Separation and Purification Technology 167 (2016) 154162

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Separation and Purification Technology

journal homepage: www.elsevier.com/locate/seppur

Synthesis and performance of iron oxide-based porous ceramsite in a

biological aerated filter for the simultaneous removal of nitrogen and
phosphorus from domestic wastewater
Teng Bao a,b, Tianhu Chen a,, Juan Tan a, Marie-Luise Wille c, Dan Zhu a, Dong Chen a, Yunfei Xi b,d,
a

Laboratory for Nanominerals and Environmental Material, School of Resource and Environmental Engineering, Hefei University of Technology, He Fei, China
Institute for Future Environments, Queensland University of Technology (QUT), 2 George Street, GPO Box 2434, Brisbane, QLD 4000, Australia
Institute of Health & Biomedical Innovation, Queensland University of Technology (QUT), Brisbane, QLD 4000, Australia
d
School of Chemistry, Physics and Mechanical Engineering, Science and Engineering Faculty, Queensland University of Technology (QUT), 2 George Street, GPO Box 2434, Brisbane,
QLD 4000, Australia
b
c

a r t i c l e

i n f o

Article history:
Received 21 January 2016
Received in revised form 9 May 2016
Accepted 9 May 2016
Available online 10 May 2016
Keywords:
Iron oxide-based porous ceramsite
Nitrogen and phosphorus removal
Nitrification
Denitrification
Biological aerated filters

a b s t r a c t
A novel iron oxide-based porous ceramsite (IPC) was synthesized and applied as a microbial biofilm carrier in a biological aerated filter (BAF) to treat domestic wastewater and compared to commercially available ceramsite (CAC). The results indicate that IPC has a higher porosity in comparison to CAC. The
uniformity and interconnectivity of pores, as well as the rough surface of the IPC are suitable for microbial
biofilm growth. Biofilm growth occurs in the internal pores of the media and promotes nitrogen and
phosphorus removal. The effect of air-water ratio (A/W) on the removal of total organic carbon, ammonia
nitrogen, total nitrogen, and phosphorus were investigated. The results show that the performance of IPC
BAF is much better than CAC BAF. For instance, at an A/W ratio of 3:1, the total nitrogen removal was
46.26% with IPC and 15.64% with CAC, and the PO34 removal was 72.25% with IPC compared with only
33.97% with CAC. An analysis of the microbial community in the IPC BAFs by polymerase chain reaction
denaturing gradient gel electrophoresis identified Dechloromonas sp., Sphaerotilus sp., and Nitrospira sp.
microbes. The diversity on microbial population, along with the attached growth benefit from the morphological properties of IPC, allows enhancement in the simultaneous nitrification and denitrification
performance in IPC BAF. Hence, IPC can be considered a very effective novel media material in BAF for
the simultaneous removal of nitrogen and phosphorus.
2016 Elsevier B.V. All rights reserved.

1. Introduction
Eutrophication can occur when excessive nitrogen and phosphorus reach water bodies through runoff or discharge of wastewater [1]. Eutrophication can lead to the deterioration in water
quality damaging aquatic ecosystems. An outbreak of harmful cyanophytes, commonly known as cyanobacteria, will eventually
result in increased mortality of aquatic organisms. Hence, the discharge of treated wastewater must meet effluent quality standards
for phosphorus and nitrogen. Novel wastewater treatments are
required for the simultaneous removal of phosphorus and nitrogen
(SPN) since conventional treatments have commonly been
designed to focus on removal of one nutrient. Traditionally, tech Corresponding authors at: School of Chemistry, Physics and Mechanical
Engineering, Science and Engineering Faculty, Queensland University of Technology
(QUT), 2 George Street, GPO Box 2434, Brisbane, QLD 4000, Australia (Y. Xi).
E-mail addresses: chentianhu@hfut.edu (T. Chen), y.xi@qut.edu.au (Y. Xi).
http://dx.doi.org/10.1016/j.seppur.2016.05.011
1383-5866/ 2016 Elsevier B.V. All rights reserved.

