Bioresource Technology 217 (2016) 113–120

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Effect of composites based nickel foam anode in microbial fuel cell using
Acetobacter aceti and Gluconobacter roseus as a biocatalysts
Rengasamy Karthikeyan a,b,c,⇑, Navanietha Krishnaraj a, Ammaiyappan Selvam c,
Jonathan Woon-Chung Wong c, Patrick K.H. Lee b, Michael K.H. Leung b, Sheela Berchmans a
a
Electrodics and Electrocatalysis (EEC) Division, Central Electrochemical Research Institute (CSIR-CECRI), Karaikudi 630 006, Tamil Nadu, India
b
Ability R&D Energy Research Center, School of Energy and Environment, City University of Hong Kong, Hong Kong, China
c
Sino-Forest Applied Research Centre for Pearl River Delta Environment, Department of Biology, Hong Kong Baptist University, Hong Kong, Hong Kong, China

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 Bio-composite anode material was

investigated in microbial fuel cells.
 Effects of conducting polymer,
biopolymer and metal carbide were
studied.
 The biocompatible composite anode
material delivers the power density of
18.8 W m 3.
 This power density is 2.3 times
higher than the bare electrode
(8.3 W m 3).

a r t i c l e i n f o a b s t r a c t

Article history: This study explores the use of materials such as chitosan (chit), polyaniline (PANI) and titanium carbide
Received 24 December 2015 (TC) as anode materials for microbial fuel cells. Nickel foam (NF) was used as the base anode substrate.
Received in revised form 24 February 2016 Four different types of anodes (NF, NF/PANI, NF/PANI/TC, NF/PANI/TC/Chit) are thus prepared and used in
Accepted 25 February 2016
batch type microbial fuel cells operated with a mixed consortium of Acetobacter aceti and Gluconobacter
Available online 3 March 2016
roseus as the biocatalysts and bad wine as a feedstock. A maximum power density of 18.8 W m 3 (2.3
times higher than NF) was obtained in the case of the anode modified with a composite of PANI/TC/Chit.
Keywords:
The MFCs running under a constant external resistance of (50 O) yielded 14.7% coulombic efficiency with
Chitosan
Polyaniline
a maximum chemical oxygen demand (COD) removal of 87–93%. The overall results suggest that the cat-
Titanium carbide alytic materials embedded in the chitosan matrix show the best performance and have potentials for fur-
Wine ther development.
Microbial fuel cell Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction

Microbial fuel cell (MFC) is a bioelectrochemical device which

⇑ Corresponding author at: Sino-Forest Applied Research Centre for Pearl River
Delta Environment, Department of Biology, Hong Kong Baptist University, Hong
Kong, Hong Kong Special Administrative Region. Tel.: +852 3411 2537; fax: +852
3411 2095.
E-mail address: karthikelectro@gmail.com (R. Karthikeyan).
generates electrical energy from the metabolic activity of living
organisms. Various parameters such as selection of substrates,
the choice of biocatalysts and their direct electron transferring
capability, the choice of mediators, the choice of terminal electron
acceptors (cathode), pH, temperature, dissolved oxygen etc will

