Our findings document the impact of acid/metalpollution from past coal mining activities on lotic salamanders in a CumberlandPlateau stream.

Keywords: stream salamanders; acid mine drainage; abandoned coal mines; pH;aluminum; Cumberland Plateau

Introduction

The Appalachian Mountains in the eastern United States are a global center for salaman-

der diversity and endemism, particularly in the family Plethodontidae (Dodd 1997; Mila-

novich et al. 2010); however, the region’s naturally low limestone-based alkalinity

makes its streams and watersheds sensitive to chemical pollution associated with acid pre-

cipitation and mine drainage (Griffith et al. 1997). Mine drainage accounts for approxi-

mately 26% of the stream acidity in acid-sensitive watersheds of the USA (Baker et al.

1991). Acid mine drainage (AMD) is a highly acidic, metal-laden discharge that emanates

from exposed mines, sometimes for decades (National Research Council 1992; Kadlec

and Knight 1996). In the Appalachian region, AMD is typically associated with aban-

doned bituminous coal mines (Powell 1988). Abandoned mines in Tennessee have

*Corresponding author. Email: Mark-Schorr@utc.edu

� 2013 Taylor & Francis

Journal of Freshwater Ecology, 2013

http://dx.doi.org/10.1080/02705060.2013.778219

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impaired water quality in an estimated 720 km of streams, generally marked by acid and

metal pollution in areas of historical coal mining (TDEC 2010).

Streams influenced by coal mine drainage typically exhibit a lowered pH, elevated

concentrations of sulfate, calcium and magnesium ions, and dissolved/precipitated trace

metals (e.g., aluminum, iron, manganese), and increased turbidity and sedimentation (Na-

tional Research Council 1992). Aquatic animal responses to acid/metal pollution may in-

clude death, avoidance behavior, and impaired growth/reproduction (e.g., Matter et al.

1978; Fromm 1980; Herrmann et al. 1993; Woodward et al. 1997; Kalff 2002). Elevated

concentrations of hydrogen and metal ions can impair osmoregulation, ion regulation,

and respiration in aquatic macrofauna, such as insect larvae, fishes, salamander larvae. In-

direct effects may result from altered food webs (e.g., prey-predator relationships) and de-

graded habitats like mining-related sedimentation and metal armoring of streambeds

(Winger 1978; Platts et al. 1979; Fromm 1980; Herrmann et al. 1993; Poleo 1995; Dodd

1997; Cherry et al. 2001; Henry et al. 2001; Kalff 2002).

Aquatic salamanders are sensitive to increased concentrations of hydrogen and metal

ions (particularly aluminum) with eggs, embryos, and larvae being the most sensitive life

stages (Dodd 1997). Acid-toxicity responses have been well documented for pond-

breeding ambystomatid salamanders (e.g., Freda and Dunson 1985; Pierce 1985; Ling

et al. 1986; Sadinski and Dunson 1992). In comparison, little is known regarding the

responses of stream-dwelling plethodontids to acid/metal pollution and other environ-

mental stressors (Petranka 1998; Rocco and Brooks 2000).

Plethodontid salamanders (e.g., Desmognathus, Eurycea spp.) play key ecological

roles in Appalachian headwater streams and riparian forests, where they may function as

both invertebrate predators and vertebrate prey (e.g., Burton 1976; Rudolph 1977; Barr

and Babbitt 2002). In weakly buffered Appalachian watersheds, acid mine/rock drainage

emanating from pyritic rock can cause acidification resulting in the local decimation of

populations of stream-dwelling plethodontids (Matthews and Morgan 1982; Gore 1983).

Plethodontid responses to pH, metal, and other environmental gradients can vary with the

species, ontogeny, and study area/design (Mushinsky and Brodie 1975; Roudebush 1988;

Rocco and Brooks 2000; Barr and Babbitt 2002; Grant et al. 2005).

