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					                                                                        Assessing Fish Community Health following the Kingston Fly Ash Spill
                                                           Anna L. George1, David A. Neely1, Shea R. Tuberty2, Robert J. Mottice1, Donna M. Lisenby3, Daniel Jackson2, Yosuke Sakamachi2
                                                             (1) Tennessee Aquarium, Chattanooga, TN; (2) Appalachian State University, Boone, NC; (3) Appalachian Voices, Boone, NC

                Fly ash has been documented to affect fishes through direct mortality from alkaline shock, chronic oxidative stress, and bioaccumulation of toxic elements such as selenium. Since January 2009, our team has conducted quarterly fish community
                sampling to begin assessment of the long-term impact of the Kingston spill on the fish community in Watts Bar Reservoir. Sampling has been conducted by boat electrofishing for 25-minute periods at five sites on the Clinch, Emory, and
                Tennessee rivers. Sampled fish are identified to species and standard measures are recorded (weight and length). A subset is retained for toxicological analysis, while the remainder is released at that site. Over the past year, ERM 2.0 has
                ranged from 4-6 species and 6-74 individuals. Of the two upstream sites, ERM 3.5 has ranged from 5-13 species and 24-71 individuals, and CRM 5.5 has ranged from 9-10 species and 113-219 individuals. Five km downstream of the spill,
                CRM 3.5 has ranged from 10-13 species and 118-148 individuals, while TRM 567.6 has ranged from 11-15 species and 96-367 individuals. Externally, fishes have displayed elevated rates of lesions and anomalies, particularly gill clubbing. In
                addition, 11.8% of the bluegills caught at Tennessee River Mile 567.6 in January 2010 (11 km downstream of the spill site) exhibited exopthalmia (popeye). Tissue from these individuals is being screened for selenium loading. As seasonality
                greatly affects community structure, we plan to continue sampling until at least January 2012.


      Introduction
      • Previous studies have documented fly ash effects on
        fish from direct mortality from alkaline shock, chronic
        oxidative stress, and bioaccumulation of toxic elements
        such as selenium (Besser et al. 1996, Lemly 2002, Ali
        2004)
      • Long-term impacts of selenium toxicity contributed to the                                                                                                                                                                              Bluegills exhibiting exopthalmia (popeye),
        extirpation of 19 of 20 species in Belews Lake, NC                                                                                                                        Opercular lesion and algal infection (Cladophora
                                                                                                                                                                                                                                               TRM 567.6, January 2010
                                                                                                                                                                                                                                                                                                        Lesion on common carp (670 mm TL), CRM
        (Lemly 2002)                                                                                                                                                              sp.) in largemouth bass (300 mm TL), TRM 567.6,                                                                       3.5, March 2009
                                                                                                                                                                                  March 2009.
      • Our objective is to begin initial studies into the long-term
        impact of Kingston ash spill on the fish community in                                                                                                                    Results
        Watts Bar Reservoir.                                                                                                                                                     • Species richness and individual abundances were always lowest at ERM 2.0, the site of the spill. Species evenness was typically
      • Our secondary objective is to determine if selenium may                                                                                                                    highest at ERM 2.0, while Shannon-Wiener indices varied (Tables 1 and 2).
        be a factor contributing to the observed morphological                                                                                                                   • Of nineteen sampled sites in the past year, seven have contained at least one fish exhibiting an anomaly, with the overall
        anomalies in sampled fishes.                                                                                                                                               percentage at that site varying from 0-13.3% (Table 2).
                                                                                                Figure 1. Location of sampling sites in Watts Bar Reservoir.

