West Virginia EPSCoR Track I: Appalachian Freshwater ... · Task Lead: Dr. Jason A. Hubbart, West Virginia University Others Involved in the Research: Dr. Elliott Kellner, Dr. Evan

West Virginia EPSCoR Track I:

Appalachian Freshwater Initiative

The Appalachian Freshwater Initiative is supported by the National Science Foundation

under Award Number OIA-1458952.

September 10, 2018

Table of Contents

About the Appalachian Freshwater Initiative ............................................................................... 1

AFI Researchers

Dr. Jim Anderson ................................................................................................................. 2

Dr. Brian Antonsen .............................................................................................................. 4

Dr. Robert C. Burns ............................................................................................................. 5

Dr. Philippe T. Georgel ........................................................................................................ 6

Dr. Jason A. Hubbart............................................................................................................ 8

Dr. David Huber ................................................................................................................. 10

Dr. Lian-Shin Lin ............................................................................................................... 12

Drs. Sridhar Malkaram & David Huber ............................................................................. 13

Dr. Sean P. McBride .......................................................................................................... 14

Dr. Michael Norton ............................................................................................................ 15

Dr. J. Todd Petty ................................................................................................................ 16

Dr. Gary E. Schultz, Jr. ...................................................................................................... 17

Dr. Jack Smith .................................................................................................................... 18

Dr. Nadja Spitzer................................................................................................................ 19

Dr. Michael Strager and Dr. Anthony Szwilski ................................................................. 20

Dr. Dorothy Vesper ............................................................................................................ 22

Dr. Nicolas Zegre ............................................................................................................... 23

September 10, 2018

About the Appalachian Freshwater Initiative

The Issue Clean, reliable water is the most fundamental natural resource, underlying everything in the

natural world and playing a vital role in nearly all aspects of society, from quality of life and health to economics, politics, and culture. The mountains of West Virginia represent the headwaters of major river networks that supply fresh water to millions of people, including Washington, DC, which lies on the Potomac River. Thus, the security and sustainability of WV water resources is of national relevance.

The Appalachian Freshwater Initiative (AFI) was established as part of National Science Foundation/EPSCoR Award No. 1458952 and represents a statewide research collaborative of biologists, ecologists, engineers, chemists, and geologists whose mission is to strengthen WV’s environmental, economic, social, and cultural well-being by promoting water security and sustainability.

Research Highlights Through unprecedented research collaborations, the AFI is developing sustainable solutions

to contemporary water quality, quantity, management, and use problems within WV. The AFI consists of three focus groups focused on:

1. Improving our ability to detection chemical, physical, and biological threats to water quality.AFI researchers are on the forefront of developing chemical- and biological-basedtechnologies for detecting pollutants and toxins. Through AFI research efforts, managementagencies are better equipped to detect, manage, and adapt to acute and chronic threats,ultimately ensuring safe water for WV residents.

2. Understanding how chemical, physical, and biological threats to water quality impact thehealth of aquatic ecosystems, as well as communities that rely on them. AFI researchactivities collectively represent the most in-depth study of how common pollutants in WVstreams affect both aquatic organisms and humans. This research is providing criticalinformation on how to effectively manage aquatic systems to maximize ecological and publichealth.

3. Developing the science and tools needed to predict future threats to aquatic resources. Criticalto ensuring aquatic resource security and sustainability is the ability to predict how waterresources (e.g., quality and quantity) will response to realistic future conditions. AFI facultyare developing datasets and models for predicting changes in water quality and quantity.

September 10, 2018 1

Environmental DNA (eDNA)

Project Objectives: 1. Test nucleic acid concentration and extraction methods.2. Improve and environmental DNA (eDNA) laboratory techniques for assessing amphibian

and fish assemblages.

Task Leader: Dr. Jim Anderson, West Virginia University

Others Involved in the Research: Yvette Halley, postdoctoral fellow

Problem: Can non-invasive environmental DNA (eDNA) technologies be used to identify fish communities?

Approach: Currently there is a paucity of native West Virginian fish species in the GenBank (NCBI’s genetic database) to use for primer design and comparative genetic analysis. We collected tissue from native fish to serve as reference sequences for comparative analysis. Environmental DNA was collected using an aquatic filtering approach. Environmental DNA was processed using next generation sequencing technologies (illumina miSeq) and the genetic sequences generated were compared to both existing sequences in the GenBank and the newly generated native species sequences. These results were compared to traditional stream surveys that were conducted in parallel to the eDNA collection.

