i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1

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Agro-food industry wastewater treatment with

microbial fuel cells: Energetic recovery issues

Daniele Cecconet, Daniele Molognoni, Arianna Callegari,

Andrea G. Capodaglio*
Department of Civil Engineering and Architecture (D.I.C.Ar.), University of Pavia, 27100 Pavia, Italy

article info abstract

Article history: Wastewater treatment, necessary for the preservation of water and environmental quality,
Received 5 June 2017 usually requires considerable energy inputs to obtain desired targets. New paradigms of
Received in revised form circular economy require that new technological approaches for energy and resource re-
24 July 2017 covery should be implemented in lieu of traditional, energy-hungry technologies. Microbial
Accepted 30 July 2017 fuel cells represent an eco-innovative technology for energy and resources recovery from a
Available online 18 August 2017 variety of wastewaters. Agrofood wastes are specially indicated due to their high biode-
gradability. The current research was conducted to: assess bioelectrochemical treatability
Keywords: of dairy wastewater by MFCs, determine operational effects on MFCs electrical perfor-
Bioelectrochemical systems mance and evaluate possible strategies to reduce overpotentials. For this purpose, two
Bioenergy parallel MFC reactors were continuously operated for 2.5 months, fed with undiluted dairy
Microbial fuel cells wastewater. The study demonstrated that these types of industrial effluents can be treated
Agrofood wastewater by MFCs with high organic matter removal, recovering a maximum power density of over
Dairy 27 W/m3. Achieved results were better than previous MFC-experiences dealing with dairy
Eco-innovative technologies (and other types of) wastewater treatment, and show that recovery of energy from treat-
ment of organic wastes is a feasible strategy on the pursuit of sustainable technologies.
© 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

new technological approaches for energy and resource re-

Introduction
Wastewater treatment, necessary for the preservation of
water and environmental quality, usually requires consider-
able energy inputs to obtain desired purification targets. On
average, the energetic consumption of wastewater treatment
with conventional processes is about 0.2e0.8 kWh/m3,
depending on various factors, including wastewater and pro-
cess type. In case of aerobic processes, the energy used for
mixed liquor aeration (oxygenation) alone can amount to
more than 50% of the total energy used by the treatment fa-
cility [1]. New wastewater cycle paradigms [2,3] suggest that
covery should be implemented in lieu of traditional, energy-
hungry technologies, such as the standard Activated Sludge
(AS) process and its modifications (including membrane bio-
reactors, MBRs). The agro-food industry, in particular, pro-
duces massive amounts of organic materials as secondary
waste streams that could be tapped for energy recovery and
resources [4]. Several alternative processes have been pro-
posed, including Granular Activated Sludge (GAS) in the aer-
obic processes realm, and Upflow Anaerobic Sludge Blanket
(UASB) in the anaerobic one. The former allows considerable
energy savings (not to mention footprint and volumetric

