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Bioresource Technology 99 (2008) 39653974

Review

Bio-electrochemical removal of nitrate from water


and wastewaterA review
Shahin Ghafari, Masitah Hasan, Mohamed Kheireddine Aroua

Department of Chemical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia
Received 24 January 2007; received in revised form 15 May 2007; accepted 15 May 2007
Available online 27 June 2007

Abstract
Nitrates in dierent water and wastewater streams raised concerns due to severe impacts on human and animal health. Diverse methods are reported to remove nitrate from water streams which almost fail to entirely treat nitrate, except biological denitrication which is
capable of reducing inorganic nitrate compounds to harmless nitrogen gas. Review of numerous studies in biological denitrication of
nitrate containing water resources, aquaculture wastewaters and industrial wastewater conrmed the potential of this method and its
exibility towards the remediation of dierent concentrations of nitrate. The denitriers could be fed with organic and inorganic substrates which have dierent performances and subsequent advantages or disadvantages. Review of heterotrophic and autotrophic denitrications with dierent food and energy sources concluded that autotrophic denitriers are more eective in denitrication.
Autotrophs utilize carbon dioxide and hydrogen as the source of carbon substrate and electron donors, respectively. The application
of this method in bio-electro reactors (BERs) has many advantages and is promising. However, this method is not so well established
and documented. BERs provide proper environment for simultaneous hydrogen production on cathodes and appropriate consumption
by immobilized autotrophs on these cathodes. This survey covers various designs and aspects of BERs and their performances.
2007 Elsevier Ltd. All rights reserved.
Keywords: Water; Wastewater; Bio-electrochemical; Biological denitrication; BER

1. Introduction
Water is of fundamental importance for life since the
mechanism of metabolism and synthesis are in close relation with the specic characteristics of water. Transport
of nutrients inside cells and interactions with the environment without water are impossible. On the other hand,
water resources are limited and only 2.66% of the total global water resources comprising groundwater, lakes and rivers, polar ice and glaciers are fresh water. Furthermore,
only a small fraction of the fresh water, about 0.6%, is
usable as drinking water. For this reason, water resources
must be necessarily treated properly and wastewater treatment must be done eciently (Shrimali and Singh, 2001).
The fast movement of humans towards urbanization,
*

Corresponding author. Tel.: +603 79675313; fax: +603 79675319.


E-mail address: mk_aroua@um.edu.my (M.K. Aroua).

0960-8524/$ - see front matter 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2007.05.026

industrialization and agricultural activities introduced various contaminants into the environment.
Nitrogen-containing compounds are examples of those
contaminants which can create serious problems when
released into the environment, such as eutrophication of
rivers (Sumino et al., 2006), deterioration of water quality
and potential hazard to human or animal health. Nitrate
is one of these compounds, though it does not pose a
human or animal health threat by itself. It is potentially
converted to nitrite in the gastrointestinal tract or to
nitrous nitrogen compounds through reduction (Foglar
et al., 2005). Up to the present time, researches are carried
out towards nitrate removal from water resources, aquaculture ponds or aquaria, and industrial wastewaters.
In this review, rstly, nitrate as a water contaminant and
problems arising from its presence are studied. Secondly,
various studies and methods applied for nitrate removal
are discussed. Finally, aiming to reveal an eective method

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S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

applicable for dierent types of nitrate-containing water


and wastewater, emphasis is on bio-electrochemical
denitrication.
2. Nitrate-containing water and wastewaters
2.1. Nitrate in water resources
Nitrate is identied as one of the hazardous contaminants in potable water that may reduce to nitrosamines
in the stomach which is suspected to cause gastric cancer
(Glass and Silverstein, 1999; Galvez et al., 2003). In addition, nitrate exposes infants and pregnant women to danger
due to the potential reduction of nitrate to nitrite inside the
stomach of foetus after digesting nitrate by the mother. The
nitrite reacts with the hemoglobin in blood and converts
the hemoglobin into methaemoglobin, which does not
carry oxygen to cell tissues. This phenomenon results in a
bluish color of infants skin so called methaemoglobinemia
or the blue baby syndrome (Shrimali and Singh, 2001).
Haemoglobin Fe2

