Progress 12/15/18 to 12/14/23
Outputs
Target Audience:Rivers and watersheds are among the most valuable natural resources. Therefore, broadly speaking, all users of river water in the Kanawha River Basin are stakeholders. Issues with the use of freshwater often involve conflicting objectives for stakeholders, particularly involving water for household consumption versus industrial manufacturing. The Kanawha River has been a heavily exploited natural resource in WV for a century. Since WWII the river has been severely damaged by industrial manufacturing, municipal pollution and transportation. However, since the 1980s, the quality of the water has greatly improved. Even so, the river represents another clear example of what has been called the 'urban river syndrome'. The research component of the grant will be useful for environmental and watershed scientists and engineers, environmental microbiologists, industrial engineers, and watershed resource managers. The target audience for the educational component of the grant was undergraduate and graduate students, WV High School science teachers and their students. Teaching supplies and in-person teaching were delivered through the grant. High school science education in West Virginia benefited by the laboratory teaching supplies that were provided for environmental science, chemistry and molecular biology instruction. The grant supported graduate and undergraduate research training in environmental science and biotechnology, and freshwater science at West Virginia State University. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?During the final period of this grant, professional development and training were provided in the PDs (Huber) laboratory for two MS degree students, one undergraduate, a Research Associate scientist, and two laboratory technicians. All of these people learned methods in freshwater analysis, hydrology, environmental chemistry, and environmental (microbial) genomics. The laboratory of Dr. Amir Hass (Co-PD) conducted chemical analysis of river water and sediment. Dr. Fernando Rojano participated in the research with hydrology expertise and assisted with data analysis and writing. Rojano is an environmental engineer employed in WVSU Land Grant Programs. A Research Associate (postdoctoral scientist) worked full-time in the Huber Lab. A Research Technician in the Huber Lab assisted part-time. Another local High School teacher in South Charleston received classroom supplies for teaching microbiology and molecular biology. Two MS degree students assisted with grant research. Four professional conferences (American Geophysical Union, Ecological Society of America, ORBCRE, UCOWR) were attended by the PD or members of his laboratory. How have the results been disseminated to communities of interest?The PD's (Huber) laboratory made fivepresentations at national and regional conferences (Ecological Society of America,UCOWR (Universities Council on Water Resources, USDA ARD). Two manuscripts are currently being written. The conference presentations were: ?? Guerrero-Toledo, F. de M., O.D. Awoyemi, A.L. Kemajou Tchamba, Amir Hass, M. del C. Gonzalez-Rangel, F. Rojano, David H. Huber. Spatial differentiation of stream geochemistry in a subwatershed affected by coal mining in Appalachia. UCOWR Conference, June 2023, Colorado. Guerrero-Toledo, Flor de Maria, Ifeoma R. Ugwuanyi, Andrielle L. Kemajou Tchamba, Amir Hass, David H. Huber. 2023. Does water flow in a large, high-discharge river homogenize microbial diversity and functions in the upper sediment layer? Ecological Society of America annual conference, Portland, Oregon, August 2023. Guerrero-Toledo, Flor, Andrielle L. Kemajou Tchamba, Maria del Carmen Gonzalez-Rangel, Amir Hass, Fernando Rojano, David H. Huber. 2023. Metagenomes show that methane, nitrogen and sulfur biogeochemical cycles are differentially affected by soil and sediment environments. Ecological Society of America annual conference, Portland, Oregon, August 2023. Sailesh Phuyal, Flor de Maria Guerrero-Toledo, Maria del Carmen Gonzalez-Rangel, Bhuparaj Bhattarai, Amir Hass, Fernando Rojano, David H. Huber. Xenobiotic Metabolism in the Kanawha-New River (WV) Watershed: Impacts of Land-Use Practices. USDA ARD (Association for Research Directors) Symposium 2024, Nashville, Tennessee. Guerrero-Toledo, F.M., M.C. Gonzalez-Rangel, B. Bhattarai, S. Phuyal, A. Hass, F. Rojano, D. H. Huber. Linking mountaintop mining to microbial diversity in a severely degraded headwater stream. USDA ARD (Association for Research Directors) Symposium 2024, Nashville, Tennessee. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The objectives of this grant were to build research and teaching capacity in the fields of environmental science, freshwater ecology and environmental (microbial) genomics at West Virginia State University. The research was done in the Kanawha-New River Basin which covers more than 31,000 square km. Kanawha River is the largest river in WV and covers many different land-use types including regions with forests, extensive coal mining, chemical industries and municipal pollution (surface runoff and wastewater effluent). This research sought to understand the role of pollutants and acid mine drainage on microbial ecosystem services provided by the river. During 2022-23, analysis of geographic variation of microbial diversity and chemical heterogeneity of Kanawha River and its tributaries was continued. Our sampling now includes temporal and geographic variation (seven locations) within the main river, as well as fifteen locations representing headwater streams in the watershed. Our focus has been river sediment because of the importance of sediment biodegradation and bioremediation processes for river health and because comparatively little is known about microbial diversity in river sediment anywhere in the world. Sediment microbiome functions were evaluated using metagenome sampling in order to evaluate the spatial distribution of biogeochemical cycle pathways. Methane and sulfur cycle genes were found to statistically vary between locations in the main river in contrast to nitrogen cycle genes which did not show similar spatial variability. Most of the xenobiotic metabolism pathways had similar geographic distributions across 80 km of the riverbed. No significant differences were found across locations or sediment layers using multiple group tests (ANOVA). The most common xenobiotic functions were benzoate, nitrotoluene and chlorocyclohexane/chlorobenzene degradation. Polycyclic aromatic hydrocarbons (PAHs) are common contaminants in rivers and soil associated with petroleum pollution. We investigated whether two common PAHs, naphthalene and phenanthrene, would be metabolized in river sediment. Sediment microcosms were created in sealed anaerobic bottles and amendments were made with these two PAHs. Methane production in the microcosms was measured with gas chromatography. Methane was used as a biosensor for PAH effects (stimulation versus suppression) of the anaerobic carbon cycle. Both naphthalene and phenanthrene were found to stimulate methane production compared to an unamended control. In addition, investigation of an acid mine drainage (AMD) headwater stream was continued. The effects of AMD on stream health involves both the movement of water and transport of sediment. Using Discriminant Analysis and PCA, it was found that spatial variation of water chemistry changed more rapidly than sediment chemistry going from upstream to downstream locations. Iron and sulfate gradients occurred in opposite directions in the water while the concentrations of both decreased in the sediment moving downstream. The most abundant bacterial species were found in families Comamonadaceae, Vicinamibacteraceae, Nitrosomonadaceae, Rhodobacteraceae and Gallionellaceae. Patterns of sediment microbiome diversity paralleled chemical variation. The most distinct microbial diversity in the stream was adjacent to the AMD seep. Resilience of microbial community structure was also found where the most downstream location was most similar to the sediment community upstream of the seep. An educational objective for the grant was to create and teach a new class at WVSU in environmental science. The new class, called Integrated Environmental Science, was taught for the second time during Spring 2023. The class was offered at the advanced undergraduate/ graduate level. The class includes lecture and laboratory components and hands-on research experience in the topics of water quality, environmental chemistry, environmental genomics, hydrology and biogeochemical cycles. The laboratory portion of the class included field measurements of water chemistry in the Kanawha River watershed. Outreach to local High School teachers was also continued. Additional teaching supplies for environmental microbiology instruction were purchased and delivered to local High School teachers. Two MS (biotechnology) graduate students conducted research that was supported by this grant during 2022-23. These research projects involved river sediment bioremediation and biogeochemical processes.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2023
Citation:
The PDs (Huber) laboratory made five presentations at national and regional conferences (American Geophysical Union, Ecological Society of America, ORBCRE (Ohio River Basin Conference), UCOWR (Universities Council on Water Resources). Two additional manuscripts are currently being written. The conference presentations were:
1. Guerrero-Toledo, F. de M., O.D. Awoyemi, A.L. Kemajou Tchamba, Amir Hass, M. del C. Gonzalez-Rangel, F. Rojano, David H. Huber. Spatial differentiation of stream geochemistry in a subwatershed affected by coal mining in Appalachia. UCOWR Conference, June 2023, Colorado.
2. Guerrero-Toledo, Flor de Maria, Ifeoma R. Ugwuanyi, Andrielle L. Kemajou Tchamba, Amir Hass, David H. Huber. 2023. Does water flow in a large, high-discharge river homogenize microbial diversity and functions in the upper sediment layer? Ecological Society of America annual conference, Portland, Oregon, August 2023.
3. Guerrero-Toledo, Flor, Andrielle L. Kemajou Tchamba, Maria del Carmen Gonzalez-Rangel, Amir Hass, Fernando Rojano, David H. Huber. 2023. Metagenomes show that methane, nitrogen and sulfur biogeochemical cycles are differentially affected by soil and sediment environments. Ecological Society of America annual conference, Portland, Oregon, August 2023.
4. Sailesh Phuyal, Flor de Maria Guerrero-Toledo, Maria del Carmen Gonzalez-Rangel, Bhuparaj Bhattarai, Amir Hass, Fernando Rojano, David H. Huber. Xenobiotic Metabolism in the Kanawha-New River (WV) Watershed: Impacts of Land-Use Practices. USDA ARD (Association for Research Directors) Symposium 2024, Nashville, Tennessee.
