Patterns in benthic macroinvertebrate assemblages in an active region of unconventional shale-gas development in the western Appalachian Plateau of West Virginia, USA.

We sampled headwater streams to characterize impacts of unconventional shale gas development (SGD) on aquatic ecosystems. The study area was relatively un-impacted by confounding activities. Intensity of SGD over the study decreased then increased again but not to levels seen the first year. Shale gas development was associated with increased, but non-impaired, water pH and specific conductance during the latter part of the study. Metrics summarizing macroinvertebrate assemblages were better on average in un-impacted reaches. A genus-level multimetric index of biotic integrity was statistically lower downstream of impacts compared to upstream, but only in the year when SGD activity was most intense. Multivariate analyses indicated that assemblages diverged in similarity downstream compared to upstream of impacts in the first and last years of the study when SGD activity was elevated. Assemblage divergence was related to variation in water quality. Indicator species analysis linked a few key taxa to un-impacted conditions in the first year of the study; tolerant taxa were indicators for impacted conditions later in the study. Our study links SGD to weak negative changes in water quality and benthic macroinvertebrates, which may have negative consequences to food quality for wildlife that rely on aquatic prey within forested systems.

Variation in DNA methylation is associated with migratory phenotypes of lake sturgeon Acipenser fulvescens in the St. Clair River, MI, USA.

Lake sturgeon Acipenser fulvescens populations show a variety of movement patterns that are poorly understood. To compare two migratory phenotypes of A. fulvescens in the St. Clair River, MI, USA, multiple data types were analysed. Individual fish were classified into migratory phenotypes based on acoustic telemetry data collected 2012-2015. Acipenser fulvescens consistently showed movement from the St. Clair River upriver into Lake Huron or downriver into Lake St. Clair. The two migratory phenotypes were then compared for differences in morphometrics, genetics and epigenetics. Morphological differences based on linear measurements were not detected between phenotypes. Microsatellite data from 11 loci suggested one population with no genetic differentiation between migratory phenotypes. Our epigenetic results indicated that the migratory phenotypes are differentially methylated (P < 0.05), thus epigenetics may be associated with migratory differences in A. fulvescens. Only one restriction site was identified to be driving the differential methylation (P < 0.05). While little evidence at neutral loci occurred for genetic differentiation of A. fulvescens, DNA methylation may play a role in the observed movement pattern variation. When combined with microsatellite and morphometric analyses, our results suggested that different migratory patterns may reflect phenotypic plasticity, allowing A. fulvescens to acclimate to short-term environmental variability. Without an integrated approach, the role of epigenetics in the migratory phenotype of A. fulvescens may have been overlooked. Further characterization of migratory phenotypes could be important for management to conserve behavioural variation across the distribution of A. fulvescens and for design of stocking guidelines.

Evaluating expected outcomes of acid remediation in an intensively mined Appalachian watershed.

Assessments of watershed-based restoration efforts are rare but are essential for the science of stream restoration to advance. We conducted a watershed scale assessment of Abram Creek before and after implementation of a watershed-based plan designed to maximize ecological recovery from acid mine drainage (AMD) impairment. We surveyed water chemistry, physical habitat, benthic macroinvertebrates, and fish community structure in three stream types: AMD-impacted (14 streams), AMD-treated (13 streams), and unimpaired reference (4 streams). We used in-stream measurements to quantify ecological loss from AMD, the amount of ecological recovery expected through remediation, and the observed degree of post-treatment recovery. Sites impaired by AMD improved in water quality with AMD treatment. Dissolved metals and acidity declined significantly in treated streams, but sulfate and specific conductance did not. Likewise, sites impaired by AMD improved in bio-condition scores with AMD treatment. EPT genera increased significantly but were lower compared to unimpaired streams. We found fish at nine treated sites that had none before treatment. Community-level analyses indicated improved but altered assemblages with AMD treatment. Analysis of pre-treatment conditions indicated that only 30% of the historic fishery remained. Remediation was expected to recover 66% of the historic fishery value, and assessment of post-treatment conditions indicates that 52% of the historic fishery has been recovered after 3 years. Developing expected endpoints for restoration outcomes provides a tool to objectively evaluate successes and can guide adaptive management strategies.

The update and optimization of an eDNA assay to detect the invasive rusty crayfish (Faxonius rusticus).