nologies such as biological and chemical treatment for phosphate

or nitrate have been widely used [2,3]. Chemical treatment
employs a metal salt to coagulate and remove phosphate by sedimentation. The excess amount of sludge produced is a major limitation to this process [2]. Adsorption is the most widely used
and suitable method for phosphorus removal. Of all the adsorbents,
iron oxides and untreated clays are considered the most effective
sorbent materials [4,5]. Previous studies have shown that raw
goethite, nano zero-valent iron (Fe0), and hematite are very effective materials adsorbing phosphorus [6,7]. Simultaneous nitrification and denitrification (SND) for the removal of nitrogen can be
achieved through biological processes. SPN removal from wastewater can be achieved by the combination of different biological
and/or chemical technologies (i.e., sequential treatment). A biological aerated filter (BAF) is a fixed-film biosystem with a small footprint that employs filter media with a high specific surface area
and porosity. The filter can promote in-growth biofilm(s) for
wastewater treatment. BAFs can reduce chemical oxygen demand

T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

(COD), suspended solids and ammonia [8,9]. Microbial biofilm supported filter media, which are made of raw clay, wastewater
sludge, construction waste and raw zeolite, have been evaluated
in recent years and have achieved good results for nitrogen
removal [2,10,11]. Dissolved oxygen (DO) plays a key role during
the BAF process. Denitrifiers are active in areas of very low DO concentration whereas nitrifiers are active in areas of high DO concentration [12]. SND has been proven to be the most effective way to
remove total nitrogen (TN) which occurs naturally inside the
microbial biofilms of filter media [13]. Porous filter media involving a mixed population and upholding oxygen diffusion limits create both anoxic and aerobic micro-environments. As such, the filter
media allows the sequential use of electron acceptors, such as oxygen and nitrate. Such mass transfer leads to the stratification of
microbial species in the microbial biofilm of filter media [15,16].
BAF technology is applied to wastewater treatment; hence, the
selection of filter media plays a significant role in maintaining a
variety of microbial populations and a high amount of active biomass. Recently, some waste materials, such as waste ceramics
and polyethylene plastic, have been utilized as filter media for biological aerobic filters, providing a new approach for disposing the
waste sawdust [2]. This strategy serves as a winwin solution
by treating wastes with wastes. Furthermore, several new filter
media for BAF have been recently investigated to enhance
biodegradation capability. For example, combined up-flow anaerobic biofilters and up-flow BAF that employ filter media prepared by
sludge, coal cinder, and straw have been used to treat synthetic
wastewater [17]. These filter media achieved favorable results for
nitrogen removal. However, only a few studies investigated in
detail SPN with BAF.
In China, commercially available ceramsite (CAC) has been
newly used in filter media for domestic wastewater treatment.
However, such material was reported to exhibit extremely low
nitrogen and phosphorus removal efficiencies in BAFs. This result
is probably due to the poor affinity of CAC to microbial biofilm biomass, which is caused by biocompatibility and suboptimal surface
hydrophilicity [2,11]. For this reason, developing a novel filter
medium as a substitute for CAC in the simultaneous removal of
nitrogen and phosphorus must be accomplished as quickly as
possible.
Previous studies reported that sawdust was treated at 700 C to
produce a desirable porous material. The formation of interior
pores improves microbial growth. It was reported that hematite
and palygorskite calcined at 700 C can effectively absorb phosphorus [2,5]. In this study, iron oxide-based porous ceramsite
(IPC) with goethite, sawdust, and palygorskite were synthesized
by calcination in an O2 atmosphere, used as microbial biofilm support in BAF and compared to commercially available ceramsite
(CAC) [14]. The properties of IPC, CAC and microbial characteristics
were characterized by scanning electron microscopy (SEM),
porosimetry analysis (PM) and micro-computed tomography
(Micro-CT). The effect of air-water ratio (A/W) on the removal of
total organic carbon (TOC), ammonia nitrogen (NH3-N), total nitrogen (TN) and phosphorus (P) was investigated using both supports.
Finally, the relationship between protozoa and metazoa as well as
the characteristics of microbial biofilms were determined using
polymerase chain reaction denaturing gradient gel electrophoresis
(PCR-DGGE) of the microbial community of the IPC BAF.