Findings: Depending on the genetic region targeted for community analysis we can either differentiate fish communities at the family level or the species level. We have more success determining fish families rather than species when the species are closely related. Multiple genetic primers have to be utilized to


assess a community because it increases the likelihood of detecting genetic material shed by various species since eDNA degrades relatively quickly.

Implications: Since eDNA has a discrete detection window it provides us with the capabilities of assessing which species recently inhabited the environment being studied. Because of its non-invasive nature it is a valuable tool to help determine if more invasive detection techniques are warranted. Our project will also contribute to the availability of non-model organism genetic sequence availability in the GenBank.


Low level manganese induces detrimental changes in behavior and physiology of crayfish

Project Objective: Characterize the impacts of low level manganese on aquatic organisms.

Task Leader: Dr. Brian Antonsen, Marshall University

Postdoc/Students Involved in the Research: Cody Lambert, Lauren Reasor, Sonia Chandi, and Brittany Short.

Problem: The toxicological and detrimental impacts of low level manganese are not well understood, although levels considered "safe" by EPA standards do induce changes in behavior and physiology of aquatic organisms.

Approach: Juvenile crayfish are exposed to levels starting two levels of magnitude below, and up to one order of magnitude above, the EPA secondary standard of 0.05 mg/L for manganese, for periods ranging from 2 days to 6 months. During and following exposure, behavior is assayed with novel environment and escape response tests, focusing on their exploratory and escape behavior, overall motility, and coordination. Following this, tissues are harvested for later histology.

Findings: Behavior becomes less predictable, suggesting a loss of control or choice making ability. Animals exposed to levels as low as 0.005mg/L for periods as short as two weeks become erratic, move less, become less cautious by moving away from shelter, and make

different choices for escape. We see increased mortality at levels as low as 0.01 mg/L, exposed for two days. Finally, muscle loss and generally poor health is evident during histological analysis.

Implications: Manganese, among other impacts, affects calcium homeostasis, which can lead to innumerable neurological. neuromuscular, and metabolic disruptions. By characterizing the changes induced by exposure to low levels of manganese, we aim to better understand how this wide ranging toxicant 1) changes behavior and the underlying neuromuscular substrates of behavior. 2) Inform how subtle changes in behavior orphysiology could impact water ecosystems.3) Inform regulatory and/or remediation efforts tobetter preserve natural water systems.

Top Left: Master’s student Cody Lambert uses the behavior analysis program, Ethovision (from Noldus), to score escape behaviors following exposure to manganese. Bottom Left: Undergraduate Student Lauren Reasor uses the Noldus Daniovision system to track motility and exploratory behavior of crayfish exposed to manganese.


Human Perceptions of Appalachian Water Quality and Associated Use and Behaviors

Project Objective: Quantify West Virginia resident and non-resident perceptions of water quality and resultant behaviors related to consumption, recreation, and other use types.

Task Leader: Dr. Robert C. Burns, West Virginia University

Others Involved in the Research: Ross Andrew and Jonas Levêque

Problem: Within Appalachia, there is a great wealth of both freshwater resources and threats. Therefore, management and understanding of water resources and their associated quality is critical. Human perceptions of water resource quality may greatly influence behaviors that help direct management and conservation actions.

Approach: The use of multiple survey instruments allowed data collection from multiple groups (resident, non-resident, student, non-student, etc.). Surveys were designed to elicit specific individual perceptions of local and regional water quality and associated use behaviors. Over 29,000 surveys were sent via both online and mail systems, with response rates ranging from 11.3-17.6%. Responses were quantified and used to model perceptions as they relate to behaviorsand other factors.

Findings: Through these data analysis and modeling efforts, several key findings have been established. First, water quality perceptions have a directly proportional effect on intention to recreate in West Virginia. Also, overall area satisfaction significantly influences water quality perceptions. Students from West Virginia tend to drink bottled water more than students who are not originally from West Virginia. The perceptions which drive water use behaviors are most commonly health risk, organoleptic (taste, odor, color), and environmental concern.

Implications: This research shows that differential management actions should be undertaken given the context of resident profiles and perceptions. Education and communication about local risks associated with water quality should be targeted for improvement throughout West Virginia and across resident groups. Such improvements could reduce unnecessary waste (both physical and financial) associated with behaviors that are perceived to be risk-averse where risks are actually relatively low.