NO
2

Can combine with O2

Methaemoglobin
Cannot combine with O2

Water resources (ground and surface water) are contaminated by nitrate in several ways. Most of the researchers reported that contamination of groundwater by nitrate is
primarily attributed to non-point agricultural sources (Feleke and Sakakibara, 2002). Nitrate originated from agriculture is increasingly growing all over the world due to
the extreme use of fertilizers. Nitrate salts reach the
groundwater as they percolate through the soil. Some other
sources of nitrate in ground and surface water are from
uncontrolled land discharges of treated or raw wastewater
from domestic and industrial wastes, landlls (Islam and
Suidan, 1998) and animal wastes particularly from animal
farms (Terada et al., 2003). Nitrate in groundwater is
increasingly an important problem, which prohibited the
direct use of the groundwater resources for human consumption in some parts of the world including India, Japan, China, Saudi Arabia, USA, UK and several parts of
Europe (Gayle et al., 1989). With the aim to protect the
consumers from adverse eects of high nitrate intake, the
United States, Canada, and the World Health Organization (WHO) have set standards to regulate the nitrate concentration in drinking water to 50 mg NO
3 /l (Sayre, 1988).
Also, the United States Environmental Protection Agency
(EPA) and WHO have set the maximum contaminant level
(MCL) of 10 mg NO
3 N=l in drinking water (Cast and
Flora, 1998). The same concern in Europe put MCL of
12 mg NO
3 N=l in drinking water (Glass and Silverstein,
1999).
2.2. Nitrate in mariculture and aquaculture systems
Cultivation of river and marine organisms in their natural or synthetic habitat is always along with the production

of ammonia (referring to NH3 or NH


4 which is considered as a serious problem. Ammonia produces a bad avor
and taste as well as harmful eects on aquatic animals.
EPA proposed that ammonia concentrations in sh culture
tanks be maintained at levels lower than 0.02 mg/l as the
criterion for non-ionic ammonia (EPA, 1976). Therefore,
indispensable need for ammonia removal complied with
applying dierent pathways such as nitrication, sludge
removal by sedimentation or mechanical ltration, and
water exchange (Rijn et al., 1996). Nitrication is reported
as one of the most common and eective methods applied
by researchers (Hagopian and Riley, 1998; Singh et al.,
1999; Zhu and Chen, 1999) in both freshwater and seawater. In this method ammonia is oxidized into nitrite and
nally nitrate as the end-product. Reported maximum
nitrate levels dier among various systems and values of
400500 mg NO
3 N=l are reported (Otte and Rosenthal,
1979). Contrary to ammonia and nitrite, nitrate is relatively
non-toxic to aquatic organisms. However, it should not be
left to accumulate, because eventually leads to some undesirable results such as phytoplankton blooms, inhibition of
nitrication and toxicity problems at certain concentrations of nitrate (Park et al., 2001; Rijn et al., 2006). As a
result, daily replacement of a fraction of the system volume
(510%) with new water may prevent accumulation of
nitrate (Masser et al., 1999), though this solution does
not sound environmentally. Increased eorts are now
directed towards nitrate control in aquaculture systems,
not only concerning toxic eect on sh, but also for complying to environmental regulations associated with permissible nitrate level in euent discharge which is as low
as 11.3 mg NO
3 N=l (European Council Directive, 1998).
2.3. Nitrate in industrial wastewaters
Concentration of nitrogen compounds in some industrial wastewaters is tremendously higher than what is in
groundwater and surface water. Ammonia and nitrate are
the most problematic nitrogen compounds in this sort of
wastewaters. Ammonia in industrial wastewater is normally eliminated by nitrication which is achieved by the
complete oxidation of ammonia to nitrate (Sumino et al.,
2006). Thus, nitrate removal from these types of industrial
wastewater is an inevitable step in treatment. Dierent
industrial wastewaters are reported to contain more than
200 mg NO
3 N=l (Almeida et al., 1995; Zayed and Winter,
1998; Peyton et al., 2001) and some contain higher nitrate
concentration. For instance, euent from some industries
producing explosives, fertilizer (Watanabe et al., 2001),
pectin, cellophane, and metals nishing industries, contain
greater than 1000 mg NO
3 N=l (Glass and Silverstein,
1999). Furthermore, nuclear industry produces nitrateloaded wastes in extremely higher concentration at many
points during the nuclear fuel cycle. The processing of
radioactive metal products at nuclear weapons production
plants and research labs reported production of wastewater
containing nitrate in excess of 50,000 mg NO
3 N=l primar-

