Bioresource Technology 123 (2012) 8691

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

Biological treatment of steroidal drug industrial efuent and electricity

generation in the microbial fuel cells
Ru Liu a, Chongyang Gao b, Yang-Guo Zhao a,c,, Aijie Wang b,, Shanshan Lu a, Min Wang a,
Farhana Maqbool a, Qing Huang a
a
b
c

College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China
School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090, China
Key Laboratory of Marine Environment and Ecology, Ocean University of China, Ministry of Education, Qingdao 266100, China

h i g h l i g h t s
3

" The MFCs reached maximum 22.3 W m

with steroidal drug industrial efuent.

" The maximum COD removal efciency of the MFCs approached 82%.
" The dominant microorganisms on anode were different from morphology to species.

a r t i c l e

i n f o

Article history:
Received 28 May 2012
Received in revised form 22 July 2012
Accepted 24 July 2012
Available online 2 August 2012
Keywords:
Microbial fuel cell (MFC)
Steroidal drug production wastewater
(SPW)
Bacterial community
Denaturing gradient gel electrophoresis
(DGGE)

a b s t r a c t
The single chamber microbial fuel cells (MFCs) were used to treat steroidal drug production wastewater
(SPW) and generate electricity simultaneously. The results indicated that the maximum COD removal
efciency reached 82%, total nitrogen and sulfate removal rate approached 62.47% and 26.46%, respectively. The maximum power density and the Coulombic efciency reached to 22.3 W m3 and 30%,
respectively. The scanning electron microscope showed that the dominant microbial populations were
remarkably different in morphology on the surface of SPW and acetate-fed anodes. PCR-denaturing gradient gel electrophoresis proles revealed that the microbial community structure fed with different concentrations of SPW presented a gradual succession and unique bacterial sequences were detected on the
SPW and acetate-fed anodes. This research demonstrates that MFCs fed with SPW achieved a high efciency of power density and simultaneous nutrient removal, and the dominant microorganisms on the
anode were related to the types and the concentrations of substrates.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
Pharmaceutical industrial efuent is difcult to treat biologically for its complex composition and high toxicity; this consequently leads to more harmful to water environment than other
types of sewage. Steroid drug mainly refers to steroids hormone,
which usually contains the complex polycyclic structure. During
the production of steroid drug, hydrocortisone is one of the important intermediates. It is commonly made from diosgenin in some
pharmaceutical factories in Shandong Province, China. Some investigations revealed that a large number of high toxic, rening
wastewater was generated during the production process of
hydrocortisone, especially in its acid hydrolysis stage (Cheng
et al., 2009; Li et al., 2010).
Corresponding authors.
E-mail addresses: yg.zhao@yahoo.com.cn (Y.-G. Zhao), waj0578@hit.edu.cn
(A. Wang).
0960-8524/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.biortech.2012.07.094

Traditional biotechnology and physicalchemical technology

had been applied to treat steroidal drug production wastewater
(SPW). Coagulationocculation pretreatment was adopted to reduce the COD and turbidity (Liang and Wang, 2010). Nevertheless,
it transferred the pollutants from the liquid to the solid phase. A
combination of modied two-phase anaerobic digestion and biological aerated lters was proposed to treat the SPW effectively
(Cheng et al., 2009; Zhao et al., 2008a). However, it consumed large
amounts of electricity and produced a lot of biochemical sludge
during the process. Nanoltration technology was attempted to reduce the discharge by reclaiming acid, sugar and water from SPW
(Peng et al., 2010), yet ultraltration membrane lifetime limits its
application in engineering widely. In summary, all of these processes have not efciently accomplished wastewater disposal and
simultaneous resource recovery.
Microbial fuel cells (MFCs), the technology developed on the basis of electrochemical techniques, is a device utilizing microorganisms as catalysts to mediate direct conversion of chemical energy

