A Comparative Study of Anaerobic Fixed Film Baffle

Yousefzadeh et al.
Biotechnol Biofuels (2017) 10:139

DOI 10.1186/s13068-017-0826-9 Biotechnology for Biofuels
RESEARCH Open Access
A comparative study of anaerobic fixed

film baffled reactor and up‑flow anaerobic
fixed film fixed bed reactor for biological
removal of diethyl phthalate from wastewater:
a performance, kinetic, biogas, and metabolic
pathway study
Samira Yousefzadeh1,2†, Ehsan Ahmadi3,4*†, Mitra Gholami5,6*, Hamid Reza Ghaffari7, Ali Azari4, Mohsen Ansari8,
Mohammad Miri9, Kiomars Sharafi10 and Soheila Rezaei11
Abstract
Background: Phthalic acid esters, including diethyl phthalate (DEP), which are considered as top-priority and hazard-
ous pollutants, have received significant attention over the last decades. It is vital for industries to select the best treat-
ment technology, especially when the DEP concentration in wastewater is high. Meanwhile, anaerobic biofilm-based
reactors are considered as a promising option. Therefore, in the present study, for the biological removal of DEP from
synthetic wastewater, two different anaerobic biofilm-based reactors, including anaerobic fixed film baffled reactor
(AnFFBR) and up-flow anaerobic fixed film fixed bed reactor (UAnFFFBR), were compared from kinetic and perfor-
mance standpoints. As in the previous studies, only the kinetic coefficients have been calculated and the relationship
between kinetic coefficients and their interpretation has not been evaluated, the other aim of the present study was
to fill this research gap.
Results: In optimum conditions, 90.31 and 86.91% of COD as well as 91.11 and 88.72% of DEP removal were achieved
for the AnFFBR and UAnFFFBR, respectively. According to kinetic coefficients (except biomass yield), the AnFFBR had
better performance as it provided a more favorable condition for microbial growth. The Grau model was selected as
the best mathematical model for designing and predicting the bioreactors’ performance due to its high coefficients
of determination (0.97 < R2). With regard to the insignificant variations of the calculated Grau kinetic coefficients (KG)
when the organic loading rate (with constant HRT) increased, it can be concluded that both of the bioreactors can
tolerate high organic loading rate and their performance is not affected by the applied DEP concentrations.
Conclusions: Both the bioreactors were capable of treating low-to-high strength DEP wastewater; however, accord-
ing to the experimental results and obtained kinetic coefficients, the AnFFBR indicated higher performance. Although
the AnFFBR can be considered as a safer treatment option than the UAnFFFBR due to its lower DEP concentrations
*Correspondence: gholamim@iums.ac.ir; ehsanahmadi_eh@yahoo.com;

e‑ahmadi@razi.tums.ac.ir
†
Samira Yousefzadeh and Ehsan Ahmadi contributed equally to this work
3
Students’ Scientific Research Center (SSRC), Tehran University of Medical
Sciences, Tehran, Iran
5
Occupational Health Research Center (OHRC), Iran University of Medical
Sciences, Tehran, Iran
Full list of author information is available at the end of the article
© The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/
publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Content courtesy of Springer Nature, terms of use apply. Rights reserved.

Yousefzadeh et al. Biotechnol Biofuels (2017) 10:139 Page 2 of 15
in sludge, the UAnFFFBR had lower VSS/TSS ratio and sludge yield, which could make it more practical for digestion.
Finally, both the bioreactors showed considerable methane yield; however, compared to the UAnFFFBR, the AnFFBR
had more potential for bioenergy production. Although both the selected bioreactors achieved nearly 90% of DEP
removal, they can only be considered as pre-treatment methods according to the standard regulations and should be
coupled with further technology.
Keywords: Biofilm, Diethyl phthalate, Wastewater, Anaerobic treatment, Organic loading rate, Hydraulic loading rate,
Biodegradation, Phthalic acid esters
Background organic matters in wastewater is an economic way for