Left: Dr. Jonas Levêque and Dr. Robert Burns present a research poster at the 2017 Society of American Foresters Annual Convention in Albuquerque, NM.


Effects of Exposure to the Opioid Buprenorphine on Brain Development

Project Objective: Determine the epigenetic changes induced by Buprenorphine exposure in brain cells.

Task Leader: Dr. Philippe T. Georgel, Marshall University

Others Involved in the Research: Tanner Bakhshi, Tanner Way, Bradley Munci, and Norman Cole

Problem: Buprenorphine (Bup), a substitute opioid, is used in medication assisted treatment programs for pregnant women. The Bup treatment period overlaps with crucial stages of embryonic Central Nervous System (CNS) development, including gliogenesis, synapto-genesis, and myelination. These developmental events are under the control of multiple epigenetic regulators, including the master regulator of brain development: Brain Derived Neurotrophic Factor (BDNF). Interestingly, BDNF itself is controlled by the epigenetic regulator Methyl CpG-binding Protein 2 (MeCP2), a protein linked with autism spectrum disorder. As the epigenetic changes induced by Buprenorphine exposure is poorly understood, we are undergoing the generation of a global mapping. Local changes at specific regulatory sequences will be investigated in the future.

Approach: Using brain cells (endothelial and oligodendrocyte precursor cells) and in vivo data from rats, we are establishing a histone modification profile of brain cells in presence and absence of Buprenorphine treatment. To complement our epigenetic profiles, we are including BDNF, MeCP2 and its partner CTCF.

Findings: Buprenorphine exposure modifies the epigenetic profile of histones in brain cells, including modifications involved in MeCP2 recruitment, which expression profile is also affected by Buprenorphine exposure, both in vitro and in vivo.

Figure 1: In vivo analysis of histone post-translational modification or PTMs (Epigenetic markers) profile of rat brain at day 7 (+/- Bup). Left Panel: Western blots. Right Panel: Summary after normalization of signals.


Implications: Changes in epigenetic profiles mediated by Buprenorphine exposure include histone PTMs important for MeCP2 recruitment (see Figure 2) and proper brain development.

Figure 2: MeCP2 regulations of BDNF expression


A quantitative comparison of fecal coliform concentration versus varying land-use types in a representative mixed-land-use

watershed of Appalachia

Project Objective: To advance understanding of watershed-scale land-use and E. coli relationships in the Appalachian region.

Task Lead: Dr. Jason A. Hubbart, West Virginia University

Others Involved in the Research: Dr. Elliott Kellner, Dr. Evan Kutta, Fritz Petersen, Parameshwor Takhache, and Rivkah Nisan

Problem: Studies are needed that advance understanding of anthropogenic influences on stream pathogen regimes. However, contemporary watershed management is increasingly complex as a result of compounding factors (e.g. climate variability, land use impacts) that confound attribution of causal mechanisms to observed hydrologic alterations and pathogen loading.

Approach: An experimental watershed study, comprising 22 monitoring sites in a mixed-land-use watershed, was established to investigate relationships between stream Escherichia (E) coli concentrations, land use, stream physicochemistry, and hydroclimate in the Appalachian region. An initial study was conducted to validate a simple practitioner method for E. coli quantification. Stream water samples have been collected weekly and analyzed for E. coli concentration. Capacity building and collaborative efforts include weekly geochemical and suspended sediment sampling, continuous climate monitoring, investigation of riparian microbial dynamics, and monthly physical habitat assessments (PHA) via traditional and drone-based methods.

Above: Dr. Elliott Kellner collects samples in the West Run Watershed in Morgantown, WV. Right: Graduate Research Assistant Fritz Petersen processes samples in the lab.


Findings: E. coli concentrations show significant (p < 0.05) spatiotemporal differences, consistently displaying correlation with agricultural land use, and thereby validating the method (paper in submission). Geochemistry results indicate significantly (p < 0.05) lower concentrations of nutrients and metals in forest tributaries, relative to developed and agricultural reaches (paper in submission). PHA findings illustrate the rapid degradation of aquatic habitat resulting from land-use-driven streamflow alterations (Hubbart et al., 2017; Hentz et al., 2018).

Implications: Results highlight the capacity of the methodology to robustly quantify pathogen regimes. The methods can facilitate targeting of remediation/restoration efforts within mixed-use watersheds, thereby improving management efficacy. Collectively, findings emphasize the importance and complexity of stream hydro-biogeochemical regimes, and thus the need for additional studies.