R. Liu et al. / Bioresource Technology 123 (2012) 8691

stored in organic matter into electrical energy. It can remove contaminant in wastewater and produce electricity simultaneously,
thus decrease the operational cost of wastewater treatment. Its potential of renewable energy from wastewater has attracted
researchers0 attention in recent years (Huang and Logan, 2008;
Rahimnejad et al., 2012). Several types of wastewater had already
been treated successfully and simultaneously generated electricity
using MFCs (Pant et al., 2010).
The population and activity of the bacteria on the anode are
very important for the electricity production capacity and SPW
treatment efciency. To date, no literatures were found addressing
the electricity-producing ora during SPW degradation. In the traditional biological treatment, the investigation of SPW degrading
bacteria was still in its infancy.
In the present study, the experiments were designed to investigate the effect of inuent SPW concentrations on performance of
electricity generation and wastewater treatment. Meanwhile, the
biolm morphology and microbial community structure on the
surface of MFC anodes were explored by using scanning electron
microscope (SEM) and PCR-denaturing gradient gel electrophoresis
(DGGE) techniques. The results provided new ideas for the SPW
control and utilization of resources.
2. Methods
2.1. Steroidal drug production efuents characteristics
Steroidal drug production wastewater was obtained from
hydrocortisone producing factory in Zibo, Shandong Province, China. The wastewater contained 6720 mg L1 COD, 760 mg L1 total
nitrogen (TN) and 2419 mg L1 sulfate. The pH and the conductivity was 9.2 and 8.63 mS cm1, respectively.
2.2. MFC conguration
The air-cathode single-chambered membrane-less MFCs were
constructed by Plexiglas vessel (working volume of 28 mL) as described by Liu and Logan (2004). Carbon brush and carbon cloth
(Ruibang Carbon Material Co., Shanghai, China) were used as anode
and cathode respectively. Air-cathode containing Pt catalyst
(0.5 mg Pt cm2) was prepared as the method described by Cheng
et al. (2006). Titanium wires were used to connect the circuit and
all wire interfaces were sealed with epoxy resin. Unless otherwise
specied, the external resistance used for these experiments was
1 kX. The MFCs were inoculated with the anaerobic sludge from
an anaerobic bafed reactor treating high sulfate wastewater. During the acclimation period, the anolyte was comprised of
500 mg L1 acetate (in COD), phosphate buffered saline (PBS)
(137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO42H2O, 2 mM KH2PO4,
pH 7.4), trace mineral and vitamin. When the biolm on the anode
matured, the acetate in chamber was replaced by 0.1, 0.2, 0.5,
and 1.0 SPW in turn. Each concentration was operated at least
two stable cycles. SPW was diluted with 1.0  PBS and water to
avoid a sudden change of conductivity (Logan, 2012). The conductivity and pH of inuents were nally maintained at 8 1 mS cm1
and 8.5 0.5. The MFCs were operated in fed-batch mode at an
ambient room temperature (2025 C).
2.3. Electrochemical measurements
Voltage (U) across the external resistor (R) in the MFC was measured using a data acquisition system (PISO-813,Hongge Taiwan)
connected to a computer. Current (I) was calculated from I = U/R,
and cell power output (P) was calculated from P = U  I. The power
density (PV and PA) was calculated using the equation:

