Microbial fuel cell (MFC): A potential system to harness bioelectricity from wastewater treatment

S Veer Raghavulu

Bioengineering and Environmental Center (BEEC) Indian Institute of Chemical Technology (IICT) Hyderabad-500 607, India

Introduction
World wide researching for carbon free power generation and neutral/positive waste water treatment MFC is a biochemical-catalyzed system which generates electrical energy through the oxidation of biodegradable organic matter in the presence fermentative bacteria. It is a renewable energy source and is an attractive source. Advantages Environmentally clean, renew ability, liberates large amount of energy and easily converted to electricity by fuel cells, only waste product being water. Dual benefits- generating a clean fuel and reducing waste. Direct generating of fuel has potential advantages- does not require the separation and purification of the gas. Currently, research on MFC is growing.

Introduction
Research is going on world wide for carbon free power generation and neutral/positive waste water treatment

Present Scenario
Increasing energy Needs Depleting fossil reserves Increasing pollution load

Possible solution
Sustainable & Efficient technology for production and utilization of energy Renewable energy sources Nonpolluting energy

Microbial Electricity Generation

Components proposed to be involved in the electron transport from cells to the anode in MFC

Some Microbes are be able to produce their own electron mediators enhancing electron transfer

MFC consists of two electrodes sandwiched around an electrolyte. Oxygen acts as a final electron accepter generating electricity, water and heat

Anodic reactions : CH3COO- + 2OH- 2CO2 + 5H+ + 8eCathodic reaction : O2 + 4e + 4H+ 2H2O

MFC is a complex system

Electrochemical activity of microorganisms Fuel cell- configuration Biofilm on the anode Fuel for anode bacteria Anodic biocatalyst Cathode/anode reaction Proton Exchange Membrane It is important to study all these aspects to make MFC

SCOPE AND OBJECTIVES

To investigate the feasibility of bioelectricity generation eukaryotic and prokaryotic microorganisms as anodic biocatalysts. To optimize physical, chemical and biological parameters. To investigate the influence of various types of proton exchange membranes (PEM) on the performance of MFC To evaluate the potential of MFC as bio-electrochemical treatment system To study microbial diversity of anodic chamber in MFC To study the effect of bioaugmentation strategy on the process performance of MFC

Scope and Objectives

Generation of carbon neutral bioelectricity as an alternative energy fuel for the sustainable environment using wastewater as substrateGreen and renewable energy Different organic wastewaters (ranging from domestic to industrial) as renewable energy resources sustainable development Simultaneous wastewater treatment dual benefit Developing an economically feasible design using low cost materials (electrodes, PEM, substrate, etc.) Non-catalyzed electrodes and mediator-less reactors Economic viability Mixed anaerobic consortia as the biocatalyst. Practical application Optimization and understanding process parameters during MFC operation Investigation of the anodic redox conditions optimum for electron transfer Analysis of microbial diversity by PCR-DGGE

Schematic overview of work

Acronyms
MFC PEM OCV AC FC ED CV TDS DSW CW OLR PDB FDB Microbial fuel cell Proton exchange membrane Open circuit voltage Aerated catholyte Ferricynide catholyte Electron discharge Cyclic voltammetry Total dissolve solvents Designed synthetic wastewater Chemical wastewater Organic loading rate Partially developed biofilm Fully developed biofilm

Anodic Biocatalysts

Characteristics of the wastewaters used as feed

Types of MFC designed and operated

Dual chambered MFCs

Single chambered MFCs

Bioelectrochemical behavior of by Prokaryotic and Eukaryotic

Evaluation of yeast biofuel cell by CV Evaluation of prokaryotic biofuel cell by CV at different feed pH and OLR
Cyclic voltammetry profiles generated during stabilized phase of biofuel cell operation at variable experimental conditions (0th black; 16th blue; 24th green; 36th megentha; 54th brown)

Bioelectricity generation by Prokaryotic and Eukaryotic

Open circuit voltage during the operation of MFC with the function of time (Mixed culture (MFCM); Pseudomonas aeruginosa (MFCP); Escherichia coli (MFCE) Shewanella putrefaciens (MFCS) and Aeromonas hydrophila (MFCA)

Voltage (open circuit) and (b) current generated during the operation of MFC at different feeding pH values and organic loading rates (OLR I, 0.91 kg COD/m3-day; OLR II, 1.43 Kg COD/ m3-day) with the function of time

Evaluation MFC configuration with mixed culture and wastewater

Three types of catholytes Ferricyanide (Double chamber) Aerated (Double chamber) Open-air cathode (Single chamber)

Among these ferricyanide having higher efficiency with respect to power. But not eco-friendly. Double chamber configuration requires higher reactor volume. Even though less power generation in single chamber compared to double chamber.. Economically more feasible Similar substrate degradation Advantages in up scaling the technology

Open circuit voltage and current variation during MFC operation using ferricyanide and aerated catholytes

Influence of anodic pH on MFC performance

Dual chamber operation

Polarization curve Acidophilic pH operation documented highest current output (5.18 mA (100 ); 0.632 V; 3.27 mW) with MFCFC ,(4.26 mA; 0.578 V; 2.46 mW) with MFCAC and 339 mV, 1.66 mA with open air cathode.

