Progress 11/19/15 to 09/30/20
Outputs
Target Audience:This research is ofinterest to two groups of professionals. First, environmental engineers and agricultural scientists who operate anaerobic digesters for waste treatment or energy production will benefit. Stakeholders also include livestock farmers, particularly poultry operations, who could benefit from these findings with regard to the improvement of anaerobic digestion applied to animal wastes and economics of the process. Research from the second major project will be relevant to environmental scientists and coal mining engineers, including the WV Department of Environmental Protection and USGS scientists, who are interested in the Appalachian Mountain region and environmental issues related to surface coal mining. This grant has provided training for graduate students in the MS degree (biotechnology) offered at WVSU, and for visiting international graduate students with specialties in agricultural engineering. This research also contributed to training in environmental microbiology and biotechnology offered by the PD in his undergraduate and graduate classes: Environmental Microbiology (BIOL 460/660), Techniques in Biotechnology II (BT572), and Current Concepts in Biotechnology (BT567). Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This grant supported the research training of WVSU M.S. degree students in biotechnology and a visiting international scientist (Ph.D. student) from Mexico. International research collaborations included one agroindustrial engineering professor from Universidad Autonoma Chapingo, Mexico. Graduate students received training in metagenomics, microbial ecology, environmental chemistry, and anaerobic digestion technology. The PD and his research group also benefited from the collaboration with the agricultural engineer who has expertise in anaerobic digestion. Two presentations based on this research were made at professional conferences (USDA Association of Research Directors and Ecological Society of America) during the final reporting period. In addition, the PD delivered an invited seminar based on this research at Universidad Autonoma Chapingo. This conference was well attended by undergraduate and graduate students at the university. The final year of the grant occurred during the pandemic of 2020 which reduced the amount of WVSU student participation (for example, student researchers) compared to normal years. How have the results been disseminated to communities of interest?During the final time period, three publications were produced, two conference presentations were made, and one invited seminar was delivered at an international university. Presentations at National Conferences: Since 2019, research presentations were made at two conferences. These are: Kemajou Tchamba, Andrielle L., Ifeoma R. Ugwuanyi, Vadesse Lhilhi Noundou, Awoyemi, Olushola D., Flor de Maria Guerrero-Toledo, Jesus E. Chavarria-Palma, Amir Hass, David H. Huber. Distinct functional diversity of river sediment microbiomes in a high-discharge Appalachian river (West Virginia) compared to forest soil in watershed. Ecological Society of American annual conference; August, 2020, Salt Lake City, Utah. Lhilhi Noundou, V., J. E. Chavarria-Palma, I. R. Ugwuanyi, T. Espinosa-Solares, D. H. Huber.Ecological Engineering of Thermophilic Anaerobic Digestion Improves Bioenergy Yield and Reduces Stress Caused by Crude Glycerol Co-Substrate. USDA ARD (Association for Research Directors) Research Symposium, Jacksonville, Fl (2019). Invited Seminar: I was also invited to present a seminar on this research at the Departmento de Ingeniería Agroindustrial, Universidad Autonoma Chapingo, Mexico (Dec, 2019). Title: Applying Ecological Resilience Principles to Anaerobic Digestion. What do you plan to do during the next reporting period to accomplish the goals?Grant is finished.
Impacts
What was accomplished under these goals?