North Chickamauga Creek (NCC), a fourth-order tributary to the Tennessee River in

the Chattanooga, Tennessee area, drains portions of the Southwestern Appalachians

(Cumberland Plateau, Plateau Escarpment) and Ridge and Valley (Southern Limestone/

Dolomite Valleys and Rolling Hills) ecoregions (Griffith et al. 1997). Drainage from

abandoned coal mines (surface and underground) has acidified (pH < 6) approximately

40.7 km in the headwaters (Cumberland Plateau) and 6.6 km of the middle reaches (Pla-

teau Escarpment, Ridge and Valley) of the NCC system (TDEC 2005). However, AMD-

related effects on lotic salamanders in the NCC system have not been documented. Our

objectives were: (1) to estimate and compare selected physicochemical conditions, in-

stream and riparian habitat features, total macroinvertebrate abundance, and salamander

abundance and species composition in AMD-impacted sites and minimally disturbed (ref-

erence) sites in the NCC study area; and (2) to examine correlations of salamander abun-

dance with selected environmental variables across all sites.

Methods

Study area

The study area included a total of seven sites in the upper watershed of NCC, Hamilton

and Sequatchie counties, Tennessee, USA (Figure 1). Stream sites (200-m sampling

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reaches) were established on third- and fourth-order reaches with drainage areas less than

35 km2.

Five mining-influenced sites were established along a 10-km section of Standifer

Creek (tributary and source of AMD; two 2nd-order reaches) and NCC proper (three 3rd-

order reaches); perennial acidification prevailed at these sites. Two reference sites were

established on Brimer Creek (2nd-order reach) and Falling Water Creek (3rd-order reach),

two tributaries which did not exhibit problematic AMD-related degradation. Selection of

AMD and reference sites was based on historical site data (OSMRE 1987), professional

consultation (T. Eagle, D. Fritz, Tennessee Department of Environment and Conservation

Figure 1. Location of acid mine drainage (AMD) sources (X’s), AMD-influenced sites (1–5) andreference sites (6–7) (filled circles) in the North Chickamauga Creek system (Tennessee Riverdrainage; Hamilton-Sequatchie counties, Tennessee).

Journal of Freshwater Ecology 3

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(TDEC) and G. Brodie, Tennessee Valley Authority), and preliminary sampling (M.S.

Schorr, unpublished data). All sites were located in the Cumberland Plateau subdivision

of the Southwestern Appalachians ecoregion (Griffith et al. 1997).

Two years of sample data (1996–1997) were gathered from six or seven sites. Sam-

pling activities at each site commenced at a downstream point and proceeded in an up-

stream direction. Most stream surveys were conducted at six sites – two in Standifer

Creek (Sites 1 and 2), two in NCC proper (Sites 3 and 5), and two in the reference streams

(Brimer, Site 6; and Falling Water Creek, Site 7) – which encompassed the study area

(Figure 1). Additional data were acquired and analyzed from another reach in NCC

proper (Site 4, located between Sites 3 and 5, which was used in a concurrent fish assem-

blage study; Schorr et al. 1997), which augmented the database on environmental condi-

tions and salamanders in the AMD-influenced section.

Physicochemical conditions

Physicochemical conditions were assessed in a shallow run (< 1 m deep) at each site.

Stream temperature, dissolved oxygen, pH, and conductivity were measured monthly,

January through December, 1996–1997 at the sampling sites using a multimeter (Hydro-

lab, Model H20, http://www.hachhydromet.com). Surface grab samples, collected

monthly in May-July (1996–1997) at six sites, were analyzed at the Tennessee Valley Au-

thority (TVA) Environmental Chemistry Laboratory (Chattanooga, Tennessee) for total

alkalinity, total hardness, calcium, magnesium, sulfate, aluminum, copper, iron, manga-

nese, zinc, and turbidity.

Habitat features

In-stream and riparian habitat characteristics were measured once per year in May-July

(1996–1997) at seven sites. Stream morphology (wetted width, mean depth) and substrate

features were quantified along 10 evenly-spaced transects spanning the stream width at

each site. Depth was recorded at 10 evenly-spaced points on each transect. Substrate com-

position (% area of fine sediment [< 2 mm], gravel [2–64 mm], cobble [64–256 mm],

boulder [> 256 mm], bedrock) and embeddedness (% gravel/cobble embedded or buried

by fine sediments) in the streambed was quantified within three or four equally-spaced

quadrats (0.3 � 0.3 m plastic grid) on each transect. Riparian vegetation (% ground

cover) was measured on ten 15-m transects (extensions of the stream width transects) on

both sides of the channel. The aforementioned habitat measurement techniques were

adapted from Platts et al. (1983) and McMahon et al. (1996)