87

PV W m3 P=V; P A mW m2 P=A

where V is the volume of the anodic chamber, A is the anode surface
area. The polarization curves were obtained by changing external
circuit resistances from 1000 to 20 X during the steady state. The
data were recorded at a time interval of 10 min. The slope of UI
curve indicated the internal resistance (Rin) of the cell (Logan
et al., 2006). The Coulombic efciency (CE) was calculated by integrating the measured current relative to the theoretical current
based on the consumed COD (Oh et al., 2004).
2.4. Chemical analysis
Quick digestion spectrophotometry (Beckman Coulter DU800,
CA, USA) was employed to analyze chemical oxygen demand
(COD). The volatile organic acids (VFAs) in the anodic solution were
quantitatively measured by a gas chromatograph (GC-2010, Shimadzu, Japan). Before measurement, the solution was ltered by
microporous membrane to remove the impurities.
2.5. Microbial community analysis
2.5.1. Scanning electron microscopy (SEM) analysis
At the end of each current production, a small pieces of SPW-fed
and acetate-fed anode electrodes were taken to examine the bacterial morphology using a SEM. Electrodes were removed from the
chambers, and prepared according to Chung and Okabe (2009).
The samples were nally observed in a scanning electron microscope (S-4800, Hitachi, Japan).
2.5.2. Microbial community structure analysis
Attached biolm samples were collected from carbon anode
surface at the end of each stage. Genomic DNA was extracted directly by the Soil DNA extraction kit (Mobio, CA, USA) according
to manufacturers instructions. PCRDGGE and 16S rRNA gene
sequencing for community analysis were conducted as previously
described (Zhao et al., 2008b).
After DGGE proles were digitized, ShannonWiener diversity
index (H0 ) analysis was computed according to formula,
P
H0  Pi log Pi, where Pi is the relative signal intensity of each
band in lane. SPSS software (SPSS Inc., Chicago IL) was used to conduct cluster analysis for the DGGE proles by Wards method.
2.6. Accession number of DNA sequence
Sequences obtained in this study had been deposited in GenBank, and the Accession Nos. were JX069268JX069294.
3. Results and discussion
3.1. Electricity generation from steroidal drug industrial efuent in
MFC
When the biolm on the anode matured, 0.1  SPW
(670 mg L1 COD) was used as the anolyte to investigate the feasibility of MFC treating SPW. Fig. 1 showed the output voltage curve
when the anolyte transferred from acetate (500 mg L1 COD) to
0.1  SPW. The rst stable cycle was acetic anolyte, the following
two cycles were fed with 0.1  SPW. The voltage presented no
obvious uctuations after anolyte replacement except that the
voltage dropped slightly.
The polarization and power density curves (Fig. 2) as a function
of current density were obtained by changing external circuit resistances from 1000 to 20 X during the steady state. The maximum
power density (MPD) of the MFC utilizing 0.1  SPW was
18.1 W m3 (Area power density was 403.5 mW m2), the internal

88

R. Liu et al. / Bioresource Technology 123 (2012) 8691

Fig. 1. Time-potential relationship in MFC during the transition of substrates. The

arrow of SPW indicates the point where the acetate (500 mg L1 COD) was replaced
by the 0.1  steroidal drug production wastewater (670 mg L1 COD).

Fig. 2. The polarization curves of the MFCs fed with the 0.1  steroidal drug
production wastewater (670 mg L1 COD) and acetate (500 mg L1 COD),
respectively.

resistance was approximately 32 X; while the acetate-fed MFC

was about 18.2 W m3 and 33 X, respectively.
The higher concentrations of SPW (1340, 2240, 3360 and
6720 mg L1 COD) were utilized as anolyte in turn. The conductiv-

ity of inow was adjusted to achieve the same level using phosphate buffer. Fig. 3A showed the output potential obtained from
SPW and acetate at a steady stage. Fig. 3B showed the maximum
voltage of different anolytes and their electricity duration chart.
As shown in Fig. 3, voltage rose rapidly after changing anolytes.
As the SPW concentration increased, the maximum voltage increased rst and then decreased. When the MFC treated 0.2  SPW,
it reached to its maximum voltage at 520 mV. And the MPD of the
MFC approached to 22.3 W m3. Voltage duration extended with
the increase of concentration of SPW. The maximum voltage fed
with 0.1  SPW was 0.49 V, lasted for about 27 h, while the maximum voltage fed with 1  SPW was 0.46 V and extended to 80 h.
We examined the content of VFAs in the raw SPW. It contained
about 400 mg L1 VFAs, most of them was acetate (Fig. 4). Therefore, 0.1  SPW contained approximately 40 mg L1 acetate. This
result would explain why no lagging phase took place when the
substrates were replaced. The MFC fed with 0.2  SPW achieved
the MPD of 22.3 W m3, which showed a better electricity generation capacity than some types of substrates in previous researches
(Ahn and Logan, 2010; Sun et al., 2009; Zhuang et al., 2009). Ahn
and Logan (2010) reported a MPD of 7.8 W m3 in a domestic sewage-fed single-chamber air-cathode MFC. And Sun et al. (2009) and
Zhuang et al. (2009) reported the MPD of 0.8 and 7.7 W m3 using
industrial efuent and brewage wastewater, respectively.
It had been found that the electricity generation efciency and
degradation rate of COD were substantially improved after adding
50 mg L1 penicillin or ceftriaxone sodium in air cathode singlechamber MFC (Wen et al., 2011a,b; Chung and Okabe, 2009). These
results concluded that some toxic and bio-refractory biomass such
as antibiotic wastewater might be available resources for electricity generation using the MFC technology. This was consistent with
the present results.
As the concentrations of SPW increased, the degradation duration of organics in SPW became longer. Accordingly, electricity production duration and power output rose. However, better
electricity generation efciency was not related with the higher
the concentration of SPW. In present study, the 0.2  SPW was
the most favorable concentration, the MPD reached to
22.3 W m3, and the potential achieved 0.52 V. In addition, microorganisms would be affected by osmotic pressure if the concentration was too high, this would further inhibit microbial metabolism
rate and decrease power production efciency.
3.2. High efcient degradation of SPW in MFC
COD, TN and sulfate removal rates in anode chambers were
monitored. The CE per cycle was also calculated (Table 1).

