Repository for Oil and Gas Energy Research (ROGER)

Abstract:
Diluted bitumen, also known as dilbit, is transported by rail and pipeline across Canada and the United States. Due to the fewer number of studies characterizing the toxicity of dilbit, a dilbit spill poses an unknown risk to freshwater aquatic ecosystems. In the following study, we compared the impact of early-life exposure to conventional and unconventional crude oils on the optomotor behavior, reproductive success, and transgenerational differences in gene expression in zebrafish and their progeny. For exposures, water accommodated fractions (WAFs) of crude oil were generated using a 1:1000 oil to water ratio for 3 different crudes; mixed sweet blend (MSB), medium sour composite (MSC) and dilbit. All three oils generated unique volatile organic compound (VOC) and polycyclic aromatic compound (PAC) profiles. Of the WAFs tested, only dilbit decreased the eye size of 2 dpf larvae, and only MSB exposed larvae had an altered behavioral response to a visual simulation of a predator. Early-life exposure to crude oil had no lasting impact on reproductive success of adult fish; however, each oil had unique impacts on the basal gene expression of the somatically exposed offspring. In this study, the biological effects differed between each of the oils tested, which implied chemical composition plays a critical role in determining the sublethal toxicity of conventional and unconventional crude oils in freshwater ecosystems.

Abstract:
Europe PMC is an archive of life sciences journal literature., Evaluating the Impact of Hydraulic Fracturing on Streams using Microbial Molecular Signatures.

Abstract:
The widespread application of directional drilling and hydraulic fracturing technologies expanded oil and gas (OG) development to previously inaccessible resources. A single OG well can generate millions of liters of wastewater, which is a mixture of brine produced from the fractured formations and injected hydraulic fracturing fluids (HFFs). With thousands of wells completed each year, safe management of OG wastewaters has become a major challenge to the industry and regulators. OG wastewaters are commonly disposed of by underground injection, and previous research showed that surface activities at an Underground Injection Control (UIC) facility in West Virginia affected stream biogeochemistry and sediment microbial communities immediately downstream from the facility. Because microbially driven processes can control the fate and transport of organic and inorganic components of OG wastewater, we designed a series of aerobic microcosm experiments to assess the influence of high total dissolved solids (TDS) and two common HFF additives—the biocide 2,2-dibromo-3-nitrilopropionamide (DBNPA) and ethylene glycol (an anti-scaling additive)—on microbial community structure and function. Microcosms were constructed with sediment collected upstream (background) or downstream (impacted) from the UIC facility in West Virginia. Exposure to elevated TDS resulted in a significant decrease in aerobic respiration, and microbial community analysis following incubation indicated that elevated TDS could be linked to the majority of change in community structure. Over the course of the incubation, the sediment layer in the microcosms became anoxic, and addition of DBNPA was observed to inhibit iron reduction. In general, disruptions to microbial community structure and function were more pronounced in upstream and background sediment microcosms than in impacted sediment microcosms. These results suggest that the microbial community in impacted sediments had adapted following exposure to OG wastewater releases from the site. Our findings demonstrate the potential for releases from an OG wastewater disposal facility to alter microbial communities and biogeochemical processes. We anticipate that these studies will aid in the development of useful models for the potential impact of UIC disposal facilities on adjoining surface water and shallow groundwater.

Abstract:
Hydraulic fracturing has become widely used in recent years to access vast global unconventional sources of oil and gas. This process involves the injection of proprietary mixtures of water and chemicals to fracture shale formations and extract the hydrocarbons trapped within. These injection fluids, along with minerals, hydrocarbons, and saline waters present within the formations being drilled into, return to the surface as flowback and produced water (FPW). FPW is a highly complex mixture, containing metals, salts and clay, as well as many organic chemicals, including polycyclic aromatic hydrocarbons such as phenanthrene. The present study sought to determine the effects of temperature on the accumulation of phenanthrene in rainbow trout (Oncorhynchus mykiss). This model organism resides in rivers overlapping the Montney and Duvernay formations, both highly developed formations for hydraulic fracturing. Rainbow trout acclimated to temperatures of 4, 13 and 17°C were exposed to either 5% or 20% FPW, as well as saline mixtures representing the exact ionic content of FPW to determine the accumulation of radiolabelled 14C phenanthrene within the gill, gut, liver and gallbladder. FPW exposure reduced the overall accumulation of phenanthrene in a manner most often similar to high salinity exposure, indicating that the high ionic strength of FPW is the primary factor affecting accumulation. Accumulation was different at the temperature extremes (4 and 17°C), although no consistent relationship was observed between temperature and accumulation across the observed tissues. These results indicate that several physiological responses occur as a result of FPW exposure and water temperature change which dictate phenanthrene uptake, particularly in the gills. Temperature (and seasonality) alone cannot be used to model the potential accumulation of polycyclic aromatic hydrocarbons after FPW spills.

Abstract:
Hydraulic fracturing flowback and produced water (FPW) is a highly complex and heterogenous wastewater by-product of hydraulic fracturing practices. To date, no research has examined how FPW exposure to freshwater biota may affect energetic homeostasis following subsequent induction of detoxification processes. Rainbow trout (Oncorhynchus mykiss) were acutely exposed for 48 h to either 2.5% or 7.5% FPW, and hepatic metabolism was assessed either immediately or following a 3-week recovery period. Induction of xenobiotic metabolism was observed with an 8.8-fold increase in ethoxyresorufin-O-deethylase (EROD) activity after 48 h exposure to 7.5% FPW, alongside a 10.3-fold increase in the mRNA abundance of cyp1a, both of which returned to basal level after three weeks. Glucose uptake capacity was elevated by 6.8- and 12.9-fold following 2.5% and 7.5% FPW exposure, respectively, while alanine uptake was variable. Activity measurements and mRNA abundance of key enzymes involved in hepatic metabolism indicated that aerobic metabolism was maintained with exposure, as was glycolysis. Gluconeogenesis, as measured by phosphoenolpyruvate carboxykinase (PEPCK) activity, decreased by ~30% 48 h following 2.5% FPW exposure and ~20% 3 weeks after 7.5% FPW exposure. The abundance of pepck mRNA activity followed similar, yet non-significant, trends. Finally, a delayed increase in amino acid catabolism was observed, as glutamate dehydrogenase (GDH) activity was increased 2-fold in 7.5% FPW exposed fish when compared to saline control fish at the 3-week time point. We provide evidence to suggest that although hepatic metabolism is altered following acute FPW exposure, metabolic homeostasis generally returns 3-weeks post-exposure.