The first objective was to understand microbiome structure and function in a thermophilic anaerobic digester that was operated with feedstock that contained plant biomass. Our experiments were conducted with two sets of replicate ten-liter thermophilic digesters. These digesters received poultry litter as the feedstock; one set was previously used in co-digestion experiments. All of the digesters were operated continuously which required measurements of biogas composition, chemical oxygen demand, ammonia and total volatile acids each week. Individual short-chain fatty acids were also measured weekly with gas chromatography. Using Illumina sequencing, time series analysis of five replicate digesters over a period of 200 days showed that the dominant phyla in this microbiome were Firmicutes (30-55%), Bacteroidetes (20-50%), Proteobacteria (2-10%), Spirochaetes (2-8%), and Thermotogae (1-8%). Order Clostridiales (one of the dominant subgroups of Firmicutes) varied from 5-15% abundance. Based on previous research, Clostridales is expected to be particularly important for the decomposition of complex carbohydrates such as plant biomass. In addition, our recently published research (see reference below) showed that a thermophilic digester stabilized on poultry litter experienced stress when subjected to co-digestion with crude glycerol for two hydraulic retention time periods. This caused a significant change in community structure which caused secondary inhibition when the feedstock was returned to mono-digestion with poultry litter. Secondary inhibition was observed as a dramatic decline in methane and CO2 production. The recovery of methane production did occur but system resilience was slow, requiring eight HRT periods. This research showed that stress caused by certain types of co-digestion (high C/N ratio co-substrates) can impede the basic digester food web, including the initial decomposition processes where hydrolysis and primary fermentation occur. The second objective was to evaluate the recovery of soil microbiome functions in soils on rehabilitated surface coal mining sites in West Virginia. Federal regulations require the restoration of soils on abandoned surface mines. Analysis of eighteen metagenomes, representing triplicate samples at three sites was done. These samples were previously collected from a time-series of mine site soils that had undergone the required restoration process over a period of several decades. The oldest renovated soils that we sampled were 30 years old; the youngest were less than 5 years since restoration. A control site representing undisturbed soil from nearby Kanawha State Forest was used. Microbial functions were classified according to SEED subsystem categories. Hierarchical clustering and multivariate ordination were used for analysis. The diversity of the genes representing the major biogeochemical processes (C, N, S cycles) were compared among the sites. For each of these cycles, the recovery of functions (genes) did occur over time. The most recently damaged soils were the most divergent from the control while the oldest restored soils (30 years since restoration) were the most similar to the control forest soil. The restored soils of intermediate age also mapped at intermediate locations in the ordination plots. Other functional categories were also evaluated for the soil microbiomes. Protein and secondary metabolism, and sporulation/dormancy functions showed a clear distinction between the most recently disturbed soils and the group representing the control and oldest restored soils. This study has shown that the major soil biogeochemical processes were damaged during surface mining but these processes did recover (in terms of gene content) during a 30-year post-restoration period of time. During 2020, one Research Technician assisted with this project full-time and a second assisted part-time. An international collaborator was Dr. Teodoro Espinosa-Solares, Department of Agroindustrial Engineering, Universidad Autonoma Chapingo (UAC), Mexico. A visiting scientist (Dr. Flor de Maria Guerrero-Toledo) from UAC also assisted part-time with data analysis.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2018
Citation:
Huber, David H., Jesus E. Chavarria-Palma, Sridhar A. Malkaram, Natalia Montenegro-Garcia, Vadesse Lhilhi Noundou, Ifeoma R. Ugwuanyi, Teodoro Espinosa-Solares. 2018. Metagenome Sequences of a Thermophilic Anaerobic Digester Adapted to a Low C/N Ratio, High-Ammonia Feedstock (Poultry Litter). Genome Announcements 6(25):e00598-18. (doi.org/10.1128/genomeA.00598-18).
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Huber, David H., Alejandro Ramirez-Garcia, Jesus E. Chavarria-Palma, Teodoro Espinosa-Solares, Vadesse Lhilhi Noundou, Natalia A. Montenegro-Garcia, Akintolami Adeleye, Chandra S. Martin. 2020. Stress induced by crude glycerol in a thermophilic digester: microbial community divergence and resilience, but slow process recovery. Appl. Microb. Biotechnol 104:10769-10781. doi.org/10.1007/s00253-020-10965-6.
- Type:
Book Chapters
Status:
Published
Year Published:
2020
Citation:
Huber, David H. Book chapter: Biogas Technology for Animal Manure Management in USA: State of the Art, Opportunities, Challenges, and Perspectives. In: Biogas Production: From Anaerobic Digestion to a Sustainable Bioenergy Industry. Nagamani Balagurusamy and Anuj K. Chandel (eds.). Springer International Publishing. (Copyright 2020). doi: 10.1007/978-3-030-58827-4.
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