* Corresponding author.
E-mail address: capo@unipv.it (A.G. Capodaglio).
http://dx.doi.org/10.1016/j.ijhydene.2017.07.231
0360-3199/© 2017 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1
reduction) compared to traditional AS processes [5], while the
latter would reduce energy consumption to the bare mini-
mum necessary for wastewater circulation and allow recovery
of a mixture of biogas, largely composed by methane [6].
Microbial electrochemical systems (MESs), also appear to
provide a potentially attractive alternative to wastewater
treatment processes with energetic recovery, within the cur-
rent global economic framework. In MESs, like microbial fuel
cells (MFCs), electroactive bacterial species (EABs) rely on the
same substances used in anaerobic biodegradation to transfer
electrons and directly produce current from the chemical
energy, contained in an organic biodegradable substrate [7].
In MFCs, EABs catalyse (one or both) oxidation and reduc-
tion reactions, each occurring in a separate chamber (anode
and cathode) and containing the respective electrodes, con-
nected by an external electrical circuit. Internally, the cham-
bers are separated by a selective ionic exchange membrane
(IEM), so that electrons and protons released by the bacteria
while degrading the substrate at the anode can travel towards
the cathode, where they combine with the terminal electron
acceptor of the reaction (usually, oxygen) [8,9]. Advantages
characterizing MFC processes, compared with other state-of-
the-art wastewater treatment processes are several: first,
these processes happen to be extremely versatile in reference
to treatable substrates. Experiences have been reported in
literature with simple liquid substrates, such as glucose [10],
volatile fatty acids [11] and alcohols [12], as well as with
complex mixtures, such as domestic wastewater [13,14],
brewery [15], dairy [16e18], winery [19,20], soybean edible oil
refinery [21], distillery [22], food-processing effluents [23].
Even when used to treat landfill leachate (usually considered
somewhat refractory to biological treatment), reported
organic removal rates reached 7 kg COD (Chemical Oxygen
demand)/m3 d [24,25], better than standard aerobic processes
(0.5e2 kg COD/m3 d), and in line with those of anaerobic
digestion (8e20 kg COD/m3 d). MFCs have been reported being
able also to remove (partially or completely) pharmaceutically
active compounds from wastewater [26]. MFC applications to
the degradation and energy recovery from solid food industry
waste have also been reported recently [27]. Additional, non-
negligible advantages over other biological processes are low
biofilm yield, possibility to operate at low temperatures,
strong reduction of aeration needs (only supplied as electron
acceptor at the cathode in dual cell systems only), direct
electricity production.
MFCs, however, are still characterized by some drawbacks,
including low specific electric production (10e100 W/m3 of
total reactor), that are still limiting the industrial applicability
of this technology [28]. Attempts of MFCs electric optimization
include development of non-Platinum-group catalysers for
oxygen reducing cathodes [29], morphologic modification of
the electrodes and combination of different electrode mate-
rials [30e32], development of biocathodes [33], hydraulic and
electric stacking of several MFC units [34], tracking methods
for the optimal external resistance [35].