Abstract:
Publicly available toxicological studies on wastewaters associated with unconventional oil and gas (UOG) activities in offshore regions are nonexistent. The current study investigated the impact of hydraulic fracturing-generated flowback water (HF-FW) on whole organism swimming performance/respiration and cardiomyocyte contractility dynamics in mahi-mahi (Coryphaena hippurus—hereafter referred to as “mahi”), an organism which inhabits marine ecosystems where offshore hydraulic fracturing activity is intensifying. Following exposure to 2.75% HF-FW for 24 h, mahi displayed significantly reduced critical swimming speeds (Ucrit) and aerobic scopes (reductions of ∼40 and 61%, respectively) compared to control fish. Additionally, cardiomyocyte exposures to the same HF-FW sample at 2% dilutions reduced a multitude of mahi sarcomere contraction properties at various stimulation frequencies compared to all other treatment groups, including an approximate 40% decrease in sarcomere contraction size and a nearly 50% reduction in sarcomere relaxation velocity compared to controls. An approximate 8-fold change in expression of the cardiac contractile regulatory gene cmlc2 was also seen in ventricles from 2.75% HF-FW-exposed mahi. These results collectively identify cardiac function as a target for HF-FW toxicity and provide some of the first published data on UOG toxicity in a marine species.

Abstract:
Unconventional methods of oil and natural gas extraction have been a growing part of North America's energy sector for the past 20-30 years. Technologies such as horizontal hydraulic fracturing have facilitated the exploitation of geologic reserves that were previously resistant to standard drilling approaches. However, the environmental risks associated with hydraulic fracturing are relatively understudied. One such hazard is the wastewater by-product of hydraulic fracturing processes: flowback and produced water (FPW). During FPW production, transport, and storage, there are many potential pathways for environmental exposure. In the current review, toxicological hazards associated with FPW surface water contamination events and potential effects on freshwater biota are assessed. This review contains an extensive survey of chemicals commonly associated with FPW samples from shale formations across North America and median 50% lethal concentration values (LC50) of corresponding chemicals for many freshwater organisms. We identify the characteristics of FPW which may have the greatest potential to be drivers of toxicity to freshwater organisms. Notably, components associated with salinity, the organic fraction, and metal species are reviewed. Additionally, we examine the current state of FPW production in North America and identify the most significant obstacles impeding proper risk assessment development when environmental contamination events of this wastewater occur. Findings within this study will serve to catalyze further work on areas currently lacking in FPW research, including expanded whole effluent testing, repeated and chronic FPW exposure studies, and toxicity identification evaluations.

Abstract:
Hydraulic fracturing of horizontal wells is a cost effective means for extracting oil and gas from low permeability formations. Hydraulic fracturing often produces considerable volumes of flowback and produced water (FPW). FPW associated with hydraulic fracturing has been shown to be a complex, often brackish mixture containing a variety of anthropogenic and geogenic compounds. In the present study, the risk of FPW releases to aquatic systems was studied using the model benthic invertebrate, Lumbriculus variegatus and field-collected FPW from a fractured well in Alberta. Acute, chronic, and pulse toxicity were evaluated to better understand the implications of accidental FPW releases to aquatic environments. Although L. variegatus is thought to have a high tolerance to many stressors, acute toxicity was significant at low concentrations (i.e. high dilutions) of FPW (48 h LC50: 4–5%). Chronic toxicity (28 d)of FPW in this species was even more pronounced with LC50s (survival/reproduction) and EC50s (total mass) at dilutions as low as 0.22% FPW. Investigations evaluating pulse toxicity (6 h and 48 h exposure) showed a significant amount of latent mortality occurring when compared to the acute results. Additionally, causality in acute and chronic bioassays differed as acute toxicity appeared to be primarily driven by salinity, which was not the case for chronic toxicity, as other stressors appear to be important as well. The findings of this study show the importance of evaluating multiple exposure regimes, the complexity of FPW, and also shows the potential aquatic risk posed by FPW releases.

Abstract:
The number of horizontally drilled shale oil and gas wells in the United States has increased from nearly 28,000 in 2007 to nearly 127,000 in 2017, and research has suggested the potential for the development of shale resources to affect nearby stream ecosystems. However, the ability to generalize current studies is limited by the small geographic scope as well as limited breadth and integration of measured chemical and biological indicators parameters. This study tested the hypothesis that a quantifiable, significant relationship exists between the density of oil and gas (OG) development, increasing stream water concentrations of known geochemical tracers of OG extraction, and the composition of benthic macroinvertebrate and microbial communities. Twenty-five headwater streams that drain lands across a gradient of shale gas development intensity were sampled. Our strategy included comprehensive measurements across multiple seasons of sampling to account for temporal variability of geochemical parameters, including known shale OG geochemical tracers, and microbial and benthic macroinvertebrate communities. No significant relationships were found between the intensity of OG development, shale OG geochemical tracers, or benthic macroinvertebrate or microbial community composition, whereas significant seasonal differences in stream chemistry were observed. These results highlight the importance of considering spatial and temporal variability in stream chemistry and biota and not only the presence of anthropogenic activities in a watershed. This comprehensive, integrated study of geochemical and biological variability of headwater streams in watersheds undergoing OG development provides a robust framework for examining the effects of energy development at a regional scale.

