Science of the Total Environment 456457 (2013) 9194

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Science of the Total Environment

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Short Communication

Enhanced electricity generation by using algae biomass and activated sludge in

microbial fuel cell
Naim Rashid a, b, Yu-Feng Cui a, Muhammad Saif Ur Rehman a, Jong-In Han a,
a
b

Dept. of Civil and Environmental Engineering, Korea Advance Institute of Science and Technology (KAIST), 373-1 Guseong dong, Yuseong-gu, Daejon 305-701, Republic of Korea
Dept. of Environmental Sciences, COMSATS Institute of Information Technology, Abbottabad, 22060, Pakistan

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

Electricity is generated using wastes.

Algae biomass and activated sludge are
used as substrate in microbial fuel cell.
A dry algae biomass of 5g/L resulted the
power output of 1.78W/m2.
Pre-treatment of activated sludge improved electricity production.
Lipid extracted algae is tested to replace whole algae for low cost electricity
production.

a r t i c l e

i n f o

Article history:
Received 16 January 2013
Received in revised form 13 March 2013
Accepted 17 March 2013
Available online 10 April 2013
Keywords:
Algae biomass
Activated sludge
Electricity
Microbial fuel cell
Pre-treatment

a b s t r a c t
Recently, interest is growing to explore low-cost and sustainable means of energy production. In this study,
we have exploited the potential of sustainable energy production from wastes. Activated sludge and algae
biomass are used as substrates in microbial fuel cell (MFC) to produce electricity. Activated sludge is used
at anode as inoculum and nutrient source. Various concentrations (15 g/L) of dry algae biomass are tested.
Among tested concentrations, 5 g/L (5000 mg COD/L) produced the highest voltage of 0.89 V and power density of 1.78 W/m2 under 1000 electric resistance. Pre-treated algae biomass and activated sludge are also
used at anode. They give low power output than without pre-treatment. Spent algae biomass is tested to
replace whole (before oil extraction) algae biomass as a substrate, but it gives low power output. This work
has proved the concept of using algae biomass in MFC for high energy output.
2013 Elsevier B.V. All rights reserved.

1. Introduction
Currently, the world is confronting with the challenges of high
energy demand and escalating fuel prices (Love et al., 2011). To cope
Corresponding author. Tel.: +82 42350 3629; fax: +82 42350 3610.
E-mail address: jihan@kaist.ac.kr (J.-I. Han).
0048-9697/$ see front matter 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.scitotenv.2013.03.067

with such challenges, researchers have paid considerable attention to

introduce sustainable and cost-effective methods of energy production
(Yinghua et al., 2013; Yuri, 1994; Scacchi et al., 2010). In this regard,
microbial fuel cell (MFC) has grasped widespread attention (Zhou
et al., 2012). MFC offers tremendous potential to produce energy in
the form of electricity (Venkata et al., 2008). MFC is a bioelectrical
chemical system for power generation based on the exploitation of

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N. Rashid et al. / Science of the Total Environment 456457 (2013) 9194