Utilization of agrofood substrates (swine manure, brewery,
dairy or winery effluents) is particularly promising for MFCs
industrialization, given their high organic content (COD
values can go up to 10 g/L) and biodegradability (generally
BOD/COD ratio is higher than 60%) [36,37]. However,
501
treatment of complex substrates increases bacterial commu-
nity complexity, and this can lead to unexpected interrelated
connections between microbial species [38]. The anode
chamber anaerobic condition may induce unwanted side-
reactions such as methanogenesis, which directly competes
with electric current production, or heterotrophic denitrifica-
tion [39]. It was demonstrated that high fermented substrates
concentrations favour methanogenic activity against exo-
electrogenesis, reducing MFCs' electrical efficiency [40].
Dairy industry occupies one of the leading positions in the
Italian agro-food sector (Italy also produces 6% of the world's
cheese production). In industrial cheese-making processes,
about 2e4 L of wastewater are produced per L of processed
milk [41], with qualitative characteristics summarized in
Table 1. The organic fraction consists usually of easily biode-
gradable organic substrates (lactose), with the presence of
some slowly biodegradable lipids and proteins [42]. Nitrogen is
mainly present in organic form, bonded in milk proteins,
while phosphorous is mainly in orthophosphate form. De-
tergents and additives used in industrial and cleaning pro-
cesses can significantly affect pH and alkalinity of
wastewater.
Bioelectrochemical recovery of electricity from dairy
wastewater with MFCs has been previously investigated in
recent years, as summarized in Table 2. Single chamber, dual-
chamber, and tubular MFCs have been tested, with electrodes
of different materials including carbon, graphite, stainless
steel or composites. The highest reported power density
(20.2 W/m3) so far was achieved by Mardanpour et al. [45],
using a tubular MFC equipped with a 0.5 mg Pt cm
2 catalyst
load on the cathode. Other studies demonstrated that cathode
catalysts may be avoided, without negative effects on organic
matter removal efficiency, but with considerable reduction of
power density (90% lower) and Coulombic Efficiency (CE, 44%
lower) [46,47]. Experiences of continuous-fed MFCs treating
dairy wastewaters achieved mixed results [17,18].
Despite all the promising results obtained mostly in short-
term studies (1 month maximum duration), MFCs scientific
literature is still lacking comparative long-term studies
related to dairy wastewater treatment in continuous condi-
tions. Molognoni et al. [18] had previously tested a single MFC
reactor for dairy wastewater treatment, achieving encour-
aging results. Before an attempt at system upscaling,
Table 1 e Physico chemical characterization of
wastewater effluents produced in a cheese factory.
(Concentration ranges elaborated from Refs. [41,43,44]).
Parameter Unit Concentration
range
Total Suspended Solids (TSS) mg TSS/L 250e2700
Chemical Oxygen Demand (COD) mg O2/L 650e3000
Biological Oxygen Demand (BOD5) mg O2/L 300e1400
Organic nitrogen (Norg) mg N/L 10e140
Ammonium nitrogen (NeNH4) mg N/L 10e20
Nitrates (NeNO3) mg N/L 10e20
Phosphate (P) mg P2O5/L 10e130
Chlorides (Cl) mg Cl/L 50e500
pH (
) pH units 4e12
Alkalinity (HCO3) mg HCO3/L 250e650
502 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1

Table 2 e Studies dealing with dairy wastewater treatment by MFCs.