Environmental and water pollution has become an issue wastewater remediation [30].
of serious international concern in recent years [1–5]. The logical reasons to use anaerobic treatment pro-
In this regard, legislation requirements for discharging cesses can be explained by considering the advantages
wastewater have recently become much stricter [6]. of these processes which are related to their low energy
Emerging synthetic chemicals have entered into the demand, less sludge production, bioenergy (methane
environment through industrial activities [6, 7]. One of and hydrogen) production, and low nutrient require-
these emerging synthetic compounds is phthalic acid ment [31–36]. Furthermore, industrial wastewaters with
esters (PAEs), commonly named phthalates [8], which are high chemical oxygen demand (COD) containing high
widely used as plasticizers [9, 10] and globally applied in concentration of pollutants cannot be easily biotreated
large quantities to make products such as plastics, pes- by aerobic bioreactors; this is while anaerobic bioreac-
ticides, adhesives, paints, and cosmetics [11–13]. PAEs tors have been preferably and successfully applied to such
are ubiquitous pollutants as they have extensive applica- wastewater [30, 31] and organic compounds of the men-
tions in industrial processes, are not chemically bonded tioned wastewaters are ideal for green bioenergy produc-
to products, and can migrate to environments [6, 14]. tion [37].
In recent years, PAEs have been considered as hazard- Attached growth (biofilm) anaerobic bioremediation
ous environmental pollutants [8]. They can have carcino- has additional advantages over suspended growth bio-
genic, mutagenic, and teratogenic potential impacts due reactors, including more stability in operation, higher
to their ability to disrupt endocrine function [15–17]. tolerance for pH, temperature, and toxic shocks, higher
Moreover, they can cause adverse health problems in utilization rate of substrate, and ability to retain high
humans’ reproductive and developmental systems [18– biomass concentration under shorter hydraulic reten-
21] and have been associated with type 2 diabetes [22]. tion time (HRT) and overcome wash-out issues [30, 31,
Therefore, some regulatory organizations includ- 37–39].
ing China National Environmental Monitoring Center As highlighted above, microbial biodegradation can
(CNEMC), the United State Environment Protection play an important role in removing PAEs and it is vital
Agency (USEPA), and the European Union have classified for industries to select the best treatment technology for
PAEs as top-priority pollutants [23]. all the wastewater with high concentration of phthalates
Diethyl phthalate (DEP) is one of the important phtha- [6]. Therefore, the main aim of the present study was
lates, the concentration of which was reported to be to evaluate the performance of two different anaerobic
about 500 mg L−1 in discharged industrial effluent [8]. biofilm bioreactors including anaerobic fixed film baf-
Although phthalates may not be significantly elimi- fled reactor (AnFFBR) and up-flow anaerobic fixed film
nated by physico-chemical processes like hydrolysis and fixed bed reactor (UAnFFFBR), which have promising
photolysis, and other advanced treatment processes such performance and have been recently used for industrial
as membrane and oxidation techniques are not advisable wastewater [40–42]. Moreover, as kinetic coefficients and
due to their high investment and operation costs, it has mathematical modeling are two crucial tools that can
been shown that microbial biodegradation can play an be used to predict the performance of bioreactors and
important role in removing these pollutants from various optimize their design [43, 44], they were used for com-
environments [8, 24–26]. paring the selected bioreactors, since they can represent
Meanwhile, a safe and cost-effective biological waste- the microbial activity in different environmental condi-
water treatment is an important issue to consider [27, tions provided by the selected bioreactors. As kinetic and
28], especially for the industries which have to find finan- mathematical modeling directly relies on removal effi-
cially feasible treatment methods to meet the permissi- ciency and microbial mass, they preferably, practically,
ble discharge levels [29]. Anaerobic biodegradation of and easily can be used by the designers and operators of