http://dx.doi.org/10.1016/j.biortech.2016.02.114
0960-8524/Ó 2016 Elsevier Ltd. All rights reserved.
114 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120
limit the performance of MFCs. One of the major issues contribut-
ing to low performance is the inefficient anodic process responsi-
ble for transferring the electrons from the bacteria to the
electrodes (Liu et al., 2015). Literature reports reveal that many
attempts has been made to prepare efficient anode materials for
MFC. Most of the MFC studies that utilize conventional carbon/-
graphite materials for anodes (Liu et al., 2010). Further improve-
ment in the performance has been achieved by modifying
carbon/graphite with functional materials, which include
polypyrrole-coated carbon, carbon felt doped with quinone deriva-
tives, ammonia-treated carbon cloth and Iron, sulfur and phenyl
group based modifications (Zhao et al., 2010). These modifications
have led to only two to three fold improvements in the current
density and the preparation of the modified electrodes requires
complex preparation procedures (Hai et al., 2009).
Graphite anode modified with ammonia treatment showed 48%
increased power output which could be due to the increased sur-
face charges of the treated carbon electrodes (Cheng and Logan,
2007). Use of carbon nanotubes along with a coating of ferrocene
on the anodes led to a foam like structure and resulted in high sur-
face area and high catalytic activity (Morozan et al., 2007). Combi-
nation of carbon nanotube and polyaniline to form a novel
composite anode exhibited an increased specific surface area and
enhanced charge transfer capacity (Qiao et al., 2007). Compared
to the use of carbon nanotubes as anode materials, the conductive
polymers show significant improvement as MFC anode materials
(Zhao et al., 2010). Among all the conductive polymers,
polyaniline- and polypyrrole-based composite materials effectively
improve the MFC performance. Recent studies on PANI and poly
(aniline-co-o-aminophenol) (PAOA) modified carbon felt anodes
resulted in about 35% and 18% improvement, respectively, when
compare to unmodified carbon felt (Li et al., 2011). The electro-
chemically active polymers such as polypyrrole on vitreous carbon
electrode could enhance the power output to a high level of
2400 W m 3 with an anolyte volume of 10 mL (Yuan and Kim,
2008). Moreover, graphite felt modified with quinone groups (C/
HNO treatment) showed three fold improvement compared to
unmodified graphite felt (Scott et al., 2007).
A conductive polyaniline nanowire network (PANI-NN) with
three-dimensional nanosized porous structures was formed on
anodes and it has been evaluated in MFCs with self-organized
microbial communities (Zhao et al., 2010). Microsized pores in gra-
phite felts would provide an increased area for microbial attach-
ment, while nanoporous structures in PANI-NN would efficiently
collect electrons from mediators excreted by microbes (Zhao
et al., 2010). On the other hand, PANI finds applications in
rechargeable batteries, corrosion protection of metals, molecular
sensors and it is relatively easy to dope PANI. Another way of
improving the efficiency of extra cellular electron transport is the
immobilization of an electron redox mediator on the anode surface
(Feng et al., 2010). The immobilization of AQDS (anthraquinone-2,
6-disulphonic disodium salt) on a carbon felt anode was accom-
plished by electro-polymerization of pyrrole with AQDS as the
dopant and this anode produced 13 times higher power than the
unmodified carbon felt (Feng et al., 2010). Ten times improvement
in the power output has been achieved by coating electron media-
tors such as Mn4+ or neutral dyes on graphite (Park and Zeikus,
2002). A high surface area graphene was synthesized as an anode
material in mediator MFC which delivered 18 times higher power
output (2668 mW m 2) than that of the unmodified anode (Zhang
et al., 2011).
It has been observed that chitosan is a biopolymer having an N-
deacetylated derivative of chitin, which could be obtained from
exoskeletons of crustacean, mollusks and insects. Since chitosan
has useful properties such as biocompatibility, nonirritant, good
film forming nature, high mechanical strength and adhesion, it
can be utilized for a wide range of applications such as sustained
releasing systems, waste water treatments, packaging, separation
membranes and biosensors (Marcasuzaa et al., 2010). The combi-
nation of PANI with chitosan has showed good electrical conduc-
tivity due to the H-bonding occurring between the chitosan
backbone (H-acceptor amide group) and PANI backbone (H-
donor imino group) (Marcasuzaa et al., 2010; Tiwari and Singh,
2007).
Reducing the activation barrier and to lower the cost of elec-
trode, researchers were attempted to interface an electro catalyst
to improve the performance of MFC. It was reported that interfac-
ing the electrocatalyst (Tungsten carbide) could significantly