155

CAC surface morphologies were examined using a Scanning Electron Microscope (SEM, Philips XL30 ESEM-Netherlands Philips
Company). The physical characteristics of IPC and CAC were measured in accordance with the sandstone pore structure method of
image analysis. Rubber casting experiments were used to generate
quantitative size and shape data from pores in the thin section
(d = 3 cm) [2,14,18]. The porosities of IPC and CAC specimens were
determined using Micro-CT (lCT 40, Scanco Medical, Brttisellen,
Switzerland). The two samples were scanned in air at 70 kV and
57 lA, with an isotropic voxel size of 12 lm. A region of interest
(ROI) was defined for each gray-value image, which corresponded
to the exact circumference of the sample. A binary threshold was
applied to each sample and Scanco built in reconstruction algorithms were used to derive the porosity and to construct the 3D
image [1921].
Microscopic observations for protozoan and metazoan population growth were carried out using a U-RFL-T Olympus Biological
Microscope (Olympus Corporation, Tokyo, Japan). The multi-point
BET surface area of IPC and CAC was measured using a Quantachrome Nova 3000e automated surface area analyzer. The
growth of biofilm was determined according methods available
in the literature [2,13]. The microbial populations in IPC BAF were
analyzed by using domain-specific F341 R907 primers and PCR
amplification of 16S rRNA gene fragments for genomic DNA from
bacteria. The PCR products were applied in the DGGE analysis
using a BioRad Dcode system (BioRad Co. Ltd., USA). A DGGE gel
of 8% polyacrylamide with a linear denaturing gradient ranging
from 35% to 60% (100% denaturing gradient contains 7 M urea
and 40% formamide) was used. Electrophoresis was conducted at
a constant voltage of 75 V in 1  TAE buffer at 65 C for 14 h. The
gels were then stained with 5% Gold view in 1  TAE buffer for
40 min and imaged using UV trans-illumination. Selected DGGE
bands were excised using the SK1131 kit (Sangon Biotech (Shanghai) Co., Ltd) and re-amplified by PCR with the aforementioned primers. The PCR products were sequenced at Sangon Biotech Co., Ltd
(Shanghai, China). The obtained sequences were submitted and
compared to the reference microorganisms in the GenBank database using the BLAST program [22,23].
2.2. Operating conditions of IPC BAF and CAC BAF
A schematic diagram of the experimental set-up is provided in
the supplementary material. Biofiltration was initiated by introducing seed sludge obtained from the Hefei City wastewater treatment plan into BAFs. Each experiment was divided into 3 stages for
each BAF. During each test stage, the operating conditions of both
BAFs were identical (see Table 1). A stable hydraulic retention time
(HRT) of 7 h was used. Three air-water (A/W) ratios of 1:1, 3:1, and
6:1, respectively, were tested. The two BAFs were monitored for
4 months after the initiation of biofiltration. Both BAFs were operated in summer at wastewater temperatures ranging from 26 C to
30 C. Samples were analyzed for NH3-N and P in accordance with
Chinese EPA standards [2,24]. A TOC/TN (Jena Multi N/C 2100) analyzer was used to measure TOC and TN. Dissolved oxygen (DO)
concentrations were measured using a portable digital DO meter
(Oxi-315, Hao-Yang Biotech Company, Shanghai, China).
3. Results and discussion

2. Materials and methods

3.1. Characteristics of IPC and CAC

2.1. Characterization and analytical methods

Table 1 shows the elemental properties of IPC and CAC. IPC possesses a higher surface area and porosity than those of CAC. Lower
apparent and bulk densities were also observed in the IPC. All of
these properties are associated with the chemical compositions