treatment plants, while the recognition of microbial type The bioreactors were operated at 25 ± 2 °C and fitted
and their number cannot be used for predicting the per- with the heaters to maintain the reactors’ temperature
formance of bioreactors [30, 45]. In addition, there are stable. The bioreactors were filled by high-density poly-
many analytical and conceptual challenges for profiling ethylene (HDPE) carriers, which acted as a fixed bed for
the diversity of microbial communities and diagnosis of biofilm (microbial) growth. The carriers had approxi-
their species in complex microbial communities such as mately 0.95–0.98 g cm−3 and 535 m2 m−3 of density and
wastewater [46, 47]. active surface area, respectively. The bioreactors were
While many studies have been conducted to find a filled by 1.6 m2 of the carriers’ available surface area (50%
practicable way for evaluating the behavior of bioreac- filling ratio).
tors applied to wastewater bioremediation, they could In the acclimation step, glucose corresponding to
not completely evaluate the bioreactors’ performance in 600 mg L−1 of chemical oxygen demand (COD) was used
terms of kinetic coefficients and mathematical modeling. as the primary substrate and sole carbon source. Then,
Moreover, other studies have been only focused on calcu- after reaching the steady-state condition, DEP was added
lating critical kinetic coefficients and computing removal stepwise and glucose concentration was reduced in paral-
efficiency, and have not investigated and evaluated their lel until DEP formed the entire carbon source. The exper-
variations [48–50]. In addition, selecting the best biore- iments were conducted in two different study steps. The
actors is traditionally done with their performance like effects of HRT and organic loading rate (different con-
COD removal, while their kinetic coefficients and bioen- centrations of DEP) on the performance of the selected
ergy production are not usually considered very well. It bioreactors were evaluated in study steps (A) and (B),
should be noted that we previously applied mathematical respectively.
modeling and kinetic evaluation for comparing the bio- To have the chemical oxygen demand/nitrogen/phos-
degradability of different PAEs as substrates [51]; to our phorous ratio of 350/5/1, ammonium chloride (NH4Cl)
best knowledge, it is the first study for comparing the per- and ammonium bicarbonate (NH4HCO3) (for nitro-
formance of the selected bioreactors by the mentioned gen source) along with mono-potassium phosphate
methods. It should also be noted that there are other (KH2PO4) (for phosphorous source) were used as nutri-
important parameters like the produced sludge quality ents in all the study steps. The composition of trace ele-
and bioenergy production which need to be considered. ments in synthetic wastewater was selected as follows:
Therefore, to fill the mentioned research gaps, this study CaCl2·2H2O (14 mg), M gSO4·7H2O (90 mg), and 0.3 mL
was mainly aimed to compare two biofilm-based reactors of trace solution per liter of synthetic wastewater. The
including anaerobic fixed film baffled reactor (AnFFBR) following compounds were dissolved per liter to pre-
and anaerobic fixed film fixed bed reactor (UAnFFFBR) pare the trace solution: KI (0.18 g), M nCl2·H2O (0.12 g),
for DEP removal from synthetic wastewater. FeCl3·6H2O (1.5 g), C uSO4·5H2O (0.03 g), H 3BO3
(0.15 g), CoCl2·6H2O (0.15 g), ZnSO4·7H2O (0.12 g),
Methods Na2MoO4·2H2O (0.06 g), and EDTA (10 g) [52]. Further-
Implementation and operation of bioreactors more, NaHCO3 was applied to adjust pH at 7.5 ± 0.2.
Two laboratory-scaled rectangular-shape bioreactors
with the identical 6 L operating volume were used in this Kinetics and mathematical modeling
study. The reactors were made of 4 mm-thick Plexiglas. Critical kinetic parameters and mathematical models
The AnFFBR was divided into three equal compartments have important impacts for predicting and designing bio-
by vertical baffles (each part contained 2 L operating vol- logical wastewater treatment plants.
ume and split by a further baffle) which were connected In the present study, three common substrate removal
internally. models, namely Stover-Kincannon, first order, and Grau
The dimensions of UAnFFFBR were 10 (length) × 10 (second order), were used to design the bioreactors and
(width) × 70 cm (height) (Additional file 1: Figure S1). predict substrate removal rate. Under steady-state con-
Both bioreactors were seeded with an anaerobic ditions (dS
dt = 0), if the first-order model prevails, the
sludge obtained from a full-scale municipal wastewa- substrate consumption rate can be predicted by Eq. (1).
ter treatment plant (Ekbatan, Iran). Synthetic waste- Furthermore, Eq. (2) is the simplified form of Eq. (1):
water was continuously pumped from the feed tank
ds Q · S0 Q·S
into the AnFFBR using a dosing pump (Etatron, Italy). − = − − K1 S (1)
Another identical dosing pump was also used to feed the dt V V
UAnFFFBR. For inoculating both the bioreactors (start-
S0 − S
up phase), further dosing pumps were used to recycle the = K1 S, (2)
washed-out sludge from the settling tank. HRT