Salamander assemblages

Benthic salamanders (along with macroinvertebrates, sediment and detritus) were col-

lected by monthly kick sampling in May and June (1996–1997) at six sites. Twenty-four

kick samples (6 sites x 2 months x 2 years) were collected in the study. At each site, three

subsamples were gathered from three different riffles (located near randomly selected

transects) on each sampling date. Each subsample was collected by kick sampling for two

minutes in a riffle area with a rectangular-frame net (500-mm mesh). Sampling areas

ranged from 1.1 m2 to 2.7 m2. Kick-net data were used to estimate overall species com-

position and site-specific abundance (index of density) in riffles. Abundance was quanti-

fied as the number of individuals per 1 m2 in riffles. (As part of a broader study, kick-net

4 M.S. Schorr et al.

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sampling was conducted primarily to collect data on benthic macroinvertebrates, Schorr

et al. 1997). Total macroinvertebrate abundance was calculated as a gross index of prey

availability for salamanders.

While sampling fish assemblages in the NCC system (Schorr et al. 1997), we collected

additional data on lotic salamanders (species composition) at four study sites. Salamanders

(incidental catch) were collected along with fishes (target fauna) in mixed habitats (riffles/

runs/pools) by monthly electrofishing in May–July, 1996–1997, at three AMD sites (NCC

proper, Sites 3–5) and one reference site (Falling Water Creek, Site 7). At each site, three-

pass electrofishing was conducted using two backpack shockers (Model 15-C 300-W,

Smith-Root, http://www.smith-root.com) and two netters within three sampling enclosures

located between randomly selected transects on each sampling date. Salamanders were

captured using dip nets or a downstream block net (6� 1.8 m seine with 0.5 cm mesh).

Kick-net samples and electrofishing samples (salamanders and other macrofauna)

were fixed in 10% formalin in the field (for processing and taxonomic identifications),

and preserved in 80% ethanol in the laboratory. Preserved specimens were identified to

species, counted, and measured in the laboratory. The total length (TL) of individual sala-

manders was measured to the nearest 0.1 mm with dial calipers.

Statistical analyses

Stream sample data were analyzed using the Statistical Analysis System (SAS 1988).

Data for several of the variables exhibited non-normal distributions and/or heterogeneous

variances, which could not be resolved with data transformations. Hence, we employed a

nonparametric approach by analyzing the ranks of the measurement data with analysis of

variance (ANOVA) (Conover and Iman 1981; SAS 1988); comparative analyses of mea-

surement data and ranks (Zar 2010) yielded the same conclusions. Statistical significance

was declared at the 0.05 level. Site-specific estimates of stream parameters are reported

as means � SE of the measurement data.

Factorial ANOVA’s (repeated measures without replication) of rank-transformed data

were used to measure site and year effects on environmental and biotic variables. Parame-

ter estimates based on site-specific, transect-based measurements of habitat features

(8–10 per site) and macroinvertebrate and salamander abundance (3 per site) were ana-

lyzed with hierarchical repeated-measures ANOVAs. Transect measurements (nested fac-

tor) were analyzed as subsamples nested within sites. Hierarchical ANOVAs and related

post-hoc tests (Dunnett’s test described below) employed the nested factor (subgroup)

mean square in the analysis (Zar 2010).

Dunnett’s test was employed (when p � 0.05 for an ANOVA-based site effect) to

compare stream conditions (mean ranks) between AMD-affected and reference reaches.

Planned comparisons between AMD and reference sites, based on the catchment location

(independent drainage basins), size (similar drainage areas) and stream order, included:

2nd-order reaches, Standifer Creek (Sites 1 and 2; AMD sites) versus Brimer Creek

(Site 6; reference site); and 3rd-order reaches, NCC proper (Sites 3–5; AMD sites) versus

Falling Water Creek (Site 7; reference site).

Spearman’s correlation analysis was used to examine monotonic relationships be-

tween salamander abundance in riffles (from kick net data; n ¼ 6 sites) and selected envi-

ronmental conditions (pH, conductivity, streambed habitat, riparian vegetation).

Environmental variables were chosen for the correlation analysis based on hypothesized

causal relationships with benthic salamanders (e.g., acidity, sedimentation), evidence of

between-site differences (ANOVA, described above), and reduced collinearity.