Details of reagents and preparation of the proposed filter

media-IPC are provided in the supplementary material. IPC and

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T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

Table 1
Operating conditions.
Sample

HRT (h)

pH

Water quality indexes and operating conditions

Stage one
7
6.7 (0.5)
Stage two
7
6.6 (0.5)
Stage three
7
7.1 (0.5)

T (C)

A/W

TOC
AIC SD

NH3-N
AIC SD

PO34
AIC SD

2730
2830
2330

1:1
3:1
6:1

26.06 0.205
26.10 0.449
25.29 0.486

9.75 0.196
9.79 0.329
9.81 0.175

0.558 0.022
0.484 0.015
0.486 0.016

Note: TOC/NH3-N/PO34 values are influent concentrations mg/L; standard deviation SD; average influent concentrations AIC; HRT hydraulic retention time; A/W air/
water ratios.

of the materials. Sawdust comprised 20% of IPC, which is propitious

for the manufacture of lightweight porous IPC because of the
decomposition of sawdust during calcination (Section 1.2, Supporting information). The lower apparent and bulk densities of IPC may
be ascribed to the addition of sawdust. The gas-forming components from the sawdust also rendered the IPC surface rougher than
the CAC surface, generating a larger surface area in the former. Furthermore, the microbial biofilm biomass was greater in amount on
the IPC than on the CAC because of the formers rough surface. For
IPC and CAC, the surface area reached over 79 m2 g 1 and 2 m2 g 1,
respectively (Table S1). The rough surface distributed with large
pores was beneficial for the microbial biofilms to grow onto the
IPC. These special surface morphology properties also demonstrated that IPC was fit as filter material.
3.2. Micro-CT based pore characterization
Micro-CT is a powerful tool for non-destructive and reproducible analysis of the architecture of porous materials based on
3D geometrical considerations [2527]. In this study, micro-CT
was used to analyze structural characteristics and porosity of IPC
and CAC. Fig. 1A shows the 3D micro-CT reconstruction of IPC.
The red spherical particles in IPC sections denote calcined molded
composite particles (calcined palygorskite, calcined sawdust, and
calcined goethite) and pores can also be observed. This 3D image
demonstrates that IPC has considerable external porosity by volume. Fig. 1B shows a 0.72 mm thick cross-section (stack of 20 scan
slices) of IPC with large pores of diameters approximately 100
500 lm. Fig. 1C and D shows a 3D reconstruction and 0.72 mm
thick cross-section (stack of 20 scan slices) of CAC and CAC filled
with epoxy, respectively. The porosity was determined to be
3.46% 2.05%. The formation of liquid phases at temperatures
P1000 C reduces the number of pores in CAC due to a higher percentage of fine particles. This characteristic in CAC hinders the initiation of cracks and results in a low porosity [28].
Porosity is a key factor in ensuring successful nitrogen removal
in a given BAF process due to the uneven distribution of DO and
substrate inside the IPCs pores or on the rough surface. This feature allows simultaneous proliferation of nitrifying and denitrifying bacteria. Micro-organisms are immobilized in the IPCs pores
which maintain a high biomass concentration while retaining a
high hydraulic loading rate [13]. Micro-CT analysis shows that
when sintered at 700 C, the IPC has a porosity of 40.86% 3.26%
with many unevenly distributed pores (100500 lm). In principle,
this characteristic is suitable for the growth of attached microorganisms on rougher surfaces [13].
3.3. Cast thin section of IPC and CAC
Fig. 2 shows the internal pore structures of IPC and CAC. The IPC
porosity (40.86% 3.26%) rendered the material ideal for microbial
biofilm in-growth (Fig. 2A). By contrast, CAC possessed much lower
porosity (3.46% 2.05%), which limits microbial growth (Fig. 2C)
[14]. Microbial biofilm growth can form on external surfaces and