5. Guerrero-Toledo, F.M., M.C. Gonzalez-Rangel, B. Bhattarai, S. Phuyal, A. Hass, F. Rojano, D. H. Huber. Linking mountaintop mining to microbial diversity in a severely degraded headwater stream. USDA ARD (Association for Research Directors) Symposium 2024, Nashville, Tennessee.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
" Rojano, F., David H. Huber, Ifeoma R. Ugwuanyi, Andrielle Larissa Kemajou Tchamba, Amir Hass. 2022. Rainstorms inducing shifts of hydrochemistry during a winter season in the Central Appalachian Region. Water 14:2687. doi.org/10.3390/w14172687
|
Progress 12/15/21 to 12/14/22
Outputs
Target Audience:The target audience for the research component of the grant is environmental and watershed scientists and engineers, environmental microbiologists, industrial engineers, and watershed resource managers. The target audience for the educational component is undergraduate and graduate students, WV High School science students, and local public school science teachers. High school science education in West Virginia will benefit through teaching provided in environmental science provided by the Co-PDs as well as the provision of supplies for science classes. The grant will support graduate and undergraduate research training in environmental science and biotechnology at WVSU. Changes/Problems:Access to WV public schools was still restricted through the first half of 2022 due to covid, but the public school component of the grant was resumed Fall 2022. WVSU student travel during the first half of 2022 was also restricted; college students were not allowed to attend the USDA ARD conference in Atlanta in April 2022. What opportunities for training and professional development has the project provided?Two MS degree students (biotechnology) began work in the Huber Lab during 2022. The laboratory of Co-PD Amir Hass performed chemical analysis of river water and sediment. Dr. Fernando Rojano participated in the research as a hydrologist, and assisted with data analysis and writing. Rojano is an environmental engineer located at WVSU. A Research Associate (postdoctoral scientist) worked full-time in the Huber Lab and participated in analysis of microbial diversity and chemistry data. A Research Technician in the Huber Lab assisted part-time. Three professional conferences (American Society for Microbiology, International Symposium for Microbial Ecology, and USDA ARD) were attended by the PD and members of his laboratory. How have the results been disseminated to communities of interest?The PD's (Huber) laboratory presented research at five national, regional and international conferences: ORBCRE (Ohio River Basin Conference), American Society for Microbiology, USDA ARD, and International Symposium for Microbial Ecology (ISME 18). Two additional manuscripts are currently being written. The conference presentations were: Fernando Rojano, Amir Hass, Robert Cantrell, David H. Huber. 2021. Long- and short-term datasets to investigate temporal variability of soil moisture of disturbed land in West Virginia. American Geophysical Union annual meeting, New Orleans, LA. Saetz-Garcia, D. R., Fernando Rojano, Flor Guerrero, Maria Gonzalez, David Huber. Potential methane production from the Kanawha River sediments in Charleston, WV. ORBCRE (Ohio River Basin Conference), October 2022, Youngstown, Ohio. Guerrero-Toledo, F. M., Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Amir Hass, Fernando Rojano-Aguilar, David H. Huber. Microbial Foundations for Riverine Ecosystem Services and Water Quality in a Large, Highly-Impacted Appalachian River, West Virginia. USDA Association for Research Directors (ARD) Conference, April 2-5, 2022, Atlanta, Georgia. Guerrero-Toledo, F.M., Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola D. Awoyemi, Maria Carmen Gonzalez-Rangel, Amir Hass, David H. Huber. Effects of Environmental Disturbances on Dynamics of Sediment Microbial Community and Geochemistry During Two Years in a High-discharge Appalachian River (WV). American Society for Microbiology Annual Meeting, World Microbe Forum, June 9-13, 2022, Washington DC. Guerrero-Toledo, F.M., Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola D. Awoyemi, Maria Carmen Gonzalez-Rangel, Amir Hass, David H. Huber. Disturbance and ecological succession in the sediment microbiome of a large Appalachian River (WV, USA) with periodic high-discharge events. International Symposium for Microbial Ecology (ISME 18), Lausanne, Switzerland, August 14-19, 2022. What do you plan to do during the next reporting period to accomplish the goals?Experiments to measure the bioremediation potential of the sediment microbiome in Kanawha River will be continued. These will test several common pollutants including agricultural fertilizers and polycyclic aromatic hydrocarbons. Methane production in sediment will be used as a proxy for the effects of chemical pollutants on the biogeochemical carbon cycle. Sediment microcosms will be created in sealed anoxic bottles where gas composition can be measured with gas chromatography. Chemical measurements of the water from the Cabin Creek watershed which has acid mine drainage damage will be continued. We are building a multi-year database for temporal and spatial variation in this highly degrade subcatchment. The WV Department of Environmental Protection began remediation of this location in Fall 2021. This provides an excellent opportunity to measure the effectiveness of standard methods for the restoration of headwater streams damaged by coal mining. This further enhances the value and impact of this grant. In addition, data analysis of sediment chemical and microbial diversity from Kanawha River and Cabin Creek will continue and manuscript writing will be done. The Co-PDs will attend several professional conferences (American Society for Microbiology, USDA ARD) during the coming year. Additional outreach to local public schools in West Virginia will be conducted. We will contact High School science teachers to assist with purchasing science teaching supplies. Two graduate students will begin their MS degree research in the Huber lab in August 2022.