Abstract:
Shale-gas production could impact freshwater quality through contamination of the physical and chemical habitat (e.g., fracturing fluids, untreated or treated effluent) or development-related impacts. Despite environmental concerns, information is lacking to support biomonitoring as a diagnostic tool to assess impacts of shale-gas production. We characterized water quality and biota in areas of high shale gas potential (Early Carboniferous bedrock in New Brunswick, Canada) and surrounding geologic areas, and we assessed patterns in benthic macroinvertebrate (BMI) and fish assemblages. Early Carboniferous stations differed primarily based on water chemistry, and BMI were associated with a gradient in conductivity and temperature across geologic classes. Concordance analysis indicated similar classification of stations by both organism groups, though fish were more related to turbidity and nutrients. Concordance among fish and BMI was strongest at high conductivity, Early Carboniferous stations. These results suggest that geology plays a strong role in driving abiotic habitats and biotic communities of streams, even at small spatial scales. Furthermore, they suggest BMI and fish can provide complementary information for biomonitoring in shale-gas development areas, with BMI responding to increased ion concentrations from surface water contamination, and fish responding to changes in nutrients and turbidity resulting from development.

Abstract:
Production of unconventional oil and gas continues to rise, but the effects of high-density hydraulic fracturing (HF) activity near aquatic ecosystems are not fully understood. A commonly used biocide in HF, 2,2-dibromo-3-nitrilopropionamide (DBNPA), was studied in microcosms of HF-impacted vs. HF-unimpacted surface water streams to (1) compare the microbial community response, (2) investigate DBNPA degradation products based on past HF exposure, and (3) compare the microbial community response differences and similarities between the HF biocides DBNPA and glutaraldehyde. The microbial community responded to DBNPA differently in HF-impacted vs. HF-unimpacted microcosms in terms of 16S rRNA gene copies quantified, alpha and beta diversity, and differential abundance analyses of microbial community composition through time. The difference in microbial community changes affected degradation dynamics. HF-impacted microbial communities were more sensitive to DBNPA, causing the biocide and byproducts of the degradation to persist for longer than in HF-unimpacted microcosms. Seventeen DBNPA byproducts were detected, many of them not widely known as DBNPA byproducts. Many of the believed to be uncharacterized brominated byproducts detected may pose environmental and health impacts. Similar taxa were able to tolerate glutaraldehyde and DBNPA, however DBNPA was not as effective for microbial control as indicated by a smaller overall decrease of 16S rRNA gene copies/mL after exposure to the biocide and a more diverse set of taxa was able to tolerate it. These findings suggest that past HF activity in streams can affect the microbial community response to environmental perturbation such as the biocide DBNPA. Importance Unconventional oil and gas activity can affect pH, total organic carbon, and microbial communities in surface water altering their ability to respond to new environmental and/or anthropogenic perturbations. These findings demonstrate that DBNPA, a common hydraulic fracturing (HF) biocide, affects microbial communities differently as a consequence of past HF exposure, persisting longer in HF-impacted waters. These findings also demonstrate that DBNPA has low efficacy in environmental microbial communities regardless of HF impact. These findings are of interest, as understanding microbial responses is key for formulating remediation strategies in UOG impacted environments. Moreover, some of DBNPA degradation byproducts are even more toxic and recalcitrant than DBNPA itself, and this work identifies novel brominated degradation byproducts formed.

Abstract:
Flowback and produced water (FPW) is a complex, often brackish, solution formed during the process of hydraulic fracturing. Despite recent findings on the short-term toxicity of FPW on aquatic biota, longer-term impacts of FPW on fish have not yet been investigated and the mechanisms of chronic effects remain unknown. The aim of the present study was to observe the effect of a diluted FPW on ionoregulatory endpoints in the rainbow trout Oncorhynchus mykiss, following a 28-d sub-chronic exposure. A salinity-matched control solution (SW), recreating the salt content of the FPW, was used to differentiate the specific effect of the salts from the effects of the other FPW components (i.e. organics and metals). Overall, fish ionoregulation was not impacted by the chronic exposure. An accumulation of strontium (Sr) and bromide (Br) occurred in the plasma of the FPW-exposed fish only, however no change of plasma ions (Na, K, Cl, Ca, Mg) was observed in SW- or FPW-exposed fish. Similarly, exposures did not alter branchial activity of the osmoregulatory enzymes sodium/potassium ATPase and proton ATPase. Finally, FPW exposure resulted in modifications of gill morphology over time, with fish exposed to the fluid displaying shorter lamellae and increased interlamellar-cell mass. However, these effects were not distinct from morphological changes that also occurred in the gills of control groups.

Abstract:
To inform sustainable energy development, it is important to understand the ecological effects of historical and current production practices and the persistence of those effects. The Williston Basin is one of North America's largest oil production areas and overlaps the Prairie Pothole Region, an area densely populated with wetlands that provide important wildlife habitat. Although historical disposal practices that released chloride-rich waters (brines) produced during oil extraction into the environment are no longer used, brine spills still occur frequently. We sampled 33 wetlands for three amphibian species in Montana and North Dakota during 2015–2017, primarily on National Wildlife Refuges, and used N-mixture models to determine how abundance varied with evidence of brine contamination. To provide insight into effects of historical versus contemporary contamination, we also estimated the association of well density and age with water quality and amphibian abundance. Abundance of boreal chorus frog (Pseudacris maculata) larvae declined most rapidly in response to increased chloride (range: 0.04–17,500 mg/L), followed by the northern leopard frog (Lithobates [Rana] pipiens) and barred tiger salamander (Ambystoma mavortium). Water quality and population- and community-level abundance of amphibians were more strongly related to nearby wells (≤800 m) installed before 1982 than to wells installed since 1982. These results suggest historical brine management practices were the primary driver of contamination and reduced amphibian abundance in wetlands we sampled, reflecting multi-decadal ecological effects. These persistent effects also underscore the critical need for tools to restore landscapes affected by brine contamination.

Abstract:
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.

Abstract:
Potential for shale gas production is linked to regional geology, which influences water chemistry of freshwater systems. However, there has been little work to establish baseline ecological conditions of rivers within areas of shale gas development. In this study, water chemistry and monitoring metrics for fish and benthic macroinvertebrates were compared among varying-sized streams in New Brunswick, Canada with different underlying geology. Water chemistry and biotic community structure differed strongly among geological age classes. Early Carboniferous stations, with highest potential for shale gas production, had the highest ions, invertebrate abundances, and richness of invertebrates and fish, with strongest differences between Early Carboniferous and older bedrock classes. A reference condition model indicated numerous sites deviated from normal, but this was not specific to geological classes, and reflected the lack of model reference site coverage. These results highlight the importance of selecti...