MFC configuration Feed Test duration Inoculum source dP h COD (%) CE Reference
Mode (W/m3) (%)
DC-MFC Batch n.a. Sludge from anaerobic digester 0.08 z100 2 [48]
DC-MFC Batch n.a. Industrial wastewater 1.1 95 14 [47]
SC-MFC Batch n.a. Sludge from anaerobic digester 0.44 82 2 [49]
Tubular Batch n.a. Sludge from activated sludge reactor 20.2 91 27 [45]
SC-MFC Batch n.a. Dairy wastewater þ 0.41 86 3 [50]
Shewanella oneidensis
DC-MFC Batch n.a. Dairy wastewater 2.7 91 17 [46]
DC-MFC Continuous n.d. Municipal & industrial wastewater 3.2 82e86 n.d. [51]
DC-MFC Continuous 30 days Dairy wastewater þ 0.02 62 n.d. [52]
Lactobacillus pentosus
SC-MFC Batch n.a. Industrial wastewater 0.038 z30 z4 [53]
DC-MFC Continuous 20 days Pre-screened municipal wastewater 1.9 63 24 [17]
DC-MFC Continuous 75 days Dairy sludge & wastewater 27 >80 46 [18]

n.a.: not applicable; n.d. not determined/not specified.

(SC-MFC) Single-chamber MFC; (DC-MFC) Dual-chamber MFC; (dP) power density; (h COD) COD removal efficiency; (CE) Coulombic efficiency.

additional verification was deemed necessary to ascertain the
performance consistency of the adopted system, and intro-
duce, if necessary, design modifications. The aim of this study
is precisely to evaluate long-term performance of duplicate
MFCs, continuously running for at least 2 and a half months in
the same conditions previously described by the Authors. A
complete comparative assessment of two parallel systems is
therefore carried out, in terms of observed electrical produc-
tion, organic matter removal and CE.
Material and methods
Two replicate, identical, dual-chamber MFCs were prepared
using a previously described design [18,54], consisting each of
an anodic and a cathodic chambers separated by a Cationic
Exchange Membrane (CMI-7000, Membranes International
Inc., USA) (Fig. 1). The chambers were filled with 800 g (each) of
granular graphite (diameter 1.5e5 mm, model 00514, EnViro-
cell, Germany), which decreased their free volume to 430 mL
net each for anodic (NAC) and cathodic compartment (NCC).
Graphite rod electrodes (250  4 mm, Sofacel, Spain) were
used to allow external electrical connection. Both cells' cir-
cuits were equipped with an external 33 U resistance,
assumed to be as close as possible to their static internal
resistance.
Dairy wastewater used as anode substrate was collected
periodically from the same source, and stored at 4  C until
actual use. Semi-continuous feeding through collapsible 10 L
plastic cans was adopted to limit the amount of wastewater
being stored and the number of trips to collect new samples.
This consisted of a cyclic feed routine pumping 3 L/d of dairy
wastewater into each cell for 20 min every hour, after which
pumping stopped for 40 min, resulting in an average feed rate
of 1 L/d per cell. Cathodes were fed with the same procedure
with oxygen-saturated phosphate buffer medium (10 mM, pH
7) containing: 819 mg/L Na2HPO4, 507 mg/L NaH2PO4, 1000 mg/
L NaHCO3, 130 mg/L KCl, 310 mg/L NH4Cl and other trace el-
ements (recipe modified from Xia et al. [33]). An internal
recirculation loop (30 L/d) was maintained in each chamber
with a separate circuit, to induce well-mixed conditions
within, minimize concentration gradients, and the chance of
partialization/clogging within the cells volume. Anodic recir-
culation was anoxic, while cathodic recirculation was
oxygenated, by the introduction of air in two external aeration
chambers, through a small fish tank air pump.
The MFCs operated at laboratory temperature (23 ± 3  C) for
the entire time. Anodic potentials were monitored with an Ag/
AgCl reference electrode (þ197 mV vs Standard Hydrogen
Electrode, Xi'an Yima Opto-electrical Technology Co., China),
recorded, together with overall cell potentials, at 1-min in-
tervals by a multifunction acquisition board (NI USB-6008,
National Instruments Italy, Milan) connected to PC.
Anodic and cathodic chambers were inoculated with 2.5 L
solution (30% aerobic AS from the dairy treatment plant, 10%
dairy wastewater, 60% distilled water). In the anodic chamber
inoculum, 2-bromoethanesulfonate (2-BES) was also added (at
10 mM concentration) to inhibit methanogens biomass
growth [55]. Inoculation was done in closed electric loop
mode, with 33 U external resistance, with low recirculation
rate (10 L/d) to promote, at the same time, adequate mixing
and bacterial fixation onto the electrodes' surface.
After 3 days, both MFCs were producing voltage in excess of
100 mV, and operation was switched to semi-continuous
mode with undiluted dairy wastewater, as described. The
collapsible bags1 containing the feed solution were replaced
approximately every 5 days, in order to maintain a quasi-
stable COD concentration in the feeding lines, with fresh
wastewater collected at the industrial WWTP every 3e5 days,
on average.
Determination of wastewater influent (one common sam-
ple every feed bag refill) and effluent COD (one sample per
MFC, variable intervals) was done regularly and analysed ac-
cording to “Standard Methods” [56]. Anodic Organic Loading
Rates (OLRs) were calculated as the daily organic matter
concentration (as COD) divided by the anode's hydraulic
retention time, adjusted every day according to observed
1
Collapsible liquid containers (plastic “jerry cans”) are used to
allow storage and wastewater feeding without leaving it directly
exposed to an oxidative atmosphere for prolonged periods, as the
volume of the container reduces as flow leaves it.
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1 503