S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

ily derived from nitric acid used for metal cleaning (Francis
and Hatcher, 1980).
3. Denitrication methods
3.1. Abiotic nitrogen removal methods
The conventional processes of water treatment method
including coagulation, ltration, and disinfection which
are applied for water potability are not ecient for the
elimination of nitrate ion from the water. Therefore, to
remove nitrate ion, a supplementary method is necessary.
Nitrate removal from water or wastewater can be achieved
using two main groups of treatment processes; physicochemical and biological treatment methods. The most common conventional treatment processes used to remove
nitrate are reverse osmosis (RO), ion exchange (IE), electro-dialysis (ED) and activated carbon adsorption in conjunction with pH adjustment (Islam and Suidan, 1998;
Ergas and Reuss, 2001; Shrimali and Singh, 2001; Feleke
and Sakakibara, 2002; Prosnansky et al., 2002). Also, a
wide interest towards some new methods such as metallic
iron-aided abiotic nitrate reduction is recently reported
by several researchers (Huang et al., 1998; Choe et al.,
2000, 2004; Devlin et al., 2000; Hu et al., 2001; Huang
and Zhang, 2002, 2004; Huang et al., 2003; Chi et al.,
2004; Ginner et al., 2004). There are some disadvantages
with these conventional methods that limit their utility,
due to their expensive operation and subsequent disposal
problem of the generated nitrate waste brine or by-products. For instance, IE process removes nitrate and sulfate
simultaneously from the water being treated. Subsequent
produced wastewater from the resin regeneration process
rich in these two anions requires a complementary method
before its discharge (Shrimali and Singh, 2001). RO,
although is able to separate and concentrate the compounds contained in water without any change in their
molecular structures, this method is limited due to its high
cost and producing concentrated waste brine that may pose
a disposal problem for further treatment (Ergas and Reuss,
2001; Shrimali and Singh, 2001). Even for the new method
of metallic iron-aided abiotic nitrate reduction, although
complete or partially nitrate removal are reported by the
researchers, formation of ammonia along with nitrogen
gas as end-products is an undesired consequence of this
method (Biswas and Bose, 2005).
3.2. Biological denitrication methods
In contrast to abiotic methods, which failed to treat
nitrate and were merely able to separate or remove it from
the stream and resulted in some problematic by-products,
biological methods provided a promising chance for real
treatment of nitrate and new novel techniques are reported
accordingly. For removing nitrate, although aerobic denitrication has been reported (Joo et al., 2005; Robertson
and Kuenen, 1984; Zart and Eberhard, 1998; Chiu et al.,

3967

2007), most of the reported denitrication investigations


are done by facultative anaerobes in the absence of oxygen
(Rijn et al., 2006). Biological denitrication is a mechanism
by which denitrifying bacteria use nitrate as terminal electron acceptor in their respiratory process in the absence of
oxygen. Denitrifying bacteria, in this method, reduce inorganic nitrogen compounds such as nitrite and nitrate into
harmless elemental nitrogen gas (Prosnansky et al., 2002)
so that no further treatment is imposed. Also, unlike some
contaminants which are in need for a certain microbe to be
treated, denitrifying bacteria are ubiquitous in nature
(Gamble et al., 1977; Szekeres et al., 2001) and numerous
researchers cultivated them using mix cultures taken from
wastewater treatment plants as seeds. Moreover, microbial
removal of nitrate may be the most economical strategy for
the reclamation of nitrate polluted waters and wastewaters
(Soares, 2000). As a result, there has been a huge interest
towards microbial removal of nitrate as the most environmentally friendly and cost-eective method, although biological denitrication is usually slow and lasts long time
particularly for industrial wastewaters containing high concentrations of nitrate (Foglar et al., 2005). Later studies
tried to speed up biological denitrication by applying
new mechanisms through which better connection provided between the nitrate and the microorganisms in the
water stream. The Inorganic Contaminants Research Committee of USA (Islam and Suidan, 1998) reported that different denitrication strategies, such as the use of granular
activated carbon (GAC), packed beds, rotating biological
contactors and soil-aquifer denitrication systems have
been undertaken by various researchers. Also, eorts are
still ongoing and some novelties in combination of biological and other abiotic methods, such as membrane biolm
reactors (MBR), were recently manifested (Terada et al.,
2003).
4. Heterotrophic versus autotrophic denitrication
Biological denitrication is normally conducted by facultative anaerobes which are in essential need for some
food and energy sources which are organic or inorganic
(Cast and Flora, 1998; Rijn et al., 2006). This fact classies
denitriers into two main groups of heterotrophs and autotrophs. Heterotrophs are microorganisms that require
organic substrates to get their carbon for growth and development. They also obtain energy from carbohydrates and
other organic material. In contrast, autotrophs are microorganisms that use inorganic substances as energy source
and carbon dioxide as a carbon source.
Heterotrophic denitrication has been applied in recirculating systems and traditional wastewater treatment
plants, whereas autotrophic denitrication is lately notied
within the last two decades. These substrates do not always
exist in the water or wastewater which is to be treated and
should be supplemented. For instance, biological removal
of nitrogen from wastewaters which contain high levels of
nitrate can be problematic due to the low concentration