Journal of Freshwater Ecology 5

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Results

Physicochemical conditions

Reaches influenced by AMD (Sites 1–5) were characterized by reduced pH and elevated

conductivity relative to reference reach conditions (Sites 6 and 7; p < 0.05; Table 1).

Physicochemical data collected at sites spanning from upper Standifer Creek to NCC

proper illustrate a downstream gradient of increasing pH (Site means ¼ 3.8–5.6) and de-

creasing conductivity (260–47 mS/cm); references reaches exhibited higher pH (6.0–6.9)

and lower conductivity (22.9–53.3 mS/cm; p < 0.05) values, although conductivity in

middle-lower reaches of NCC proper was similar to reference levels. Temperature and

DO levels were similar at AMD and reference sites (p > 0.10). Between-year differences

were detected (p < 0.05) for temperature, DO, pH, and conductivity.

AMD-influenced sites exhibited low levels of alkalinity and high levels of sulfate,

hardness, aluminum, and manganese compared to reference sites (p < 0.05; Table 2).

Mean conditions at AMD sites reflected a downstream gradient of increasing alkalinity

and decreasing hardness, sulfate, aluminum, and manganese. Iron and turbidity were rela-

tively high at AMD Site 1, but a longitudinal gradient was not observed for these constitu-

ents. Between-year differences were detected (p < 0.05) for alkalinity, sulfate,

manganese, and turbidity. Zinc was not detected (detection limit ¼ 0.01 mg/L) in more

than 50% of the samples and, thus, was not tested for between-Site differences and copper

was not detected (detection limit ¼ 0.01 mg/L) in any of the samples.

Habitat characteristics

Stream habitat features were generally similar at AMD and reference sites (Table 3), and

did not exhibit a longitudinal pattern. However, between-Site differences (AMD versus

references reaches) were observed at two sites in the AMD section (p < 0.05). Upper

Standifer Creek (Site 1) exhibited relatively low levels of riparian plant cover and high

levels of streambed sedimentation and embeddedness. The middle-reach Site in NCC

proper (Site 4) had a relatively high level of cobble in the streambed.

Table 1. Mean physicochemical conditions (SE in parentheses, n ¼ 23–24) at AMD-influencedsites and reference sites in the North Chickamauga Creek system, January-December 1996–1997.Estimates are based on two years of data (11–12 monthly measurements per site per year). Asterisksdenote significant differences (rank-transformed data) between AMD and reference sites withinstream-order groups (p < 0.05, ANOVA and Dunnett’s test).

2nd-order streams 3rd-order streams

AMD reachesin Standifer Creek

Referencereach

AMD reaches in NorthChickamauga Creek proper

Referencereach

Parameter Site 1 Site 2 Site 6 Site 3 Site 4 Site 5 Site 7

Temperature (�C) 11.88 11.66 11.52 11.55 12.46 12.84 12.51(0.85) (1.08) (1.14) (1.10) (1.24) (1.40) (1.28)

DO (mg/L) 9.54 9.81 9.57 9.83 9.62 9.66 9.50(0.22) (0.26) (0.34) (0.28) (0.29) (0.29) (0.41)

pH 3.75� 4.59� 6.01 4.88� 5.41� 5.64� 6.94(0.05) (0.07) (0.08) (0.07) (0.09) (0.09) (0.06)

Conductivity(mS/cm)

266.2� 139.6� 22.88 93.1� 62.9 47.2 53.3(18.9) (9.7) (0.44) (8.1) (5.7) (3.6) (3.5)

6 M.S. Schorr et al.

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Macroinvertebrate abundance

Benthic macroinvertebrate abundance was consistently lower in the AMD-influenced sec-

tion than in the reference streams (p < 0.05). Total macroinvertebrate abundance (all taxa

combined; number of individuals per m2) ranged from 74.6 � 4.2 to 134.4 � 74.1 (Sites 1

Table 2. Mean physicochemical conditions (SE in parentheses, n ¼ 6) at AMD-influenced sitesand reference sites in the North Chickamauga Creek system, Tennessee, May-July 1996–1997.Estimates are based on two years of data (two monthly measurements per site per year). Asterisksdenote differences (rank-transformed data) between AMD and reference sites within stream-ordergroups (p < 0.05, ANOVA and Dunnett’s test).