improve the performance of microbial anode since it can oxidize
the microbial fermentation products (eg. Formate) (Rosenbaum
et al., 2006). Carbon electrode coated with tungsten carbide
showed two fold increase in the power output in a glucose fed
MFC (Rosenbaum et al., 2006). Carbide materials such as Nickel–
tungsten carbide, Tungsten carbide, and Molybdenum carbide
can be interfaced along with biocatalyst and they can also be
increase the MFC performance not only due to the oxidation of fer-
mentation product also it can improve the reaction kinetics by
lowering the activation barrier of the electrode (Nagai et al.,
2007; Rosenbaum et al., 2007; Zeng et al., 2010, 2012; Wang
et al., 2014). As far as our knowledge goes, the synergistic effects
of conducting polymer, biopolymer and metal carbide for the
development of anode material in microbial fuel cells has not been
investigated.
In the present study, the influence of incorporating titanium
carbide along with the conducting polymer PANI is studied using
nickel foam as a base substrate. The effect of chitosan matrix on
the composite catalyst PANI + Titanium carbide is also investigated
to increase power out from bad wine. Two compartments batch
type MFCs were constructed with the aforesaid materials and the
MFC performance was evaluated. Acetobacter aceti and Gluconobac-
ter roseus were used as biocatalysts, bad wine was used as feed-
stock and ferricyanide was used as the catholyte.
2. Methods
2.1. Anode modification and electrode preparations
Nickel foam was chosen as a base material for MFC anode due to
the porous structure can acts as a good support for the incorpora-
tion of the composite material. The nickel foam (5  5  0.2 cm,
MarkeTech International, Inc.) was pretreated by sonication in ace-
tone for 10 min followed by immersion in 5% HCl for 1 min to
remove any surface impurities and finally rinsed thoroughly with
distilled water and dried at room temperature (Dai et al., 2008).
After that the electrode was modified by brush coating with the
composite slurry obtained by the mixer of activated carbon – 5%,
polyaniline-1% (PANI, Aldrich, emeraldine salt, grafted with lignin),
polyvinylenedifluoride – 10% (PVDF, Aldrich) in N-methyl-2-
pyrrolidone (NMP) (Alfa Aesar, HPLC grade, 99.5%). For further
modification, titanium carbide (1%) and chitosan (2%) (from shrimp
shells, Sigma Aldrich) were used. Then the coated anodes (coating
thickness 0.5 mm) were dried at room temperature for about 48 h
(see Supplementary information S1, S2). The total composite layer
is around 100–180 mg/cm2 of nickel foam. The nickel foam anode
and the modified nickel foam of every modification was separately
evaluated as anode material. Four different types of anodes viz.,
were evaluated for their performance in the MFCs (Fig. S2).
I. unmodified nickel foam (NF)
II. NF/PANI: NMP = 50 mL, PANI = 0.5 g (1%/v), PVDF-5g (10%/v),
Activated carbon: 2.5 g (5%/v)
 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120
III. NF/PANI/Titanium carbide (NF/PANI/TC): NMP = 50 mL,
PANI = 0.5 g (1%/v), PVDF-5g (10%/v), TiC = 0.5 g (1%/v), Acti-
vated carbon: 2.5 g (5%/v)
IV. NF/PANI/Titanium carbide/Chitosan (NF/PANI/TC/Chit): 2%/v
chitosan solution (1 g of chitosan was dissolved in 50 mL of
diluted acetic acid (1:1)) was coated thin layer over on dried
NF/PANI/TC electrode
2.2. Selection of inoculum, feedstock, MFC setup and characterizations
Acetic acid bacteria namely G. roseus and A. aceti are primarily
responsible for spoilage of wine. These two genera can be used
to oxidize sugars, sugar alcohols, and ethanol with the production
of acetic acid as the major end product. Acetic acid bacteria are
present at all stages during the wine making, from the mature
grape through vinification to conservation (Joyeux et al., 1984). It
was reported for the first time the mixed culture of Acetobaceter
aceti and G. roseus is a suitable biocatalyst for treating bad wine
cum electricity production (Karthikeyan and Berchmans, 2012).
This phenomenon has been proven by the presence of membrane
bound enzymes (alcohol dehydrogenase, ADH: glucose dehydroge-
nase, GDH) in the acetic acid bacteria. Bioelectrocatalytic activities
and their direct electron transfer process of microbe/electrode in
presence of glucose, ethanol, acetate and bad wine has been
reported on carbon based material and nickel deposited carbon
material in our previous publications (Karthikeyan et al., 2009,
2012; Karthikeyan and Berchmans, 2012). In general, the quality
of wine can be assessed based on the production of lactic and acetic
acids by residual bacterial populations during the storage period in
the wine bottle. It has been reported that the residual bacteria can
be increased from 0.3  102 cells/mL to 3.4  102 cells/mL after
3 months. Hence, the conversion of wine into waste wine or bad
wine is controlled mainly by two factors. Firstly, the oxidation of
wine on exposure to air and secondly, the bacterial proliferation