Abstract:
We conducted a large-scale assessment of unconventional oil and gas (UOG) development effects on brook trout (Salvelinus fontinalis) distribution. We compiled 2231 brook trout collection records from the Upper Susquehanna River Watershed, USA. We used boosted regression tree (BRT) analysis to predict occurrence probability at the 1:24,000 stream-segment scale as a function of natural and anthropogenic landscape and climatic attributes. We then evaluated the importance of landscape context (i.e., pre-existing natural habitat quality and anthropogenic degradation) in modulating the effects of UOG on brook trout distribution under UOG development scenarios. BRT made use of 5 anthropogenic (28% relative influence) and 7 natural (72% relative influence) variables to model occurrence with a high degree of accuracy [Area Under the Receiver Operating Curve (AUC) = 0.85 and cross-validated AUC = 0.81]. UOG development impacted 11% (n = 2784) of streams and resulted in a loss of predicted occurrence in 126 (4%). Most streams impacted by UOG had unsuitable underlying natural habitat quality (n = 1220; 44%). Brook trout were predicted to be absent from an additional 26% (n = 733) of streams due to pre-existing non-UOG land uses (i.e., agriculture, residential and commercial development, or historic mining). Streams with a predicted and observed (via existing pre- and post-disturbance fish sampling records) loss of occurrence due to UOG tended to have intermediate natural habitat quality and/or intermediate levels of non-UOG stress. Simulated development of permitted but undeveloped UOG wells (n = 943) resulted in a loss of predicted occurrence in 27 additional streams. Loss of occurrence was strongly dependent upon landscape context, suggesting effects of current and future UOG development are likely most relevant in streams near the probability threshold due to pre-existing habitat degradation.

Abstract:
Production of natural gas using unconventional technologies has risen as demand for alternative fuels has increased. Impacts on the environment from waste generated from these processes are largely unexplored. In particular, the outcomes of organismal exposure to hydraulic fracturing waste have not been rigorously evaluated. We evaluated the effects of exposure to surrogate hydraulic fracturing waste (HF waste) on mucosal bacterial community structure of the brook trout (Salvelinus fontinalis) epidermis. Brook trout are fish native to streams at risk to HF waste exposure. Here, fish were exposed to four treatments (control, 0.00%; low, 0.01%; medium, 0.10%; and high, 1.0% concentrations) of surrogate HF waste synthesized to mimic concentrations documented in the field. Epidermal mucus samples were collected and assessed 15 days post-exposure to determine if the associated bacterial community varied among treatments. We observed differences in epidermal mucosal bacterial community composition at multiple taxonomic scales among treatments. These community changes reflected compositional differences in taxa dominance and community similarity rather than losses or gains in taxonomic richness. The dominant bacterial genus that explained the greatest variation in community structure between exposed and unexposed fish was Flavobacterium. Two genera associated with salmonid diseases, Flavobacterium and Pseudomonas, were statistically more abundant in high treatments than controls. These results suggest that exposure to low levels of HF waste influences bacterial colonization and may lead to a disruption that favors bacterial populations associated with fish disease.

Abstract:
The environmental impacts of hydraulic fracturing, particularly those of surface spills in aquatic ecosystems, are not fully understood. The goals of this study were to (1) understand the effect of previous exposure to hydraulic fracturing fluids on aquatic microbial community structure and (2) examine the impacts exposure has on biodegradation potential of the biocide glutaraldehyde. Microcosms were constructed from hydraulic fracturing-impacted and nonhydraulic fracturing-impacted streamwater within the Marcellus shale region in Pennsylvania. Microcosms were amended with glutaraldehyde and incubated aerobically for 56 days. Microbial community adaptation to glutaraldehyde was monitored using 16S rRNA gene amplicon sequencing and quantification by qPCR. Abiotic and biotic glutaraldehyde degradation was measured using ultra-performance liquid chromatography--high resolution mass spectrometry and total organic carbon. It was found that nonhydraulic fracturing-impacted microcosms biodegraded glutaraldehyde faster than the hydraulic fracturing-impacted microcosms, showing a decrease in degradation potential after exposure to hydraulic fracturing activity. Hydraulic fracturing-impacted microcosms showed higher richness after glutaraldehyde exposure compared to unimpacted streams, indicating an increased tolerance to glutaraldehyde in hydraulic fracturing impacted streams. Beta diversity and differential abundance analysis of sequence count data showed different bacterial enrichment for hydraulic fracturing-impacted and nonhydraulic fracturing-impacted microcosms after glutaraldehyde addition. These findings demonstrated a lasting effect on microbial community structure and glutaraldehyde degradation potential in streams impacted by hydraulic fracturing operations.

Abstract:
Horizontal drilling and hydraulic fracturing extraction procedures have become increasingly present in Pennsylvania where the Marcellus Shale play is largely located. The potential for long-term environmental impacts to nearby headwater stream ecosystems and aquatic bacterial assemblages is still incompletely understood. Here, we perform high-throughput sequencing of the 16 S rRNA gene to characterize the bacterial community structure of water, sediment, and other environmental samples (n = 189) from 31 headwater stream sites exhibiting different histories of fracking activity in northwestern Pennsylvania over five years (2012–2016). Stream pH was identified as a main driver of bacterial changes within the streams and fracking activity acted as an environmental selector for certain members at lower taxonomic levels within stream sediment. Methanotrophic and methanogenic bacteria (i.e. Methylocystaceae, Beijerinckiaceae, and Methanobacterium) were significantly enriched in sites exhibiting Marcellus shale activity (MSA+) compared to MSA− streams. This study highlighted potential sentinel taxa associated with nascent Marcellus shale activity and some of these taxa remained as stable biomarkers across this five-year study. Identifying the presence and functionality of specific microbial consortia within fracking-impacted streams will provide a clearer understanding of the natural microbial community’s response to fracking and inform in situ remediation strategies.