biocatalytic reactions with active microbial fuels (Rosenbaum and

Angenent, 2010). MFC is composed of two components, anode and
cathode. At anode, organic compounds are oxidized and electrons are
liberated (Wen et al., 2011). These electrons move through an external
circuit to the cathode. At cathode, electrons combine with an electron
acceptor (Rosenbaum and Angenent, 2010; Wang et al., 2010) to
generate electricity. Generally, pure organic compounds such as acetate, glucose and cysteine are used as substrate at anode. However,
their use is economically unviable (Vlyssides and Karlis, 2004). Alternatively, the mixture of biodegradable materials, such as domestic
wastewater, piggery wastewater, and animal wastewater can be used
as substrate (Martin et al., 2010; Rodrigo et al., 2007). Such types of substrates have some limitations. For example, the electricity generation
from pure substrates is economically unviable (Velasquez-Orta et al.,
2011). On the other hand, power output of mixed organic compounds
is quite low (Wen et al., 2011). To improve the power output and to
make MFC technology cost-effective, new substrates should be introduced (Yuri, 1994). In this perspective, the use of algae biomass as a
substrate can provide an opportunity for sustainable and low-cost production of electricity (Velasquez-Orta et al., 2009). Algae are potential
pollution vector in lakes and ponds causing eutrophication (Bubrick,
1991). A large amount of algae residues is produced during water treatment process. These residues are discharged into the sewer system
without pre-treatment, which increases the pollution load in downstream wastewater plants (Bringmann and Khn, 1980). Therefore,
algae residues have to be removed before being discharged into the environment. Algae residues can be used to produce a wide variety of
biofuels such as methane, hydrogen, biodiesel and bioethanol (Karube
et al., 1986; Scott et al., 2010). However, the biofuel production
from algae biomass is not cost-effective (Lehr and Posten, 2009).
To use algae biomass in MFC is a promising approach to generate
electricity. This approach serves dual purpose, waste treatment and
electricity generation. Certain bacteria such as Alcaligenes faecalis,
Enterococcus gallinarum, Pseudomonas aeruginosa, and Shewanella
use algae biomass as nutrient and transfer electrons to the electrodes.
Activated sludge is reported to contain various types of electricity
producing bacteria. Therefore, activated sludge is used as an inoculum
at anode. However, the bacteria can't consume algae biomass directly
due to its strong cell wall. Pre-treatment is a necessary step to break
the cell wall and to make it digestible for the microbes. Various
pre-treatment techniques can be used to make algae biomass digestible for the microorganisms.
In this study, we have proposed to use algae biomass as a substrate
for high power output. A mixture of activated sludge and algae biomass
is used as anolyte. Algae biomass, which is composed of proteins, carbohydrates and lipids, serves as nutrient source for the microbes present
in activated sludge. Thermal and sonication pre-treatments are employed
to improve net power output.
2. Material and methods
2.1. Algae biomass and activated sludge pre-treatment
Scenedesmus was selected as algae specie due to its high lipid contents and its use as nutrient source for the microbes, at anode. It was
obtained from Korea Research Institute of Bioscience and Biotechnology
(KRIBB), Republic of Korea. Scenedesmus was grown in an open pond
system. Its biomass was collected after 15 day cultivation and dried at
40 C until constant weight. For pre-treatment experiments, dry algae
biomass (after mixing with the medium, described in Section 2.3.)
was sonicated at 20 KHz at maximum power output of 2.2 kW for
2 min (Choi et al., 2011). Two types of pre-treatment were applied to
activated sludge: (1) sonication and (2) thermal pre-treatment. The
sonication conditions applied for algae biomass were used for activated
sludge as well. For thermal pre-treatment, activated sludge was kept at
70 C for 24 h (Vlyssides and Karlis, 2004).

2.2. MFC design

Cubical, dual-chamber reactors made by acrylic with each electrode
chamber holding a volume of 112 mL (7 cm 4 cm 4 cm) were
used in this study. Both, anode and cathode had working volumes of
100 mL. The reactor was separated by cation exchange membrane
(CEM, CMI-7000, Membrane International, Inc. USA) with an effective
surface area of 16 cm2 (4 cm 4 cm). A carbon brush of same conguration (L = 2.5 cm, D = 2.5 cm) connected with titanium wire was
used as cathode electrode. 0.4 mg/cm2 Pt catalyst (20 wt% Pt/C, Alfa
Aesar) was coated on both sides of carbon cloth. All experimental stuff
was autoclaved at 121 C for 20 min (Velasquez-Orta et al., 2009).
2.3. Operation
A mixture of activated sludge 50% (w/w), collected from Daejeon
Sewage Treatment Plant, Daejeon (Metropolitan City, Republic of Korea)
and media 50% (w/w) containing KH2PO4 5356 mg, K2HPO43H2O
164 mg, Na2HPO4 12H2O 11867 mg, MgCl26H2O 100 mg, NaHCO3
1000 mg, CaCl22H2O 66 mg, NH4Cl 500 mg, MnSO46H2O 15 mg,
FeSO47H2O 25 mg, CuSO45H2O 5 mg, CoCl25H2O 0.0125 mg, NiSO4
32 mg, ZnCl2 23 mg, and (NH4)6Mo7O24H2O 14 mg (per liter) (Yang
et al., 2011) was pumped into the anode compartment. Activated sludge
served as anodic inoculum (Patil et al., 2009). Aeration at a ow rate of
50 mL/min was provided in cathode during the MFC start-up operation.
The MFC start-up operation was carried out in fed-batch mode at 25
2 C. After three acclimation cycles, a replacement of total solution only
with substrate and medium was conducted and afterwards reproducible voltages were successfully obtained. Subsequently, power densities at an external resistance of 1000 at different concentrations of
substrate were measured.
Electrochemical properties were monitored with a data acquisition
system (Model 2700, Keithley instrument) connected to a personal
computer at 10 min intervals. The study-state voltage and current
were measured as I = V/R, where V is the measured voltage, R is the
external resistance and I is the current. The polarization curve was
obtained by plotting voltage vs current density. The power density (P)
was calculated from the measured voltage as P = V2/RA, where A is
the projected cathode surface area (Yuan et al., 2011). For discharge
operation, an external resistance of 1000 was connected to the circuit
(Logan and Regan, 2006). All polarization experiments were conducted
three times.
2.4. Analysis
Chemical oxygen demand (COD), protein, and carbohydrate were
measured by standard methods (American Public Health Association,
APHA, 1995) (Velasquez-Orta et al., 2009). Total solids (TS) and volatile
solids (VS) were measured by standard method (Method 5220, APHA
1995) (Cheng et al., 2006). Samples were ltered through 0.45 m
(pore diameter). The nitrate and phosphate contents of ltrates were
measured with reaction kit TNT plus HR (535 mg/L NO3-N, TNT 836,
Hach) and HR (1.0100 mg/L, TNT 10127, Hach), respectively.
3. Results and discussion
3.1. Electricity generation using activated sludge
Activated sludge was used at anode to generate voltage. The voltage produced by pre-treated and without pretreated activated sludge
(as control) was compared. In pre-treatment, thermal and sonication
techniques were employed according to the method mentioned in
Section 2.1. The basic characteristics of activated sludge are given in
Table 1. A voltage of 0.289 V was produced in un-pre-treated, 0.242 V
by thermal pre-treated and 0.400 V by sonicated sludge (Fig. 1). This
shows the positive effect of sonication on the voltage. The increase in