Dr. Jason Hubbart (center) meets with Research Assistant Angela Hentz, Charles Yuill (in red), and Dr. Paul Kinder from the WVU Natural Resource Analysis Center (NRAC) to develop a plan for monthly physical habitat assessments (PHA) using unmanned aerial vehicles (UAVs).

References: Hentz, A., Kinder, P., Hubbart, J.A., Kellner, E. 2018. Accuracy and Optimal altitude for

Physical Habitat Assessment (PHA) of stream environments using Unmanned Aerial Vehicles (UAV). Drones 2(20): 1-15.

Hubbart, J.A., Kellner, E., Kinder, P., Stefan, K. 2017. Challenges in Aquatic Physical Habitat Assessment: Improving Conservation and Restoration Decisions for Contemporary Watersheds. Challenges 8: 31.


Spatial Structure of Sediment Microbial Diversity in a Heavily Impacted Appalachian River, West Virginia

Project Objective: To determine whether microbial diversity shows spatial variation, and to test whether sediment chemistry drives spatial patterns.

Task Leader: Dr. David Huber, West Virginia State University

Others Involved in the Research: Ifeoma R. Ugwuanyi, Andrielle Larissa Kemajou, Dr. Sridhar A. Malkaram, Dr. Amir Hass, Vadesse Lhilhi Noundou, Natalia Montenegro-Garcia, and Jesus E. Chavarria-Palma

Problem: Healthy watersheds depend on ecosystem services provided by sediment microbiomes. However, surprisingly little is known about the structure, distribution and functional potential of these microbiomes. Potential drivers of spatial diversity patterns include watershed discharge, local geochemistry, land-use practices, and stressors. The Kanawha River is a tributary of the

Ohio River and serves the Charleston (WV) metropolitan area. The watershed encompasses 12,000 sq. miles and includes surface mining, acid mine drainage, logging, municipal inputs and an 80-year chemical industry.


Approach: Sediment samples were collected from six locations along a 60km region of the river. Two upper layers of sediment (1-5 and 6-10 cm) were collected. Illumina sequencing using Earth Microbiome Project protocols was used to target 16S rRNA gene diversity. Coordinated chemical analysis of 20 variables was done with ICP-OES and Ion Chromatography. 15 million sequences were obtained from 56 sediment samples.

Findings: Geographic variation in sediment chemistry was found. Beta diversity analysis (Bray-Curtis and UniFrac) also showed that bacterial OTU diversity displayed spatial differentiation among sites. Abundance of major phyla was most strongly associated with total organic carbon, sulfate, K, Mg, Mn, and Al. Correlation analysis also showed that bacterial phylogenetic groups had distinct geochemical profiles implying phylogenetic niche coherence.

Implications: We found that the high-flow and high sediment load did not homogenize microbial diversity and that spatial structure of diversity was strongly associated with local sediment geochemistry.


Innovative iron-based technologies for value creation: turning coal wastes management into revenue generating operations

Project Objective: Our research goal is to turn coal waste management into revenue-generating operations through engineering innovative technologies and networking waste infrastructure.

Task Leader: Dr. Lian-Shin (Lance) Lin, West Virginia University

Others Involved in the Research: Dr. Karen Buzby, Musfique Ahmed, Rifat Anwar, Dongyang Deng, Nashid Mirza, Franklin Cavallo, Carney Quek, Jack Bajerski, Danielle Schlapo, and Madison Haddix

Problem: In coal producing regions, the current piecemeal and linear approach of coal wastes management practices has resulted in a plethora of issues including acidification and elevated total dissolved solids in receiving waters, toxic chemicals leaching, and storage structure failures posing serious threats to the environment and public health.

Approach: Our research has a focus on developing innovative technologies for extracting iron and other valuable elements from coal wastes and utilize them in various beneficial applications to create a novel economy. In particular, we have been developing a suite of innovative iron-based technologies for various applications in food, energy, and water sectors.

Findings: An innovative Fe-based anaerobic wastewater treatment process has successfully developed and operated at WVU. This innovative treatment method was designed to provide multi-faceted benefits including energy efficiency, high levels of phosphorus removal, and low biological sludge yields compared to existing aerobic treatment methods. The research has recently been expanded to investigate ammonium removal through a reaction pathway termed FEAMMOX. In addition, an iron sorbent has been developed and used in nutrient management for tomato growth. Greenhouse experiments have demonstrated its benefits in reducing runoff nutrient loads and fertilizer application rate while providing a constant source of phosphorus for food production.