within internal pores of the IPC; a rich supply of organic matter

and nutrient substance allows these microbial biofilms to thrive.
The microbial biofilm excretes slimy, glue-like exopolymeric substances that further improve microbial anchoring to the IPC [29].
DO affects the distribution, nitrification, and denitrification rates
of the microbial biofilm of the IPC as follows. Autotrophs are
mainly located on the outer layer of the microbial biofilm to meet
a fundamental requirement for converting NH+4N. By contrast, the
location of heterotrophs in the microbial biofilm is less restricted
because the microorganisms can occupy the microbial biofilm center. In this case, the heterotrophs may grow with an organic substrate during the feast phase with the use of NO3 N as electron
acceptor [30]. Heterotrophs may also grow on the outer layers of
the microbial biofilm where DO can be used as electron acceptor
[30]. The concentration of DO first decreased rapidly within the
microbial biofilm thickness and then decreased gradually to zero
at the outer region of the microbial biofilm [31]. Nitration occurred
outside the microbial biofilm. Given the limited oxygen diffusion,
gradients of DO concentrations within the microbial biofilm
became possible. Deep within the microbial biofilm, oxygen transfer may be blocked, and large external oxygen consumption may
occur, thus producing anoxic zones. Denitrifying bacteria predominantly enabled anoxic denitrification within the microbial biofilm.
Hypoxic microbial biofilms were found within the pores. SND was
also observed both inside and outside of the flocs [3235].
Previous research has shown that staining epoxy resin can be
used to impregnate the pore space of rock samples. This method
is a very useful technique for identifying mineral components of
sedimentary rocks [36,37]. However, few reports have investigated
the porosity of filter media by using staining epoxy resin. The thin
sections of CAC and IPC, as depicted in Fig. 2, were analyzed to
obtain direct evidence of microorganisms in the porous structure
of both materials. The rubber casting experiments were used to
generate quantitative size and shape data from pores in the thin
section [36,37]. The intergranular pore textures observed in a thin
section and the red netted texture sections in Fig. 2A are identified
as calcined palygorskite, goethite (hematite) and sawdust (biomass
residues), while the gray area (arrowed) represents internal pores
of the IPC. IPC have a greater variety of internal pores than guest
microbial biofilm [2]. In addition, the size of those interconnected
pores is about 100500 lm (Fig. 2A), which is consistent with the
results of Micro-CT analyses. It is well known that most microbes
are approximately 0.5 lm in diameter. Thus, microbes can achieve
a sustained growth of bacterial population in these open pores. In
Fig. 2B, the blue section was obtained by vacuum impregnation
with blue-dyed epoxy (arrowed) in IPC. Fully interconnected pores
with a diameter of 100500 lm between the narrowest parts can
be observed. In fact, Fig. 2C and D shows that blue-dyed epoxy in
the porosity system cannot overcome the capillary resistance of
the throat(s) and enter into porosity space [2]. There are also some
closed pores in CAC (closed pore - arrowed), where microbes cannot penetrate the internal pores of the CAC. Previous studies have
shown that the biofilm biomass was 63.4 mg TN/g for IPC BAF and
2.2 mg TN/g for CAC BAF [14].

T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

157

(A) 3 dimensional reconstruction of IPC;

(B) 20 slices (0.72 mm) cross-section of IPC;
(C) 3 dimensional of CAC;
(D) 20 slices (0.72 mm) cross-section CAC;

Fig. 1. Micro-CT of IPC and CAC.