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S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

of carbon source required for the denitrifying bacteria


(Kim et al., 2004). Hence, a number of researchers have
found that denitrication activity in organic carbon-limited
waters or wastewaters can successfully be enhanced by adding some carbon sources such as methanol, ethanol, acetic
acid, glucose, or starch (Killingstad et al., 2002; Kim et al.,
2002). The choice of substrate depends on a number of
considerations such as cost, capacity and conguration of
reactor, and post-treatment of the denitried water (Szekeres et al., 2002).
In case of heterotrophic denitrication, both liquid and
solid forms of organic carbon sources are conventionally
used though the aqueous type is more common for treatment of water and wastewater. Among liquid carbon
sources, the most common ones are methanol, ethanol
(Gomez et al., 2003) and acetic acid which have been used
for heterotrophic denitrifying processes in full-scale plants
of drinking water treatment (Feleke and Sakakibara, 2002).
However, there is some concern regarding these external
organic carbon sources in the heterotrophic denitrication.
Some organic carbon sources are toxic and their excess will
remain in water. In addition, by-products built-up from
organic carbon sources seems to be unavoidable which dictates some further treatment methods for their removal as
well as disinfection in case of water treatment (Shrimali
and Singh, 2001; Feleke and Sakakibara, 2002). Besides,
denitrication rate is strongly susceptible to type of carbon
source, concentration of carbon source (Galvez et al., 2003;
Gomez et al., 2003), and C/N ratio (Rijn et al., 2006). This
would vary for dierent microorganisms, water streams,
and environmental conditions (Chiu and Chung, 2003).
Hence, some researchers have investigated the inuence
of dierent types and magnitude of carbon sources to
achieve the optimum removal (Kim et al., 2002; Chiu and
Chung, 2003; Joo et al., 2005). A poor C/N ratio leads to
improper denitrication while a high C/N ratio may cause
accumulation of nitrite or extra production of nitrous other
than nitrogen gas (Kim et al., 2002; Chiu and Chung,
2003).
In contrast, autotrophic denitriers utilize inorganic carbon substrates (carbon dioxide or bicarbonate) as the food
source and rely on electron donors like hydrogen or
reduced sulfur compounds for energy (Biswas and Bose,
2005). Autotrophic denitrication using carbon dioxide
gas as an inorganic carbon source and some electron
donors like hydrogen (Kurt et al., 1987; Feleke et al.,
1998; Islam and Suidan, 1998; Ergas and Reuss, 2001; Sakakibara and Nakayama, 2001; Feleke and Sakakibara,
2002; Prosnansky et al., 2002; Park et al., 2005), thiosulfate
(Claus and Kutzner, 1985a,b), sulde (Kleerebezem and
Mendeza`, 2002), and sulfur stone (Flere and Zhang,
1999) have been reported in literature. Some advantages
of autotrophic over heterotrophic denitrication are; evasion of poisoning eect of some organic carbon, low biomass build-up and less sludge production which result in
reduction of reactor clogging and easier post-treatment
(Shrimali and Singh, 2001; Rijn et al., 2006).

5. Hydrogenotrophic denitrication
Hydrogenotrophic denitriers refer to the denitrifying
microorganisms utilize hydrogen as energy source which
are ubiquitous in nature (Till et al., 1998). In addition to
the general advantages of autrotrophic denitrication,
using hydrogen as electron donor brings more advantages
into hydrogenothropic denitrication method compared
to other autotrophic methods using other electron donors.
This is attributed to the properties of hydrogen gas such as
being harmless, naturally clean, and low solubility (1.6 mg/
l at 20 C) in water. Thus, in contrast to other substrates
and electron donors, no further steps are needed to remove
the residues. Furthermore, higher eciency is obtained
with hydrogen rather than with organic electron donors;
for example, Gentzar (1995) reported an 8.5-fold greater
eciency with hydrogen when compared to organic electron donors. Besides, hydrogen is not as expensive as some
other organics. As a result, a number of laboratory and
eld systems have been reported in which hydrogen was
sparged into the water to be treated hydrogenotrophically
(Gros and Treutler, 1986; Gros et al., 1986; Kurt et al.,
1987; Dries et al., 1988).
Nevertheless, there are some drawbacks with sparging
hydrogen. For instance, low solubility of hydrogen in water
may hamper the rate of autotrophic denitrication by dissolution or poor mass transfer rate of hydrogen from gas
phase to liquid phase. To overcome this obstacle, bioreactors with large volumes to increase the contact time, may
be required to achieve the complete denitrication (Prosnansky et al., 2002). In addition, the hazardous (explosive)
nature of hydrogen during use, transportation and storage
may limit the use of hydrogen gas in denitrication reactors. Furthermore, the same as traditional biological processes, control of pH by introducing an acid equivalent is
still required (Feleke and Sakakibara, 2002; Prosnansky
et al., 2002). These disadvantages would result in higher
initial costs as well as higher operating costs of hydrogenotrophic denitrication in comparison with heterotrophic
denitrication.
An appropriate method has to be found to overcome the
above mentioned problems. As such, some researchers
tried to produce hydrogen in situ to remove the risk in
transfer, storage and operation in using hydrogen. Additionally, bacteria can utilize hydrogen at the time of generation more eectively since there is no need to dissolve it
prior to feeding into the reactor. Consequently, the internal
production of hydrogen has been recently studied extensively through hydrogen production from either chemical
reaction with water or electrolysis of water (Prosnansky
et al., 2002).
The production of hydrogen within the reactor using
chemical reactions has some drawbacks. Metallic ironassisted hydrogenotrophic denitrication process is an
example of denitrication applying chemically produced
hydrogen from corrosion of iron in water according to
Eqs. (1) and (2) (Biswas and Bose, 2005).