Implications: These innovative technologies are expected to enable new interlinkages of material flows among food, energy, and water sectors and to improve resource utilization efficiencies. We anticipate that an integrated waste management at a regional scale can create a novel regional economy while providing environmental, economic, and social benefits.


Model the microbial population levels with respect to physical and chemical changes in Kanawha River

Project Objective: Evaluate multiple regression models to predict populations of microbial (pathogenic) communities using the physical and chemical factors.

Task Leaders: Dr. Sridhar Malkaram and Dr. David Huber, West Virginia State University

Others Involved in the Research: Ifeoma Ugwuanyi, Shaka Wilkerson

Problem: Chemical pollutants can have immediate and long-term effects on the river water and sediment microbial populations and change the normal nutrient cycling. It is necessary to study the responses and changes of microbial community profiles in relation to the chemical stressors.

Approach: In year 3, data from 7 locations on Kanawha river was used, to analyze the effect of environment variables on abundance profiles of specific microbial taxa. Simple linear regression and multiple regressions are used to find the effects of specific chemicals or combination of chemicals on the abundances of specific taxonomic or sub-taxonomic groups.

Findings: We found that few specific taxonomic groups/sub-groups were correlated with certain environmental (chemical) gradients. Currently this data is being analyzed using multiple regression methods to identify specific stressors and specific indicator micro-organisms.

Implications: An outcome of this work included the development of a Jupiter notebook workflow which is available online for exploratory analysis of metagenomic data. The workflow was developed in coordination with the thesis work of a graduate student. During the summer of year 3, a summer student was involved in developing an application for comparatively analysis of metagenomic profiles. It is currently being added functionality for general usage.


Ultrathin Porous Self-Assembled Nanoparticle Membranes for Water Filtration

Project Objective: Demonstrate the rejection capabilities of novel ultrathin porous self-assembled highly ordered nanoparticle membranes intended for water purification. Eventual contaminants that will be passed through these membranes and tested for rejection will range from micron to sub-micron organic matter, nanometer sized molecular dyes, down to dissociated salt ions such as those found in streams, lakes, rivers, and ocean water.

Task Leader: Dr. Sean P. McBride, Marshall University

Others Involved in the Research: Ryan Vincent

Problem: With only 2.5 percent of water on earth being fresh water and with only 0.3 percent being accessible on the surface of the Earth, having access to fresh water is a global problem (Shiklomanov, 1993). The 0.3 percent does not distinguish between polluted and non-polluted ‘fresh water’ sources. Developing innovative cost-effective membranes that addresses this large-scale problem of fresh water scarcity is the main focus of this project.

Approach: This research will focus on using ultrathin porous self-assembled highly ordered nanoparticle membranes (SHO NPMs), which when layered together make a composite nanoparticle (NP) filter with high permeability.

Findings to Date: It has been previously shown that such self-assembled nanoparticle membranes can be used to modify porous materials and increase ionic rejection for low concertation salt water solutions (Barry et al, 2014) The work at Marshall will continue to focus on both the steric and ionic rejection capabilities from such self-assembled membranes.

Implications: Results from this proposed research could help to determine the feasibility of these types of innovative filters and if they could develop into the next generation of water filtration or desalination technologies.

References Shiklomanov, I.A. 1993. World Fresh Water

Resources in Water in Crisis, A guide to the World's Fresh Water Resources, Oxford University Press, pp. 13-24.

Barry, E., McBride, S., Jaeger, H., Lin, X.M. 2014. Ion Transport Controlled by Nanoparticle-Functionalized Membranes. Nature Communications 5: 5847.


Sensor Platform Development

Project Objective: Improve detection of chemical, physical, and biological threats to water quality.

Task Leader: Dr. Michael Norton, Marshall University

Others Involved in the Research: Kathryn Pitton and Zachary Boggs

Problem: Transparent substrates with high binding to DNA are required in order to implement DNA nanostructures (DNANS) as sensor platforms. No optimal transparent substrates which enable these sensing systems to retain their structures, yet bind to solid surfaces, a requirement in order to best utilize DNANS as organizing substrates, have been identified to date.

Approach: The interaction of DNANS with several candidate transparent substrates, including sapphire, were characterized using Atomic Force Microscopy. Sapphire had to be processed at high temperature in order to perform this characterization.