                                                                                                                                                                                 • Under Jaccard’s Index, seven site comparisons from January 2010 were of low similarity, while 3 were moderate. Under the
                                                                                                                                                                                   Morisita Index, comparisons of the community at ERM 2.0 with any other site were of low similarity, while all six other community
      Methods                                                                                                                                                                      comparisons were of high similarity (Table 3).
      • Sampling was conducted at a total of eight sites in Watts Bar Reservoir from January 2009-January 2010 (Figure 1).
                                                                                                                                                                                 • The incidence of bluegills exhibiting popeye has risen from 1.5% (July 2009, CRM 3.5) to 11.8% (January 2010, TRM 567.6).
        Fishes were collected by boat electrofishing for 25-minute set periods. Most sites were visited at least three times over the
        sampling period. After collection, fishes were examined for evidence of morphological anomalies, identified to species,                                                  • The proportion of maximum eye width : interorbital width was significantly higher in fish identified as popeye. Whole body
        and measured (mm TL). A subset were retained for toxicological analysis; the remainder were released at the site.                                                          selenium concentrations among the popeye fish ranged from 2.34-6.06 mg/kg dry weight, with a mean of 3.87 mg/kg.
                                                                                                                                                                                   Concentrations among the control group ranged from 3.36 to 5.23 mg/kg, with a mean of 3.98 mg/kg. There was no significant
      • Species richness, evenness, and the Shannon-Wiener index were calculated for each sampling site. Jaccard’s and
                                                                                                                                                                                   difference in either selenium concentrations or condition between these two groups. There was also no correlation between extent
        Morisita similarity indices were calculated for pairwise comparisons of all sites sampled during January 2010.
                                                                                                                                                                                   of popeye and selenium content or condition index and selenium content.
      • During the January 2010 sampling at TRM 567.6, a random subset of 12 bluegills exhibiting exopthalmia (popeye) were
        retained for comparison with a random subset of 12 bluegills not exhibiting the condition. These fishes were weighed, and                                                                                                                                                                         ERM          ERM            CRM         CRM        TRM
        measurements of total length, head length, interorbital width, and maximum eye width were taken with dial calipers. Fish                                                 Discussion                                                                                                               2.0          4.0            5.5         3.5        567.6
        were freeze dried, whole fish homogenized, (75% solids, 25% water), digested 0.5 g with 10 ml OmniTrace 70% nitric acid,                                                 • The fish community at the site of the spill is least similar from                                         ERM
        microwaved for digestion, and analyzed on a Barian ICPAES.                                                                                                                                                                                                                           2.0                   -      0.125             0.3     0.36          0.21
                                                                                                                                                                                   all other, demonstrating decreased species richness,
                                                                                                                                                                                   evenness, and abundance.                                                                                  ERM
      • Differences in whole body selenium concentration and condition (weight/length) between these two groups were analyzed                                                                                                                                                                4.0               0.11               -     0.17        0.33           0.2
        using a t-test. Correlations between the extent of popeye or condition index and selenium content were examined                                                          • Selenium cannot be eliminated as a cause of popeye in                                                     CRM
        separately for the two groups.                                                                                                                                             sampled bluegills at TRM 567.6.                                                                           5.5             0.083         0.92               -     0.54          0.47
                                                                                                                                                                                                                                                                                             CRM
                                                                                                                                                                                 • Due to seasonality, fish movement, large population sizes, and
                                                                                                                                                                                                                                                                                             3.5               0.27        0.95         0.97             -         0.5
                                                                                                                                                                                   reduced sampling effort, this study will need to continue over
                                Jan-09                                   Mar-09                                    Jul-09                                      Jan-10                                                                                                                        TRM
                                                                                                                                                                                   many years to fully document the impact of fly ash on the fish
                                                                                                                                                                                                                                                                                             567.6           0.026         0.93         0.95        0.94             -
                      R             H             E            R             H             E             R            H             E             R              H       E         community in Watts Bar Reservoir.
                                                                                                                                                                                                                                                                                              Table 3. Jaccard’s Index (above diagonal) and Morisita Index (below diagonal)
ERM 2.0               5            1.3         0.808           9           1.65         0.751            6           1.64         0.915           4             1.33    0.959                                                                                                                 of similarity for all collections made during January 2010 sampling.


ERM 4                 9           1.38         0.628            -            -             -            13           1.35         0.526           5             1.2     0.746                                      Jan-09                                      Mar-09                                       Jul-09                                      Jan-10

CRM 5.5               -             -             -            9           1.56         0.710           10           1.33         0.578           9             1.22    0.555                       Total         With                          Total          With                          Total         With                          Total           With
                                                                                                                                                                                                   Number        Anomaly         Percent       Number         Anomaly        Percent        Number        Anomaly        Percent        Number          Anomaly       Percent
CRM 3.5               -             -             -            10          1.36         0.591           13           1.69         0.659          11             1.56    0.651
                                                                                                                                                                                ERM 2.0                13              0             0%            74              0            0%              15             2          13.33%              6              0            0%

TRM 567.6             -             -             -            11          1.79         0.746           15           1.7          0.628          14             1.03    0.390   ERM 4                  55              0             0%             -              -              -             71             0             0%              24              0            0%

ERM 14                -             -             -            16          1.76         0.635            -             -             -            -              -        -     CRM 5.5                 -              -              -            135             0            0%             113             1           0.88%             219             2           0.91%

ERM 0.1               6           0.69         0.385            -            -             -             -             -             -            -              -        -     CRM 3.5                 -              -              -            148             1          0.68%            118             2           1.69%             125             0            0%

ERM 1.0               8           1.19         0.572            -            -             -             -             -             -            -              -        -
                                                                                                                                                                                TRM 567.6               -              -              -            96              1          1.04%             99             0             0%              367             33          8.99%

Table 1. Calculated species richness (R), Shannon-Wiener Index (H), and species evenness (E) at eight total sites in Watts Bar Reservoir over four sampling periods.
                                                                                                                                                                                ERM 14                  -              -              -            91              0            0%               -             -              -               -              -             -

 Literature Cited:                                                                                                                                                              ERM 0.1               111              0             0%             -              -              -              -             -              -               -              -             -

     Ali M, Parvez S, Pandey S, Atif F, Faur M, Rehman H, Raisuddin S. 2004. Fly ash leachate induces oxidative stress in freshwater fish                                       ERM 1.0                32              0             0%             -              -              -              -             -              -               -              -             -
     Channa punctata (Bloch). Environ. Int. 30(7):933-8.
                                                                                                                                                                                Table 2. Total number of individuals, number exhibiting an anomaly, and the percentage of anomalies in the population at eight total sites in Watts Bar Reservoir over four sampling periods.
     Besser J, Glesy J, Brown R, Buell J, Dawson, G. 1996. Selenium bioaccumulation and hazards in a fish community affected by coal fly
     ash effluent. Ecotox. and Env. Safety 35(1):7-15.
     Lemly A. 2002. Symptoms and implications of selenium toxicity in fish: the Belews Lake case example. Aqua. Tox. 57(1-2):39-49.                                             Acknowledgements: Heather DeGaetano, Lee Friedlander, Matt Hamilton, Dave McKinney, Lyndhurst Foundation, TWRA, World Wildlife Fund Southeast
                                                                                                                                                                                Rivers and Streams Program

				
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