Fig. 1 e Experimental scheme, showing liquid, electric and monitoring circuits: anodic and cathodic circuits e continuous
lines; electric and monitoring circuits e dashed lines. Legend: (A) anode chamber; (C) cathode chamber; (Rext) external
resistance; (1) aeration tanks; (2) anode electrode; (3) cathode electrode; (4) Ag/AgCl reference electrode.

parameter values. Organic matter removal efficiency (hCOD e
%) was determined as described in Molognoni et al. [35].
Values of pH and conductivity were measured at least once
every 5 days for both anode and cathode influents and efflu-
ents (IntelliCAL™ probes þ HQd™ Digital Meter, Hach Lange,
Italy).
Current (I) and power (P) were determined from MFCs
continuous voltage measurement (V). Power and current
densities (dP and dI) were evaluated dividing the respective
values of P and I by the NAC volume of each cell. Polarization
curves were periodically obtained using a potentiostat (NEV3,
Nanoelectra, Spain) and imposing a linear decrease of
0.5 mV s
1 from the Open Circuit Voltage (OCV) down to 0 mV.
From these curves, internal resistance was calculated by the
power density peak method [57]. Anodic CE was calculated
using daily average data of current intensity and flow-rate.
Energy losses (anode and cathode overpotentials -hAn and
hCat-, ionic -Eionic-, pH gradient eEDpH- and membrane trans-
port losses -Et) were then determined according to the prac-
tical methodology reported in Molognoni et al. [35] in
correspondence to each obtained polarization curve.
Results and discussion
Both MFCs were operated according to the above listed speci-
fications for 72 days. From the 3rd day after inoculation the
cells started to be operated in semi-continuous mode, with
undiluted dairy wastewater. Even though pumping rates cycles
were constant in time and identical for the two systems, anodic
OLRs varied considerably among the two systems (0.2e6.6 kg
COD/m3 d). The intrinsic variable nature of dairy wastes, from
one collected sample to the other (as shown in Table 1) caused
cells loads variation in time, allowing the study of non-steady
process conditions. In addition, “random” hydraulic response
variations among the two systems of identical construction,
and operated with the same pumps (with multiple heads), were
attributed to different dynamic head losses in the circuit (air
bubbles or temporary clogging due to occasional agglomeration
of solids in feeding tubes, observed by checking cells' effluent
volumes) that were occasionally detected, causing temporary
differentiation of the electric production of the two systems.
Such “unplanned” OLR perturbations allowed to study the in-
fluence of this operating parameter on MFCs performance,
both in relation with wastewater treatment efficiency and
power generation capacity.
Fig. 2 shows the temporal evolution of influent COD, and
resulting effluent COD concentrations from the two cells
systems (indicated as MFC1 and MFC2). OLRs for each system
are also shown, calculated from the average daily effluent
volumes: even though the same pump was used to feed both
cells, MFC1 showed a slightly higher feeding rate than MFC2
(by 13%, on average), that could be in part attributed to
different connecting pipes length, slightly different fitting of
504 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1
Fig. 2 e Evolution of influent and effluent COD concentrations (CODin, CODout1, CODout2) and organic loading rates (OLR1,
OLR2) in the two-cells system.
pipe joints, and partial tubes clogging, resulting in different
head losses (in time) on the two feed circuits.
The temporal evolution of voltage generation by the two
cells, in terms of daily average and (daily) standard deviation
values is shown in Fig. 3. During inoculation phase, the voltage
increased gradually (reaching 71 mV for MFC1 and 36 mV for
MFC2, on day 3). Once the semi-continuous feeding strategy
was initiated, generated voltage increased exponentially,
reaching a quasi-stable condition at day 6 (601 mV for MFC1,
498 mV for MFC2). From this point on, continuing significant
differences were observed between cells, with MFC1 achieving
on average consistently higher values than MFC2
(583 ± 47 mV, vs. 422 ± 44 mV, respectively). This difference
may be only partly explained by the higher feeding rate (and
thus higher OLR) of MFC1, as mentioned, and could be due to
differential anodic colonization in the two cells by a micro-
biome of different composition. On day 21, data from both
MFCs exhibited an abrupt voltage decrease, due to a relatively
long disconnection (few hours) needed to solve technical
problems in the data acquisition system. Several intervals
were characterized by higher variability of operations not only
between systems: MFC1 (525 ± 63 mV) and MFC2 (232 ± 51 mV),
but also within the same system, with higher than usual
temporal variability (up to ± 20%). These phenomena could
have been originated by different factors, including: opera-
tional failures (on day 29, MFC1 feeding rate decreased to
0.3 L d
1, and MFC2's to 0.2 L d
1 due to tubing failure, with
Fig. 3 e Evolution of the voltage generated by the two MFCs (daily averages ± standard deviations).
ordinary conditions re-established only after one day), tech-
nical problems (on day 44 there was an operational delay due
to erroneous reading of polarization curves). During the final
period (day 60e68), performances of the two cells became
comparably low (395 ± 81 mV for MFC1, 334 ± 43 mV for MFC2),
most likely due to very similar, low OLRs. The temporal evo-
lution of current (Fig. 4) and power densities follows the same
trends of voltage readings.
Maximum average daily power density values recorded
were: 27.3 W/m3 NAC for MFC1 (day 27) and 17.1 W/m3 NAC for
MFC2 (day 6). Comparing these with the values reported in
Table 2, it can be observed that the obtained data are higher
than the highest values obtained in batch mode by Mardan-
pour et al. [45]. Usually, values obtained in batch mode are
higher than those observed in continuous running mode,
therefore the obtained results confirm once more the tech-
nical suitability of dairy effluents treatment by MFCs tech-
nology with bioenergy recovery.
Analysis of average anodic and cathodic potentials reveals
a stable anode potential in both cells (
180 ± 55 mV vs. SHE for
MFC1, -195 ± 33 mV vs. SHE for MFC2), meaning that EABs
were effectively able to colonize the electrodes of both cells,
and transferred the released electrons from substrate oxida-
tion with some, although variable, efficiency. On the contrary,
cathodic potentials were more variable, a likely reason of the
observed voltage fluctuations in both cells. A similar conclu-
sion can be made observing open circuit electrode potentials,
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1 505

Fig. 4 e Temporal trend of current density in the two cells.

