The Clinch River (CR) and North Fork Holston River (NFHR) contain some of the most diverse freshwater mussel assemblages in the United States; however, both rivers are experiencing declines in mussel populations. The first component of this study used histological evaluations and water quality data to determine whether mussels were negatively impacted in the CR zone of decline (ZD) and to inform future management of freshwater mussels in the river. In the 91 kilometer (km) section from Carbo, Virginia (CRKM 431) downstream to Speers Ferry, Virginia (CRKM 340), referred to as the ZD, mussel density decreased >90% from 1979 to 2014 at key sites such as Semones Island (CRKM 378.3) and Pendleton Island (CRKM 364.2). Laboratory propagated mussels were placed in cages in the river for one year from June 2012 to May 2013 at four sites within the ZD and four sites in reaches where mussel populations remain stable or are increasing, a zone of stability (ZS). The survival, growth and histological results indicated that there are continuing impacts to mussels in the ZD. Research investigating impacts to the ZD and methods to improve water quality in this zone are needed.

The laboratory component of this study examined sublethal effects of potassium (K⁺), chloride (Cl⁻), and un-ionized ammonia (NH₃-N) on mussel tissues at concentrations relevant to those found in the NFHR. Historical industrial activities at Saltville, Virginia, as well as continued pollution of the NFHR from chemical waste ponds at this location, are believed to be significant contributors to mussel declines. Contaminant seepages from the waste ponds that include Cl⁻, K⁺, and NH₃-N have been shown to be toxic to adult and juvenile mussels.

A three-month laboratory study was conducted to assess impacts to organ tissues (gills, digestive glands, kidneys, and gonads) of adult Villosa iris exposed to environmentally relevant concentrations of K⁻ (4 and 8 mg/L), Cl⁻ (230 and 705 mg/L), and NH₃-N (0.014 and 0.15 mg/L) using histological evaluations. No detectable differences were observed among the histological endpoints from mussels held in treatments and control (p>0.05). The study design was modified and repeated using increased concentrations of K⁺ (8, 16, and 32 mg/L) and Cl⁻ (705, 1410, and 2820 mg/L) for a two-month exposure period. Due to issues with maintaining NH3-N in mussel holding chambers, the second study did not the second study did not include NH₃-N exposures. Control mussels in both studies had a higher abundance of lipofuscin in kidneys and degraded cytoplasm in the digestive gland diverticula compared to baseline mussels, indicating that captivity influenced mussel tissues. Future studies are needed to more thoroughly address these captivity effects. Both survival and histological data in the second test showed a significant negative effect of the increased concentrations of Cl⁻ and K⁻, which were representative of those found at some sites in the NFHR downstream of Saltville, Virginia.