Fig. 3. The potential and duration in the MFCs fed with acetate and different concentrations of the steroidal drug production wastewater (SPW) (A) and the duration of
maximum voltages with different substrates (B). 0.1  SPW, 0.2  SPW, 0.5  SPW, 1.0  SPW mean the corresponding concentration of SPW.

R. Liu et al. / Bioresource Technology 123 (2012) 8691

Fig. 4. The relationship between voltage output and volatile organic acids (VFAs) in
a batch MFC fed with the raw steroidal drug production wastewater (1.0  SPW),
Error bars indicate the standard error from two neighboring cycles in one MFC.

Table 1
The Coulombic efciency and nutrient removal rates with different substrates in
MFCs.
Inuent COD
(mg L1)

COD
removal rate
(%)

Coulombic
efciency (%)

TN
removal
rate (%)

Sulfate
removal
rate (%)

Acetic acid, 500

0.1  SPW, 670
0.2  SPW, 1340
0.5  SPW, 3360
1.0  SPW, 6720

84
72
70
82
78

60
30
24
12
10

62
56
48
39

26
24
21
16

COD removal efciency of the MFC fed with different SPW uctuated from 70% to 82%, and 0.5  SPW achieved its maximum removal rate of 82%. In this MFC, the similar COD removal rate (84%)
was also obtained with acetate. In addition, TN and sulfate removal
rate declined obviously as the SPW concentration increased. TN removal efciency decreased from 62% to 39%, and sulfate removal
rate decreased from 26% to 16% with the increasing of SPW concentrations. According to the amount of sulde detected in the MFC
(data not shown), most of removed sulfate was reduced to sulde,
which will impact heavily on the environment for its odor and corrosion. Therefore, further recovery of sulde by biological or chemical methods is often required.
During stabilized period of 1.0  SPW, the anodic solution was
taken every three hours to measure VFAs. One cycle of voltage
and VFAs content was shown in Fig. 4. As SPW degraded, VFAs concentration gradually increased, this indicated that the SPW degradation was accompanied by the production of VFAs. Afterwards,
the VFAs gradually decreased, and was fully degraded by anode microbe at the end of the cycle. This means that, when microorganisms produced electricity, they degraded small molecular weight
organic matter rst and then gradually degraded macromolecular
organic matter. The same trend of VFA in anode chamber was also
discovered in former research (Liu et al., 2012). They found that
lactate was used directly by some bacteria, some other zymophyte
degraded lactate and generated intermediate species, which made
the concentration of VFAs high and then lower down afterwards.
In the eld of wastewater treatment, degradation and effective
utilization of substrates in wastewater are practically important.
Steroidal drug production wastewater contained methanol, acetate, ethanol, benzene, toluene, acetone, chloroform, pyridine,
ethyl acetate, chlorobenzene, cyclohexanone, dichloroethane, fermentation medium, reaction intermediate and the lost products