Findings: High temperature water vapor treatment, a pseudo-hydrothermal method, was demonstrated to lead to terrace development while maintaining necessary transparency. In addition, compatibility of DNA based nanostructures with this surface was demonstrated for the first time.

Figure 1. AFM images of the process of transforming optically flat sapphire surface into near atomic flatness, compatible with high resolution AFM and Optical imaging.

Implications: Although single molecule scale sensing will not require ultraflat substrates in the long term, necessary “troubleshooting” and structure validation of nanoscale assembly will be required for initial studies throughout the development phases. The methods developed here constitute an enabling technology and the materials compatibility demonstrated will be valuable to researchers using DNA structures for species organization for the foreseeable future.


Cumulative effects of landscape stressors on freshwater environments in West Virginia

Project Objective: Develop predictive models quantifying the effects of landscape stressors on abiotic and biotic characteristics of freshwater environments.

Task Leader: Dr. J. Todd Petty

Others Involved in the Research: Dr. Brock M. Huntsman, Josh Ankeny, and Brian Gordon

Problem: Freshwater ecosystems are exposed to numerous anthropogenic stressors throughout the United States, with climate change, urban development, and mineral extraction especially concerning for aquatic resources within West Virginia. The recent expansion of unconventional oil and gas (UOG), on top of legacy effects of the coal and timber industries, has the potential to exacerbate the deterioration of already stressed freshwater ecosystems of West Virginia.

Approach: We are currently developing spatiotemporally robust models to elucidate patterns of complex interactions between landscape stressors and biota within aquatic environments. The cumulative impacts of temperature and chemical contaminants within stressed landscapes will be used to predict responses of ecological communities to climate and landscape change.

Findings: Preliminary findings have identified the health of aquatic communities are most sensitive to the legacy effects of historic coal mining practices, with impacts of UOG on water chemistry and community structure being mild in comparison. However, elevated stress levels in creek chub populations indicated the cumulative impacts of UOG in concert with other landscape stressors may escalate deleterious effects on even the most resilient aquatic taxa. Distribution of sensitive taxa like brook trout were similarly constrained by the cumulative effects of multiple landscape stressors. However, remediation of stressful water temperatures within at least one watershed suggests brook trout productivity may benefit by improved thermal conditions.

Implications: Preliminary results emphasize that multiple levels of biological organization are to some degree sensitive to landscape stressors, with the potential of deleterious effects exacerbated by compounding disturbances on the landscape.


Lotic water column microbial community dynamics including the influence of environmental and toxicological factors

Project Objective: To determine whether large lotic systems such as the Ohio River contain core communities of bacteria that are persistent, abundant, and biologically relevant, particularly with respect to the introduction of disturbances such as exposure to chemicals and to changes in temperature.

Task Leader: Dr. Gary E. Schultz, Jr., Marshall University

Others Involved in the Research: Yasamine Sadeghian and Sydney Harry

Problem: Determine the impact of potential toxic spills into lotic systems on bacterial communities.

Approach: Two approaches were attempted. In the first, my lab used micro- and meso- cosms to attempt to replicated the bacterial community of the Ohio River. The second approach was to determine whether there is a CORE bacterial community in the Ohio River that is persistent, abundant, biologically relevant, AND predictable.

Findings: In all cases, the bacterial community in the artificial environments failed to mimic the Ohio River, either in metabolic potential or in community structure. Thus, these attempts were abandoned. However, my lab determined that the Ohio River does exhibit a Core Community that is persistent, abundant, and biologically relevant. This Core appears to be most directly controlled by temperature (and salinity when encountered) and therefore may also be predictable (study ongoing).

Implications: If we know and can predict the CORE community of bacteria at any moment in a large river system, we can use that information in many different ways: stop hypoxia; stop harmful algal blooms; determine best way to treat a toxic spill; manipulate bacteria to provide optimal habitat; be aware of and potentially remove potential pathogens; provide safer drinking water; and predict effect of climate change.

This work can be expanded to cover other large lotic systems with the hope of forming collaborations with other researchers as well as managers of such systems in order to help preserve the quality and function of these important assets.


Aquavit: A Virtual Collaboration Portal for Water Quality and Watershed Research

Project Objective: Modelling of Toxicity and Biological Impacts

Task Lead: Jack Smith, Marshall University

Problem: Develop a virtual collaboration portal for water quality and watershed research.

