FY2016 Annual Report

Biological Systems Unit
Professor Igor Goryanin


Our research in the Biological Systems Unit focuses on wastewater treatment applications for microbial fuel cells (MFCs). Within MFCs complex interactions between microbial populations result in removal of organics and generation of electricity. We seek to understand and manipulate these interactions so as to improve MFC performance characteristics. For instance, differences in anode community composition, as determined by metagenomic sequencing and analysis, are monitored with the ultimate goal of building idealized communities for treatment of specific substrates. In FY2016 we continued monitoring three pilot scale MFC to our fourth-year collaboration with a local awamori distillery to increase our treatment capacity and establish automatic monitoring system. In addition, the second tubular-style MFC was added to winery wastewater treating MFC system in Santa Rosa, California, USA.  This will allow us to compare the relative effectiveness of MFC type on removal of organics from various agro-industrial wastewaters.

1. Staff

  • Dr. Michael Cohen, Group Leader
  • Dr. Larisa Kiseleva, Staff Scientist
  • Dr. Slava Fedorovich, Staff Scientist
  • Dr. Jovan Rebolledo-Mendez, Staff Scientist
  • Dr. Lukasz Szydlowski, Postdoctoral Scolar
  • Mr. David Simpson, Technical Staff
  • Mr. Geoffry Schaffer-Harris, Technical staff
  • Ms. Koharu Okada, Technical Staff
  • Mr. Nattawet Sriwichai, Research Intern
  • Ms. Nazgul Sakenova, Research Intern
  • Mr. Askarbek Orakov, Research Intern
  • Ms. Shizuka Kuda, Administrative Assistant

2. Collaborations

  • Theme: Biology of Microbial Fuel Cells
    • Type of collaboration: Joint research
    • Researchers:
      • Professor Anatoly Sorokin, Institute of Biophysics, RAS, Pushchino, Russia
      • Professor Mikhail Gelfand, Institute for Information Transmission Problems, RAS, Russia
  • Theme: Improvement of Microbial Fuel Cell performance using nanotechnologies
    • Type of collaboration: Joint research
    • Researchers:
      • Professor Mukhles Sowwan, Nanoparticles by Desighn Unit, OIST
  • Theme: Microbial Fuel Cell treatment of whisky distillery wastewater
    • Type of collaboration: Joint research
    • Researchers:
      • Professor Alan Harper, Heriot Watt University, Edinburgh, UK
  • Theme: Microbial Fuel Cell treatment of winery wastewater
    • Type of collaboration: Joint research
    • Researchers:
      • Prof. Prof. Farid Farahmand, Department of Engineering Science, Sonoma State University, California, USA
  • Theme: Microbial Fuel Cell treatment of starch processing wastewaters
    • Type of collaboration: Joint research
    • Researchers:
      • Prof. Saowalak Kalapanulak, Bioinfromatics and Systems Biology Program, School of Bioresources and Technology, King Mongkut’s University of Technology Thonburi, 10150 Bangkok, Thailand
  • Theme: Microbial Fuel Cell recycling of ethanol fermentation wastewater
    • Type of collaboration: Joint research
    • Researcher:
      • Dr. Bo Chen, Assistant Director of Biotechnology Center, COFCO Nutrition & Health Research Institute Co., Ltd. Beijing, China 

3. Activities and Findings

3.1 Crude oil degradation by Thalassospira HJ

The ability of the previously isolated strain of Thalassospira sp HJ to degrade crude oil with simultaneous electricity generation was evaluated in half-MFC systems inoculated with Marine medium supplemented with 1% or 2.5% crude oil. The microbial culture optical density, voltage and oil residue were observed for pure culture of Thalassospira, multi-species electrogenic culture and the mixture of both during 10 days of incubation under 37oC and 100 rpm. The results demonstrated that Thalassospira HJ biodegraded crude oil in all tested concentrations. However, the tested electrogenic community showed best results in both electrical power production and crude oil degradation.

3.2 Multi-step electrogenic bacteria selection in half-MFC systems

Electrogenic bacteria (or electricigens) are capable of producing electrical power from organic-rich substances such as domestic and food production wastewaters. Electricity producing species may belong to distinct bacterial phyla and to be differently abundant in complex electrogenic communities. Development of methods for selective enrichment and accurate identification of electricigens would facilitate synthetic design of optimal electrogenic microbial consortia for efficient wastewater treatment.

Electrogenic bacteria were selectively enriched by applying 100mV potential to the anode of half-MFC system containing complex electrogenic microbial biofilm as initial inoculum. After a week of operation, the biofilm comprising of selected microorganisms was transferred to the fresh media half-MFC with applied 100mV potential. This step was repeated several times. Whole community shotgun sequencing was applied to track the changes in anodic communities during step-to-step selection process. MG-RAST and Kaiju, the tools that utilize different algorithmic approaches in sequencing data annotation, were used for taxonomic analysis of sampled microbial communities. The results obtained by different bioinformatics tools were similar in terms of microorganism abundance on all taxonomic levels.