Abstract:
To date, most studies focusing on the effects of high volume hydraulic fracturing on freshwater systems have been at the small watershed scale (i.e. <130 km2). Here we review the findings from the small watershed studies and present a case study of a larger watershed, the South Fork Little Red River (SFLRR) watershed (127–193 km2). Hydraulic fracturing within the SFLRR watershed resulted in no detectable changes in physicochemical parameters or algal biomass. However, sensitive macroinvertebrate taxa densities were reduced downstream of hydraulic fracturing relative to upstream. Increased distance from the hydraulic fracturing, in-stream processing of physicochemical properties, and additional landscape disturbances, likely made it more difficult to assess the effects of hydraulic fracturing in most stream response variables at the larger watershed scale.

Abstract:
This study aims to investigate macroinvertebrate assemblage structure and composition across the three major waterbody types (temporary rivers, depression wetlands and semi-permanent dams) of the Eastern Cape Karoo, and to identify important environmental and spatial correlates of macroinvertebrate assemblage composition in the region. A total of 33 waterbodies (9 dams, 13 depression wetlands and 11 rivers) were sampled. Altogether, 91 taxa were recorded in November 2014 and 82 in April 2015. Twenty-seven taxa were common to all three waterbody types (across both sampling occasions), with 17 of these observed in November and 19 in April. The ANOSIM tests revealed significant differences in assemblage composition between the depression wetlands and rivers for both sampling occasions, but dams did not differ from the other waterbody types. SIMPER analyses indicated that the notonectid Anisops varia and the corixid Micronecta scutellaris were abundant across all three waterbody types during both sampling occasions. The mayfly Cloeon africanum and the damselfly Pseudagrion sp. were abundant in river habitats during both sampling occasions, while the gastropod mollusc Bulinus tropicus and the copepod Lovenula falcifera best characterised depression wetlands on both occasions. Non-metric multidimensional scaling ordination highlighted a clear separation of assemblages between November and April, while distance-based Redundancy Analysis revealed that conductivity, altitude, turbidity and pH were the most important variables explaining the variation in macroinvertebrate assemblage patterns. These results provide baseline information which is important for future biological monitoring of impacts associated with hydraulic fracturing activities and climatic changes in the region.

Abstract:
Hydraulic fracturing fluid are complex mixtures containing high concentrations of salts (up to 330,000 ppm), organic, and metal contaminants. However, little data exist on the potential mechanisms of toxicity of these flowback and produced wastewaters (FPW) on aquatic biota.

Abstract:
Few studies have examined the relationship between natural gas development and surface water quality using well pad density (the number of well pads/km2) as an indicator of potential impacts to aquatic life, specifically fish, salamander, and crayfish assemblages. We tested the hypothesis that animal assemblages, assessed by densities of ecologically important species, will differ among groups of watersheds with different well pad densities. In 2011, sites were sampled in Pennsylvania between July 13 and September 14 (N = 28; 10 = no well pads, 8 = low density, 10 = moderate density). We did not detect evidence of impact on fish, salamander, and crayfish assemblages. It should be noted that this study primarily assessed infrastructure-related impacts. Watersheds under investigations in this study area included no stream withdrawals for drilling and relatively low well pad density (≤0.541 well pads/km2), which reduces the amount of roads and pipeline as well as sediment runoff reaching the stream from this and other infrastructure during development. In addition, setback and landscape limitations also kept well pads relatively far from surface waters, and these watersheds remained highly forested after development because most were in state forests. Therefore, these findings represent the least intrusive scenario for impacts to aquatic communities while extracting natural gas.

Abstract:
The effects of hydraulic fracturing (HF) flowback and produced water (HF-FPW), a complex saline mixture of injected HF fluids and deep formation water that return to the surface, was examined in rainbow trout (Oncorhynchus mykiss). Exposure to HF-FPWs resulted in significant induction of ethoxyresorufin-O-deethylase (EROD) activity in both liver and gill tissues. Increased lipid peroxidation via oxidative stress was also detected by thiobarbituric acid reactive substances (TBARS) assay. The mRNA expressions of a battery of genes related to biotransformation, oxidative stress, and endocrine disruption were also measured using quantitative real-time polymerase chain reaction (Q-RT-PCR). The increased expression of cyp1a (2.49 ± 0.28-fold), udpgt (2.01 ± 0.31-fold), sod (1.67 ± 0.09-fold), and gpx (1.58 ± 0.10-fold) in raw sample exposure group (7.5%) indicated elevated metabolic enzyme activity, likely through the aryl hydrocarbon receptor pathway, and generation of reactive oxygen species. In addition, the elevated vtg and era2 expression demonstrated endocrine disrupting potential exerted by HF-FPW in rainbow trout. The overall results suggested HF-FPW could cause significant adverse effects on fish, and the organic contents might play the major role in its toxicity. Future studies are needed to help fully determine the toxic mechanism(s) of HF-FPW on freshwater fish, and aid in establishing monitoring, treatment, and remediation protocols for HF-FPW.

Abstract:
Quantitative flow–ecology relationships are needed to evaluate how water withdrawals for unconventional natural gas development may impact aquatic ecosystems. Addressing this need, we studied current patterns of hydrologic alteration in the Marcellus Shale region and related the estimated flow alteration to fish community measures. We then used these empirical flow–ecology relationships to evaluate alternative surface water withdrawals and environmental flow rules. Reduced high-flow magnitude, dampened rates of change, and increased low-flow magnitudes were apparent regionally, but changes in many of the flow metrics likely to be sensitive to withdrawals also showed substantial regional variation. Fish community measures were significantly related to flow alteration, including declines in species richness with diminished annual runoff, winter low-flow, and summer median-flow. In addition, the relative abundance of intolerant taxa decreased with reduced winter high-flow and increased flow constancy, while fluvial specialist species decreased with reduced winter and annual flows. Stream size strongly mediated both the impact of withdrawal scenarios and the protection afforded by environmental flow standards. Under the most intense withdrawal scenario, 75% of reference headwaters and creeks (drainage areas <99 km2) experienced at least 78% reduction in summer flow, whereas little change was predicted for larger rivers. Moreover, the least intense withdrawal scenario still reduced summer flows by at least 21% for 50% of headwaters and creeks. The observed 90th quantile flow–ecology relationships indicate that such alteration could reduce species richness by 23% or more. Seasonally varying environmental flow standards and high fixed minimum flows protected the most streams from hydrologic alteration, but common minimum flow standards left numerous locations vulnerable to substantial flow alteration. This study clarifies how additional water demands in the region may adversely affect freshwater biological integrity. The results make clear that policies to limit or prevent water withdrawals from smaller streams can reduce the risk of ecosystem impairment.