Single chamber operation

Alternative material to PEM

Function of various types of proton exchange membranes studied

The experiments depict replacing Nafion117 with glass wool and cellulose material as proton exchange membrane which is cost effective and utilizing wastewater as substrate for in situ power generation

Treatment of Cellulosic material

MFC with 0.75M H2SO4 treated cellulose membrane (CM) as PEM showed maximum OCV (334 mV) and current (1.37 mA at 100 ) Plant based cellulosic material prepared and used as a PEM in MFC

OCV during operation of MFC with the function of time and treatments

Biofilm growth on anode influencing MFC performance

Influences the direct electron transfer Age of the biofilm Biofilm growth Environment Electron discharge and power generation

SEM images of the biofilm developed on anode

The biofilm formed on the anode was subjected to scanning electron microscopy (SEM). a) PDB and b) FDB on graphite anode

Electrochemical influence of bio-augmentation on MFC

Pseudomonas auriginosa

Aeromonas hydrophila

E.coli

Shewanella putrifiecience

CV of anode generated from MFCP, MFCM and MFCE fuel cell operations using Ag/AgCl as reference electrode(Block- 0th h ,Pink - 12th ,Cyan - 24th , Blue -36th and Brown - 48th )
Before augmentation equal electron discharge (ED) (1.04 0.16 mA). Higher ED (11.73 mA) was observed with S. Putrificiens augmented system followed by P. aeruginosa e (8.42 mA), A .hydrophila (6.32 mA) and E. coli (3.17 mA) in the CV

Performance of fuel cell with bio-augmentation

Open circuit voltage during the operation of MFC P. aeruginosa augmented system yielded higher power output (OCV, 418 mV; 3.87 mA at 100 ) followed by S. putrifiencs (OCV, 378 mV; 2.73 mA at 100 ) and A. hydrophila (OCV, 296 mV; 2.26 mA at 100 ). E.coli augmented system registered lower power generation (OCV, 216 mV; 1.76 mA at 100 ).

Bioaugmented strains traced by fluorescent molecular probing

Survival of augmented strains was traced by FISH technique using cy3 labeled fluorescent probes which was important prerequisite for success of bioaugmentation

Microbial Diversity analysis

DGGE was performed by the PCR amplified product of 16S rDNA at variable V3 region using universal primers (341F, 517R) for both dual and single chamber MFC. Phylogenetic sequence affiliation and similarity to the closet relative of amplified 16 rDNA sequence excised from DGGE gels observed dual and single chamber MFCs

Phylogenetic tree
Sequences were submitted to the Nucleotide Sequence Database to the GeneBank public database under the accession numbers from FR670602 to FR670610. The phylogenetic distribution showed significant diversity in microbial community.

Neighbor-joining trees constructed using Mega 4.0 from MFCDC to closely related sequences from Gene Bank

Neighbor-joining trees constructed using Mega 4.0 from MFCSC to closely related sequences from Gene Bank

MFC function as Bio-electrochemical treatment system apart from power generation

Performance of MFC as BET

Conclusions
MFC operated with mixed culture was more
effective in power generation, wastewater treatment and industrial applicability

Performance of MFC influenced by

Reactor configuration (Double and Single chamber) Operating conditions (pH, Organic loading rate, waste composition)

Bio-electrochemical treatment was achieved in MFC due to in situ biopotential of MFC

Anodic biofilm development and bioaugmentation strategies were used to

enhance the electron transfer from bacterial cell to electrode

The study evaluate the different operational parameters required for

optimizing towards scaling up of bioelectricity by MFC

Publications from the reported work

1. Veer Raghavulu S., Suresh Babu P., Kannaiah Goud R., Srikanth S., Venkata Mohan S.
Bioaugmentation of electrochemically active strain to enhance the electron discharge of mixed culture: Process evaluation through electrokinetic analysis.