Overall, this study provides a snapshot of step-to-step microbial community evolvement in multi-step selection process, and the results provide insight to control electrogenic biofilm development.

3.3 Parallel electrogenic bacteria selection in half-MFC systems

Following the multi-step electrogenic microorganisms enrichment we designed and performed a parallel enrichment experiment, where we observed formation and content of anodic biofilms under -50 mV,  -150 mV, -250mV and -350 mV applied voltage. 

3.4 Continued on-site operations of microbial fuel cell treatment systems

Two MFC-based treatment systems, one in Okinawa and one in California, continued to be monitored for COD removal and power generation across a range of COD loadings.

Mizuhou Awamori Distillery, Naha, Okinawa: Three pilot-scale tray-type MFC bioreactors, each with 50 L working volume and containing 8 electrode pairs, were operated in continuous-flow mode at the Mizuho distillery using rice-wash wastewater as the substrate (Figure 2). 

D’Argenzio Winery, Santa Rosa, California: A second 49-L working volume tubular MFC reactor was added to an operating pilot system at the winery (Figure 2). Effluent from this new MFC cycles back to the mixing tank to help maintain alkalinity and active microbes in the system (Figure 3).


Figure 3. The operational MFC-based system located at Vintners Square in Santa Rosa, CA. Sonoma State University undergraduate research student Gabriel Sacher pictured.

3.5 Biological consilience of hydrogen sulfide and nitric oxide in plants

One of us (M.C.) co-authored with Prof. Hideo Yamasaki of the University of the Ryukyus review article that covered recent progress on H2S studies, highlighting plants and bacteria, and linkages between H2S, NO and O2 biology. Although plant H2S-producing activities have long been known, our knowledge of H2S biology in plant systems has not been updated to the extent of mammalian studies. Hydrogen sulfide is produced in the mammalian body through the enzymatic activities of cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE) and 3-mercaptopyruvate sulfurtransferase (3MST). A growing number of studies have revealed that biogenic H2S produced in tissues is involved in a variety of physiological responses in mammals including vasorelaxation and neurotransmission. It is now evident that mammals utilize H2S to regulate multiple signaling systems, echoing the research history of the gaseous signaling molecules nitric oxide (NO) and carbon monoxide (CO) that had previously only been recognized for their cytotoxicity. In the human diet, meats (mammals, birds and fishes) and vegetables (plants) containing cysteine and other sulfur compounds are the major dietary sources for endogenous production of H2S. Plants are primary producers in ecosystems on the earth and they synthesize organic sulfur compounds through the activity of sulfur assimilation (Figure 4). 

Figure 4. Exogenous and endogenous sources of H2S production in plants. (Published in Nitric Oxide, 2016, 55:91-100).

4. Publications

4.1 Journals

  1. Yamasaki, H., Cohen, MF. Biological consilience of hydrogen sulfide and nitric oxide in plants: Gases of primordial earth linking plant, microbial and animal physiologies. Nitric Oxide, 55-  (2016)

4.2 Books and other one-time publications

  1. Yamasaki, H., Watanabe NS, Sakihama Y, Cohen, MF. (2016) An overview of methods in plant NO research: Why do we always need multiple methods? Methods in molecular biology: Plant nitric oxide research: methods and protocols. KJ Gupta Ed. Humana Press, Totowa, NJ. 1424:1-14

4.3 Oral and Poster Presentations

  1. Cohen MF.   “Nitric oxide-responsive rapid root abscission in the symbiotic water fern Azolla,” The 9th International Conference on the Biology, Chemistry, and Therapeutic Applications of Nitric Oxide, Sendai, Japan. May 22, 2016.
  2. Cohen MF.   “Development of a hybrid anaerobic digestion-microbial fuel cell system for treating winery wastewater” Sustainable Enterprises Conference, Rohnert Park, April 7, 2016.
  3. Alvarez O, Marquez A, Cohen MF, Simpson D, Fedorovich S, Goryanin I, Farahmand F. “MFC Remote Monitoring (MRM) System,” 3rd Meeting of the North American branch of the International Society for Microbial Electrochemistry and Technology. Stanford, CA. October 5-7, 2016.
  4. Sacher G, Monforte F, Sternfeld E, Kozlowski J, Simpson DJW, Goryanin I, Farahmand F, Cohen MF. “Pilot testing of a microbial fuel cell-based system for treating winery wastewater,” 8th Annual Water Resources and Policy Initiatives Conference. Long Beach, CA. April 28, 2016.

5. Intellectual Property Rights and Other Specific Achievements

Nothing to report

6. Meetings and Events

6.1 Microbial electrochemistry opens new doors for sustainable environmental technology

  • Date: February 16, 2017
  • Venue: OIST Campus Lab1
  • Speaker: Professor. Stefano Fregula (The University of Queens)

7. Other

Popular press and outreach

  1. Grizzle N, “Water into Wine (and Back Again),” Sonoma Insights. Fall 2016.
  2. “Utilizing microbial electrical partnerships for treating wastewaters,” Sonoma State Osher Lifelong Learning Science Club, September 29, 2016.