Approach: (1) Install an instance of the HUBzero open source web-based platform (from Purdue University) on a publicly accessible server at Marshall University, join the HUBzero Foundation, engage with the Science Gateways Community Institute, and with their assistance develop a sustainable collaboration platform (called Aquavit) for sharing data, building and sharing computational and visualization tools, and providing access to compute resources for all the researchers and collaborators on this project and beyond. (2) Work with and train researchers to develop and share/publish tools, datasets, and otherresources on Aquavit. (3) Work with the EPA WQX/STORET and Purdue University GABBsteams to develop a WQX-compatible data portal as an open source plug-in for the HUBzeroframework to facilitate the collection, analysis, publication, and exchange of water quality datawith the EPA/USGS Water Quality Portal.

Findings: Adoption of Aquavit as a collaboration platform by researchers has been slower than expected, and development of the WQX-compatible data portal has been somewhat delayed due to some (positive) changes in the EPA's strategy for interfacing with the WQX, but the recent promotion by the leadership, improved Single-Sign-On capabilities, and the addition of Jupyter Notebooks and the RStudio IDE to the HUBzero framework for developing and sharing web-based tools has dramatically raised the interest in and usability of Aquavit among researchers.

Implications: Uniform collection and sharing of data, collaborative publication, leveraging of tools and models, participation in the federation of data management for water quality data, and a valuable extension to the HUBzero framework for broader adoption and potential adaptation to other research data.

Aquavit Workshop in Marshall University's Visualization Lab.


Effects of emerging environmental contaminants on the nervous system

Project Objective: Assess changes in neural function and morphology in the presence of contaminants at the cellular and molecular level.

Task Leader: Dr. Nadja Spitzer, Marshall University

Others Involved in the Research: Daniel Kipps and Monica Stanwick

Problem: Environmental contaminants can alter physiological function of cells at exposure levels below those that are cytotoxic, and many of these effects are not well understood. We investigate such physiological changes in brain cells as a result of exposure to silver nanoparticles or manganese. Silver nanoparticles are incorporated in consumer products such as clothing, plastics, and appliances to confer antibacterial properties. They are shed during use, inhaled or ingested, and bioaccumulate in tissues including brain. Manganese levels are elevated in water downstream of soil disturbances and in the air near steel mills and other industry. Children living in these communities exhibit cognitive and motor development defects, and overexposure leads to a Parkinson’s like condition, manganism.

Approach: We use cultured adult neural stem cells as an accessible model system to investigate changes in brain cell physiology when exposed to environmental contaminants. Cultured neural stem cells are induced to differentiate in the presence of contaminants and evaluated using immunocytochemistry, immunoblot, and time lapse microscopy approaches.

Findings: Previously, we found that low levels of silver nanoparticles induced cytoskeletal dysfunction and neurite collapse in differentiating neural stem cells. More recently, we reported a reduction of neurite length and changes in gene expression in cells exposed to manganese.

Implications: Our findings indicate that the physiological function of neural cells is altered at low levels of contaminants. Studies at the cellular level will help understand the mechanisms underlying manganism and neurodevelopmental defects associated with manganese overexposure. Finally, our work may inform regulation and use of these substances.


Applying Geospatial Technologies to Improve the Modeling of Toxicity and Biological Impacts

Project Objective: Modeling of Toxicity and Biological Impacts

Task Leaders: Dr. Michael Strager, West Virginia University, and Dr. Anthony Szwilski, Marshall University

Others Involved in the Research: Pariya Pourmohammadi, Ritika Khurana

Problem: The challenge exists to better characterize and map the distribution and types of land cover to improve the accuracy of modeling toxicity and biological impacts. The Appalachian region is experiencing rapid landscape change due to energy expansion (oil and gas pads, pipelines) as well as agriculture conversion to residential and developed impervious surfaces.

Only with spatially accurate and recently mapped landscape features can we expect to model the receiving stream conditions.

Approach: We are using publicly available National Aerial Image Program data to demonstrate the utility of object based classification using machine learning for accurate mapping of land cover and use classes that are no older than two years and have a minimal mapping unit of 1 meter. Drones are also being used to create high resolution and timely training data for use in classification.

Findings: We have found that object based classification requires server based processing and large amounts of training data to classify the objects in which the machine based classifiers identify.

Implications: We now have the ability to quickly inventory the changing Appalachian landscape specifically due to energy extractive industries to better understand the impacts on water quality and quantity to help guide policies.