where K1 is the kinetic constant for the first-order model In steady-state conditions, the substrate concentration
(day−1), S0 and S are the influent and effluent substrate changes in Eq. (8) can be ignored ( dS
dt = 0) and Eqs. (8) and
concentrations (mg L−1), respectively, Q is the inflow rate (9) given above can be combined as the following equation:
(L day−1), V is the reactor volume (liter), and HRT is the
1 k A · X (A ) 1
hydraulic retention time (day). = − . (10)
In the modified Stover–Kincannon model, the sub- S Ks Q(S0 − S) Ks
strate utilization rate for the biofilm-based bioreactors Finally, KS as the half saturation constant (mg L−1) and
can be determined by Eq. (3) which is based on organic (K) as the overall reaction rate (day−1) can be calculated
loading rate. Equation (4) is also the linearized form of from the linear regression of plotting S1 versus Q(SX0att−S) line.
Eq. (3): Furthermore, Xatt which is the attached mass of biofilm
(g VS) can be calculated by multiplying A (total available
area, m−2) by XA:

S0
dS Umax × Q × V
= (3) Xatt = XA × A. (11)

dt KB + Q × V S0
Moreover, (Y) and (Kd) as the biomass yield coefficient

(g VS produced/g substrate consumed) and biomass
V KB V 1
= × + (4) decay rate (day−1), respectively, can be calculated by mass
Q × (S0 − S) Umax Q × S0 Umax balance equation (Eq. 12) and Monod growth kinetic
where Umax is the maximum removal rate of the sub- (Eq. 13) [8]:
strate (mg COD L−1 day−1) and KB is the saturation value
dX
constant (mg L−1 day−1). After the linear regression V = QX0 − QXe + A(rg ) (12)
of Eq. (4), these coefficients can be calculated from the dt
intercept and slope of the linear graph, respectively.
rg = Y (rsu ) − Kd · A · X(A) (13)
Equations (5) and (6) represent the second-order sub-
strate removal model proposed by Grau. Equation (6) is where rg and Xe are the specific growth rate (g
also the integrated form of Eq. (5): VSS m−2 day−1) and sloughed VS from the reactor
(g VS day−1). If steady-state conditions are achieved
(dX
dt = 0), Eq. (14) can be obtained by combining Eqs. (12)
2
−dS S
= KG × X × (5) and (13) as follows:
dt S0

(S0 − S) Kd Xatt 1
HRT
= + . (14)
Xe Y Q · Xe Y
= (n × HRT) + m (6)
E Finally, by the linear regression of Eq. (14), (Y) and (Kd)
where n (dimensionless) and m (day−1) are the constants can be determined.
for a second-order model which can be determined from Subsequently, (µm) coefficient which is the maximum
the intercept and slope of the plotted line of HRT versus specific growth rate (maximum specific growth rate,
(S0 ×HRT)
(S0 −S) . Moreover, (X) is the suspended biomass con- day−1) can be calculated by multiplying (K) and (Y) coeffi-
centration (mg VSS L−1), (E) is the fractional substrate cients expressed in Eq. (15). Eventually, influent substrate
removal efficiency (dimensionless), and KG is the Grau utilization rate can be calculated from Eq. (16), obtained
second-order substrate removal rate constant (day−1). KG by combining Eq. (9) with Eq. (14) [30, 49]:
can be obtained from the following equation [53]:
µm = K · Y (15)
S0
m= . (7) µm · X · S0
KG × X
rsu = . (16)
Other kinetic coefficients including (KS) and (K) in the Y (Ks + S0 )
biofilm-based reactors can be obtained by combining Analytical methods
mass balance equation [Eq. (8)] with Monod equation COD and biofilm characteristics including total solids
[Eq. (9)]. rs is the substrate utilization rate (g m−2 day−1). (TS) and volatile solids (VS) were analyzed by the analyti-
cal techniques of standard methods [54].
dS
V = QS0 − QS − A(rs ) (8) Total organic carbon (TOC) samples were filtered by
dt a 0.45 µm filter and, subsequently, analyzed by TOC-
ds K · S · Xatt Vcsh (Shimadzu, Japan) to measure the extend of
rs = − = . mineralization.
dt Ks + s (9)