leads to bad wine production. For our study, we exposed the wine
(Golconda Ruby wine, United spirits Ltd., India. 16% v/v ethanol) to
air for 1 h and used it as fuels (hereafter referred to as bad wine).
Then the wine sample was stored in refrigerator for further use.
The batch type MFC experiments were conducted by using four
anodes (5  5  0.2 cm) as described in the last section. Cleaned
graphite cathodes (5  5  0.5 cm) were used and 0.1 M ferri-
cyanide in phosphate buffer (pH 7, 0.1 M) was used as the catho-
lyte. Two chamber MFCs separated by a proton exchange
membrane (Nafion 117, Aldrich) was constructed using plexi glass
plates. The volume of the chamber was 125 mL. A. aceti (NCIM No.
2116) and G. roseus (NCIM No. 2049) were purchased from
National Chemical Laboratory (NCL), Pune, India. Sub-culturing
was carried out using the following media: tryptone (1 g), yeast
extract (1 g), glucose (1 g) and CaCO3 (1 g) in 100 mL of distilled
water). About 0.5 g each of the centrifuged cells (centrifugation
at 12,000g for 5 min) of A. aceti and Gluconobacter roeseus were
used as inoculums in the anode chamber. The anolyte consisted
of phosphate buffer (pH 7, 0.1 M) and bad wine with a chemical
oxygen demand (COD) of 7.8 g L 1. The graphite cathode was kept
in the cathode chamber containing 0.1 M K3Fe(CN)6 in phosphate
buffer (pH 7, 0.1 M). The anode chamber was kept under anaerobic
conditions and the MFC was run under ambient temperature.
The half-cell electrode potential (vs. normal calomel electrode,
NCE, Hg/Hg2Cl2 in 1 M KCl) and fuel cell voltage were monitored
by connecting to the data acquisition unit (Agilent 34970 A)
attached to a personal computer. The Eanode was recorded up to
110 h in order to develop the biofilm on the anodes in the presence
of suspended inoculums (A. aceti and Gluconobacter roseus). At the
end of 110 h the medium was replenished with fresh phosphate
buffer to reach steady state electrode potential of biofilm. The com-
mon practice of fresh catholyte (ferricyanide, 0.1 M K3Fe(CN)6)
115
addition was also followed during polarization test for each MFC.
Once the biofilm reach their steady state, bad wine were supplied
as feed stock to conduct the polarization curve analysis. The polar-
ization of the fuel cell was carried out by applying a range of exter-
nal resistances (10 K O to 10 O). The resulting voltage could be
used for the analysis of current density and power density by using
the Ohm’s law (Yazdi et al., 2015). To conduct constant polariza-
tion all MFCs were externally connected to 50 X for further
130 h and the derived current was used for coulombic recovery .
After operating at constant resistance (50 X), the anolyte sample
(centrifuged at 12,000g for mins and filtered through 200 lM)
was analyzed and chemical oxygen demand (COD, g/L) level was
estimated by a COD reactor kit (spectroquant 320, Merck). The
coulombic efficiency was calculated from the derived current at
fixed time interval (Cobs) with the theoretically calculated charge
obtained from COD removal (Ccal).
2.3. SEM and Epifluorescence studies
The biocompatibility and surface morphology of the anode were
analyzed by the epifluorescence and scanning electron microscopy
(SEM). A small portion of MFC anode was removed and stained
with 1 mL of 0.1% (w/v) acridine orange for 15 min. Green fluores-
cent cells were viewed with a Nikon epifluorescence microscope
(E200; Nikon, Tokyo; equipped with a 10/0.25 numerical aper-
ture and 100/1.25 numerical aperture) using a 100 objective
and a 10 eyepiece. Microscopic images were captured with a
short exposure time (<0.2 s) using a digital charge-coupled device
(CCD) camera. Another sample of the anode portion was treated
with 100 lL of phosphotungstic acid (0.5%) solution and then
rinsed with ethanol and water and dried under nitrogen and ana-
lyzed by scanning electron spectroscopy (Hitachi model S-3000H).
3. Results and discussion
3.1. Measurements of half-cell potentials and open circuit voltage of
MFC
The time required to grow the bacteria on the anode is based on
the surface charge associated with the anode material and it can be
determined through the open circuit voltage and half-cell anode
potential. It was reported that the affinity for the microbes/anode
can differ by surface charge property of the anode material
(Franks et al., 2010). The positively charged molecules on the
anode can accelerate the bio-interface such as ammonia treated
carbon felt, carbon foam modified with polyethyleneimine (PEI)
and their fuel cell voltage (Kramer et al., 2012), chitosan based
anode (He et al., 2012; Higgins et al., 2011), nitrogen doped anode
to increase active site (Ci et al., 2012).
Fig. 1(a) shows the open circuit voltage of MFC as a whole. The
measured open circuit voltage (OCV) values are in the following
order NF/PANI/TC/Chit (0.838V) > NF/PANI/TC (0.773V) > NF/PANI
(0.750V) > NF (0.734V). The pattern of variation is same as that of
the half-cell anode values under open circuit conditions. Fig. 1(b)