Origins, fluxes, fate and transport of DIC from forested, mined and agricultural landscapes

Project Objective: Assess origins, fluxes, fate and transport of dissolved inorganic carbon (DIC) and the generation of alkalinity from forested, mined and agricultural landscapes.

Task Leader: Dr. Dorothy Vesper, West Virginia University

Others Involved in the Research: Jonney Mitchell, Kyle Lee, and Emily Bausher

Problem: The export of carbon – in both dissolved (alkalinity) and gaseous (CO2) form – from Appalachian Watersheds is poorly constrained. These fluxes are critical for understanding carbon mass balances in terrestrial aquatic systems.

Approach: We are conducting field and laboratory measurements to determine fluxes. Field data is being collected from mined, forested and agricultural landscapes. Several abandoned coal mines are sampled ~ monthly; mountain springs are sampled on a quarterly basis. Laboratory activities include developing an improved measurement method for dissolved CO2 and analyzing types of alkalinity present in mine waters.

Findings: Carbon fluxes from forested and agricultural landscapes are primarily in the form of dissolved alkalinity due to the dissolution of carbonate rocks and driven by carbonic acid. The mine waters often have much higher carbon export than the other sites and it can be dominated by either export of dissolved carbon or degassing of CO2. The carbon released from the mine sites is due to rock dissolution driven by sulfuric acid. The CO2 concentrations change over time but the controlling processes are yet unclear.

Implications: (1) The field studies contribute to our understanding of carbon cycling in Appalachian springs and streams and the importance of terrestrial evasion of carbon; (2) the mine water studies document a significant release of geologically bound carbon due to human activities and have implications for the treatment of mine waters using limestone; (3) the laboratory tests provide a means for better measuring CO2 in water and a better understanding of mine water chemistry.

Left: Geology graduate students Kyle Lee and Miles Reed survey the stream of acid mine drainage in preparation for a CO2 flux study. Lee’s research was assisted by NSF-EPSCoR. Right: Volunteers form Indian Creek Watershed Association and WVU geology graduate students collecting data in a creek in Monroe County WV.


Modeling hydrologic intensification under various climate scenarios

Project Objective: Generate water yield, water availability and water quality visualizations tools across watersheds and counties to identify vulnerable and resilient watersheds and communities throughout the region.

Task Leader: Dr. Nicolas Zegre, West Virginia University

Postdoc/Students Involved in the Research: Dr. Luis Andres Guillen, Dr. Brandi Gaertner, and Justin Earle

Problem: Water resources are critical to Appalachian ecosystems, communities, and economies, but climate change, land use/land cover changes, and human water use are impacting water quantity and quality, hence, regional water security.

Approach: We are developing high resolution (daily, 4-km), gridded historic and future climate and water resources datasets for the Appalachian region that are being used to quantify fresh water availability, and community/ecosystem vulnerability to change. Furthermore, we are studying forest ecosystem response to climate change using field based methods and remotely sensed data to quantify ecosystem drivers of change.

Findings: Future climate is expected to become warmer and wetter throughout the region over the 21st century. Importantly, precipitation is expected to increase in the northeastern part of the region, and decrease in the southwestern part of the region, with a transition zone over the central Appalachian region. Potential evapotranspiration is also expected to increase throughout the region at a rate greater than precipitation, increasing the aridity and potentially offsetting increases in precipitation. Furthermore, growing season throughout the region’s forest has increased, on average, by 20 days over the last 30 years, and by as much as 60 days in some parts of West Virginia. Growing season changes are dominated by earlier springs and to a lesser degree by lengthening autumn. Increases in humidity we shown to be a larger driver of growing season length as opposed to warmer air temperatures.

Implications: Changes in precipitation, potential evaporation, and transpiration have important implications of for fresh water security in the region. Despite expected increases in precipitation,

atmospheric demand and forest water use may limit freshwater availability to the regional economy and ecosystems.

Undergraduate students measuring streamflow at the U.S. Forest Service Fernow Experimental Forest in support of growing season change and water balance study .


The Appalachian Freshwater Initiative is supported by the National Science Foundation under Award Number OIA-1458952.

Documents

West Virginia EPSCoR Track I: Appalachian Freshwater ... · Task Lead: Dr. Jason A. Hubbart, West Virginia University Others Involved in the Research: Dr. Elliott Kellner, Dr. Evan