Impacts
What was accomplished under these goals?
Broadly speaking, this grant concerns the drivers of water quality (chemistry), ecosystem services, and riverine microbiome functions in a major Appalachian river that has been severely impacted for a hundred years by many different anthropogenic processes and land-use changes. During 2022 we continued a study that concerned the effects of rainstorms on the water quality of Kanawha River. The Kanawha-New River Basin is a large watershed of about 31,000 square km. Rainstorms across the watershed can disrupt water chemistry although the specific chemical changes and the dynamic responses to river discharge levels have not been known. Data collected during two previous years was analyzed and published in the MDPI journal, Water. Water chemistry was measured with multiprobe sondes deployed at two locations which provided high frequency data as well as grab samples. The sondes collected hourly measurements of temperature, pH, turbidity, and nitrate and chloride ions. The dynamic responses in the concentrations of anions and cations during changes in water discharge were calculated. Piper diagram analysis showed that the water type was calcium-chloride. Using Gibbs diagram analysis, the origin of the dissolved solids (anions and cations) was found to be primarily due to rock weathering. Soil moisture was found to be very important for determining the water chemistry dynamics of HCO3 and NO3. This work was done in collaboration with Dr. Fernando Rojano who is an environmental engineer at WVSU. In addition, water chemistry measurements were continued in the Cabin Creek watershed which has been severely damaged by acid mine drainage. Field measurements were taken with a portable YSI meter. These measurements contributed to a time series dataset and analysis of this subcatchment that spans the time before and after remediation. Association network analysis was continued to understand how the river microbiome responds to sediment turnover. It was found that community structure and the network associations within the sediment microbiome changed over time, implying that secondary succession occurs in the sediment microbiome following disturbances.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2022
Citation:
The PDs (Huber) laboratory presented research at five national, regional and international conferences: ORBCRE (Ohio River Basin Conference), American Society for Microbiology, USDA ARD, and International Symposium for Microbial Ecology (ISME 18). Two additional manuscripts are currently being written. The conference presentations were:
1. Fernando Rojano, Amir Hass, Robert Cantrell, David H. Huber. 2021. Long- and short-term datasets to investigate temporal variability of soil moisture of disturbed land in West Virginia. American Geophysical Union annual meeting, New Orleans, LA.
2. Saetz-Garcia, D. R., Fernando Rojano, Flor Guerrero, Maria Gonzalez, David Huber. Potential methane production from the Kanawha River sediments in Charleston, WV. ORBCRE (Ohio River Basin Conference), October 2022, Youngstown, Ohio.
3. Guerrero-Toledo, F. M., Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Amir Hass, Fernando Rojano-Aguilar, David H. Huber. Microbial Foundations for Riverine Ecosystem Services and Water Quality in a Large, Highly-Impacted Appalachian River, West Virginia. USDA Association for Research Directors (ARD) Conference, April 2-5, 2022, Atlanta, Georgia.
4. Guerrero-Toledo, F.M., Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola D. Awoyemi, Maria Carmen Gonzalez-Rangel, Amir Hass, David H. Huber. Effects of Environmental Disturbances on Dynamics of Sediment Microbial Community and Geochemistry During Two Years in a High-discharge Appalachian River (WV). American Society for Microbiology Annual Meeting, World Microbe Forum, June 9-13, 2022, Washington DC.
5. Guerrero-Toledo, F.M., Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola D. Awoyemi, Maria Carmen Gonzalez-Rangel, Amir Hass, David H. Huber. Disturbance and ecological succession in the sediment microbiome of a large Appalachian River (WV, USA) with periodic high-discharge events. International Symposium for Microbial Ecology (ISME 18), Lausanne, Switzerland, August 14-19, 2022.
