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 research for carbon free power generation and

neutral/positive waste water treatment

The Microbial Fuel Cell or 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 friendly, renewable, 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 ongoing world wide for carbon free power generation and
neutral/positive waste water treatment

Present Scenario Possible solution

Increasing energy Needs Sustainable & Efficient technology for
production and utilization of energy
Depleting fossil reserves Renewable energy sources
Increasing pollution load Nonpolluting energy
 Microbial Electricity Generation

Components proposed to be involved in the electron Some Microbes are be able to produce their own
transport from cells to the anode in MFC electron mediators enhancing electron transfer

MFC consists of two electrodes sandwiched around an

electrolyte. Oxygen acts as a final electron accepter Anodic reactions : CH3COO- + 2OH- → 2CO2 + 5H+ + 8e-
generating electricity, water and heat Cathodic reaction : O2 + 4e− + 4H+→ 2H2O
 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
Schematic overview of work
 Acronyms
MFC Microbial fuel cell
PEM Proton exchange membrane
OCV Open circuit voltage
AC Aerated catholyte
FC Ferricynide catholyte
ED Electron discharge
CV Cyclic voltammetry
TDS Total dissolve solvents
DSW Designed synthetic wastewater
CW Chemical wastewater
OLR Organic loading rate
PDB Partially developed biofilm
FDB Fully developed biofilm
DGGE Denaturing gradient gel electrophoresis
Anodic Biocatalysts
Characteristics of the wastewaters used as feed
 Types of MFC designed and operated

Dual chambered Single chambered MFCs

MFCs
 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 Ω); 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 Ω)

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

a b
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 Ω) followed by S. putrifiencs (OCV, 378 mV; 2.73 mA at 100 Ω) and A.
hydrophila (OCV, 296 mV; 2.26 mA at 100 Ω). E.coli augmented system registered
lower power generation (OCV, 216 mV; 1.76 mA at 100 Ω).
 Bioaugmented strains traced by fluorescent
molecular probing

Survival of augmented strains was traced by FISH technique using cy3 labeled
fluorescent probes which was important pre-requisite 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 Neighbor-joining trees constructed using

Mega 4.0 from MFCDC to closely related Mega 4.0 from MFCSC to closely related
sequences from Gene Bank 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 bio-

potential 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

• MFC was opportunistic source for alternative fuel for future generations
Evaluate the influence of inorganic phosphate
and micronutrients on biomass growth rate and
lipid production using with microalgae+

Department of Environmental Engineering, Yonsei University, Wonju,

Gangwon-do, 220-710, South Korea
 Objective of the Study
The present study focused on the demonstrate
a key factor such as inorganic phosphate and
micronutrients in enhancing biomass growth
and lipid production of the microalgae
 Experimental Methodology
Media preparation
 Influence of inorganic phosphate PO4 and

micronutrients: zinc (ZnSO4 ), Manganese

(MnCl2), cupper (CuSO4), cobalt
(Co(NO3)2) upon the biomass production
and lipid productionwere evaluated using
Micractinium pusillum algae with Bold
Basel metabolic (BBM) media.
 Experimental Methodology

Algae strain and culture condition

 Micractinium pusillum was isolated
from effluent of municipal sewage
treatment plant (Wonju, South Korea).
It was preserved in Bold basal medium
(BBM) (Bischoff and Bold, 1963)
 Algae were inoculated at 10% (v/v) in

250 mL Erlenmeyer flasks containing

100 mL liquid medium.
Result and Discussion
 Conclusion:

 The study revealed that inorganic phosphate

and micronutrients play major role in
microalgae biomass production and lipid
production. Depletion of nutrients especially
phosphate which showed more adverse
affect on both the biomass production and
lipid production of microalgae.
 Microbial interactions in batch and
continuous culture-influence of signalling
molecule

Department of Neuroscience, Karolinska

Institutet,Sweden
Outlines of Research
Evaluation of MIC for Staphylococcus
aureus based OD
Evaluation of MIC for Staphylococcus
aureus based CFU

Cont…
Evaluation of MIC for Staphylococcus
aureus based CFU
MIC for E coli
Microbial interactions in batch culture
Microbial interactions in continuous culture
 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
electro-kinetic 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, 666–674

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. Anodic pH microenvironment

influence on microbial fuel cell (MFC) performance in concurrence with aerated and ferricyanide catholytes. Electrochemical
Communications. 2009. 11, 371-375

7. Venkata Mohan, S., Veer Raghavulu, S., Dinakar, P., Sarma, PN. Integrated function of microbial
fuel cell (MFC) as bio-electrochemical treatment system associated with bioelectricity generation under higher substrate load.
Biosensors and Bioelectronics. 2009. 24, 2021-2027

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, 2240–2246
 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, 3284–3292
Thank You