S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

Fe 2H2 O ! H2 Fe2 2OH


anaerobic iron corrosion


2NO
3 5H2 ! N2 4H2 O 2OH

biological denitrification

The relatively slow rate of anaerobic in situ corrosion process (Eq. (1)) with consequent low hydrogen production
rate slows denitrication process (Eq. (2)). The rate also depends on the type of metallic iron as well as on the surface
area (Biswas and Bose, 2005). Another drawback of in situ
chemical production of hydrogen is the presence of probable by-products and their undesirable reactions with other
materials in the treated water or wastewater.
On the contrary, in situ hydrogen production through
electrolysis of water revealed some advantages due to the
benecial characteristics of electricity. The most signicant
advantages of electricity are that it is dirt free without any
residual by-product as well as easy to control which means
easy handling of hydrogen production. Hence, autotrophic
denitrication using internal production of hydrogen by
means of electricity inspired a promising process which
entailed generation of electro-assisted bioreactors or bioelectro reactors (BERs).
6. Bio-electrochemical denitrication in BERs
Biological denitrication assisted by electricity would
theoretically be performed through in situ hydrogen production on the cathode surface (not on the anode surface).
Hydrogen and a lower oxidation reduction potential
(ORP) environment produced through the cathodic reactions would be utilized by hydrogenotrophs to reduce
nitrate nitrogen into nitrogen gas (Zhang et al., 2005).
A proper contact between microorganisms and hydrogen improves biological denitrication. BERs involving

3969

direct immobilization of denitriers on the surface of the


cathode, where hydrogen is being produced, facilitates
appropriate access of hydrogen to the microorganisms
(Park et al., 2005). BERs provide immobilized microorganisms with a compulsory supply, rather than accidental contact, of electron donors which results in faster
denitrication. This is in addition to the advantages of
the hydrogenotrophic method and in situ hydrogen production by electrolysis of water. BERs may be fed by
organic or inorganic carbon sources which serve heterotrophic and autotrophic hydrogenothrops, respectively. Table
1 lists the reactor type, type of microorganisms, treated
inuent and the range of nitrate for bio-electrochemical
method of denitrication found in the literature.
In general, utilizing autotrophs in a BER ideally serves
all the advantages attributed to autotrophic denitrication,
hydrogenotrophic electron donation, and electrolytic
in situ hydrogen production in a single method. Consequently, researchers focused on denitrication by autotrophic microorganisms in a BER as a prospective
pathway for nitrate removal from any nitrate-containing
water or wastewater.
6.1. Theory of denitrication in BERs
The presumed pathway of complete reduction of
nitrate NO
3 to nitrogen gas (N2) which involves four
consecutive steps is illustrated below. The oxidation state
of the nitrogen atoms in each substance is presented in
parentheses (Feleke and Sakakibara, 2002; Killingstad
et al., 2002):
5


NO
3 ! NO2 ! NO ! N2 O ! N2

Decomposition of nitrate in BERs can be illustrated


through seven equations including electrolysis of water
which are shown below.

Table 1
List of some previous studies applying bio-electrochemical method to denitrify water or wastewater containing various doses of nitrate
Reactor

Microorganism

Treated inuent

Nitrate
concentration
(mg NO
3 N =l

Ref.