- Type:
Journal Articles
Status:
Published
Year Published:
2022
Citation:
" Rojano, F., David H. Huber, Ifeoma R. Ugwuanyi, Andrielle Larissa Kemajou Tchamba, Amir Hass. 2022. Rainstorms inducing shifts of hydrochemistry during a winter season in the Central Appalachian Region. Water 14:2687. doi.org/10.3390/w14172687
|
Progress 12/15/20 to 12/14/21
Outputs
Target Audience:Two former MS degree students from the PDs (Huber) laboratory were hired for several months to continue research with the Kanawha River watershed. The laboratory of Co-PD Amir Hass conducted chemical analysis of river water and sediment. Dr. Fernando Rojano participated in the research as a hydrologist, and assisted with data analysis and writing. Rojano is an environmental engineer located at WVSU. A Research Associate (postdoctoral scientist) worked full-time in the Huber Lab. Another Research Technician in the Huber Lab assisted part-time. A local teacher from Nitro High School participated in research during summer 2021. An MS degree graduate student began a research project during Spring 2022. Changes/Problems:The delivery of the hands-on science educational component to WV public schools was further delayed Fall semester 2021 because of the new surge in covid cases in WV (and nationally). In-person delivery of environmental science teaching to WV High School teachers and students was still not possible through the end of 2021. However, the covid cases peaked in December and have been declining in the early months of 2022 (although WV has been recovering more slowly than nearby states). Travel was also restricted. For example, WVSU Administration did not allow university students to attend in-person the USDA ARD conference in Atlanta in April 2022. However, the WV public school educational objective in the grant will resume Fall 2022. What opportunities for training and professional development has the project provided?Two former MS degree students from the PDs (Huber) laboratory were hired for several months to continue research with the Kanawha River watershed. The laboratory of Co-PD Amir Hass conducted chemical analysis of river water and sediment. Dr. Fernando Rojano participated in the research as a hydrologist, and assisted with data analysis and writing. Rojano is an environmental engineer located at WVSU. A Research Associate (postdoctoral scientist) worked full-time in the Huber Lab. Another Research Technician in the Huber Lab assisted part-time. A local teacher from Nitro High School participated in research during summer 2021. An MS degree graduate student began a research project during Spring 2022. Two professional conferences (American Society for Microbiology, Ecological Society of America) were attended by the PD and members of his laboratory. How have the results been disseminated to communities of interest?The PD's (Huber) laboratory made five presentations at national conferences (American Society for Microbiology, Ecological Society of America). The presentations were: Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Flor de M. Guerrero-Toledo, Amir Hass, David H. Huber. Distribution and Diversity of Phylum Desulfobacterota in the Sediment of a Large, Urbanized Appalachian River. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021 Olushola D. Awoyemi , Andrielle Larissa Kemajou Tchamba, , Flor de M. Guerrero-Toledo, Amir Hass, David H. Huber. The Impact of Acid Mine Drainage as a Heterogeneous Stressor on Chemical Gradients and Microbial Community Structure in an Appalachian Headwater Stream in WV. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021 Flor de Maria Guerrero-Toledo, Ifeoma R. Ugwuanyi, Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Amir Hass, David H. Huber. Unexpected Microbial Diversity Associated with Flow Regime Changes in a High-Discharge Appalachian River, WV. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021 Flor de Maria Guerrero-Toledo, Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola Deborah Awoyemi, Amir Hass, David H. Huber. 2021. Impact of urbanization and industrialization on Kanawha River (West Virginia): Network analysis reveals habitat differentiation. Ecological Society of America annual conference, August 2021. Andrielle Larissa Kemajou Tchamba, Flor de Maria Guerrero-Toledo, Jesus E. Chavarria-Palma, Olushola D. Awoyemi, Ifeoma R. Ugwuanyi, Amir Hass, David H. Huber. Spatial distribution of Bacteroidota, Myxococcota, Spirochaetota and Verrucomicrobiota in sediment of the urbanized Kanawha River (West Virginia). Ecological Society of America annual conference, August 2021. What do you plan to do during the next reporting period to accomplish the goals?During the next year, additional outreach to local public schools will be conducted. We will continue to contact local teachers in order to assist them by purchasing science teaching supplies. We will broaden the participation to include Junior High and Elementary Schools as well. We will also deliver teaching modules on environmental science and microbiology through in-person instruction. Public schools in WV are now open to having visitors in the classrooms. Additional High School science teachers will work in the PDs (Huber) laboratory. One or two graduate students will begin their MS degree work in the Huber lab in August 2022. Chemical sampling of the Cabin Creek watershed which has the acid mine drainage site and extensive coal mining will continue in order to incorporate temporal as well as spatial variation in this important location. The WV Department of Environmental Protection began remediation of this location during Fall 2021. This is an exciting development because measurements of Cabin Creek can now be used for comparisons of river quality both before and after remediation. This further enhances the value and impact of our study which will be able to directly analyze the effectiveness of traditional watershed environmental restoration methods. Our sediment microcosm experiment is not completed yet and will be continued. In addition, data analysis of sediment chemical and microbial diversity will continue and manuscript writing will be done during 2022. The new GC/MS system that was purchased with this grant will be used for measuring organic pollutants in Kanawha River. The Co-PDs will attend several professional conferences (American Society for Microbiology, International Society for Microbial Ecology [ISME], USDA ARD, and possibly others) during the coming year.
Impacts
What was accomplished under these goals?