reported in Table 3 that summarizes the parameters observed COD removal efficiency reached an average value of
during recording of the polarization curves. 80 ± 10% for MFC1 and 83 ± 11% for MFC2, referred to the
Fig. 5 shows wastewater treatment efficiency of the cells, monitoring period from day 14e65 (when cells were assumed to
both in terms of COD removal and Coulombic Efficiency (CE). be well settled in pseudo steady-state). These results are in line
CE represents the ratio of recovered electrons (i.e. electricity with, or better than previously conducted studies (Table 2). COD
production) and the total amount of electrons made available removal efficiency is an index of the efficiency of a wastewater
by substrate oxidation (i.e. the organic removal rate) in a cell, treatment process in removing organic load from the liquid
and therefore represents its electric efficiency. stream. On the other hand, CE, index of electric efficiency of the
system, is influenced by the different electrical performances
of the two cells. The relationship between the two is nonlinear,
as it depends also on the system energy losses, which can also
Table 3 e Polarization curves recorded for MFC1 and vary in time. On average, MFC1 showed a CE of 20 ± 16%, while
MFC2. MFC2 exhibited a lower value (14 ± 11%), although showing
Day Rint OCV EAn oc ECat oc Pmax Iscc better average COD removal. Fig. 5 shows OLR effects on COD
(U) (mV) (mV vs SHE) (mV vs SHE) (mW) (mA) removal and CE, calculated by averaging the data collected
MFC1 from both cells. A clear dependency between OLR and hCOD
14 21.2 NA n.a. n.a. 13.0 41.8 cannot be easily inferred (Fig. 6), although previous studies
21* 41.4 NA n.a. n.a. 7.1 26.4 showed that better carbon removal efficiency can be achieved
29 9.3 762
247 515 20.3 67.3
in substrate limiting conditions (OLR < 1 kg COD/m3d), as these
44* 29.8 790
254 536 9.7 38.8
48 15.2 723
213 510 11.8 48.4
favour EAB's growth versus methanogenic population compe-
51 12.4 743
217 526 14.7 51.0 tition [54]. On the other hand, CEs of both cells appears to
65 17.2 698
206 492 9.2 33.8 decrease exponentially with the increase of applied OLRs. In
Average 15.0 732
221 511 13.8 48.4 this experiment, CE varied from 46% at low OLR (0.4 kg COD/
MFC2 m3d), to less than 5% at high OLR (5.8 kg COD/m3d). High OLRs
14 23.9 NA n.a. n.a. 7.6 27.5
cause increased microbial competition between EABs and side-
21* 27.9 NA n.a. n.a. 3.8 21.9
populations (methanogens, heterotrophs) in the MFC anodic
29 26.1 713
251 462 8.9 34.9
44* 13.3 279
259
10 1.7 21.0 chamber. EABs usually outcompete other microbial pop-
48 33.0 587
219 368 4.5 24.8 ulations more easily at low COD concentration, as shown by
51 20.4 637
207 43 7.3 28.5 Pinto et al. [40], and strategies to improve predominance of EAB
65 20.5 704
199 505 8.4 28.4 population (or inhibition of methanogens) under high organic
Average 24.8 660
219 345 7.4 28.8 loads should be investigated to improve general MFCs effec-
Note the variability over time of these parameters. (Rint) internal tiveness as wastewater treating systems.
resistance; (OCV) open circuit voltage; (Ean OC) open circuit anode Fig. 7 summarizes the main electrical losses parameters
potential; (Ecat OC) open circuit cathode potential; (Pmax) maximum gathered by the polarization curves obtained during the
power point; (Iscc) short circuit current. The analyses affected by
experiment (an example of polarization and power curves is
known operational issues and are marked with*.
shown in Fig. 8). MFC1 showed a mean internal resistance of
506 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1

Fig. 5 e Efficiency of MFC1 and MFC2, in terms of organic matter removal (hCOD) and Coulombic efficiency (CE). Organic
loading rates (OLR) are also reported.

Fig. 6 e Organic loading rate (OLR) effect on COD removal (h COD) and Coulombic efficiency (CE) in MFC1 (A) and MFC2 (B).
Shown data are calculated as the average of the data previously reported in Fig. 6.

Fig. 7 e Average energy losses distribution of MFC1 and MFC2.