Multi-cathode BER

Autotrophic

Synthetic contaminated water

13.820.3

Multi-cathode BER combined with


microltration (MF)
BER

Autotrophic

Synthetic contaminated
groundwater
Synthetic contaminated
groundwater
Synthetic contaminated water
Aquaria water

1540

Sakakibara and Nakayama


(2001)
Prosnansky et al. (2002)

20

Feleke et al. (1998)

20
20

Islam and Suidan (1998)


Grommen et al. (2006)

w/wa
IPTb contaminated

22.6492
24

w/w
contaminated water
w/w

50300
100
200

Park et al. (2005)


Feleke and Sakakibara
(2002)
Beschkov et al. (2004)
Cast and Flora (1998)
Watanabe et al. (2001)

Autotrophic

BER
Separate reactors for electrolysis and
denitrication
BER
BER combined with adsorption

Autotrophic
Autotrophic

BER
BER
BER

Heterotrophic
Heterotrophic
Heterotrophic

a
b

w/w: wastewater.
IPT: isoprothiolan.

Autotrophic
Heterotrophic

Synthetic
Synthetic
water
Synthetic
Synthetic
Synthetic

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S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

Electrolysis of water:

of BER. Table 2 lists some of these specications in previous studies.


In previous studies, various types of metals and carbons
were used as cathode and anode materials. Carbon in the
form of GAC and graphite as well as metals such as stainless steel, titanium, nickel and copper has been studied.
Some researchers tried to study eciency of dierent electrode materials; for example, Cast and Flora (1998) investigated metal cathode performance in comparison with
graphite as the most common electrode because use of
graphite electrodes in large-scale processes may be limited
by its brittleness and bulk.
The shape of electrodes and their arrangements are
other important conditions in the construction of the reactor. In the case of the bio-electrochemical method, electrode rods, plane and cylindrical shapes are applied. A
plane parallel conguration of electrodes is the most common electrode system, which grants many advantages such
as availability of materials and uniform current. However,
this conguration is only eective for very high concentrations of contaminants such as in the case of electrochemical
reductions (or depositions) of metal ions (Sakakibara and
Nakayama, 2001). In contrast, porous material such as
packed beds, mesh or reticulated planes, screen metals,
and expanded metals have been investigated in both plane
and cylindrical shapes in BERs. Using numerous cathodes
in BERs resulted in higher eciency due to providing extra
surface area for immobilized denitriers and hydrogen
production as well as expanded contact area with contaminants. In this case, arrangement of cathodes is signicantly

On the anode:
5H2 O ! 2:5O2 10H 10e

On the cathode:
10H2 O 10e ! 5H2 10OH

Stepwise autotrophic denitrication of nitrate to nitrogen


gas is illustrated below. The main role of hydrogen gas as
an electron donor is shown in this decomposition:

2NO
3 2H2 ! 2NO2 2H2 O

2NO
2 2H2 ! N2 O H2 O 2OH

6
7

N2 O H2 ! N2 H2 O

Net reaction of denitrication on cathode obtained from


Eqs. (5)(8):


2NO
3 6H2 O 10e ! N2 12OH

The overall reaction in bio-electrochemical reactor is a


combination of Eqs. (4) and (9):

2NO
3 H2 O ! N2 2:5O2 2OH

10

6.2. Design and specications of BERs


Specications of electrodes and design of reactors play a
signicant role in denitrication rate. The main parameters
in constructing the reactor are materials, shape, number of
electrodes and their arrangement as well as conguration

Table 2
Dierent materials and numbers of cathodes used in various shapes and congurations in bio-electrochemical reactors
Electrode
shape
Plane
Plane

Plane

Number and material of used electrodes

Surface area
of cathode
(cm2)

Conguration

Ref.

Cathode

Anode

Stainless steel
Five cathodes made by contacting
GAC with stainless steel
expanded metal
One graphite felt cathode

One anode made of


platinum-coated
titanium
One mesh-type
dimensionally stable
anode
One amorphous
carbon anode

750.0 (Each
cathode: 150)

Cathodes set in parallel at


upstream of anode

Beschkov et al. (2004)


Prosnansky et al.
(2002)

105.0

Vertical anode and horizontal


cathode

Park et al. (2005)

251.0

Anode between cathodes in


parallel

Feleke et al. (1998),


Feleke and
Sakakibara (2002)
Islam and Suidan
(1998)

Plane

Two stainless steel


cathodes + porous matrix

Cylindrical
cathode and
anode rod
Cylindrical

One cylindrical graphite cathode

One graphite anode


rod

Anode at the center surrounded by


cylindrical cathode

Eight expanded metal cathodes


made of titanium + porous
matrix
Twelve Carbon cathode rods

Two expanded
metal Pt-coated
anodes
One carbon anode
rod

Rod

Four stainless steel cathode rods/


Four graphite cathode rods

One graphite anode


rod

Tubing
electrodes

One stainless steel mesh cathode

One stainless steel


anode

7.6

Anodes at the center surrounded


by cylindrical cathodes
concentrically
Anode at the center surrounded by
cathodes in parallel (attached to
the cap)
Anode at the center surrounded by
cathodes in parallel (attached to
the cap)
In parallel and attached to the cap