Journal of RSC Advances , 2012, 2, 677-688

2. Veer Raghavulu, S., Sarma, PN., Venkata Mohan, S., Bioelectrochemical behavior of Pseudomonas aeruginosa and Escherichia coli with the function of anaerobic consortia during biofuel cell operation. Journal of Applied Microbiology, 2011. 110, 666674 3. Venkata Mohan, S., Veer Raghavulu, S., Goud, RK., Sarma, PN. Microbial diversity analysis of long term operated biofilm configured anaerobic reactor producing hydrogen from wastewater under diverse conditions. International Journal of Hydrogen Energy, 2010. 35, 12208-12215 4. Veer Raghavulu, S., Venkata Mohan, S., Goud, RK., Sarma, PN. Saccharomyces cerviceae as anodic biocatalyst in non-catalyzed aerated biofuel cell: influence of redox condition andsubstrate load on power generation. Bioresource Technology, 2011. 102, 2751-2757 5. Veer Raghavulu, S., Venkata Mohan, S., Reddy, MV., Sarma, PN. Behavior of single chambered
mediatorless microbial fuel cell (MFC) at acidophilic, neutral and alkaline microenvironments during chemical wastewater treatment.

International Journal of Hydrogen Energy. 2009. 34, 7547-7554

Publications from the reported work

6. Veer Raghavulu, S., Venkata Mohan, S., Goud, RK., Sarma, PN.
Communications. 2009. 11, 371-375
Anodic pH microenvironment influence on microbial fuel cell (MFC) performance in concurrence with aerated and ferricyanide catholytes.

Electrochemical

7. Venkata Mohan, S., Veer Raghavulu, S., Dinakar, P., Sarma, PN.
Biosensors and Bioelectronics. 2009. 24, 2021-2027

Integrated function of microbial fuel cell (MFC) as bio-electrochemical treatment system associated with bioelectricity generation under higher substrate load.

8. Venkata Mohan, S., Veer Raghavulu, S., Sarma, PN. Influence of anodic biofilm growth on bioelectricity production in single chambered mediatorless microbial fuel cell using mixed anaerobic consortia. Biosensors and Bioelectronics. 2009 24, 41-47 9. Venkata Mohan, S., Srikanth, S., Veer Raghavulu, S., Mohanakrishna, G., Kiran Kumar, A., Sarma, PN. Evaluation of the potential of various aquatic eco-systems in harnessing bioelectricity through benthic fuel cell: Effect of electrode assembly and water characteristics. Bioresource Technology. 2009. 100, 22402246

Publications from the reported work

10. Venkata Mohan, S., Veer Raghavulu, S.,Sarma, PN. Biochemical evaluation of bioelectricity production process from anaerobic wastewater treatment in a single chambered microbial fuel cell (MFC) employing glass wool membrane. Biosensors and Bioelectronics. 2008 23, 1326-32. 11. Venkata Mohan, S., Sarvanan, R., Veer Raghavulu, S., Mohankrishna, G., Sarma PN.
Bioelectricity production from wastewater treatment in dual chambered microbial fuel cell (MFC) using selectively enriched mixed microflora: Effect of catholyte. Bioresource

Technology.2008. 99(3), 596-603

12. Venkata Mohan, S., Veer Raghavulu, S., Srikanth, S., Sarma, PN. Bioelectricity production by
meditorless microbial fuel cell (MFC) under acidophilic condition using wastewater as substrate: influence of substrate loading rate.

Current Science. 2007. 92(12), 1720-1726

Other Publications
1. Min-Kyu Ji, Veer Raghavulu S, Hyun-Shik Y,Reda A.I, Jaeyoung C, Wontae Le, Thomas C. Timmes, Inamuddin, Byong-Hun Jeon. Simultaneous nutrient removal and lipid production from pretreated piggery wastewater by Chlorella vulgaris YSW-04 Applied Microbiology and Biotechnology 2012 (Accepted) 2. Venkata Mohan, S., Veer Raghavulu, S., Mohanakrishna, G., Srikanth, S., Sarma, PN. Optimization and evaluation of fermentative hydrogen production and wastewater treatment processes using data enveloping analysis (DEA) and Taguchi design of experimental (DOE) methodology. International Journal of Hydrogen Energy. 2009. 34, 216-226 3. Reddy, BS., Reddy, BP., Veer Raghavulu, S., Ramakrishna, S., Venkateswarlu, Y., Diwan, PV. Evaluation of antioxidant and antimicrobial properties of Soymida febrifuga leaf extracts. Phytotherapy Research. 2008 22 (7), 943-947 4. Venkata Mohan, S., Mohanakrishna, G., Veer Raghavulu, S., Sarma, PN.
Enhancing biohydrogen production from chemical wastewater treatment in anaerobic sequencing batchbiofilm reactor (AnSBBR) by bioaugmenting with selectively enriched kanamycin resistant anaerobic mixed consortia.

International Journal of Hydrogen Energy. 2007. 32, 32843292

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