The objectives of this grant are to build both research and teaching capacity in the fields of freshwater and molecular environmental science at West Virginia State University. The research is focused on the Kanawha River watershed. Kanawha River is the largest river in WV, traversing regions with forests, coal mining, chemical industries and municipal pollution. We are seeking to understand the role of landscape variation and regional stressors on microbial diversity and ecosystem services provided by the river. During 2021, analysis of geographic variation of microbial diversity and chemical heterogeneity of Kanawha River and its tributaries was continued. Our sampling now includes temporal and geographic variation (seven locations) within the main river, as well as fifteen locations within the watershed. Our focus is river sediment because of the importance of sediment biodegradation processes for river health and because little is known about microbial diversity in river sediment anywhere in the world. We found spatial patterns and temporal variation in sediment microbial diversity. Spatial patterns have been identified at the levels of phylum and class for bacteria and archaea. For example, Bacteroidota and Verrucomicrobiota exhibited a wider geographic distribution while Myxococcota and Spirochaetota showed a more localized distribution. The abundance and diversity of bacterial and archaeal phyla also varied between sediment layers. Geographic and temporal variation in sediment chemistry was found and associated with spatial variation of microbial diversity. These patterns are suggestive of phylogenetic niche conservatism which has not previously been shown in river sediment microbiomes. Overall chemical profiles correlated with microbial diversity patterns but no single chemical variable was most important. For example, zinc which is a chemical proxy for urbanization, was positively correlated with Spirochaetota but negatively with Bacteroidota. A sediment microcosm experiment was begun to measure variations in biogeochemical processes in sediment from different regions of the watershed that have different chemical profiles. In order to understand the structure and dynamics of the sediment microbiome, association network analysis was conducted for six river locations. Spearman correlations of Illumina sequence data were used for network analysis. We found that networks can be used to gain insight into the environmental factors that control microbiome structure. This analysis is ongoing. A second research objective was to continue the analysis of a tributary stream (sub-watershed) called Cabin Creek. This stream has been damaged by Acid Mine Drainage and coal mining for decades and flows into Kanawha River. Our analysis continued to measure changes in water chemistry over time as well as including a nearby stream in the same drainage area as a control. We measured water and sediment chemistry in both streams. Linear Discriminant Analysis was found to be a good method for distinguishing subtle differences in water chemistry between locations in a single stream. The impact of AMD was evident in the water and sediment of Cabin Creek for more than 3 km downstream. Water upstream of the AMD seep was also found to be impacted by regional coal mining. Correlations of variation in microbial diversity with sediment chemistry were continued with additional sampling using sequencing of 16S rRNA genes. Sediment and water chemistry were measured with ICP-OES (inductively coupled plasma spectroscopy) and ion chromatography. Field measurements of water chemistry were made with a portable YSI meter. ICP and ion chromatography measurements were made in the lab of Co-PD Amir Hass. Microbial diversity sampling was done with Illumina sequencing and bioinformatics analysis was done in the PD's (Huber) laboratory. When completed, our analysis of this sub-watershed will make a valuable contribution to environmental monitoring in the Appalachian region. A major educational objective for the grant was to create and teach a new class at WVSU in environmental science. Preparation for the class was done during Fall 2021, and the new class, titled Integrated Environmental Science, is being taught by the PD for the first time during Spring semester 2022. The class is offered at the advanced undergraduate/graduate level. The class includes research experience for students regarding water quality, environmental chemistry and biogeochemical cycles in the Kanawha River watershed. A WV High School teacher worked in the PDs laboratory during summer 2021 on a watershed environmental science project in conjunction with this research. Teaching supplies for environmental science were purchased and delivered to local High School teachers. One MS graduate student began a research project involving river sediment biogeochemical processes.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
1. Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Flor de M. Guerrero-Toledo, Amir Hass, David H. Huber. Distribution and Diversity of Phylum Desulfobacterota in the Sediment of a Large, Urbanized Appalachian River. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021
2. Olushola D. Awoyemi , Andrielle Larissa Kemajou Tchamba, , Flor de M. Guerrero-Toledo, Amir Hass, David H. Huber. The Impact of Acid Mine Drainage as a Heterogeneous Stressor on Chemical Gradients and Microbial Community Structure in an Appalachian Headwater Stream in WV. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021
3. Flor de Maria Guerrero-Toledo, Ifeoma R. Ugwuanyi, Andrielle Larissa Kemajou Tchamba, Olushola D. Awoyemi, Amir Hass, David H. Huber. Unexpected Microbial Diversity Associated with Flow Regime Changes in a High-Discharge Appalachian River, WV. American Society for Microbiology Annual Meeting, World Microbe Forum, June 2021
4. Flor de Maria Guerrero-Toledo, Andrielle Larissa Kemajou Tchamba, Ifeoma R. Ugwuanyi, Olushola Deborah Awoyemi, Amir Hass, David H. Huber. 2021. Impact of urbanization and industrialization on Kanawha River (West Virginia): Network analysis reveals habitat differentiation. Ecological Society of America annual conference, August 2021.