Biogas measurements were performed as described in Results and discussion

the study of Lay et al. [55]. Evaluating performance of bioreactors
Moreover, the analytical measurement of diethyl Experimental results for the AnFFBR and UAnFFFBR
phthalate was performed using a gas chromatograph performance in terms of removing diethyl phthalate
(GC) which was coupled with a flame ionization detector under different operation conditions including differ-
(FID) and equipped with a HP-5 capillary column. ent HRTs and influent DEP concentrations are shown in
To measure the DEP concentration, 10 mL of the Tables 1 and 2, respectively. It should be noted that data
wastewater sample was filtered through a glass fiber fil- presented in Table 2 [study step (A)] which obtained
ter with 0.7 µm pore size and, subsequently, extracted by under different HRTs have previously been used for com-
2 mL of n-hexane solution. Finally, 2 µL of the extracted paring biodegradability of different PAEs [51]. The con-
sample was injected into GC-FID. Concentration of the centrations of COD, TOC, and DEP in the effluent of the
samples was measured by comparing them to the stand- bioreactors were determined as the bioreactor responses.
ard curve prepared at five points. Naphthalene was added The initial analysis for the COD and TOC experiments
as the internal standard. showed that each 1 g DEP L−1 produced 1.85 g COD L−1
GC temperature program was as follows: the oven’s and 0.62 g TOC L−1, respectively.
initial temperature was set at 70 °C for 1 min and, then,
followed by a 10 °C min−1 ramp to 250 °C as its final Analyzing impacts of hydraulic retention time
temperature and was maintained at this temperature In study step (A), the influent concentration of DEP
for 2 min. The injector and detector temperatures were was maintained constant to 300 mg L−1 and HRTs were
set at 250 and 260 °C, respectively. To quantify the DEP changed from 12 to 36 h. At this stage, the obtained
concentration in effluent sludge (sloughed biofilm), 1 g results showed that the DEP removal rate for the AnFFBR
of the freeze-dried sludge sample was ground and, subse- can increase from 73.46 to 90.26% with the retention
quently, extracted by 10 mL of n-hexane. time increase from 12 to 36 h. Pirsaheb et al. [56] found
To detect the metabolites of biodegradation, a gas similar results for the suspended growth anaerobic baf-
chromatograph system equipped with a mass spectrom- fled reactor (ABR) and realized that higher HRTs resulted
eter (MS) detector and liquid chromatography tandem– in more COD removal. The related COD and TOC
mass spectrometry (LC–MS/MS) were used. removal efficiencies which showed similar behaviors and
Table 1 Performance of anaerobic fixed film baffled reactor (AnFFBR) in removing DEP
Study step A B
Study phase 1 2 3 4 5 1 2 3 4
−1
Influent DEP concentration (mg L ) 300 300 300 300 300 400 500 600 700
HRT (h) 12 18 24 30 36 36 36 36 36
Lorg (g COD m−2 day−1)a 4.162 2.775 2.081 1.665 1.387 1.85 2.312 2.775 3.237
Attached mass [TS (mg) of biofilm] 6180 4860 4180 3880 3710 4560 5400 6260 7170
Attached mass [VS (mg) of biofilm] 4500 3300 2820 2520 2400 3120 3900 4560 5340
VS/TS ratio 0.728 0.679 0.674 0.649 0.647 0.684 0.722 0.728 0.744
Effluent total suspended solids 334.3 205 137.7 111.4 90 141.6 168.8 239.5 276.2
(mg day−1)
Effluent volatile suspended solids 224.4 130.2 84.5 64.2 48.4 81 98.6 144.3 168.9
(mg day−1)
DEP concentration in TSSe (mg g−1)b 8.5 8.2 6.6 6.2 4.1 7.3 7.5 7.5 7.9
SRT (day) 18.48 23.7 30.35 34.83 41.22 32.2 31.99 26.13 25.96
Methane production (L/g CODrem) 0.21 (0.19)c 0.28 (0.25) 0.32 (0.29) 0.37 (0.34) 0.44 (0.4) 0.38 (0.35) 0.35 (0.32) 0.34 (0.31) 0.31 (0.28)
Methane percentage (%) 42.2 44.2 52.5 56.4 64.8 63.3 62.1 61.7 61.1
DEP removal (%) 73.46 78.43 81.4 86.33 90.26 90.62 90.64 90.83 91.11
COD removal (%) 66.66 70.1 77.33 82.29 87.01 88.04 89.48 89.6 90.31
TOC removal (%) 51.82 57.95 66.82 75.7 83.17 83.06 81.90 82.5 82.57
a
Organic loading rate
b
Effluent total suspended solids
c
Numbers in brackets are methane yield at STP (1 bar and 273.15 °K)

