FY17_Kulp_SOW_Tennessee_Tech.pdf

STATEMENT OF WORK

Assessment of Benthic Macroinvertebrate Response to Antimycin During Brook Trout

Restoration in Little Cataloochee Creek, Great Smoky Mountains National Park

Statement of Work: The goal of the study is to implement a before-after, control-impact (BACI) design to provide data on potential for antimycin and potassium permanganate effects on aquatic macroinvertebrates. Four 200-m sample sites will be established in the Little

Cataloochee Creek sub-watershed in 2nd and 3rd order streams and corresponding control sites of the same stream order will be established in the adjacent Correll Branch sub-watershed at similar elevations (Figure 1). The control sites will provide baseline data to compare with treatment sites to determine the influence of antimycin exposure on the benthic macroinvertebrate community in Little Cataloochee Creek. Aquatic macroinvertebrates will also be sampled downstream of the potassium permanganate neutralization site below the confluence with Correll Branch to monitor the effects of the neutralizing agent on aquatic invertebrates. Samples will be collected during the same time periods before treatment as after (i.e., 23 m, 11 m, 1m, 1 w, and 1 d before and 1 d, 1 w, 1 m, 13 m, 25 m after) to provide a balanced view of macroinvertebrate community changes. A Hess sampler with a defined area of 0.1 m2 will be used to take three replicate samples per site per time period to obtain density estimates for benthic macroinvertebrate taxa over time in response to antimycin exposure.

Each 200m site will be subdivided into 20-m sections to ensure no local substrate is sampled more than once for the duration of the study. Collection will include 3 replicates at 9, 20m sites during 10 sample events for a total of 270 samples. To prevent macroinvertebrate escape, the sampler will be completely embedded within the streambed, after which the substrate within the sampler will be disturbed for approximately 60 seconds to allow organisms to dislodge and collect in a 500-μm plankton bucket. Each sample will be bagged separately after all dislodged material in the capture net was rinsed into the plankton bucket. The brook trout restoration treatment is slated to begin in September 2017.

All samples will be preserved in 10% formalin and processed in the laboratory where all individual organisms were separated from debris and identified to the lowest possible taxonomic status. Common aquatic macroinvertebrate keys will be used to identify insects collected during this study. Additional regional and order-specific keys were used, when necessary, to facilitate species-level identification of final instars. Due to fragility of terrestrial adult stages, some specimens are commonly incomplete or deteriorated; therefore, terrestrial adults will only identified to order. Chironomids (midge larvae) and oligochaetes will be permanently slide-mounted using CMCP-10 mounting media to allow proper identification of microscopic structures enabling genus-level identification. High quantities (>50) of chironomids or oligochaetes within a single quantitative sample will be subsampled to ensure timely identification. All samples have been processed, organisms have been separated from debris, and individuals have been sorted and identified to taxonomic order.

Figure 1. Map of macroinvertebrate sampling locations in the Little Cataloochee Creek watershed.

Statistical community analysis as well as repeated measures analysis of the major aquatic insect orders (i.e., Coleoptera, Diptera, Ephemeroptera, Plecoptera, and Trichoptera) will be completed for all sites. Multivariate techniques will be used to analyze changes among macroinvertebrate communities, whereas univariate data, such as ordinal abundances or taxa richness, will be analyzed with a mixed-model approach to determine effects over time. These analyses can provide greater insight into short-term and long-term effects to specific taxonomic groups and the macroinvertebrate communities as a whole. Data will be analyzed using

Preliminary Analysis of Similarity (ANOSIM) of major taxonomic orders using Primer Statistical

Software (2016) to determine if there are any significant differences between control and treatment sites. Non-metric multidimensional scaling will be used to determine if there is any separation between control and treatment sites in support of ANOSIM results. All results will be provided in a summary report within 12 months of the end of the last sample collection.

The summary report should include summary tables of species collected throughout the study, statistical comparisons over time and an evaluation of the effects of antimycin and potassium permanganate treatment, if any, on aquatic macroinvertebrate samples at each site.