Abstract:
Unconventional natural gas (UNG) extraction (fracking) is ongoing in 29 North American shale basins (20 states), with ~6000 wells found within the Fayetteville shale (north-central Arkansas). If the chemical signature of fracking is detectable in streams, it can be employed to bookmark potential impacts. We evaluated benthic biofilm community composition as a proxy for stream chemistry so as to segregate anthropogenic signatures in eight Arkansas River catchments. In doing so, we tested the hypothesis that fracking characteristics in study streams are statistically distinguishable from those produced by agriculture or urbanization.

Abstract:
Unconventional natural gas development and hydraulic fracturing practices (fracking) are increasing worldwide due to global energy demands. Research has only recently begun to assess fracking impacts to surrounding environments, and very little research is aimed at determining effects on aquatic biodiversity and contaminant biomagnification. Twenty-seven remotely-located streams in Pennsylvania’s Marcellus Shale basin were sampled during June and July of 2012 and 2013. At each stream, stream physiochemical properties, trophic biodiversity, and structure and mercury levels were assessed. We used δ15N, δ13C, and methyl mercury to determine whether changes in methyl mercury biomagnification were related to the fracking occurring within the streams’ watersheds. While we observed no difference in rates of biomagnificaion related to within-watershed fracking activities, we did observe elevated methyl mercury concentrations that were influenced by decreased stream pH, elevated dissolved stream water Hg values, decreased macroinvertebrate Index for Biotic Integrity scores, and lower Ephemeroptera, Plecoptera, and Trichoptera macroinvertebrate richness at stream sites where fracking had occurred within their watershed. We documented the loss of scrapers from streams with the highest well densities, and no fish or no fish diversity at streams with documented frackwater fluid spills. Our results suggest fracking has the potential to alter aquatic biodiversity and methyl mercury concentrations at the base of food webs.

Abstract:
A survey of the large branchiopod fauna of the Eastern Cape Karoo region of South Africa was undertaken to provide baseline biodiversity information in light of impending shale gas development activities in the region. Twenty-two waterbodies, including nine dams and thirteen natural depression wetlands, were sampled during November 2014 and April 2015. A total of 13 species belonging to four orders were collected, comprising five anostracans, one notostracan, six spinicaudatans and one laevicaudatan. Cyzicus australis was most common, occurring in 46% of the waterbodies. Species co-occurred in 87% of the waterbodies, with a maximum number of six species recorded from the same waterbody. Our new distribution records for Lynceus truncatus, Streptocephalus spinicaudatus and Streptocephalus indistinctus represent substantial expansions of the previously known ranges for these species. Tarkastad is now the westernmost record for Streptocephalus spinicaudatus, while Jansenville now constitutes the southernmost record for Streptocephalus indistinctus. Large branchiopod distribution data from previous Eastern Cape records were combined with our current data, demonstrating that a total of 23 large branchiopod species have been recorded from the region to date. As the Karoo is one of the few major shale basins in the world where the natural baseline is still largely intact, this survey forms a basis for future reference and surface water quality monitoring during the process of shale gas exploration/extraction.

Abstract:
Because shale gas development is occurring over large landscapes and consequently is affecting many headwater streams, an understanding of its effects on headwater-stream faunal communities is needed. We examined effects of shale gas development (well pads and associated infrastructure) on Louisiana waterthrush Parkesia motacilla and benthic macroinvertebrate communities in 12 West Virginia headwater streams in 2011. Streams were classed as impacted (n = 6) or unimpacted (n = 6) by shale gas development. We quantified waterthrush demography (nest success, clutch size, number of fledglings, territory density), a waterthrush Habitat Suitability Index, a Rapid Bioassessment Protocol habitat index, and benthic macroinvertebrate metrics including a genus-level stream-quality index for each stream. We compared each benthic metric between impacted and unimpacted streams with a Student's t-test that incorporated adjustments for normalizing data. Impacted streams had lower genus-level stream-quality index scores; lower overall and Ephemeroptera, Plecoptera, and Trichoptera richness; fewer intolerant taxa, more tolerant taxa, and greater density of 0–3-mm individuals (P ≤ 0.10). We then used Pearson correlation to relate waterthrush metrics to benthic metrics across the 12 streams. Territory density (no. of territories/km of stream) was greater on streams with higher genus-level stream-quality index scores; greater density of all taxa and Ephemeroptera, Plecoptera, and Trichoptera taxa; and greater biomass. Clutch size was greater on streams with higher genus-level stream-quality index scores. Nest survival analyses (n = 43 nests) completed with Program MARK suggested minimal influence of benthic metrics compared with nest stage and Habitat Suitability Index score. Although our study spanned only one season, our results suggest that shale gas development affected waterthrush and benthic communities in the headwater streams we studied. Thus, these ecological effects of shale gas development warrant closer examination.

Abstract:
Hydraulic fracturing (fracking) poses significant threats to freshwater resources and stream ecosystems. Little research exists to quantify the ecological impact and in Pennsylvania alone over 10,000 wells have been permitted. This study aimed to determine if hydraulic fracturing is having any impacts on stream ecosystem health by measuring stream pH and temperature, macroinvertebrate index of biological integrity (IBI), and the gill morphology of individuals in the Hydropsychidae Diplectrona taxa. Six streams in northwestern Pennsylvania were selected as study sites (three with fracking occurring in their watershed and three without fracking). IBI scores were significantly higher at non-fracked sites and were also correlated with stream pH. Macroinvertebrate gill width did not vary between fracked and non-fracked sites but was correlated with percent hydric soils, suggesting that hydric soils may be a good long-term indicator of stream dissolved oxygen. While our results did not indicate differences in Hydropsychidae Diplectrona gill widths between fracked and non-fracked sites, we did observe that fracked sites had more acidic stream water and lower IBI scores. These results indicate the need for further study to assess the potential impacts of hydraulic fracturing on stream ecosystems.