shows the half-cell potential (V vs. NCE) of the four different
MFC anodes with respect to time. Initially, the half-cell anode
potential of NF ( 0.181 V) was more negative than the other mod-
ified anodes namely NF/PANI (0.057), NF/PANI/TC (0.009) and NF/
PANI/TC/Chit ( 0.109) in the presence of bacterial inoculums with
bad wine as the feedstock. NF anode showed a constant negative
voltage of 0.190 V up to a period of 85 h and then steeply
switches over to 0.458 V (at 110 h). The reduction of anodic
potential due to the electrons released from the substrate (cat-
alyzed by the bacteria), could still be captured by the anode result-
ing in a reduction of electrode potential even though the circuit
116 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120
Fig. 1. Open circuit voltage of MFCs (Ecell) (a) and half-cell potential of anodes
(Eanode) (b) of nickel foam (NF) and modified NF composite anodes during a period
of 130 h. The portion beyond 112th h indicates anode with established biofilm.
was open. The time lag for switching the Eanode from low negative
to higher negative potential is related to the time taken for the cat-
alytic activity of bacteria on the anode surface. The lagging period
was reduced in the case of modified anodes. The potential of NF/
PANI anode reached 0.478 V after 41 h and NF/PANI/TC anode
took only 32 h after inoculation. NF/PANI/TC and NF/PANI/TC/Chit
anodes required a lagging period of 59 h even though they possess
higher anode potential when compared to other anodes. The
increased negative anode potentials indicate the influence of bio-
film growth on the anode. After obtaining the anode steady-state
potential (after 120 h for each anode), the anolyte media was
removed (along with the suspended bacteria) and replaced with
fresh phosphate buffer (pH 7, 0.1 M) along with bad wine as the
feedstock. At the end of 130 h, the anodes with the biofilms only
reached the maximum half-cell steady-state potential and their
values (Eanode,i=0) are in the order NF/PANI/TC/Chit ( 0.538 V) >
NF/PANI/TC ( 0.521 V) > NF/PANI (-0.500 V) > NF ( 0.473 V). This
indicates that biofilms on composite electrodes have their higher
negative potential ( 0.538 V) and thermodynamically feasible for
anodic oxidation of the substrates in a more efficient manner com-
pared to NF anode ( 0.473 V).
3.2. Polarization and power generation of MFCs
Polarization of MFCs helps us to understand the ability of sus-
tainable power production for practical applications. Fig. 2 shows
Fig. 2. Polarization curve analysis; cell voltage (a) and power density (b) as a
function of current density derived under various external load resistances (closed
circuit conditions).
the polarization of MFCs carried out by connecting different resis-
tors (10 K O to 10 O) in the external circuit. The current density and
power density of MFCs have been derived from their resulting
steady-state voltage using Ohm’s law. The maximum power den-
sity of 18.82 W m 3 (at 54.9 A m 3) derived from the MFC with
the anode NF/PANI/TC/Chit was 2.3 times higher than that of the
MFC with NF anode (i.e., 8.29 W m 3 vs. 25.8 A m 3). The power
density obtained from NF/PANI and NF/PANI/TC show a similar
output of 15.17 W m 3 (49.28 A m 3) and 15.59 W m 3
(45.6 A m 3), respectively. This indicates that in the case of tita-
nium carbide, only a marginal improvement of power density
was achieved, while combining TiC with chitosan matrix, a higher
power output was observed. This can be due to the good biocom-
patible microenvironment provided by chitosan for proteins or
enzyme present in the bacterial cells within the matrix containing
TiC and PANI were eventually participate in the anodic reaction as
we see from the power output curve(Fig. 2) (Kang et al., 2009). The
performance of A. aceti and G. roseus has previously been investi-
gated in MFCs and proved their effectiveness using bad wine as
feedstock and carbon felt as anode (Karthikeyan et al., 2009;
Karthikeyan and Berchmans 2012). The cyclic voltammetry behav-
ior of these bacteria on carbon paper, glassy carbon electrode and
electrodeposited nickel surface have been reported (Karthikeyan
 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120
et al., 2012). But as shown here, the same mixed culture can also be
used with composite anode material. Furthermore, the power den-
sity derived from composite anode is 18.8 W/m3, which is 5 times
higher than carbon felt anode (3.8 W/m3) using similar fuel and
biocatalyst (Karthikeyan and Berchmans, 2012). Hence, the electri-
cally conductive materials such as PANI, TC and ionic conductive
biopolymer play a major role in improving the performance of
MFC output.
3.3. Over potential of modified anodes in MFC under polarizations
During the polarization study, the individual potential of
anodes and cathodes with respect to NCE were recorded at differ-
ent current densities (Min and Angelidaki, 2008; Zhang et al.,