89

(Li and Ni, 2011). In this research, the CE reached 24% when inow
was 0.2  SPW, which was better than brewery wastewater as the
substrates (in which substrate was COD 1600 mg L1, CE achieved
15%) (Feng et al., 2008), and equal to straw hydrolyzate (in which
COD was 2501000 mg L1 and CE was about 2030%) (Zuo et al.,
2006). As shown in Fig 1, the CE of MFCs fed with 0.1  SPW was
much lower than that of acetate-fed MFC, and the CE decreases
remarkably with the increase of SPW concentration. This might
be due to the fact that the SPW contained high complicated organic
pollutants and sulfate. The pollutants needed to be decomposed
into small molecule before used by microorganisms to produce
electricity and this consumed large of electrons (Huang and Logan,
2008).The sulfate concentration became higher as the concentration of the wastewater increased. Sulfate reduction would consume some electron in anaerobic environment and this resulted
in the drop of CE (Feng et al., 2008; Zhang et al., 2006; Zhao
et al., 2008b; Schroder, 2007). In the present study, the raw SPW
contained 25 mM sulfate. According to Table 1, approximately
4 mM sulfate was reduced to sulde, which would at least capture
32 mM electrons. According to the COD removal, the COD provided
654 mM electrons. Thus, the sulfate reduction would capture about
5% of provided electrons. This consequently decreased the CE.
The maximum COD removal rate was 82% in the present MFCs,
which reached the similar level as acetate-fed MFC. Previous researches (Wen et al., 2011a,b) found that a small amount of antibiotics accelerated degradation of COD. The COD removal rate rose
from 88% to 96% when 3050 mg L1ceftriaxone were dosed. It inferred that a little quantity of steroid might improve organic degradation rate by microorganisms, this was further investigated.
3.3. Characteristics of microbial community on the surface of MFC
anodes
The SEM analysis showed that the rough surface of the anode
electrode was almost covered by different types of bacteria, while
the obvious differences in bacterial morphology of the two types of
anodes were observed by the SEM (Supporting Information,
Fig. S1). The anode surface of the SPW-fed MFC was covered with
a substantial number of the coccoid cells (5 lm in diameter) and
a small amount of rod-shaped cells. However, on the surface of
the acetate-fed MFC anode, the biolm heavily inhabited with
homogeneous rod-shaped, about 12 lm long bacteria.
The structures of microbial communities at the anodes of acetate-fed and SPW-fed MFCs were investigated using the DGGE
(Fig. 5). ShannonWiener diversity analysis was conducted for
DGGE proles to make quantitative description about the changes
of populations and numbers of microorganisms in different stages.
Diversity evaluated the richness of species within a microbial community, as well as the number of microorganisms. According to
Fig. 5, there were more bands on the acetate-fed lane, and the index of microbial diversity was 3.0. This indicated that the microbial
community was abundant in this period. After the replacement of
substrate, the DNA bands are signicantly reduced, and the index
of microbial diversity decreased to about 2.8. In the SPW environment, the diversity index gradually increased to the level of acetate-fed MFC when the MFC treated 0.5  SPW. When inuent
changed to the 1.0  SPW, diversity index dropped nally to 2.5.
Species and the number of microorganisms presented signicant uctuations in the process of the cascade changes of the inuent substrates. After 20 days, when the acetate (A1) was replaced
with 0.1  SPW (S1), the community structure separated approximately 7% in distance (Fig. 5B). Compared with that, the distance
prolonged to 20% (about 70 days later) when inuent was changed
to the 1.0  SPW. The gradual community succession process was
due to the changes of carbon substrates of anolyte. However, the
community succession process presented obvious hysteresis; this

90

R. Liu et al. / Bioresource Technology 123 (2012) 8691

A1 S1 S2 S3 S4

A1 S1 S2 S3 S4

10

15

20

3.00 2.89

2.76

2.99

2.51

25

Fig. 5. DGGE proles of microbial communities of the MFCs (A) and cluster analysis
for DGGE proles (B). A1, S1, S2, S3 and S4 are the anodic biolms fed with acetate,
0.1 , 0.2 , 0.5 , 1.0  steroidal drug production wastewater (SPW), respectively.
The numbers under each lane in pattern A are their ShannonWiener indices; the
numbers on the right of bands indicate the bands to be sequenced; the scale on the
right of pattern B is the separation distance (%).