3.4. Comparison of the influence of A/W ratio on TOC removal in the

IPC BAF and CAC BAF
Fig. 3A, Tables 1 and 2 show the IPC BAF and CAC BAF performance for TOC removal with a HRT of 7 h at different A/W ratios
(i.e., 1:1; 3:1; 6:1). These results confirm that for steady organic
input loading, microbe bioactivity is enhanced with increase of
A/W ratio (1:1 ? 6:1). On the other hand, diffusivity is strengthened with increase of A/W ratio and contributes to TOC removal.
Fig. 3A and Table 2 also indicate that the IPC has higher performance due to enhanced physicochemical properties of IPC as confirmed by the analyses of Micro-CT and porosimetry. The surface
and internal pore structures of IPC are uniform and well developed
so that organic matter and nutrient substances are able to reach
the internal pores of IPC. The available space for microorganism
growth, mass transfer rate of DO and the dispersion potential of
biofilm biomass are also increased by soluble pollutants extending
to the internal pores of the IPC. These attributes result in a more
favorable development of the filter layer and improve the effectiveness of biofiltration [13].
3.5. Influence of A/W ratio on NH3-N and TN removal
The data in Fig. 3B, Tables 1 and 2 also show optimum removal
of NH3-N. The removal rate is higher when the A/W ratio increases

from 1:1 to 3:1 and then decreases at higher A/W ratio to 6:1. A
higher air-flow inhibits the microbial biofilm resulting in
decreased NH3-N removal. IPC BAF had a slightly higher NH3-N
removal compared to the CAC BAF. A/W is one of the key parameters responsible for nitrification; therefore, it is the limiting factor
for nitrification when the A/W ratio in the BAF is lower than
required for bio-reactions with an A/W ratio of 1:1. Studies by
Lazarova and Manem [38] have reported that autotrophic bacteria
compete with heterotrophic bacteria for oxygen, substrates and
habitation area within a microbial biofilm. This outcome is because
slow growing autotrophic bacteria inherently compromises the
nitrification process [38]. The main cause of this difficulty is a
decrease in nitrifying activity that affects the competition between
aerobic heterotrophic bacteria and autotrophic bacteria [12].
From Fig. 3C, Tables 1 and 2, it can be seen that TN removal efficiencies first increase (1:13:1) and then decrease with an increase
of A/W ratio (3:16:1). The results confirm that TN removal is
affected by the A/W ratio in the two BAFs. Denitrification produces
less energy yield than oxygen respiration. Hence, a bacterial cell
growing in aerobic conditions will choose to use oxygen as a terminal electron acceptor. In addition to this competitive effect, oxygen
controls denitrification at two levels: (i) reversible inhibition of the
activities of the denitrification enzymes and (ii) regulation of gene
expression [39]. It can be seen that the IPC BAF has a slightly higher
TN removal rate compared with the CAC BAF (Fig. 3C). Normally, a

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T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

(A) IPC; (B) IPC filled with epoxy (light blue = open pores);
(C) CAC; (D) CAC filled with epoxy;

TOC (mg/L) Removal efficiency (%)

A
A/W=6:1

A/W=3:1

A/W=1:1

80

40
CAC removal
IPC removal

0
20
Influent
CAC effluent
IPC effluent

10

25

50

75

NH 3 -N (mg/L) Removal efficiency (%)

Fig. 2. Internal porosity of CAC and IPC (cross-section). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this
article.)

B
120
80
CAC removal

40

IPC removal

10
Influent

CAC effluent

IPC effluent

0
0

25

Time (d)

75

D
A/W=6:1

A/W=3:1

A/W=1:1

CAC removal
IPC removal

60
30
12
0
9
Influent
CAC effluent
IPC effluent

P (mg/L) Removal efficiency (%)

TN (mg/L) Removal efficiency (%)

50

Time (d)

90

A/W=6:1

A/W=3:1

A/W=1:1

150
A/W=3:1

A/W=1:1

A/W=6:1

CAC removal

100

IPC removal

50
0
0.9

Influent
CAC effluent
IPC effluent

0.6
0.3
0.0

25

50

Time (d)

75

25

50

Time (d)

Fig. 3. Removal efficiency of TOC, NH3-N, TN, and PO34 over time.