 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1
Fig. 8 e Polarization and power curves for MFC 1 and MFC 2 recorded on day 51.
15 ± 8 U, lower than that obtained for MFC2 (25 ± 6 U). This
result is in agreement with the higher voltage and power
production achieved by the former. Values of open circuit
voltage, maximum power and short circuit current are in
agreement with previous observations: MFC1 achieved
maximum power of 20 mW (day 29) with average value of
13.8 mW; MFC2 exhibited maximum power of 9 mW, with
average value of 7.4 mW (taking into account the analysis of
all polarization curves). These average data correspond to
maximum power densities of 32 and 17 W/m3 NAC, respec-
tively for MFC1 and MFC2. Both reactors exhibited a
constantly lower internal resistance than the fixed external
one chosen. Thus, both MFCs were operating on the “left side”
of their respective power curves, rather than at their
maximum (optimal) value. Based on a rough estimation,
therefore, power recovery could have been 20e30% higher if
external resistance loads were periodically adjusted to match
the instantaneous value of Rint, possibly achieving 41.6 and
22.1 W/m3 NAC in MFC1 and MFC2, respectively. This type of
control was not applied in this experiment, since it was the
first authors' attempt at dairy wastewater treatment with the
apparatus used, and it may not have been possible to distin-
guish a posteriori between effects of changing OLRs and of a
continuously-varying resistance. However, maximum power
point tracking (MPPT) for external resistance adjustment,
already successfully used for MFC start-up time reduction [35]
is a likely strategy to adopt in future studies for electric pro-
duction maximization.
Table 4 summarizes average energy losses distribution in
the two MFCs, calculated from individual polarization curves
analysis. Energy losses represent the difference between
MFCs electromotive force (i.e. theoretical maximum voltage
achievable) and the actual voltage measured at the electrodes,
and depend on a number of factors [58,59]. It was not unex-
pected to calculate energy losses of MFC2 higher than those of
MFC1, due to the lower electric performance of the former. In
507
both cells, the sum of cathodic and membrane overpotentials
(hCat and Et) was the main energy loss contributor, summing
up to 35e39% (MFC1) and 27e33% (MFC2) to total energy los-
ses, respectively. pH gradients between anode and cathode
(around 2 pH-units in MFC1, 1.6 in MFC2), and anodic over-
potentials affected cells' electric behaviour in a minor way.
Ionic transport through electrolytes could be considered
negligible in this case, due to the relatively high conductivity
of both anodic and cathodic media (1.6 ± 1.3 mS/cm for the
anolyte, 4.5 ± 2.9 mS/cm for the catholyte).
Discussion
The long-term experiment herein reported was successful in
treating real dairy wastewater with MFC systems, with better
results, in terms of power density, of any other reported in
literature using this substrate. Still, there is space for further
process improvement. The calculated internal resistance of
both parallel systems showed (Table 3) that this was different
than the external resistance of 33 U applied. MFCs theory says
that power generation in a cell is maximum when the internal
resistance equals the applied external resistance. A MPPT
control method such as that applied by Molognoni et al. [35]
should therefore be able to achieve maximum energy pro-
duction from either cell. Furthermore, the study of the elec-
trical losses during operation of the process has shown that
cathodic, membrane and pH losses were the most important
loss categories in the system studied. A variety of strategies
can be adopted to reduce the energy losses of MFCs, therefore
improving electricity recovery from substrate. Cathode over-
potentials may be reduced by intervening at different levels:
(i) developing new, more efficient electrode and catalyser
materials (best if avoiding the economically unsus-
tainable use of Pt). Cathode electrode materials have a
508 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1

Table 4 e Time-varying energy losses distribution of MFC1 and MFC2 (average values and percentage contribution).
Day 29 48 51 55 65 Average
mV % mV % mV % mV % mV % mV %
MFC 1
Electrolytes (Eion) 11 3% 25 4% 19 4% 21 4% 32 5% 22 ± 7 4%
Cathode (hcat) 154 35% 188 34% 173 39% 191 40% 197 29% 181 ± 15 35%
Anode (han) 112 26% 75 13% 26 6% 23 5% 62 9% 60 ± 33 11%
Membrane (Et) 85 19% 161 29% 76 17% 94 20% 280 42% 139 ± 77 27%
pH gradent (EDpH) 74 17% 112 20% 151 34% 146 31% 101 15% 117 ± 29 23%
Total loss 436 559 446 476 673 518 ± 89
MFC 2
Electrolytes (Eion) 6 1% 9 1% 20 4% 19 4% 27 4% 16 ± 8 2%
Cathode (hcat) 207 29% 359 44% 290 50% 220 42% 214 30% 258 ± 59 39%
Anode (han) 108 15% 79 10% 64 11% 34 6% 65 9% 70 ± 24 10%
Membrane (Et) 313 44% 291 36% 100 17% 113 21% 291 42% 222 ± 94 33%
pH gradent (EDpH) 74 11% 74 9% 104 18% 144 27% 105 15% 100 ± 26 15%
Total loss 708 812 578 529 702 666 ± 101