2nd-order streams 3rd-order streams

AMD reachesin Standifer Creek

Referencereach

AMD reaches in NorthChickamauga Creek proper

Referencereach

Parameter Site 1 Site 2 Site 6 Site 3 Site 5 Site 7

Alkalinity (mg/L CaCO3) 0.00� 1.25� 2.42 1.67� 2.42� 15.83(0.00) (0.96) (0.71) (0.76) (0.76) (1.11)

Hardness (mg/L CaCO3) 81.48� 52.77� 7.47 31.38� 18.80 19.35(14.54) (7.88) (0.45) (5.32) (1.98) (1.22)

Sulfate (mg/L) 87.17� 52.17� 5.83 25.17� 15.33� 6.17(19.26) (9.56) (0.40) (4.43) (2.14) (0.65)

Aluminum (mg/L) 2.94� 0.92� 0.23 0.48� 0.16 0.10(0.52) (0.25) (0.06) (0.15) (0.03) (0.04)

Iron (mg/L) 1.02� 0.23 0.26 0.18 0.13 0.13(0.29) (0.11) (0.09) (0.05) (0.04) (0.02)

Manganese (mg/L) 1.32� 0.57� 0.04 0.34� 0.13� 0.01(0.31) (0.13) (0.03) (0.07) (0.04) (<0.01)

Turbidity (NTU) 0.67� 1.42 3.17 2.25 1.83 1.83(0.11) (0.44) (1.01) (0.48) (0.53) (0.31)

Table 3. Mean habitat conditions (SE in parentheses, n ¼ 2) at AMD-influenced sites and refe-rence sites in the North Chickamauga Creek system, Tennessee, May–July 1996–1997. In-streamsubstrate (cobble, sediment, embeddedness) and riparian vegetation estimates are based on twoyears of data (recorded at 16–20 transects per site per year). Asterisks denote significant differences(rank-transformed data) between AMD and reference sites within stream-order groups (p < 0.05,ANOVA and Dunnett’s test).

2nd-order streams 3rd-order streams

AMD reachesin Standifer Creek

Referencereach

AMD reaches in NorthChickamauga Creek proper

Referencereach

Parameter Site 1 Site 2 Site 6 Site 3 Site 4 Site 5 Site 7

Cobble (%) 21.90 19.10 28.15 21.22 44.75� 19.69 26.54(0.14) (1.50) (10.09) (4.82) (4.03) (3.38) (4.86)

Fine sediment (%) 17.68� 6.37 3.52 3.95 4.76 4.77 15.86(6.43) (2.95) (1.57) (1.08) (0.91) (2.17) (0.50)

Embeddedness (%) 73.92� 43.62 32.77 68.43 43.04 46.20 46.14(4.59) (7.49) (20.21) (3.32) (2.10) (9.18) (0.16)

Riparian plants (%) 16.78� 36.51 35.62 46.32 44.08 61.17 48.65(5.28) (0.32) (10.79) (0.91) (0.08) (6.86) (2.10)

Journal of Freshwater Ecology 7

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and 2) in Standifer Creek, from 190.1 þ 83.7 to 201.6 � 50.4 in NCC proper (Sites 3

and 5), and from 434.2 � 117.9 to 550.1 þ 213.7 in the reference streams (Sites 6 and 7,

respectively). Abundances were statistically similar in 1996 and 1997 (p > 0.10).

Salamander assemblages

Riffle samples (six sites, 99 individuals, Figure 2) contained mostly D. fuscus (92%) with

a small number of E. cirrigera (8%). Mixed-habitat samples (four sites; 113 individuals,

Figure 2) were comprised mostly of D. fuscus (81%), followed by E. cirrigera (13%),

Gyrinophilus porphyriticus (4%), and P. ruber (2%); all four species and 58% of the catch

were found at the Falling Water Creek reference Site (Site 7) versus two species (D. fus-

cus and G. porphyriticus) and 42% of the catch at the NCC proper sites (Sites 1–3). Virtu-

ally all of the salamanders collected (> 90%) were premetamorphic larvae with external

gills and fully developed, functional limbs (Petranka 1998).

