Histological evaluations of organ tissues reveal sublethal effects in a freshwater mussel (Villosa iris) exposed to chloride and potassium concentrations below benchmark estimates.

Salinization of freshwater ecosystems due to anthropogenic sources will increasingly impact biodiversity. An example of point-source industrial salinization has occurred from historical activities at a U.S. Environmental Protection Agency Superfund Site near Saltville, Virginia USA and its associated chemical waste ponds adjacent to the North Fork Holston River. These point source discharges are documented contributors to mussel declines, partially due to high concentrations of chloride (Cl-, ≤ 26,000 mg Cl-/L) and potassium (K+, ≤ 97 mg K+/L). During a chronic 61-day laboratory study, Rainbow mussels, Villosa iris, were exposed to concentrations of Cl- (0, 416, 831, and 1,663 mg/L) and K+ (0, 4, 8, and 17 mg/L) to determine effects on survival and organ tissues. All test mussels died by day-2 in the 1,663 mg Cl-/L exposure, and 50% of mussels died by day-13 in the 17 mg K+/L concentration. Significantly greater abundances of tissue abnormalities were observed in digestive glands and kidneys with exposures to the 4 and 8 mg/L concentrations of K+ versus the control, and significantly greater abundances of lesions in kidneys were observed in the 416 and 831 mg Cl-/L concentrations compared to the control. The sublethal effects to digestive glands and kidneys were below reported effect (EC50, 20, 10 and LOEC) concentrations. Significant histological differences between control and baseline (day-0 sample) mussels were observed, suggesting the need for further study on the effects of captivity during longer-term laboratory experiments.

Understanding the influence of multiple pollutant stressors on the decline of freshwater mussels in a biodiversity hotspot.

The Clinch River watershed of the upper Tennessee River Basin of Virginia and Tennessee, USA supports one of North America's greatest concentrations of freshwater biodiversity, including 46 extant species of native freshwater mussels (Order Unionida), 20 of which are protected as federally endangered. Despite the global biological significance of the Clinch River, mussel populations are declining in some reaches, both in species richness and abundance. The aim of this study was to evaluate the exposure of adult resident mussels to a suite of inorganic and organic contaminant stressors in distinct sections of the Clinch River that encompassed a range of mussel abundance and health. To provide insight into the potential role of pollutants in the decline of mussels, including within a previously documented "zone of mussel decline", the mainstem Clinch River (8 sites) and its tributaries (4 sites) were examined over two consecutive years. We quantified and related metals and organic contaminant concentrations in mussels to their associated habitat compartments (bed sediment, suspended particulate sediment, pore water, and surface water). We found that concentrations of organic contaminants in resident mussels, particularly the suite of 42 polycyclic aromatic hydrocarbons (PAHs) analyzed, were related to PAH concentrations in all four habitat (media) compartments. Further, PAH concentrations in mussel tissue (range 37.8-978.1 ng/g dry weight in 2012 and 194.3-1073.7 ng/g dry weight in 2013) were negatively related to the spatial pattern in mussel densities (rs = -0.64, p ≤ 0.05 in 2012 and rs = -0.83, p ≤ 0.05 in 2013) within the river, and were highest in the "zone of mussel decline". In contrast, the suite of 22 metals analyzed in resident mussels were largely unrelated to the spatial pattern of variation of metals in the four habitat compartments except for Manganese (Mn; range 3630.5-23,749.2 µg/g dry weight in 2012 and 1540.4-12,605.8 µg/g dry weight in 2013) in surface water (rs = 0.58, p < 0.1) and pore water (rs = 0.76, p ≤ 0.05). This study revealed that PAHs and Mn are important pollutant stressors to mussels in the Clinch River and that they are largely being delivered through the Guest River tributary watershed. Accordingly, future conservation and management efforts would benefit by identifying, and ideally mitigating, the sources of PAHs, Mn, and other current or legacy mining-associated pollutants to the mainstem river and its tributaries.

Assessment of growth, survival, and organ tissues of caged mussels (Bivalvia: Unionidae) in a river-scape influenced by coal mining in the southeastern USA.

The Clinch River contains one of the most diverse freshwater mussel assemblages in the United States, with 46 extant species, 20 of which are listed as federally endangered. In a 91 km section of the Clinch River, mussel densities have severely declined at key sites from 1979 to 2014 (zone of decline), compared to other river sections that support high density and stable mussel populations (zone of stability). Coal mining has occurred in tributary watersheds that drain to the zone of decline since the late nineteenth century. To determine differences in survival, growth, and organ (gills, digestive glands, kidneys, and gonads) tissue health between the river zones, laboratory-propagated mussels were deployed to cages for one year at four sites each within the zones of mussel stability and decline. Means of growth and survival of mussels in the zone of stability were significantly greater than in the zone of decline, and mean fractions of kidney lipofuscin in mussels in the zone of decline was significantly greater than in the zone of stability. Water concentrations of K+, Na+, Al, and Mn were significantly greater in the zone of decline than in the zone of stability. Statistical correlation results indicated that mussel survival was positively associated with concentrations of Mn in water, and kidney lipofuscin was negatively associated with concentrations of dissolved K+, SO42-, and Mg2+. Most concentrations of contaminants were below published benchmark criteria, yet our results suggest that conditions continue to exist in the zone of decline that promote impacts to mussels that are at least partially associated with low concentration coal-related contaminants. More research is needed to determine the additive, synergistic, or antagonistic effects of these complex ionic mixtures on freshwater mussels from river environments, such as in the Clinch River, where constituent concentrations are relatively low.

Identification of mutants of Arabidopsis defective in acclimation of photosynthesis to the light environment.

In common with many other higher plant species, Arabidopsis undergoes photosynthetic acclimation, altering the composition of the photosynthetic apparatus in response to fluctuations in its growth environment. The changes in photosynthetic function that result from acclimation can be detected in a noninvasive manner by monitoring chlorophyll (Chl) fluorescence. This technique has been used to develop a screen that enables the rapid identification of plants defective at ACCLIMATION OF PHOTOSYNTHESIS TO THE ENVIRONMENT (APE) loci. The application of this screen to a population of T-DNA-transformed Arabidopsis has successfully led to the identification of a number of mutant lines with altered Chl fluorescence characteristics. Analysis of photosynthesis and pigment composition in leaves from three such mutants showed that they had altered acclimation responses to the growth light environment, each having a distinct acclimation-defective phenotype, demonstrating that screening for mutants using Chl fluorescence is a viable strategy for the investigation of acclimation. Sequencing of the genomic DNA flanking the T-DNA elements showed that in the ape1 mutant, a gene was disrupted that encodes a protein of unknown function but that appears to be specific to photosynthetic organisms, whereas the ape2 mutant carries an insertion in the region of the TPT gene encoding the chloroplast inner envelope triose phosphate/phosphate translocator.