great impact on power performance of MFCs: in addi-
tion to high conductivity and surface area, they should
have high redox potential to facilitate oxygen reduction
on their surface. Carbon based materials like cloth,
paper, graphite and metallic electrodes were utilized as
cathodic materials so far. Santoro et al. [60] found that
the electric production performance of MFCs was line-
arly correlated with the nano-scale roughness of the
cathode's electrode materials: since higher roughness
originated better contact between cathode and
membrane.
(ii) improving oxygen transfer kinetics in the cathodic
chamber. Not efficient oxygen reduction reaction (ORR)
on the cathode is one of the most important problems of
MFC technology in practical implementation.
(iii) growing a biocathode (relying on bacterial, rather than
chemical, catalysis of oxygen reduction reaction). Uti-
lization of biocathodes, can improve power perfor-
mance and simultaneously eliminate many toxic
pollutants [61e63].
Recently, ceramic membranes were indicated as a viable
option for practical application in MFC technology due to low
production cost, availability, very good stability and high
structural strength and capability to solve some of the issues
above reported [64,65]. Regarding energy losses associated with
membrane ionic transport, these could be reduced by the
development of new membranes with lower internal resis-
tance, or less subject to biofouling, or by avoiding their use
altogether. pH gradient losses may be reduced by increasing pH
buffer strength, or by reducing the hydraulic residence time of
MFCs' electrolytes. Application of ion-selective membranes as
a separator in MFCs can lead to pH splitting and acidification of
the anodic chamber, which may inhibit development and ac-
tivity of EABs at the anode, deteriorating MFCs' power perfor-
mance. In contrast, size-selective separators like ceramic
membranes, transferring protons based on porosity, do not
exhibit any pH drop in the anolyte [64]. Of course such correc-
tive measures must be considered in relation with their costs
and efficiency, in terms of wastewater treatment and not just
aiming at maximizing electricity recovery.
Another impairment against optimal electric performance
of MFCs is undesired methanogenesis that may considerably
diminish the CE of a system, although it may not directly
affect COD removal from substrate. Improving system per-
formance by suppressing methanogenesis is an ongoing issue
in the field of bioelectrochemical systems, including MFCs, as
this may ultimately compete against electrogenic activity of
EABs reducing the coulombic energy of a working cell. Injec-
tion of 2-bromoethanesulfonate (at concentration of
0.1e0.27 mM) in a cell can even double CE by suppressing
methanogens (such addition in fact was made while starting
up the MFCs in this study), however, 2-BES is an expensive
chemical, and its use for large scale operations would be
economically problematic. To increase overall MFC perfor-
mance, substrate oxidation by metabolism other than that
leading to electrogenesis should be reduced; elimination or
suppression of methanogens in the anode compartment is
therefore required, without affecting the exo-electrogenic
population. Unfortunately, similar growth conditions for both
populations make it difficult to suppress only methanogens by
altering operating condition. Some success in controlling
methanogenic populations was achieved by Kaur et al. [66]
investigating the effects of various duty cycles of open cir-
cuit (OC) and closed circuit (CC) MFC operation on the inhibi-
tion of methane production, and consequently on the
preponderance of methanogens in the cells' population.
Extended starvation of the cells for 12 days before re-starting
normal operations was clearly able to suppress any meth-
anogenic activity.
Conclusions
This study aimed at demonstrating the feasibility of dairy
wastewater treatment with bioenergy recovery through MFCs,
quantify the amount of power extractable, and highlight the
major sources of energy loss in the system. Two replicate,
identical laboratory-scale cells were built and continuously
fed with dairy wastewater for 72 days, longer than any other
literature-reported application to this substrate. Both pro-
cesses proved to be stable, with an average COD removal
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y 4 3 ( 2 0 1 8 ) 5 0 0 e5 1 1
efficiency of 80%, irrespective of OLR fluctuations over time. A
maximum power density of 27 W m
3 NAC was observed and
recovered during this study, the highest observed so far in a
long-term, continuously running application. CE was found to
decrease exponentially by increasing the OLR, with an optimal
value of 46% recorded at a load of 0.4 kg COD/m3 d. Achieved
results were mostly better than previously reported experi-
ences. This study confirms that dairy wastewater treatment
may represent an ideal application niche for MFCs technology.
The study also analysed and highlighted the major sources of
energy losses in MFC systems, and suggested direct or indirect
solutions for their reduction.
Most MFC studies are focusing on the undoubtedly inter-
esting feature of recovering electric energy directly from the
ongoing process. This study showed that some non-negligible
amount of energy could in fact be recovered, and that this
could perhaps have been further increased with more
aggressive operative process management. However, in
addition to the generation and recovery of electric energy,
MFCs have some additional advantages over traditional bio-
logic treatment processes, whether aerobic or anaerobic.
Compared to the former, MFCs offer a lower energy demand,
mainly in the form of lower aeration requirements (some
cathode aeration is needed, however to a much lesser degree
than a regular activated sludge plant, that could be altogether
eliminated by using an air-cathode configuration), faster
process kinetics, and lower sludge production. Compared to
the latter, MFCs still offer faster degradation rates and energy
recovery in the form of electricity or biogas production that,
although harmful to the direct electricity generation process,
may still be collected and usefully employed.
Although MFC technology is still in an experimental phase
compared to more traditional wastewater treatment tech-
nologies, it definitely offers some aspects of great interest for
future, sustainability-oriented applications. Current chal-
lenges for achieving industrial MFC application include:
finding new materials or surface modification techniques to
reduce cathodic losses and other internal losses, and mini-
mization of non-exo-electrogenic reactions in the system.
Acknowledgements
At the time of this work, Dr. Molognoni was supported by a
research grant from the Department of Civil Engineering and
Architecture of the University of Pavia. He is now employed at
the Leitat Technological Center in Terrassa, Spain.
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