2011). Higher current density leading to a lowering of overvoltage
indicates an efficient anode. Fig. 3 shows the steady-state half-cell
anode potential and half-cell cathode potential at various external
loads and the respective current density in each MFC. The potential
of the anodes were observed to be NF ( 0.498 V), NF/PANI
( 0.501 V), NF/PANI/TC ( 0.517 V), and NF/PANI/TC/Chit
( 0.535 V). The respective cathodic potentials were found to be
0.302 V, 0.288 V, 0.318 V and 0.255 V. From these results, it is pos-
sible to measure the over potential of anode and cathode at fixed
current density. For comparison, a constant current density of
25.8 A m 3 (3.22 mA) was chosen from the optimal operating cur-
rent density of control anode (NF-MFC). At the current density of
25.8 A m 3, the difference in the open and closed circuit potential
Fig. 3. Half-cell potential of anode (a) and cathode (b) under variable external
resistance (under closed circuit conditions).
117
(CCP) of the four anodes are (OCPanode–CCPanode) 170 mV,
108 mV, 121 mV, and 97 mV whereas in the case of cathodes were
47 mV, 33 mV, 80 mV, and 18 mV. It should be noted that anode
and cathode were separated by proton exchange membrane. Dur-
ing the polarization curve analysis at different external resistance,
the anode and cathode potentials were recorded simultaneously.
The charge transfer from bioanode to cathode (ferricyanide) at dif-
ferent external resistance causes change of anode and cathode
potential from its open circuit potential. Moreover, Fig. 3 indicates
that over potential of anode is significantly larger than over poten-
tial of cathode.
It can be seen that a decrease anode over potential is observed
with all the modified anodes compared to the unmodified NF
anode. The anodes have the vital role on the power delivery. The
lowering of anode potential as observed in the case of NF/PANI/
TC/Chit-MFC is likely due to the acceleration of electron supply
to cathode or the kinetics of electron transfer from the electricigens
to the anode is additionally enhanced. The overall observation of
the results obtained from the polarization experiments indicate
that NF/PANI/TC/Chitanode gives rise to a higher anodic limiting cur-
rent density (Ilim 13.7 mA, 109.6 A m 3) which was nearly four
times higher than that obtained from the control anode (NFanode,
Ilim 3.42 mA, 27.4 A m 3). The other electrodes like NF/PANIanode
(Ilim 8.6 mA, 68.8 A m 3) and NF/PANI/TCanode (Ilim 9.9 mA,
79.2 A m 3) also yielded only lower current densities. Also, the
internal resistance (slope at mid-point region of polarization
curves) of MFCs during the polarization was calculated. It could
be found that the higher ohmic resistance was attributed to the
unmodified nickel foam MFC (116 O) whereas the MFCs with the
NF/PANI and NF/PANI/TC anodes showed a lower value (<70 O).
NF/PANI/TC/Chitanode–MFC exhibited the lowest ohmic resistance
of 46 O, leading to a higher output.
3.4. Electricity generation under closed circuit condition
Electricity generation from the MFCs under closed circuit condi-
tions provides an indication on the efficiency of anodic electron
transfer reactions. Fig. 4 shows closed circuit voltage (CCV) of wine
fed (COD of 7.8 g L 1) MFCs under a constant external resistance
(50 O) during a period of 130 h. Starting CCV of NF-MFC (0.140 V)
was low when compared to modified anode MFCs like NF/PANI-
MFC (0.292 V), NF/PANI/TC-MFC (0.309 V) and NF/PANI/TC/Chit-
MFC (0.383 V). Further, the voltage of NF-MFC was steadily
decreasing until the cut off voltage (voltage at which the maximum
discharge were reached).
The voltage of NF/PANI-MFC was higher initially compared to
NF-MFC and it started decreasing after 60 h. Similar behavior
was noted in the case of other modified electrodes and in these
cases voltage started decreasing quickly beyond 80 h. This was
explained by the continual exhaustion of wine substrates due to
oxidation reaction on the anode. In the case of chitosan with tita-
nium carbide electrode (NF/PANI/TC/Chit-MFC), the starting CCV
was the highest and the voltage declined faster after a period of
100 h. Hence, it is clear that the performance of the NF/PANI/TC/
Chit anode is better than the other electrodes. Fig. 5 shows the
derived current density and their corresponding power density
over a period of 130 h for wine fed MFCs. Here in the case of NF-
MFC, the derived current decreased from 22.4 A m 3 (6.2 W m 3)
to 7.3 A m 3 (0.67 W m 3). Similarly, in the case of NF/PANI/TC/
Chit-MFC, the current decreased from 46.72 A m 3 (13.6 W m 3)
to 3.7 A m 3 (0.09 W m 3).
Decline of cell voltage (Fig. 4) and current density (Fig. 5) under
closed circuit is related to depletion of fuel content with respect to
time. In this stage, bioanode continuously transfer the electron to
cathode as soon as the fuel is oxidized at the anode. Naturally,
the fuel concentration (COD value) is lowered as the cell voltage
118 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120