75

TOC
A/W
Material
IPC
CAC

1:1
AEC SD
19.93 1.21
21.87 2.63

NH3-N
ARR SD
23.51 4.88
16.07 10.13

3:1
AEC SD
13.53 3.68
16.91 2.32

ARR SD
47.95 14.96
35.06 9.81

6:1
AEC SD
9.98 2.52
12.45 1.34

1:1
AEC SD
9.11 0.76
9.97 0.62

ARR SD
20.02 7.70
15.33 7.46

3:1
AEC SD
0.57 0.61
4.62 2.54

ARR SD
94.22 5.97
52.62 27.02

6:1
AEC SD
1.25 1.81
1.98 1.20

ARR SD
87.19 18.65
79.81 12.39

ARR SD
72.25 5.74
33.97 10.73

6:1
AEC SD
0.12 0.08
0.33 0.07

ARR SD
76.11 15.95
32.55 14.32

PO34

TN
A/W
Material
IPC
CAC

1:1
AEC SD
7.80 0.75
8.25 0.69

ARR SD
60.50 10.25
50.77 5.29

ARR SD
15.86 8.74
8.09 5.09

3:1
AEC SD
5.55 1.84
8.76 0.88

ARR SD
46.26 18.95
15.64 10.56

6:1
AEC SD
7.22 0.98
8.99 0.87

ARR SD
31.73 8.94
14.96 8.44

Note: Standard deviation SD; average removal rates ARR (%); average effluent concentrations AEC (mg/L).

1:1
AEC SD
0.16 0.048
0.37 0.07

ARR SD
73.70 9.01
34.21 12.75

3:1
AEC SD
0.13 0.03
0.32 0.05

T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

Table 2
Average removal rates for IPC and CAC in BAF at different air/water ratios.

159

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T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

(A) CAC; (B) CAC with microbial loading;

(C) IPC; (D) IPC with microbial loading;

Fig. 4. SEM images of IPC and CAC before and after microbial loading.

denitrification process occurs in anoxic conditions as there is

enough organic matter to be used as an electron donor. In two
BAFs, the denitrification process is limited by diffusion, mixing,
biofilm thickness and substrate availability [40]. It was suggested
by Laursen et al., that nitrate removal in the IPC BAF takes place
deep in the biofilm [41]. The depth to which oxygen can penetrate
into the biofilm is determined by the bulk liquid DO concentration
and the diffusion rate of oxygen. In the aerobic layer, nitrate does
not take part in any reactions and diffuses through the biofilm
inactively. Nitrate is utilized by microorganisms for cell synthesis
and growth in the deeper layers [12]. In Section 3.8, we evaluate
the microbial community and confirm that nitrifying bacteria
and denitrifying bacteria are present among other forms. This communal mix of bacteria is an indicator of good performance in
wastewater treatment systems.
3.6. Removal of PO34 on both IPC and CAC BAF
The rates of PO34 removal at different A/W ratios (1:16:1) for
both BAF systems are shown in Fig. 3D and Table 2. A higher
removal rate of PO34 is observed in IPC compared to CAC. Previous
work from this group shows that exposed iron due to dehydroxylation is able to combine with hydroxyl ions (derived from
hydrolyzation) to maintain charge balance after saturation of
annealed goethite due to an increase in the number of adsorption
sites [5]. In addition, thermal activation increases phosphate sorption capacity. The highest phosphate sorption capacity occurred at
700 C due to an increase in dolomite in palygorskite. As a naturally occurring mineral, dolomite is a double salt of calcium and
magnesium carbonate (CaMg(CO3)2). Dolomite decomposes into