Rod

1800.0

Sakakibara and
Nakayama (2001)
Watanabe et al. (2001)

Cast and Flora (1998)

Skadberg et al. (1999)

S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

considered and some pathways such as concentrically


arrangement of cylindrical cathodes (Sakakibara and
Nakayama, 2001) and successive parallel installment of
plane cathodes (Prosnansky et al., 2002) have been
examined.
6.3. Denitrication performances in BERs
Cast and Flora (1998) applied heterotrophic denitriers
to compare denitrication eciency of two cathode materials in water treatment. They examined stainless steel rods
wrapped with stainless steel mesh (SS/SS) and graphite
rod wrapped with polypropylene mesh (Gr/PP). The
impact of copper on water denitrication was investigated.
They utilized a reactor consisting of a graphite anode rod
surrounded by four cathode rods. This bio-electrochemical
reactor switched to continuous ow operation after 5 days
of batch operation aiming to culturing biolm on the cathodes surface (for over 1 month). Initial concentration of
100 mg NO
3 N=l was tested applying electric current of
1 mA with ow rate of 640 ml/d (correspond to a hydraulic
residence (HRT) time of 5.5 days). Nitrite was produced by
denitrication and accumulated in the electrochemical cell
during all experiments and complete denitrication was
not achieved in any of the tests. The dierence between
reduction rates for two types of cathode materials was
insignicant. There was no mention of the exact removal
rate using each cathode material. They also found out that
the denitrifying process would not be possible when a
wastewater stream contained heavy metals due to deposition of heavy metals on the surface of cathodes, which
inhibited biolm activity.
In 2001, Watanabe and co-workers studied denitrication of high acidic wastewater of copper metal pickling
using heterotrophic denitriers. They applied one carbon
rod along the axial center of the cylindrical reactor as
anode, which was surrounded by 12 carbon cathode rods
wrapped with carbon ber felt. The bio-electrochemical
reactor was continuously operated with a synthetic wastewater similar to real copper metal pickling wastewater in
major compositions, adding acetate to support heterotrophic denitrifying bacteria. Dierent ranges of hydraulic
retention time (HRT = 536 h), electric current intensity
(1523 mA), and C/N ratio (0.71.2) were examined.
Nitrate and copper removal at dierent concentrations of
copper (135 mg/l) was also investigated. The results demonstrated that copper ion removal, denitrication and neutralization could be achieved simultaneously by using a
single bio-electrochemical reactor. Inuence of copper ions
on the performance of denitrifying bacteria was evaluated
by means of the denitrication rate and the duration of
lag time before denitrication started. Denitrication was
eective and almost complete as long as the concentration
of copper was less than 30 mg/l. Higher copper concentration led to the accumulation of nitrate while copper was
almost removed implying that electric current contributed
to copper removal by chemical precipitation instead of

3971

hydrogen production. Nitrite was not produced and pH


remained around neutral at all concentrations of copper.
Islam and Suidan (1998) used a recycle biolm-electrode
reactor consisting of a cylindrical graphite cathode located
along the reactor wall and a graphite anode rod located in
the center of the reactor. This electrode conguration was
used to achieve a uniform current distribution throughout
the reactor. Groundwater treatment was examined using a
synthetic wastewater containing 20 mg NO
3 N=l and
autotrophic denitrifying microorganisms immobilized on
the inner surface of cylindrical cathode. Phosphate was
used as the buer primarily, but was replaced with carbonate to simulate real groundwater in the absence of phosphate. The experiments were conducted in two phases.
During the forward phase, electric current was increased
from 0 to 100 mA, while during the reverse phase, current
was decreased from 100 to 0 mA. Flow rate and recycle
rate in both phases were 2.65 l/d and 1.14 l/min, respectively. Nitrate removal increased as electric current intensity increased to 20 mA. However, there was a decline in
nitrate removal when the current was increased from 25
to 100 mA, which is attributed to hydrogen inhibition.
Excess hydrogen along with methane started to appear
when the current was beyond 60 mA. A stable nitrate
removal eciency of 8287% was achieved for over 2 years.
Besides, the highest 98% nitrate removal eciency was at
current 20 mA using phosphate as a buer.
Prosnansky et al. (2002) studied a combined BER and
membrane ltration (MF) performance. Autotrophic denitrication using CO2 in a BER consisted of ve porous
electrodes acting as multiple cathodes and an inert anode
was applied. Installing a membrane after a BER can avoid
microorganism escaping from the BER. Cathodes made by
contacting GAC with stainless steel expanded metals and
were connected in series with the platinum anode coated
by titanium. Experimental results revealed that multi-cathode BER was capable of operating at high denitrication
rates with low hydraulic retention time of 20 min
(HRT = 0.33 h). They reported an enhancement denitrication rate of 16.4 mg NO
3 N=h, which is 360 times
higher than previous studies. This implied that they overcame the slow operation of biological denitrication, the
most signicant drawback of this method. They attributed
this superior performance to the large eective surface area
and low ORP zone formation in the multi-cathode region.
A novel multi-electrode system was proposed by Sakakibara and Nakayama (2001) for biological water treatments
by mean of autotrophic denitrifying biolm located on the
surface of cathodes. A long term (over 500 days) continuous experiment was conducted to enhance treatment of
dilute solutions such as groundwater or surface water containing nitrate, about 20 mg NO
3 N=l. Eight cathodes and
two anodes in cylindrical shapes made by expanded metal
were set concentrically in that BER. Polyurethane foam
was used on the surface of the cathodes for supporting denitrifying biolms. They applied individual electrode potential or electric current to each part of the multi-electrode in