indicated that the enrichment of electricity-producing microbes

might take longer time. With different concentrations of SPW
(Fig. 5, S1S4), the structures of microbial community have higher
similarity, especially, in the adjacent lanes. This revealed the obvious community succession with the series of substrates.
SEM images showed that the main bacteria on the anode surface were coccoid cells in the SPW-fed MFC, while the main bacteria were rod-shaped cells in the acetate-fed MFC. DGGE proles
also indicated that the varieties and microbial number presented
great changes during the carbon substrates switching process.
The difference was further conrmed by comparing the band sequences with the database (Table 2). For example, the clones similar to Acidovorax sp., Holophaga sp., Anaerolum sp., Spirochaeta sp.
and Propionibacterium sp. (Band Nos. 10, 17, 20, 15, 19, 24 and 25)
were only found on the SPW-fed anode, while the clones similar to
Flavobacterium sp., Fusobacterium sp. and Comamonas sp. (Band
Nos. 9, 12, 18 and 23) were discovered in the acetate-fed MFC.
Velasquez-Orta et al. (2011) found that the bacterial morphology
had great differences between acetate-fed MFC and starch-fed
MFC. Zhang et al. (2006) also found that microorganisms differed
from each other in MFCs enriched with acetate and glucose. The results conrmed that different substrates lead to signicant difference of dominant microbial species on the MFC anode surface.
The communities on the anodes shared a portion of species found
previously in the MFC, such as Desulfovibrio sp., Dysgonomonas sp.,
Pseudomonas sp., and Desulfobulbus sp., which were proved to produce electricity in MFCs (Holmes et al., 2004; Rabaey et al., 2005;
Zhao et al., 2008b).
Some microorganisms in SPW-fed MFC anode were previously
reported to be able to metabolize macromolecular organic matter.

Table 2
Sequence similarities to closest relatives and phylogenetic afliations of DNA recovered from DGGE gel.
Band

Closest relative

No.

Length
(bp)

Accession
No.

Organism

Accession
No.

Similarity
(%)

Phylogenetic
afliation

Isolation source

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

189
189
194
194
195
194
169
194
189
194
194
189
194
172
172
194

JX069268
JX069269
JX069270
JX069271
JX069272
JX069273
JX069274
JX069275
JX069276
JX069277
JX069278
JX069279
JX069280
JX069281
JX069282
JX069283

Dysgonomonas gadei strain JCM 16698

Dysgonomonas gadei strain JCM 16698
Comamonadaceae bacterium
Desulfobulbus rhabdoformis (T)
Pseudomonas sp.
Desulfovibrio sp.
Rhodopseudomonas sp.
Pseudomonas sp.
Flavobacterium sp.
Acidovorax caeni strain R-24608T
Desulfovibrio sp. H1
Flavobacterium sp.
Bacterium DR.Y13
Synergistes genomosp. C1
Anaerolum pentosovorans (T)
Desulfovibrio desulfuricans strain Ser-1

AB548675
AB548675
AB114618
U12253
HQ003454
FJ225426
AB272323
AY303276
AF368756
AM084006
FJ225426
AF368756
DQ226214
AY278615
X97852
EU980605

100
100
99
96
99
95
99
100
99
99
95
99
100
99
99
99

Bacteroidetes
Bacteroidetes
b-Proteobacteria
d-Proteobacteria
c-Proteobacteria
d-Proteobacteria
a-Proteobacteria
c-Proteobacteria
Bacteroidetes
b-Proteobacteria
d-Proteobacteria
Bacteroidetes
c-Proteobacteria
Pyramidobacter
Firmicutes
d-Proteobacteria

17
18
19
20
21

194
173
194
194
194

JX069284
JX069285
JX069286
JX069287
JX069288

Acidovorax delaeldii strain DSM 50263

Fusobacterium sp.
Spirochaeta caldaria (T) strain DSMZ7334
Acidovorax delaeldii strain DSM 50263
Desulfovibrio desulfuricans strain Essex 6