N. Rashid et al. / Science of the Total Environment 456457 (2013) 9194

TS
VS
Total COD
Soluble COD
Total carbohydrate (soluble)
Total protein
Total nitrogen
Total phosphate
pH

mg/L
mg/L
mg/L
mg/L
mg/L
mg/L
mg/L
mg/L

7980
6380
10110
15.3
32.4
93.06
11.23
5
6.2

50
50
460
5.3
0.26
3.06
3.66
1
0.05

voltage is probably because sonication disintegrates the activated

sludge and increases the dissolved carbohydrate concentration and
chemical oxygen demand (COD) in the electrolyte (Choi et al., 2011).
Denaturing Gradient Gel Electrophoresis (DGGE) analysis of anolyte
showed that Desulfovibrio sp., P. aeruginosa, Cytophaga xylanolytica,
Dechloromonas sp., Thiomonas perometabolis, and Cytophaga sp. were
dominant bacteria to produce electricity (data not shown). In our experiment, the soluble COD was increased from 15.3 5.3 in un-pretreated to 500 5 in pre-treated activated sludge mg/L (data not
shown). Increased carbohydrate and COD allowed the bacteria to
grow promptly and generate high voltage (Min et al., 2005). Decrease
in voltage in thermal pre-treated activated sludge could be due to the
suppression of bacterial activity at high temperature. Similar fashion
of voltage decrease was shown by Min et al. (2005).
3.2. Electricity generation using algae biomass as substrate
A mixture of activated sludge (as an inoculum), medium (as nutrient
source) and algae biomass (as substrate) was rst fed to the system, for
conditioning purpose. Different concentrations (1, 2 and 5 g/L) of algae
biomass were used to nd the optimal concentration (Fig. 2). A xed
concentration of sludge (50% w/w) and the medium (50% w/w) was
used. Open circuit voltage was quantied at each concentration. The initial voltage was accompanied with biomass concentration. The highest
initial voltage of 0.89 V was quantied at 5 g/L (5000 mg COD/L) concentration followed by 0.86 V at 2 g/L (2000 mg COD/L) and 0.84 V at
1 g/L (1000 mg COD/L), respectively. The increased voltage at high
biomass concentration is because the potential to release electrons is
accompanied with biomass concentration. The voltage increased when
the microbes had enough food to use in the form of algae biomass. The
maximum power density was 1.78 W/m2 under 1000 using 5 g/L of
algae biomass. The power densities were 0.85 W/m2 and 0.56 W/m2
at algae biomass of 2 g/L and 1 g/L. The cell voltage dropped to zero at
1.93 A/m2 and 1 g/L biomass, 2.27 A/m2 at 2 g/L biomass and 4.6 A/m2