Abstract:
Exploration and extraction of oil and natural gas have increased in recent years and are expected to expand in the future. Reduction in water quality from energy extraction may negatively affect water supply for agriculture and urban use within catchments as well as down river. We used non-invasive genetic techniques and capture–recapture modeling to estimate the abundance and density of North American river otters (Lontra canadensis), a sentinel species of aquatic ecosystems, in Southwestern Wyoming. While densities in two of three river reaches were similar to those reported in other freshwater systems in the western US (1.45–2.39 km per otter), otters appeared to avoid areas near energy development. We found no strong difference in habitat variables, such as overstory cover, at the site or reach level. Also, fish abundance was similar among the three river reaches. Otter activity in our study area could have been affected by elevated levels of disturbance surrounding the industrial gas fields, and by potential surface water contamination as indicated by patterns in water conductivity. Continued monitoring of surface water quality in Southwestern Wyoming with the aid of continuously recording devices and sentinel species is warranted.

Abstract:
Oil and gas extraction in shale plays expanded rapidly in the U.S. and is projected to expand globally in the coming decades. Arkansas has doubled the number of gas wells in the state since 2005 mostly by extracting gas from the Fayetteville Shale with activity concentrated in mixed pasture-deciduous forests. Concentrated well pads in close proximity to streams could have adverse effects on stream water quality and biota if sedimentation associated with developing infrastructure or contamination from fracturing fluid and waste occurs. Cumulative effects of gas activity and local habitat conditions on macroinvertebrate communities were investigated across a gradient of gas well activity (0.2-3.6wells per km(2)) in ten stream catchments in spring 2010 and 2011. In 2010, macroinvertebrate density was positively related to well pad inverse flowpath distance from streams (r=0.84, p<0.001). Relatively tolerant mayflies Baetis and Caenis (r=0.64, p=0.04), filtering hydropsychid caddisflies (r=0.73, p=0.01), and chironomid midge densities (r=0.79, p=0.008) also increased in streams where more well pads were closer to stream channels. Macroinvertebrate trophic structure reflected environmental conditions with greater sediment and primary production in streams with more gas activity close to streams. However, stream water turbidity (r=0.69, p=0.02) and chlorophyll a (r=0.89, p<0.001) were the only in-stream variables correlated with gas well activities. In 2011, a year with record spring flooding, a different pattern emerged where mayfly density (p=0.74, p=0.01) and mayfly, stonefly, and caddisfly richness (r=0.78, p=0.008) increased in streams with greater well density and less silt cover. Hydrology and well pad placement in a catchment may interact to result in different relationships between biota and catchment activity between the two sample years. Our data show evidence of different macroinvertebrate communities expressed in catchments with different levels of gas activity that reinforce the need for more quantitative analyses of cumulative freshwater-effects from oil and gas development.

Abstract:
Mercury (Hg) is a persistent element in the environment that has the ability to bioaccumulate and biomagnify up the food chain with potentially harmful effects on ecosystems and human health. Twenty-four streams remotely located in forested watersheds in northwestern PA containing naturally reproducing Salvelinus fontinalis (brook trout), were targeted to gain a better understanding of how Marcellus shale natural gas exploration may be impacting water quality, aquatic biodiversity, and Hg bioaccumulation in aquatic ecosystems. During the summer of 2012, stream water, stream bed sediments, aquatic mosses, macroinvertebrates, crayfish, brook trout, and microbial samples were collected. All streams either had experienced hydraulic fracturing (fracked, n = 14) or not yet experienced hydraulic fracturing (non-fracked, n = 10) within their watersheds at the time of sampling. Analysis of watershed characteristics (GIS) for fracked vs non-fracked sites showed no significant differences (P > 0.05), justifying comparisons between groups. Results showed significantly higher dissolved total mercury (FTHg) in stream water (P = 0.007), lower pH (P = 0.033), and higher dissolved organic matter (P = 0.001) at fracked sites. Total mercury (THg) concentrations in crayfish (P = 0.01), macroinvertebrates (P = 0.089), and predatory macroinvertebrates (P = 0.039) were observed to be higher for fracked sites. A number of positive correlations between amount of well pads within a watershed and THg in crayfish (r = 0.76, P < 0.001), THg in predatory macroinvertebrates (r = 0.71, P < 0.001), and THg in brook trout (r = 0.52, P < 0.01) were observed. Stream-water microbial communities within the Deltaproteobacteria also shared a positive correlation with FTHg and to the number of well pads, while stream pH (r = −0.71, P < 0.001), fish biodiversity (r = −0.60, P = 0.02), and macroinvertebrate taxa richness (r = −0.60, P = 0.01) were negatively correlated with the number of well pads within a watershed. Further investigation is needed to better elucidate relationships and pathways of observed differences in stream water chemistry, biodiversity, and Hg bioaccumulation, however, initial findings suggest Marcellus shale natural gas exploration is having an effect on aquatic ecosystems.

Abstract:
Fracking fluids were released into Acorn Fork, KY, a designated Outstanding State Resource Water, and habitat for the threatened Chrosomus cumberlandensis (Blackside Dace). As a result, stream pH dropped to 5.6 and stream conductivity increased to 35,000 µS/cm, and aquatic invertebrates and fish were killed or distressed. The objective of this study was to describe post-fracking water quality in Acorn Fork and evaluate if the changes in water quality could have extirpated Blackside Dace populations. Semotilus atromaculatus (Creek Chub) and Lepomis cyanellus (Green Sunfish) were collected from Acorn Fork a month after fracking in lieu of unavailable Blackside Dace. Tissues were histologically analyzed for indicators of stress and percent of fish with lesions. Fish exposed to affected Acorn Fork waters showed general signs of stress and had a higher incidence of gill lesions than unexposed reference fish. Gill lesions observed were consistent with exposure to low pH and toxic concentrations of heavy metals. Gill uptake of aluminum and iron was demonstrated at sites with correspondingly high concentrations of these metals. The abrupt and persistent changes in post-fracking water quality resulted in toxic conditions that could have been deleterious to Blackside Dace health and survival.