Environmental DNA (eDNA) is nuclear or mitochondrial DNA shed into the environment, and amplifying this DNA can serve as a reliable, noninvasive way to monitor aquatic systems for the presence of an invasive species. Assays based on the collection of eDNA are becoming increasingly popular, and, when optimized, can aid in effectively and efficiently tracking invasion fronts. We set out to update an eDNA assay to detect the invasive rusty crayfish, Faxonius rusticus. We tested for species specificity compared to other stream crayfish and field tested the assay at sites with known presence (N = 3) and absence (N = 4) in the Juniata River watershed in central Pennsylvania, USA. To maximize sensitivity, we field tested different storage buffers (Longmire's buffer and ethanol), DNA extraction methods (Qiagen's DNEasy and PowerWater kits), and quantitative polymerase chain reaction (qPCR) chemistries (TaqMan and SYBR green). Our assay confirmed the presence data and performed optimally when filter samples were stored in Longmire's buffer, DNA was extracted with DNeasy Blood and Tissue Kit, and TaqMan qPCR chemistry was utilized. With proper sample processing, our assay allows for accurate, noninvasive detection of F. rusticus in streams.

Demographic characteristics of an avian predator, Louisiana Waterthrush (Parkesia motacilla), in response to its aquatic prey in a Central Appalachian USA watershed impacted by shale gas development.

We related Louisiana Waterthrush (Parkesia motacilla) demographic response and nest survival to benthic macroinvertebrate aquatic prey and to shale gas development parameters using models that accounted for both spatial and non-spatial sources of variability in a Central Appalachian USA watershed. In 2013, aquatic prey density and pollution intolerant genera (i.e., pollution tolerance value <4) decreased statistically with increased waterthrush territory length but not in 2014 when territory densities were lower. In general, most demographic responses to aquatic prey were variable and negatively related to aquatic prey in 2013 but positively related in 2014. Competing aquatic prey covariate models to explain nest survival were not statistically significant but differed annually and in general reversed from negative to positive influence on daily survival rate. Potential hydraulic fracturing runoff decreased nest survival both years and was statistically significant in 2014. The EPA Rapid Bioassessment protocol (EPA) and Habitat Suitability Index (HSI) designed for assessing suitability requirements for waterthrush were positively linked to aquatic prey where higher scores increased aquatic prey metrics, but EPA was more strongly linked than HSI and varied annually. While potential hydraulic fracturing runoff in 2013 may have increased Ephemeroptera, Plecoptera, and Trichoptera (EPT) richness, in 2014 shale gas territory disturbance decreased EPT richness. In 2014, intolerant genera decreased at the territory and nest level with increased shale gas disturbance suggesting the potential for localized negative effects on waterthrush. Loss of food resources does not seem directly or solely responsible for demographic declines where waterthrush likely were able to meet their foraging needs. However collective evidence suggests there may be a shale gas disturbance threshold at which waterthrush respond negatively to aquatic prey community changes. Density-dependent regulation of their ability to adapt to environmental change through acquisition of additional resources may also alter demographic response.

Water chemistry-based classification of streams and implications for restoring mined Appalachian watersheds.

We analyzed seasonal water samples from the Cheat and Tygart Valley river basins, West Virginia, USA, in an attempt to classify streams based on water chemistry in this coal-mining region. We also examined temporal variability among water samples. Principal component analysis identified two important dimensions of variation in water chemistry. This variation was determined largely by mining-related factors (elevated metals, sulfates, and conductivity) and an alkalinity-hardness gradient. Cluster analysis grouped water samples into six types that we described as reference, soft, hard, transitional, moderate acid mine drainage, and severe acid mine drainage. These types were statistically distinguishable in multidimensional space. Classification tree analysis confirmed that chemical constituents related to acid mine drainage and acid rain distinguished these six groups. Hard, soft, and severe acid mine drainage type streams were temporally constant compared to streams identified as reference, transitional, and moderate acid mine drainage type, which had a greater tendency to shift to a different water type between seasons. Our research is the first to establish a statistically supported stream classification system in mined watersheds. The results suggest that human-related stressors superimposed on geology are responsible for producing distinct water quality types in this region as opposed to more continuous variation in chemistry that would be expected in an unimpacted setting. These findings provide a basis for simplifying stream monitoring efforts, developing generalized remediation strategies, and identifying specific remediation priorities in mined Appalachian watersheds.