AJ420323
DQ355179
EU580141
AJ420323
AF192153

100
100
96
99
100

b-Proteobacteria
Fusobacteria
Spirochaetes
b-Proteobacteria
d-Proteobacteria

22
23
24

195
194
194

JX069289
JX069290
JX069291

Smithella sp.
Comamonas testosterone strain Q10
Holophaga foetida (T) DSM 6591

EU888819
AF519533
X77215

100
100
100

d-Proteobacteria
b-Proteobacteria
Acidobacteria

25

173

JX069292

AJ704571

93

Actinobacteria

26
27

176
172

JX069293
JX069294

Propionibacterium jensenii (T) strain DSM

20535
Atopobium sp.
Lachnospiraceae bacterium strain DSM
11261

Infected gall bladder of a man

Infected gall bladder of a man
Rice plant
Wateroil separation system
Gurudongmar Lake
Wastewater digestor in Tunisia
Upland eld Soil
Upland and paddy soils in Korea
Ultrapure water systems
Activated sludge
Wastewater digestor in Tunisia
Ultrapure water systems
Soils
Human mouth
Anaerobic bioreactors
UASB reactors of the brewery
industries
Soil
Digestive tract
Freshwater hot spring
Soil
Abdominal or brain abscesses or
blood
UASB reactor
Activated sludge
Black anoxic freshwater mud
sample
Butter milk

HQ616367
AF550610

97
95

Actinobacteria
Firmicutes

Human oral cavity

Polluted pond in Hamilton New
Zealand

R. Liu et al. / Bioresource Technology 123 (2012) 8691

Although they did not produce electricity, these syntrophic

microbial species also participated in the degradation process.
For example, Acidovorax delaeldii (Band Nos. 17 and 20) degraded
biodegradable phenol and formaldehyde (Maneeld et al., 2005),
Holophaga foetida (Band No. 24) metabolized the methyl groups
of methoxylated aromatic compounds either to acetate or to dimethyl sulde (Kappler et al., 1997), Anaerolum pentosovorans
(Band No. 15) reduced the sulfate and produce H2S using the
hydrosoluble polyacrylamide as the sole carbon source. Hence,
they were break a variety of compounds in SPW down into small
molecules and then these complex compounds were utilized by
electricigens such as Dysgonomonas sp., Pseudomonas sp.(Band
Nos. 1, 2, 5 and 8), which potentially represented the possible reasons for variation tendency of VFAS in anode chamber (Fig. 4).
However, these bacterial consortia would decrease CE in MFCs
due to their competition with electricigens for electron donors.
ShannonWiener diversity analysis showed that the index rst
decreased and then increased as inow changed. The microbial
diversity decreased, possibly ascribing to the fact that there were
less microorganisms able to use the complicate pollutants in
SPW, and the sudden switching of SPW limited the growth of part
of microorganisms. In the SPW environment, after a period of acclimation, microbial diversity and density began to increase. At the
same time, the diversity index gradually returned to the acetate
value in the 0.5  SPW. It indicated that the microorganisms able
to degrade SPW increased gradually. At last, at the raw SPW stage,
the number of microorganisms has a signicant decline, indicating
that high concentration of SPW was not conducive to the growth of
the electricigens and this conclusion was further conrmed by
voltage output (Fig. 3) and wastewater treatment (Table 1).
4. Conclusion
The single chamber MFCs were used to treat steroidal drug
industrial efuent and generate electricity simultaneously. They
achieved a high efciency of power generation and simultaneous
nutrient removal. The maximum COD and total nitrogen removal
efciency reached 82% and 62.47%, respectively. And the maximum
power density approached to 22.3 W m3.
The electricigens and non-electricigens were both detected on
the anode and the dominant microorganisms were remarkably different from morphology to species in MFCs fed with different concentrations of steroidal drug industrial efuent.
Acknowledgements
We thank Dr. Xin Wang in Nankai University, China, for his
technical support. This work was supported by the Scientic Research Foundation for the Excellent Middle-Aged and Youth Scientists of Shandong Province of China (BS2011NJ018) and the
National Natural Science Foundation of China (Grant No.
40801193).
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.2012.07.
094.
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