Abstract:
Expansion of natural gas drilling into the Marcellus Shale formation is an emerging threat to the conservation and restoration of native brook trout (Salvelinus fontinalis) populations. Improved drilling and extraction technologies (horizontal drilling and hydraulic fracturing) have led to rapid and extensive natural gas development in areas overlying the Marcellus Shale. The expansion of hydraulic fracturing poses multiple threats to surface waters, which can be tied to key ecological attributes that limit brook trout populations. Here, we expand current conceptual models to identify three potential pathways of risk between surface water threats associated with increased natural gas development and life history attributes of brook trout: hydrological, physical, and chemical. Our goal is to highlight research needs for fisheries scientists and work in conjunction with resource managers to influence the development of strategies that will preserve brook trout habitat and address Marcellus Shale gas development threats to eastern North America's only native stream salmonid.

Abstract:
Sediment toxicity tests were conducted to assess potential effects of contaminants associated with coal mining or natural gas extraction activities in the upper Tennessee River basin and eastern Cumberland River basin in the United States. Test species included two unionid mussels (rainbow mussel, Villosa iris, and wavy-rayed lampmussel, Lampsilis fasciola, 28-d exposures), and the commonly tested amphipod, Hyalella azteca (28-d exposure) and midge, Chironomus dilutus (10-d exposure). Sediments were collected from seven test sites with mussel communities classified as impacted and in proximity to coal mining or gas extraction activities, and from five reference sites with mussel communities classified as not impacted and no or limited coal mining or gas extraction activities. Additional samples were collected from six test sites potentially with high concentrations of polycyclic aromatic hydrocarbons (PAHs) and from a test site contaminated by a coal ash spill. Mean survival, length, or biomass of one or more test species was reduced in 10 of 14 test samples (71%) from impacted areas relative to the response of organisms in the five reference samples. A higher proportion of samples was classified as toxic to mussels (63% for rainbow mussels, 50% for wavy-rayed lampmussels) compared with amphipods (38%) or midge (38%). Concentrations of total recoverable metals and total PAHs in sediments did not exceed effects-based probable effect concentrations (PECs). However, the survival, length, or biomasses of the mussels were reduced significantly with increasing PEC quotients for metals and for total PAHs, or with increasing sum equilibrium-partitioning sediment benchmark toxic units for PAHs. The growth of the rainbow mussel also significantly decreased with increasing concentrations of a major anion (chloride) and major cations (calcium and magnesium) in sediment pore water. Results of the present study indicated that (1) the findings from laboratory tests were generally consistent with the field observations of impacts on mussel populations; (2) total recoverable metals, PAHs, or major ions, or all three in sediments might have contributed to the sediment toxicity; (3) the mussels were more sensitive to the contaminants in sediments than the commonly tested amphipod and midge; and (4) a sediment toxicity benchmark of 1.0 based on PECs may not be protective of mussels. Environ. Toxicol. Chem. 2013;32:207–221. © 2012 SETAC

Abstract:
Shale gas development may involve trade-offs between energy development and benefits provided by natural ecosystems. However, current best management practices (BMPs) focus on mitigating localized ecological degradation. We review evidence for cumulative effects of natural gas development on brook trout (Salvelinus fontinalis) and conclude that BMPs should account for potential watershed-scale effects in addition to localized influences. The challenge is to develop BMPs in the face of uncertainty in the predicted response of brook trout to landscape-scale disturbance caused by gas extraction. We propose a decision-analysis approach to formulating BMPs in the specific case of relatively undisturbed watersheds where there is consensus to maintain brook trout populations during gas development. The decision analysis was informed by existing empirical models that describe brook trout occupancy responses to landscape disturbance and set bounds on the uncertainty in the predicted responses to shale gas development. The decision analysis showed that a high efficiency of gas development (e.g., 1 well pad per square mile and 7 acres per pad) was critical to achieving a win-win solution characterized by maintaining brook trout and maximizing extraction of available gas. This finding was invariant to uncertainty in predicted response of brook trout to watershed-level disturbance. However, as the efficiency of gas development decreased, the optimal BMP depended on the predicted response, and there was considerable potential value in discriminating among predictive models through adaptive management or research. The proposed decision-analysis framework provides an opportunity to anticipate the cumulative effects of shale gas development, account for uncertainty, and inform management decisions at the appropriate spatial scales.Environmental Practice 14:1–16 (2012)

Abstract:
Over the summer and fall seasons, 2006–2010, we surveyed the fish and macroinvertebrate communities of the Tenmile Creek basin in southwestern Pennsylvania, an area undergoing accelerated extraction of energy resources—historically coal and more recently natural gas associated with the Marcellus formation. Tenmile Creek, its major South Fork (SF), and numerous tributaries drain a basin of 875 km2. The drainage network is characterized as warm-water, low-gradient, and net alkaline. The purpose was to provide synoptic baseline data on water quality and the resident aquatic communities in terms of species richness, stress tolerance, and trophic structure. Overall, we sampled 20 stations on the 2 main branches and 1 on each of the 12 tributaries. We collected 26,375 fishes representing nine families and 54 species/hybrids along with 989 macroinvertebrates from 25 separate taxa. The parameter which defines water quality here is specific conductance which ranges from natural background levels of about 400 μS/cm on the minimally impaired Tenmile mainstem to 4,500 μS/cm on its SF. Diverse fish and macroinvertebrate communities were documented at levels of specific conductance exceeding 1,000 μS/cm, well above the 300 μS/cm criterion to protect aquatic life proposed by the US Environmental Protection Agency for streams in the Central Appalachian Region. South Fork fish communities exhibit impairment at levels of specific conductance approaching the maximum observed here.