Figure 2. Salamander species (n ¼ number of individuals, TL ¼ total length (mean � SE) col-lected by kick-netting in riffles (May-June) and electrofishing in mixed habitats (pools, riffles, andruns; May–July) in the North Chickamauga Creek system, Tennessee, 1996–1997.

8 M.S. Schorr et al.

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Both total salamander and D. fuscus abundance in riffles (Figure 3) was consistently

lower in the AMD-influenced reaches of Standifer Creek and NCC proper than in the

respective reference reaches (p < 0.05); abundances were similar in 1996 and 1997

(p > 0.10). Dusky salamander (D. fuscus) abundance (number of individuals per m2)

ranged from: 0 to 0.1 � 0.1 in the AMD reaches and 1.3 � 0.5 to 2.8 þ 0.7 in the refer-

ence reaches. Abundance of two-lined salamanders (E. cirrigera), which were found at

only two sites (Site 5, < 0.1 � 0.04; Site 6, 0.4 � 0.2), was not tested statistically.

Ecological correlations

Salamander abundances in riffles (D. fuscus and total) were strongly correlated with pH

(r ¼ 0.943) and conductivity (r ¼ �0.943; p < 0.05); these physicochemical parameters

Figure 3. Salamander abundance (mean number of individuals per 1 m2, SE bars, n ¼ 4) based onkick-net sampling in riffles in AMD-influenced (2nd-order sites 1 and 2; 3rd-order sites 3 and 5) andreference reaches (2nd-order site 6; 3rd-order site 7) in the North Chickamauga Creek system, May-June, 1996–1997. Asterisks (�) denote significant differences (rank-transformed data) in order-spe-cific comparisons between AMD and reference sites (p < 0.05; ANOVA and Dunnett’s tests).

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were correlated with alkalinity, hardness, and most minerals and metals (Ca, Mg, Al, Mn;

p < 0.05), but were not correlated with the other environmental variables (p > 0.10). In

1996 and 1997, D. fuscus abundance was very low in reaches with a pH of < 5.5 and con-

ductivity >100 mS/cm (Figure 4). Correlations of salamander abundances with other en-

vironmental variables were not statistically significant (p > 0.10).

Discussion

Findings from the present study contribute to the small but growing body of work regard-

ing the effects of cultural acidification on plethodontid salamander assemblages in Appa-

lachian Mountain streams. In this manuscript we (1) characterize environmental

conditions and lotic salamander assemblages in a Cumberland Plateau watershed; (2) doc-

ument the effects of AMD from abandoned coal mines on stream water chemistry and sal-

amander abundance; and (3) illustrate the response of riffle-dwelling salamander larvae to

gradients of acid/metal pollution.

Salamander assemblage data (species composition and abundance) collected in the

NCC study area are generally consistent with findings from other studies of stream-dwelling

Figure 4. Scatter plots depicting relationships of dusky salamander (Desmognathus fuscus) abun-dance in riffles with stream pH and conductivity in the North Chickamauga Creek system, May-June, 1996 (circles) and 1997 (squares).

10 M.S. Schorr et al.

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salamanders in Appalachian watersheds. Biotic sampling in riffles and other habitats in

the NCC system yielded four species of plethodontids. Desmognathus fuscus was by far

the most abundant species (92% in riffles; 80% in mixed habitats), followed by E. cirri-

gera (8% in riffles; 14% in mixed), G. porphyriticus (4% in mixed), and P. ruber (2% in

mixed). Similar salamander assemblages (1–7 plethodontid species; predominantly

Desmognathus and/or Eurycea spp.) have been documented in other studies of Appala-

chian streams (Blue Ridge, Cumberland Plateau, Ridge and Valley ecoregions; Gore

1983; Rocco and Brooks 2000; Barr and Babbitt 2002; Grant et al. 2005).

In-stream habitats in the Cumberland Plateau portion of the NCC system were utilized

by larval plethodontids. Other investigators have reported mostly plethodontid larvae in

aquatic habitats: riffles (Gore 1983), stream banks (Rocco and Brooks 2000), pools/riffles

(Barr and Babbitt 2002), whereas adults were found in terrestrial stream banks (Grant

et al. 2005).