5. Andrielle Larissa Kemajou Tchamba, Flor de Maria Guerrero-Toledo, Jesus E. Chavarria-Palma, Olushola D. Awoyemi, Ifeoma R. Ugwuanyi, Amir Hass, David H. Huber. Spatial distribution of Bacteroidota, Myxococcota, Spirochaetota and Verrucomicrobiota in sediment of the urbanized Kanawha River (West Virginia). Ecological Society of America annual conference, August 2021.
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Progress 12/15/19 to 12/14/20
Outputs
Target Audience:The target audience for the educational component of the grant is undergraduate and graduate students, WV High School science students, and local public school teachers. High school science education in West Virginia will benefit through teaching provided in environmental science provided by the Co-PDs as well as the provision of supplies for science classes. The grant will support graduate and undergraduate research training in environmental science and biotechnology at WVSU. The target audience for the research component is environmental and watershed scientists, environmental microbiologists, industrial engineers and watershed resource managers. Changes/Problems:Delivery of the hands-on science educational component to public schools has been delayed by one year due to the 2020-21 pandemic. In-person teaching of local High School teachers and students will begin summer 2021. What opportunities for training and professional development has the project provided?Undergraduate students have been conducting research on nitrogen cycle gene diversity and antibiotic resistant bacteria in the Kanawha River watershed. The undergraduate research projects have helped us to define appropriate microbiology projects that we will use for environmental science education with WV High School students. During year two, one MS degree student worked on grant objectives and received training in microbial genomics and environmental chemistry. The professional development of one full-time technician in the areas of environmental chemistry and statistics applications for data analysis was also accomplished. The graduate student participated in a national conference (American Geophysical Union). How have the results been disseminated to communities of interest?One presentation was made by a graduate student at a national conference (American Geophysical Union, 2020), and one presentation was made by an undergraduate student at a local science conference. The presentations were: Awoyemi, Olushola D., Andrielle L. Kemajou Tchamba, Amir Hass, Flor de Maria Guerrero-Toledo, Fernando Rojano, David H. Huber.American Geophysical Union. December 2020. Surprisingly high diversity of the cosmopolitan, metabolically diverse bacterial family Comamonadaceae in an Appalachian (WV) headwater stream impacted by Acid Mine Drainage. Chinyere Elizabeth Ugwuanyi, Olushola D. Awoyemi, Andrielle Larissa Kemajou Tchamba, David H. Huber. Detecting genes from the nitrogen biogeochemical cycle in Kanawha river sediment. Undergraduate Research Day at the Capital, West Virginia State Capital, Charleston. (Feb, 2020) What do you plan to do during the next reporting period to accomplish the goals?Educational outreach will be expanded during 2021. We expect that local public schools will have (near) normal operations with regard to the problems caused by the pandemic by Fall semester 2021. West Virginia has been recovering fairly quickly during the winter-spring season. This will enable us to deliver teaching modules on environmental and aquatic science, and microbiology to science students. We much prefer in-person science education, but if this is still limited by school regulations in the Fall semester, we will convert the teaching modules to online instruction. We further expect to have High School science teachers working in the Huber Lab during summer 2021. A new Integrated Environmental Science class will be offered at WVSU. One or two new graduate students will be trained with the grant. Additional field sampling will be done of the Cabin Creek tributary which has been damaged by acid mine drainage in order to clarify the relationships between chemical gradients and microbial ecology. Additional microbial and chemical sampling will also be done in the mainstem (Kanawha) river for regions with unusual microbial diversity in order to understand specific landscape drivers of diversity patterns.
Impacts
What was accomplished under these goals?