0.45
0.4
Sonicate pretreated sludge

Cell voltage, V

Thermal pretreated sludge

0.8

1.8
1.6

0.7

1.4

0.6

1.2

0.5

0.4

0.8

0.3

0.6

0.2

0.4

0.1

0.2

Power density, W/m2

Value

Cell voltage, V

Unit

0
0

Current density, A/m2

Fig. 2. Voltage, current and power output as a function of algae biomass in MFC operation.

at 5 g/L of algae biomass (Fig. 2). A positive correlation between current

density and open circuit voltage was observed. A negative relationship
between the cell voltage and current density was found. The power
density decreased with increase in current density. Both, the power
density and current density were also the function of algae biomass
concentration.
High voltage using algae biomass as substrate proves its potential
to be used in MFC. A number of studies have been carried out showing
the potential of various substrates in MFC; however, the use of algae
biomass seems the best choice because of its ability to produce high
voltage (Min and Logan, 2004; Powell et al., 2010; Strik et al., 2008;
Bucksch and Egeback, 1999; Muhammad et al., 2013; Mehwish
et al., 2013). Table 2 shows a comparison of voltage produced by
various substrates.
3.3. Electricity generation using pre-treated algae biomass
We tested the effect of algae biomass pre-treatment on the voltage.
An optimized concentration (5 g/L) of algae biomass was mixed together
with activated sludge (50% w/w) and medium. The mixture was pretreated via sonication. The maximum voltage of 0.604 V was produced
in un-pre-treated algae biomass and 0.290 V in pre-treated mixture
(Fig. 3). It shows the negative effect of sonication. The plausible reason
for this phenomenon could not be known yet.
We also investigated to replace whole algae (without lipid extraction)
with spent algae (lipid extracted) to reduce the overall cost of bioelectricity production as lipid extracted algae contains protein (32.4%),
lipid (6.5%), carbohydrate (24.7%) and ash (10%) (Yang et al., 2011).
These retaining nutrients could be used by electricity generating bacteria. The voltage produced by lipid extracted algae was 0.021 V only
(Fig. 3). Based upon these results, it would be premature to conclude

Untreated sludge

0.25

Table 2
A comparison of power densities using different substrates in MFC.

0.2
0.15
0.1
0.05
0

1g/L
2g/L
5 g/L
Voltage
Power density

0.9

Parameters

0.3

Table 1
Characteristics of activated sludge.

0.35

93

10

20

30

40

Time, hrs
Fig. 1. Voltage prole of un-treated and pre-treated sludge in MFC operation.

Substrate

Power density, mW/m2

References

Glucose
Acetate
Acetate
Butyrate
Butyrate
Dextran
Starch
Swine wastewater
Dry algae biomass

212
286
661
220
349
150
242
225
1780

Min et al. (2004)

Min et al. (2004)
Min et al. (2005)
Min et al. (2004)
Min et al. (2005)
Min et al. (2004)
Min et al. (2004)
Min et al. (2005)
This study

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N. Rashid et al. / Science of the Total Environment 456457 (2013) 9194

0.7
0.6

Cell voltage, V

0.5
0.4

Algae biomass (w/o pretreatment)

0.3

Spent algae biomass

0.2

Sonicated algae biomass

0.1
0
0
-0.1

10

15

20

Time, hrs

Fig. 3. Voltage produced by pre-treated, un-treated and spent algae biomass in MFC.

that lipid extracted algae can't be used as substrate to generate bioelectricity. It is likely that toxic chemical retains in lipid extracted algae biomass while extracting lipids, which inhibit the growth of microbes in
MFC. Therefore, we suggest further exploring the chemical toxicity
effect during lipid extraction and its subsequent use in MFC. Recognizing
the potential of lipid extracted algae as MFC substrate would be a
meaningful contribution in MFC eld.
4. Conclusions
This work veries the concept of using algae biomass as a substrate
in MFC. It produces much higher power density than other reported
substrates. The use of algae biomass with activated sludge serves dual
purpose, the waste mitigation and electricity generation. To introduce
algae biomass as a substrate, further developments are required to
ensure its cost-effectiveness. Investigating the potential of lipid extracted
algae and by exploiting pre-treatment techniques would warrant high
energy output at low cost.
Acknowledgments
This work was nancially supported by the National Research
Foundation of Korea (NRF) grant funded by the Korea government Ministry of Education, Science and Technology (MEST) (NRF2012M1A2A2026587).
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