Table 2 Performance of up-flow anaerobic fixed film fixed bed reactor (UAnFFFBR) in removing DEP
Study step A B
Study phase 1 2 3 4 5 1 2 3 4
Influent DEP concentration (mg L−1) 300 300 300 300 300 400 500 600 700
HRT (h) 12 18 24 30 36 36 36 36 36
Lorg (g COD m−2 day−1) 4.162 2.775 2.081 1.665 1.387 1.85 2.312 2.775 3.237
Attached mass [TS (mg) of biofilm] 5840 4370 4110 3880 3720 4340 5440 6080 6840
Attached mass [VS (mg) of biofilm] 4080 2940 2700 2460 2280 2880 3660 4320 4980
VS/TS ratio 0.698 0.672 0.657 0.634 0.613 0.663 0.672 0.71 0.728
Effluent total suspended solids (mg day−1) 326.8 227.3 169.3 129.6 112.2 144.5 181.4 220 270
Effluent volatile suspended solids 164.4 102.4 74.4 55.7 48 76.8 96 117.6 151.8
(mg day−1)
DEP concentration in TSSe (mg g−1) 9.7 9.3 8.6 8.1 5.2 7.4 7.5 8.4 9.1
SRT (day) 17.87 19.22 24.27 29.94 33.15 30.03 29.98 27.63 25.33
Methane production (L/g CODrem) 0.22 (0.2) 0.24 (0.22) 0.28 (0.26) 0.34 (0.31) 0.42 (0.38) 0.34 (0.31) 0.32 (0.29) 0.30 (0.27) 0.27 (0.24)
Methane percentage (%) 41.1 42.3 52.8 56.1 61.5 61.0 59.5 59.6 57.3
DEP removal (%) 67.9 72.5 76.46 80.96 87.86 88.15 88.3 88.36 88.72
COD removal (%) 57.91 60.86 71.85 78.79 83.55 84.25 86.02 86.27 86.91
TOC removal (%) 45.32 53.97 62.58 70.05 78.33 78.34 78.25 78.79 79.65
Lorg organic loading rate, TSSe total solids, VS volatile solids, TSSe effluent concentration of total suspended solids
maximum removal of COD and TOC was measured as The effluent quality of the UAnFFFBR, especially in
87.01 and 83.17%, respectively, in 36 h of HRT. The DEP study phases (A-1) and (A-2), was significantly less than
removal in the AnFFBR was considerably higher than its that in the AnFFBR. This result revealed the AnFFBR
mineralization (COD and TOC removal), which can be could tolerate higher organic and hydraulic loading rates
related to the presence of benzene ring (aromatic struc- than the UAnFFFBR. Greater performance and toler-
ture) known to be more refractory to biodegradation [8, ance of the AnFFBR can be the result of phase separation
57]. The experimental results of study phase (A-1) which known as the significant advantage of anaerobic baffled
compared DEP and TOC removal confirmed this theory. reactors (ABR) [58]. This phenomenon may cause acido-
Nearly 21.64% higher removal rate of DEP than TOC genesis and methanogenesis bacteria to be divided into
showed that the side chains of DEP were biodegraded two different phases in the AnFFBR, which subsequently
faster than its benzene ring. This difference reduced to make it possible for these bacteria to grow and synthe-
7.1% by increasing HRT to 36 h. size under their favorable conditions. In this regard, the
Similar observations were recorded for the UAnFFFBR. growth rates of the attached biofilm mass (as mg VS and
The improved effluent quality of both bioreactors at TS) were higher for the AnFFBR than UAnFFFBR. Fur-
higher HRTs can be related to the higher contact time thermore, as the volatile solids (VS)-to-total solids (TS)
of microbial mass to DEP. Another critical factor is solid ratio of the biofilm were higher for the AnFFBR than
retention time (SRT), which shows the presence time of UAnFFFBR, it can be concluded that the microorganisms
microbial mass to fulfill the organic substrate biodeg- had greater activity in the AnFFBR.
radation. In study step (A), the SRTs of both reactors Moreover, higher SRTs in the AnFFBR are due to
notably increased by reducing the organic loading rate greater persistence of microbial mass to the attained con-
and increasing HRT. The highest SRTs for study step (A) ditions, which consequently leads to more effective bio-
were observed in 36 h HRT, which were equal to 41.22 reaction in the AnFFBR. However, both the bioreactors
and 33.15 (days) for the AnFFBR and UAnFFFBR, respec- performed well at 300 mg influent DEP concentration per
tively. These obtained SRTs were considerably higher L and 36 h HRT, resulting in 90.26 and 87.86% of DEP
than other suspended growth processes (e.g., conven- removal to be achieved for the AnFFBR and UAnFFFBR,
tional aerobic activated sludge) ranging from 3 to 30 days. respectively.
It was also known that SRTs between 5 and 50 days are
ideal for xenobiotic compounds to be biodegraded which Analyzing effect of organic loading rate
can be considered as an advantage for both of the studied The bioreactors’ performance under different diethyl
reactors [30]. phthalate concentrations and organic loading rates