The objectives of this grant are to build both research and teaching capacity in the fields of freshwater and molecular environmental science at West Virginia State University. During 2020, analysis of the microbial diversity and chemical heterogeneity of Kanawha River and its tributaries was continued. The first research objective was to measure the spatial variability of microbial diversity and chemical gradients in an Acid Mine Drainage (AMD) headwaters stream. This work is being done in the Cabin Creek watershed which is a tributary of Kanawha River, West Virginia. Kanawha River is the primary industrialized river in WV and its watershed provides both drinking water and industrial water for the region. Cabin Creek, a tributary of Kanawha River, has been damaged by acid mine drainage for many years. During 2020, extensive sampling of both chemical and microbial variability was done in this watershed. Water and sediment chemistry were measured with ICP-OES (spectroscopy), ion chromatography, and field measurements were made with a portable YSI meter. The ICP and ion chromatography measurements were made in the laboratory of Co-PD Amir Hass. Microbial diversity sampling was done with Illumina sequencing. Cabin Creek was sampled with respect to microbial diversity and relevant chemical variables (metals and freshwater ions). Sediment samples were collected in locations that represent an AMD-impact gradient and extended from upstream of the seep to more than 20 km downstream. This stream showed bacterial diversity gradients that paralleled the chemical gradients created by the AMD seep. Upstream of the seep the dominant bacteria were Anaerolineaceae, Terrimonas, and Vicinamibacteriaceae. Dominant genera at the seep were Sideroxydans and Gallionella which are iron oxidizers. Downstream of the seep the dominants were Nitrosomonadaceae, Pirellula, Rokubacteriales, and Polaromonas. In addition, microbial and chemical sampling of Kanawha River was continued to investigate spatial variation of urbanization effects. The diversity of sediment was found to vary between regions of the main river with a high-flow regime compared to those with low flow. The main river was dominated by Proteobacteria while a low-flow area was dominated by Chloroflexi and Crenarchaeota. Unexpectedly, this region of the river area has substantially different diversity, including many unnamed and uncharacterized major groups of bacteria. The most abundant orders were Bathyarchaeia, GIF9, Spirochaetales, MSBL9, Methanomicrobiales, and Burkholderiales. Bacterial diversity within the intensely urbanized region of Kanawha River was also analyzed and correlation analysis was done to relate diversity to environmental chemistry. Correlation analysis with 20 chemical variables was done. Phylum Desulfobacterota was abundant here (3-18%) and diverse. Of 18 known classes in this phylum, 11 were found in the urban area. Although many representatives of the classes are known to be sulfate reducers, only Desulfobulbaceae was positively correlated with sulfate. In contrast, at the family level, the majority of positive and negative correlations were due to Zn, Al, Br, Cl, Na, Mg, NH4 and NO3 which are representative of urbanization. Educational outreach was done to a local high school. An online presentation was made to Calvary High School Academy by Huber and Fultz where they presented an overview of their research. The feedback from the students and teacher was very positive and encouraging. An MS degree student who was supported by this grant completed her research and graduated in November, 2020.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
1. Awoyemi, Olushola D., Andrielle L. Kemajou Tchamba, Amir Hass, Flor de Maria Guerrero-Toledo, Fernando Rojano, David H. Huber. American Geophysical Union. December 2020. Surprisingly high diversity of the cosmopolitan, metabolically diverse bacterial family Comamonadaceae in an Appalachian (WV) headwater stream impacted by Acid Mine Drainage.
- Type:
Other
Status:
Accepted
Year Published:
2020
Citation:
2. Chinyere Elizabeth Ugwuanyi, Olushola D. Awoyemi, Andrielle Larissa Kemajou Tchamba, David H. Huber. Detecting genes from the nitrogen biogeochemical cycle in Kanawha river sediment. Undergraduate Research Day at the Capital, West Virginia State Capital, Charleston. (Feb, 2020)
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Progress 12/15/18 to 12/14/19
Outputs
Target Audience:The target audience is undergraduate and graduate university students and high school science students, academic environmental scientists, industrial engineers and environmental resource managers. High school science education in West Virginia will benefit through teaching in environmental science and the provision of supplies for science classes. The grant will support graduate and undergraduate research training in environmental science and biotechnology at WVSU. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two undergraduate students have been conducting research on identifying antibiotic resistant bacteria in the Kanawha River. This project is helping us to define appropriate microbiology projects that we will use for environmental science education with WV high school student. At this time one MS degree student is working on grant objectives. How have the results been disseminated to communities of interest?No dissemination yet; research is still in the early stages. Conference presentations will be made next year. What do you plan to do during the next reporting period to accomplish the goals?The mass spectrometer that is funded by the grant will be purchased in 2020. The functional diversity of the microbiome in the AMD-affected river will be measured using metagenomics. High school teachers and students will be engaged in laboratory-based training in molecular environmental science and microbiology. Protocols will be developed for using the new mass spectrometer to measure the breakdown of organic pollutants in the laboratory microcosms. The development of the curriculum for the new class, Molecular Environmental Science, will be done. The research will be used to train two MS degree students during 2020.
Impacts
What was accomplished under these goals?
This is a progress report for an 1890 Research Teaching Capacity Building Grant. The objectives of this grant are to build both research and teaching capacity in the fields of freshwater and molecular environmental science at WVSU. Our first research objective is to investigate the microbial diversity and chemical variability found in an Acid Mine Drainage (AMD) stream that occurs in the watershed of Kanawha River, West Virginia. During 2019, we began the analysis of Fifteenmile Fork and Cabin Creek rivers which form a tributary that enters Kanawha River. Fifteenmile Fork is directly impacted by an AMD seep (pH3.8). This region of West Virginia has considerable surface mining which affects the watershed, particularly the headwaters streams that originate here. Our initial sampling of Fifteenmile fork using Illumina sequencing of 16S rRNA gene diversity has shown that microbial diversity is quite different from the mainstem (Kanawha) river. For example, the dominant bacteria near the AMD seep are iron-oxidizers, Gallionella and Betaproteobacteria SBla14. We also designed a flow-through sediment microcosm (bioreactor) that can be used to measure the breakdown of chemical pollutants by microorganisms.
Publications
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