polarization at 50 Ω
0.8 a - NF
b - NF/PANI
0.7

polarization curve analysis

c - NF/PANI/TC
0.6 d - NF/PANI/TC/Chit
Cell voltage (V)

0.5

0.4

0.3
c

0.2 d
a
0.1
c
b
0.0
0 20 40 60 80 100 120 140
Time (h)
Fig. 4. Cell voltages variation with time under a constant load resistance (50 O).

Fig. 5. Current density and power density variation with time at constant load resistance (50 O). (a) NF, (b) NF/PANI, (c) NF/PANI/TC and (d) NF/PANI/TC/Chit.

becomes lowered with respect to time. Under the closed circuit
conditions the product of current over the time was calculated as
experimental coulombic charge (Cobs) which was then compared
to theoretically available coulombic charge. At the end of 130 h,
the COD removal was in the range of 87–93%. The values of
coulombic efficiency increases in the order 6.95% (NF-MFC)
< 9.55% (NF/PANI-MFC) < 10.5% (NF/PANI/TC-MFC) < 14.7% (NF/
PANI/TC/Chit-MFC). The performance of the modified anode with
respect to unmodified anode used in wine fed MFCs is presented
in Table 1. Herein the investigated MFC parameters such as lagging
period (h), half-cell anode potential (V), power density (W/m3),
over potential (mV), limiting current density (mA/m3), internal
resistance (O), discharge capacity (mA h) and coulombic efficiency
(%) are compared in order to understand the comparative perfor-
mance of the modified anodes (Table 1). In each parameter the
composite electrode have showed advantages over control NF
 R. Karthikeyan et al. / Bioresource Technology 217 (2016) 113–120 119