Physicochemical conditions in the reference reaches varied from slightly acidic to cir-

cumneutral (mean pH ¼ 6.0–6.9) with fairly low levels of alkalinity, conductivity, hard-

ness, sulfate, and trace metals; these conditions approximate those in relatively

undisturbed streams draining sandstone-based Appalachian watersheds (Kalff 2002;

Merovich et al. 2007; Bernal 2010). Mean abundances of D. fuscus at the reference sites

(generally 2–3 individuals/ m2) approximate densities reported for Desmognathus larvae

in other North American streams. Petranka (1998) reported > 1–2 individuals/m2 of D.

fuscus in “optimal habitats” and Davic and Welsh (2004) reported 1.7–2.8 individuals/m2

of D. quadramaculatus.

Mining-impacted reaches in the NCC system exhibited reduced pH and alkalinity val-

ues, increased conductivity and hardness levels, and elevated concentrations of calcium,

magnesium, sulfate, and trace metals (aluminum, iron, manganese). These conditions are

characteristic of Appalachian streams acidified by coal mine drainage (Powell 1988;

National Research Council 1992; Kadlec and Knight 1996). High conductivity and hard-

ness downstream of coal mines in upper NCC reflect greater concentrations of dissolved

constituents, which correspond with elevated levels of minerals and metals.

Benthic salamander abundance was exceedingly low at the mining-impacted sites in

Standifer Creek (mean pH ¼ 3.8–4.6) and NCC proper (pH ¼ 4.9–5.6) where average pH

conditions approximated respective acidity thresholds at which embryonic/larval ambys-

tomatids exhibit high rates of mortality (pH < 4.0–4.5) and sublethal effects (pH ¼ 4.5–

5.5; Freda and Dunson 1985; Pierce 1985; Ling et al. 1986; Sadinski and Dunson 1992;

Petranka 1998). Similarly, larval plethodontids (D. quadramaculatus, D. marmoratus,

E. wilderae) were virtually eliminated from a contaminated section of a Blue Ridge

stream following exposure to an acidic (pH ¼ 4.5) pyrite-rich drainage from Anakeesta

rocks used in a road construction project (Huckabee et al. 1975); this event resulted in

long-term acidification and pronounced declines in stream-breeding plethodontids (and

other macrofauna), which altered the population density and structure of terrestrial-breed-

ing plethodontids (Kucken et al. 1994). In a related study, Matthews and Morgan (1982)

documented 100% mortality of D. marmoratus larvae after 96-hour exposure to Ana-

keesta leachates with pH ¼ 3.9–5.0 and elevated concentrations of metals.

Aluminum toxicity, in addition to acidity, probably contributed to the reduced abun-

dance of salamanders in the AMD-influenced section. The toxicity of aluminum to

aquatic amphibians is complex and influenced by pH, hardness, dissolved organic carbon

(DOC), and the species and ontogenetic stage (McCahon and Pascoe 1989; Freda 1991).

Aluminum, elevated at Sites 1–3 where the mean pH was < 4–5, forms potentially toxic

free aluminum cations (Alþ3) at pH < 5.5 (Kalff 2002). Moreover, aluminum hydroxide

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cations (Al[OH]2þ, Al[OH]þ2) can cause acute toxicity at pH of 5.5–6.0 in ‘mixing

zones’ (e.g., confluence of acidic and neutral streams, Poleo 1995), which likely occurred

in upper NCC proper immediately downstream of the Standifer-Brimer confluence.

Total aluminum levels (mg/L) in Standifer Creek and upper NCC proper (mean

[range]: Site 1 ¼ 2.94 [0.92–4.4]; Site 2 ¼ 0.92 [0.15–1.80]; Site 3 ¼ 0.48 [0.14–1.0])

exceeded the widely used acute aquatic life standard of 0.75 mg/L (EPA-based critical

maximum criterion; U.S. Environmental Protection Agency 2009), which has been

adopted in studies of mined Appalachian watersheds (e.g., Freund and Dodd 2007; Mero-

vich et al. 2007). Additionally, total aluminum recorded at the AMD sites approximated

or exceeded levels known to cause mortality or impair growth/development in aquatic

amphibians (tadpoles or larval salamanders) in pH-DOC-controlled laboratory experi-

ments (Blem and Blem 1991; Freda 1991; Freda and McDonald 1993).