(stable hydraulic loading rate of 4 L day−1) was evalu- It can be observed that these bioreactors achieve more
ated in study step (B). According to the bioreactors’ removal rates as they increase the HRT and SRT, which
best performance observed in study phase (B-4), 90.31 are in agreement with the observed results of the present
and 86.91% of COD and 91.11 and 88.72% of DEP were study [8, 60, 64]. In addition, there is a diverse relation
removed by the AnFFBR and UAnFFFBR, respectively. In between the length of alkyl-side chains of phthalates and
this step, the variation of removal rate slightly increased their biodegradability [8, 61].
as the substrate and organic loading rate increased from
400 to 700 mg DEP L−1 and 1.85 to 3.237 (g COD/m2carrier Biogas production
day), respectively. It can be concluded that HRT had a Methane, carbon dioxide, hydrogen, and hydrogen sulfide
greater impact than organic loading rate on these biofilm were observed as the main gases in biogas. Both meth-
reactors. Although the bioreactors’ removal efficiencies ane production rate and its percentage for bioreactors
increased in study step (B), the COD and DEP concentra- were affected by HRT and organic loading rate. Accord-
tions slightly increased in the effluent. Moreover, VS/TS ing to the results, the methane production increased by
ratio and active biofilm mass are two parameters which increasing HRT (study step A) and, then, decreased by
can affect the bioreactors’ performance in study step (B). increasing organic loading rates (study step B). It is com-
It can be concluded that, by increasing organic loading monly known that operational parameters can critically
rate, both the attached microbial masses and their vola- affect the biogas production rates [66]. The maximum
tile portion increased and, consequently, led to higher methane production rates were observed in study phase
biodegradation capacity of the bioreactors (Tables 1, 2). (A-5) with 36 h of HRT as 0.44 and 0.42 L CH4 g COD−1 rem
It should be noted that better effluent concentra- at 1 bar and 298.15 °K (or 0.4 and 0.36 L CH4 g COD−1 rem
tion and performance of the AnFFBR than those of the at 1 bar and 273.15 °K) for the AnFFBR and UAnFFFBR,
UAnFFFBR in study step (B) can be related to its higher respectively.
biofilm mass and VS/TS ratio. Another factor is phase For most of the study phases, the methane produced
separation which was previously mentioned. Moreover, by the AnFFBR was considerably higher than that pro-
it has been known that anaerobic reactors’ performance duced by the UAnFFFBR, which may be the result of
in higher organic loading rates and for strong wastewa- phase separation provided by the baffles of AnFFBR that
ters (more than 1000 mg L−1 of influent COD) is better consequently help the acetogenic bacteria to utilize the
compared to when they are used for low strength waste- volatile fatty acids (VFAs) produced by acidogenic bacte-
water treatment [31]. The experimental results of this ria before these metabolites reach methanogenic bacteria
study (which indicated stable performance of the anaero- and inhibit their activities. This issue can be confirmed by
bic bioreactors) along with Farzadkia et al.’s investigation the results obtained in the study by Wang et al., indicat-
can demonstrate the advantage of these anaerobic biofilm ing that some VFAs like propionic acid can significantly
reactors over aerobic bioreactors [59]. In the study by inhibit the methanogenesis process [67]. This hypoth-
Farzadkia et al., high organic loading rate of the substrate esis can be supported when the methane yield of the
had an adverse effect on the biodegradation and meta- UAnFFFBR in study step B more significantly decreased
bolic activity of aerobic fixed bed activated sludge hybrid than that of the AnFFBR by increasing organic load-
reactor, and the removal efficiency of the reactor dropped ing rate. This phenomenon can be attributed to greater
from about 96–79% when organic loading rate was accumulation of some inhibitory compounds (e.g.,
increased from about 1 to 4.5 kg COD m−3 day−1 [59]. propionic acid) in the UAnFFBR. Moreover, the study
It should be also noted that the statistical t test analyses results of step (A) demonstrated that methanogenic bac-
for DEP, COD, and TOC removal efficiencies showed the teria needed more time to convert the VFAs and other
mentioned parameters to be statistically different in most metabolites into methane. It can be also observed that
of the study phases when compared between the bioreac- increasing HRTs from 12 to 36 h for each of the biore-
tors (P value <0.05). actors can lead to the increase of the methane produc-
A summary of some studies conducted on the evalu- tion for about twice. These results could be attributed to
ation of biological removal of PAEs is presented in slow growth and synthesis rate of methanogen bacteria,
Table 3. It should be noted that, although all these bio- which results in more SRTs requirement for optimum
logical methods indicate that bioremediation can play an growth and the substrate concentration increase needed
important role in removing PAEs, the type and condition by these bacteria (e.g., acetic acid) in higher HRTs. How-
of bioreactors including (anaerobic, aerobic, and anoxic ever, some of the obtained methane yields in the present
conditions), their operational conditions (particularly study, especially in study phase (A-5), were higher for
HRT and SRT), and finally type of the selected phthalate both of the bioreactors compared to some other studies
can impact the performance of bioreactors [8, 60–65]. which have commonly reported the methane yields of