Table 1 Hence the results indicate that the conducting polymer (PANI)
Comparative performance of composite anode with unmodified anode under composite electrode has significantly improved MFC performance.
operative parameters.
The presence of protonated group ‘‘green imines” in PANI is highly
Parameters MFC-Anode Results conductive and it strongly conducts the electrons from the bacte-
Lagging period (h) NF 85 ria. The interaction of bacteria with conducting polymer (PANI)
NF/PANI 41 can be explained by either electron transfer mediation or electro-
NFPANI/TC 32 catalysis (Li et al., 2011). The backbone of the PANI with activated
NF/PANI/TC/Chit 59
Half-cell anode NF 0.473( 193 m V vs. SHE)
carbon creates the high surface area for bacterial adhesion. While
potential (V vs. NCE) NF/PANI 0.500 ( 220 mV vs. SHE) doping the TC to the PANI matrix, the coulombic efficiency was
NFPANI/TC 0.521 ( 241 mV vs. SHE) slightly higher. The enhancement observed is attributed to the tita-
NF/PANI/TC/Chit 0.538( 258 mV vs. SHE) nium carbide. Furthermore, the positive charges on the chitosan
3
Power density (W m ) NF 8.29
and the positive charges on PANI lead to higher adhesion of bacte-
NF/PANI 15.17
NFPANI/TC 15.59 ria, which are negatively charged yielding higher power density
NF/PANI/TC/Chit 18.82 and coulombic efficiency (Zhang et al., 2011; Tiwari and Singh,
Over potential at NF 170 2007; Higgins et al., 2011).
25.8 A m 3 (mV) NF/PANI 108
NFPANI/TC 121
NF/PANI/TC/Chit 97 4. Conclusion
Limiting current density NF 27
(A m 3) NF/PANI 68.8
NFPANI/TC 79.2 In conclusion, the biocompatible chitosan matrix embedded
NF/PANI/TC/Chit 109.6 onto the composites of conductive PANI-TC network could serve
Internal resistance (O) NF 116 as an efficient current collector in MFCs. The individual effects of
NF/PANI 68
chitosan, titanium carbides and PANI on nickel foam were investi-
NFPANI/TC 63
NF/PANI/TC/Chit 46 gated. The lower ohmic resistance was attained for all modified
Discharge capacity at NF 227 anodes compared with that of unmodified NF anode. The surface
50 O for 130 h (mA h) NF/PANI 312 morphology of NF/PANI/TC/Chit composite exhibited better bacte-
NFPANI/TC 343 rial adhesion, which combined with its fibrillar structure, made the
NF/PANI/TC/Chit 479
anodic reaction more facile for the transfer of electrons to the com-
Coulombic efficiency (%) NF 6.95
NF/PANI 9.55 posite interface.
NFPANI/TC 10.5
NF/PANI/TC/Chit 14.7
Acknowledgements
Note: NF – nickel foam, PANI – polyaniline, TC – titanium carbide, Chit – chitosan.

The authors would like to thank Dr. K. Manimaran, Senior Scien-

anode. For example the power density composite electrode have
improved 26–33% when compared to NF anode. This can be com-
parable to the studies on PANI and poly (aniline-co-o-
aminophenol) (PAOA) modified carbon felt anodes resulted in
about 35% and 18% improvement, respectively, when compare to
control carbon felt (Li et al., 2011). Moreover when compare to
control NF anode the composite electrode showed their reduction
of internal resistance into 16–23% and discharge capacity increased
into 23–35%.
tist, Electrochemical Power sources division, Central Electrochem-
ical Research Institute (CSIR-CECRI), India for providing us Nickel
foams to carry out this study.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.biortech.2016.02.
114.

3.5. Biocompatibility and surface morphology of the anodes References

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