In an extensive survey of 78 streams in the southern Cumberland Plateau, Gore (1983)

documented one salamander species (D. fuscus) in benthic habitats (Surber samples) of 17

coal-mined watersheds; D. fuscus larvae were commonly found in reaches with a pH of

6–7 and a conductivity of < 100 mS/cm and rarely found in reaches with a pH of < 5.

Similarly, we found that the abundance of benthic D. fuscus larvae (kick-net samples)

was high at a pH of 6.0–7.0, variable at a pH of 5.5–6.0, and low at a pH of < 5.5 or a

conductivity of >100 mS/cm (Figure 4). However, in low-conductivity (< 100 mS/cm)

reaches, D. fuscus abundance was quite variable (range ¼ 0–3.6 individuals per m2) and

appeared to be determined more by pH (range ¼ 4.7–7.0).

Macroinvertebrate abundance was reduced at the AMD sites, suggesting that there

was a reduced trophic base for benthic salamanders in the mining-influenced portion of

the NCC study area. Lotic salamanders feed primarily on benthic macroinvertebrates

(Petranka 1998), particularly feeding guilds of larval plethodontids (e.g., Desmognathus,

Eurycea; Burton 1976; Rudolph 1977) such as those observed in the NCC system.

Habitat features (streambed, riparian plant cover), which did not exhibit an overall

pattern of differences between the AMD and reference reaches, were not correlated with

salamander abundance. Most sites were characterized by rocky substrates, shaded stream

channels, and forested riparian zones. Pronounced acid/metal contamination in the AMD

section probably masked the effects of other environmental factors. However, reduced ri-

parian forest cover and streambed sedimentation (characteristic of mined watersheds,

Starnes and Gasper 1995) were observed in the uppermost reach on Standifer Creek

(closest to abandoned coal mines).

In summary, localized declines in the abundance of plethodontid salamanders in the

AMD-influenced stream reaches (Standifer Creek-NCC proper) corresponded to tren-

chant gradients of acid and metal pollution emanating from abandoned coal mines. Spe-

cifically, our data indicate that AMD from abandoned coal mines in the Standifer Creek

watershed appears to have severely decimated lotic salamander populations (D. fuscus

and probably other species) in the NCC headwaters.

Recent data collections (M.S. Schorr 2008, 2010–2012) indicate that acidification issues

persist in Standifer Creek (Sites 1–2: pH ¼ 3.4–4.6; conductivity ¼ 131–400 mS/cm).

Problematic AMD point sources in the NCC system (six in Standifer Creek, Cumberland

Plateau and one in Hogskin Branch, Plateau Escarpment) include abandoned surface coal

mines and deep coal mines (G. Vickrey, North Chickamauga Creek Conservancy, unpub-

lished report). Pollution-abatement systems (e.g., aerobic limestone wetlands, anoxic

limestone drains, successive alkalinity producing systems) have been constructed at five

AMD sources in the Standifer Creek watershed (T. Eagle, TDEC, unpublished report),

which could improve stream water quality and, therefore, salamander numbers and

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diversity. Reference-reach data from the present study suggest that the NCC headwaters

have the potential to support healthy assemblages of lotic salamanders.

Acknowledgements

This study was funded by the Tennessee Wildlife Resources Agency. We thank the TennesseeAquarium and Southeast Aquatic Research Institute (now the Tennessee Aquarium ConservationInstitute), particularly George Benz for providing laboratory space and facilities and Chris Coco,Robert Mottice, and Tom Tarpley for assistance with the field work. Many students at the Univer-sity of Tennessee at Chattanooga (UTC) assisted with field and/or laboratory work, particularlyJohn Beck, Jeannie Cuervo, David Fielder, Katherine Freeman, Kristie Jenkins, Dan Mathis, MaryJane Middlekoop, Sabrina Novak, and Tiffany Watts. Erika Tierce helped with some of the data en-try and graphics. Requested information and/or services were provided by the North ChickamaugaCreek Conservancy, Tennessee Department of Environment and Conservation, Tennessee ValleyAuthority, and U.S. Office of Surface Mining Reclamation and Enforcement. Andy Carroll (UTC)created the map of the study area. Landowners in the upper watershed of North Chickamauga Creekgraciously allowed us access to the sites.

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