During the REU you will be assigned to work with a faculty mentor on your research project.

The upper Clark Fork (UCF) river has its headwaters in Butte, Montana, which is home to the largest Superfund site in the United States. A century of intensive hard rock mining activities in numerous tributaries throughout the watershed has resulted in extant pollution of arsenic, lead, copper, zinc, and cadmium in surface waters and sediments in the UCF despite ongoing remediation efforts. Sources of metals to the UCF include aquifers in former mining areas that emerge as springs and seeps that flow into UCF tributaries. Microbes affect metal concentration and speciation in fluids and sediments of these springs, tributaries, and the UCF by a variety of mechanisms, including detoxification and metabolic processes. These processes can lead to major changes in the composition of both sediments and overlying fluids. The overarching goal of this research is to determine the role of microbes in controlling the transport, speciation, and overall flux of metals in mine impacted waters in and around Butte to inform restoration efforts in the broader UCF watershed. Methods include geochemical and microbial community characterization, microcosm experiments, laboratory-based microbiology experiments, and geochemical analysis. Preliminary results have shown that iron and sulfate metabolism, which are associated with numerous covariant metal transformations, are particularly prevalent in these systems. Undergraduate participation is appropriate and encouraged in all aspects of this project. The interdisciplinary project allows students to contribute to the project by focusing on either microbiology, geochemistry, or analytical chemistry, depending on their interests and career goals. Students will be guided as they design a hypothesis, experiments, and methodology to evaluate the magnitude and environmental consequences of a process of their choosing.

Climate change is predicted to raise temperatures, affect rainfall patterns, and increase the number of extreme weather events. Of particular global concern is how these environmental changes will affect the incidence and distribution of infectious disease particularly zoonotic diseases (diseases that are transmitted from animals to humans). Although the potential impacts of environmental change on vector-borne zoonotic diseases have been highly studied, directly-transmitted zoonotic pathogens have been neglected, in particular in regards to how weather affects population dynamics and physiological stress of reservoir hosts. There is evidence that Sin Nombre hantavirus (SNV, reservoired in the deer mouse) is affected by environmental factors (temperature, precipitation, food supply), but the interactions between these factors, reservoir host density, virus transmission, and infection prevalence are not fully understood.

Our group has been studying SNV in deer mice since 1999. We have monitored deer mice populations in sylvan and peridomestic settings, including duplicate 1-ha grids at field sites across Montana including a site approximately 15 miles from campus that has been impacted by mining. The close proximity to Montana Tech makes it logistically easy to involve undergraduates in field sampling. Students will be assisted in developing hypotheses related to the project and learn methodology appropriate for their own project. The various field aspect of the project including gathering demographic information on captured rodents, collecting samples for performing stress assays, PAGEIA ELISAs, determining parasite loads, and blood smears to measure immune response. The long term dataset associated with this project will also allow students opportunities for comparing their data with past data as well as opportunities for statistical analysis and data exploration.

An ongoing project in the Laboratory Exploring Geobiochemical Engineering and Natural Dynamics (LEGEND) is producing the first view of microbial diversity and activity in headwaters of the Clark Fork, providing both an indication of metal contamination from past mining on the overall health of the system and serving as a baseline for evaluating the effects of future climate change on microbial and chemical processes in this ecosystem (Cox et al., unpublished). It links microbial activity to geochemistry, informing how the microbial activity relates to water quality at this vital location on the Clark Fork watershed. Little information is available on microbial activity and how it may be affected by and influencing the system, although interactions between water chemistry and insects and vertebrates have been investigated and modeled (Nimick et al., 2007; Luoma et al., 2009; Balistrieri, 2012). The ongoing research is addressing the following basic questions: What is the baseline microbial community and activity in the headwaters of the Clark Fork? How does the community relate to the geochemistry? What is the level of metal contamination reached in these headwaters and does the microbial community reflect that? How will microbial activity change with the climate (lower water flow, higher CO2 available for photosynthesis)? The microbial community and activity are expected to correlate with the geochemistry and reflect the level of metal concentrations in these waters. These results are expected to contribute to water quality and remediation solutions both now and in the future. Undergraduates working on this project will learn how to sample and perform field experiments elucidating specific questions about the presence of sulfate reduction and the cycling of metals.

For over a century, the Clark Fork River and its headwaters were contaminated with sediments rich in copper, arsenic, cadmium, lead, and other metals derived from the mining activities that took place in the Butte area. Today the Clark fork River has been subject to a major cleanup effort that impacts approximately 120 river miles. Our group has been involved in the riparian and in-stream habitat monitoring of the completed phases. This work has led to a good understanding of how plant communities recover after the cleanup and restoration. Ecological restoration also helps the recovery of soil microbial communities; however, we have limited knowledge about that recovery process. Based on that the proposed research focuses on finding answers to the following questions: 1.) How is cleanup and restoration along the Clark Fork River helping/changing the soil microbial communities compared to control reaches? 2.) How are microbial communities change with time? 3.) How are microbial communities changing with changing soil conditions? 4.) What is the relationship between cover types and microbial communities?

Soil microbial comminutes will be evaluated by molecular techniques for functional groups and taxonomical characteristics. We would like to assess the targeted use of beneficial soil microbes in the area in the frame of a controlled greenhouse experiment. The use of beneficial soil microbes (bacteria and fungi) is a novel approach in ecological restoration, that can help vascular plants to increase nutrient availability and they can also defend them from pathogens, which would result in healthier individuals of plants and healthier functioning plant communities. The experiments will be conducted with the involvement of undergraduate and graduate students, who are involved in the Ecological Restoration Certificate or the Ecological Restoration MS Program at Montana Tech, directed by Dr. Pal. The students will perform all activities in the frame of their field practicum and capstone projects. Graduate students will prepare their theses from the proposed research. Several students are expected to find internships and employment in the area as over 30 miles of the river still require remediation and restoration. That way there is a straightforward pathway for the students to be involved in future research along the Clark fork River.