CaO and MgO as temperature is increased. Therefore, phosphate

ion is preferred when adsorbed by Ca and Mg in natural and activated palygorskite [42]. As shown in Table S1, SiO2 and CaO constitute 60.24% and 0.41% of CAC composition, respectively. CAC with a
high SiO2 content, has a low efficiency for PO34 retention [43].
Alternatively, the exposed iron combines readily with phosphate
as previously reported [5].
3.7. SEM analysis of IPC and CAC
The morphology of IPC and CAC was determined with SEM as
shown in Fig. 4. Fig. 4A clearly shows that for CAC small pores
are irregularly distributed on the smooth surface. Fig. 4B also
shows that a small number of microbes are immobilized on the
exterior surface of the pores in CAC. Fig. 4C shows that large pores
and coarse surfaces are common for the IPC sample(s). These features are caused by the release of accumulated gases generated
by the decomposition of sawdust in the mixture of goethite and
palygorskite during the sintering process [2]. Fig. 4D shows that
the microbial biofilm is apparently spread over the surface of IPC
as indicated by the connectivity of extracellular secretions.
Attached cells, morphologically similar to bacillus, spheres, and
spirals are also observed on the surface of IPC [13,44,45].
3.8. Microbial community analysis of IPC BAF
As shown in Fig. 5, the identity of the diverse bacterial species
populating the IPC BAF were determined using PCR-DGGE analysis.
A detailed classification of the microbial community can be found
in the supplementary material. Briefly, in band 3, the dominant

T. Bao et al. / Separation and Purification Technology 167 (2016) 154162

161

IPCBAF for line 1, line 2, and line 3.

Line 1

Line 2

Line 3

Fig. 5. DGGE profiles of bacterial and archea 16S rRNA genes from the IPCBAF for line 1, line 2, and line 3.

bacteria, Dechloromonas sp. (No. 6) is a genus in the phylum proteobacteria. Dechloromonas sp. is facultative denitrifying bacteria
[46]. No. 2 demonstrates 99% similarity with Azonexus sp., a genus
in the phylum proteobacteria [47]. No. 3 demonstrates 99% similarity with Hydrogenophaga sp., a genus of comamonad bacteria. Several of these bacterial species were previously classified in the
genus Pseudomonas [48]. No. 7 and No. 8 demonstrate 99% similarity with Sphaerotilus sp., an aquatic periphyton organism associated with polluted water that forms colonies commonly known
as sewage fungus. However, an aquatic periphyton organism
Sphaerotilus sp. was later identified as a tightly sheathed filamentous bacteria [49]. No. 16 demonstrates 100% similarity with Nitrospira sp. which is a phylum of bacteria. This phylum contains only
one class, nitrospira, which itself contains one order (Nitrospirales)
and one family (Nitrospiraceae) [50]. This analysis demonstrates
the biological diversity of microbes found in the biofilm and confirms that nitrifying bacteria and denitrifying bacteria are present.
This diversity of both bacterial types enhances performance in
wastewater treatment systems.

enhanced microbial growth for IPC compared to CAC. A higher

microbial biofilm production rate is observed in IPC compared to
CAC when used in the BAF system. The performance efficiency of
the IPC BAF is higher than that for CAC BAF, particularly for TOC,
TN, P, and NH3-N removal. This efficiency also improves when
the A/W ratio is increased from 1:1 to 6:1. These results indicate
that IPC is a promising material for the simultaneous removal of
nitrogen and phosphorus in wastewater treatment. The biological
diversity of microorganism communities in the IPC BAF plays a
key role in the simultaneous removal of nitrogen and phosphorus.
Finally, a large quantity of protozoan and metazoan organisms
observed in IPC BAF are indicators of good performance in wastewater treatment systems.
Acknowledgements
We gratefully acknowledge the support by the National Natural
Science Foundation of China China (41572029 and 41130206).
Appendix A. Supplementary material

4. Conclusion
This study reports the performance of synthesized IPC and CAC
as filter media in a BAF process. SEM and PM results suggest that
uniform and interconnected pores in IPC are more suited to microbial growth compared with the compact and closed pore structure
of CAC. Three-dimensional rendering using micro-CT of the pores
also reveals critical details of the pore structure and is an accurate
method to characterize porosity. These observations support

Supplementary data associated with this article can be found, in

the online version, at http://dx.doi.org/10.1016/j.seppur.2016.05.
011.
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