3972

S. Ghafari et al. / Bioresource Technology 99 (2008) 39653974

order to obtain uniform cathodic or anodic reactions without the limitation caused by low electrolytic conductance.
The euent nitrite concentration was almost kept below
0.5 mg NO
3 N=l. Sakakibara and Nakayama (2001) used
HRT range of 26 h which was two times smaller than
those in previous BERs using monopolar electrode systems. They believed that the complete and enhanced denitrication in this long term experiment was attributed to
using the multi-electrode system. The system provided a
large eective surface area, the charge transfer mechanism
2
by dissociative electrolytes HCO
3 and CO3 , and the formation of low ORP zone in the multi-electrode system.
Park et al. (2005) denitried a synthetic wastewater by
autotrophic microorganisms which gained electrons
directly from the cathode. In contrast to the other researchers, they treated high nitrate laden wastewater which was
not able to be treated if hydrogen was to be the electron
donor. They acclimatized the denitrifying biolm present
with 200 mA current for 60 days and achieved 98% nitrate
reduction in applying the same current during experiments.
With less or higher electric current, lower eciencies were
obtained. At higher than 200 mA, electricity was consumed
for hydrolysis of water and there was production of hydrogen on cathode. Inuent nitrate range of 20492 mg
NO
3 N=l was examined at 200 mA. In the case of 492 mg

NO
3 N=l inuent nitrate, there was 434.78 mg NO3 N=l
removal. Hydrogen produced was 100-fold lower than the
required amount for such reduction, implying that microorganisms had taken electron directly from the cathode.
Therefore, unlike other studies, an extremely high inuent
nitrate rate was converted to nitrogen gas because denitriers activities were not limited by the amount of produced
hydrogen as electron donor. This is meaningful because
nitrate levels are usually high in wastewater, up to as much
as 5001000 mg NO
3 N=l. Park and co-workers (2005)
made a comparison with other researchers results in terms
2
of mg NO
3 N=cm (biolm surface area) day). Their max2
imum denitrication rate was 0.17 mg NO
3 N=cm d,
which was higher than the maximum rate achieved by Kurt
2
et al. (1987) (0.031 mg NO
3 N=cm d, Macdonald
2
(1990) (0.083 mg NO
3 N=cm d, Liessens et al. (1993)
2
(0.023 mg NO
3 N=cm d, and Sakakibara and Kuroda

(1993) (0.038 mg NO3 N=cm2 d.
7. Conclusion
The available water resources are increasingly diminishing and we must preserve them through treating wastes
before discharging. Nitrate removal in drinking water supply or treatment of wastewaters, cannot be carried out
through conventional abiotic methods and this review
showed the capability of bio-electrochemical method in
denitrication. Some baseline information about this
method and signicant principles, which inuence BERs
performances are illustrated in this survey. There are many
advantages of this method compared to other existing

methods in nitrate removal. Some results from previous


researches illustrated the direct eect of electric current
and reactor design on the nitrate removal. This can provide
some new ways for researchers in the application of this
method for improving denitrication. Since this method
is relatively new and there is neither sucient documentation nor widely implementation in full scale, further studies
are needed. Future research directions in this important
eld should focus on studying the eects of electrode material and shape, surface area of cathode, and design of the
BERs. Development of correlations and mathematical
models for the simulation and optimization, and the economic evaluation of the process should be also considered
in future research work.
Acknowledgements
This study was carried out with the aid of a research
Grant of Kuok Foundation.
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