Table 3 Summary of some studies conducted for biological removal of PAEs
Treatment method Substrate type (PAEs) Experimental conditions and removal effi- Important observation Refs.
ciency
Anaerobic/anoxic/oxic (AAO) treatment system DMP The optimal HRT and SRT for DMP and nutrients The biodegradation process of DMP by the [60]
removal were 18 h and 15 days, respectively, selected method was in accordance with the
and the degradation rates of anaerobic, anoxic first-order kinetics model. Under the optimal
and aerobic zones for DMP were 13.4, 13.0, and conditions, about 73.8, 5.8, 19.3, and 1.1% of
67.7%, respectively DMP was biodegraded, released in the effluent,
accumulated in the system, and remained in
the waste sludge, respectively
Trickling filter DEP and DEHP Trickling filter achieved 94–99% of DEP and 44% DEHP was the most recalcitrant among the [61]
of DEHP removal selected phthalates and DEP with less molecu-
Yousefzadeh et al. Biotechnol Biofuels (2017) 10:139
lar weight was biodegraded with higher rate

Cyclic activated sludge technology (CAST), DEP, DMP, DnBP, BBP, DEHP and DOP The overall removal efficiency of all the selected The better performance of the CAST process was [62]
anoxic/oxic (AO), and anaerobic/anoxic/oxic PAEs was more than 72% in CAST while AO and attributed to its better indoor-conditions for
(AAO) processes AAO only achieved about 30% of PAEs removal bacterial community
Moving bed biofilm reactor (MBBR) DAP and DEP In optimum conditions with HRT of 9 h, about MBBR tolerated both of the selected phthalates [8]
95 and 94% of DEP and DAP were removed, with the influent concentrations of 100–
respectively. In addition, more than 92% of COD 300 mg L−1. Sludge yield constants (Y) varied
removal was achieved for both phthalates from 0.3 to 0.54. The maximum specific growth
rates were 0.37 and 0.34 day−1 for DEP and DAP,
respectively
Up-flow anaerobic sludge blanket reactor (UASB) DMP More than 99% of DMP and 93% of COD were The sludge yield was estimated as 0.08 g [63]
removed from the wastewater containing VSS g COD−1. DMP was first de-esterified to
600 mg L−1 of DMP (corresponding to 3 g mono-methyl phthalate (MMP) and, then, to
COD L−1 day−1) of organic loading rate of ) at phthalate (or phthalic acid) and, subsequently
8 h of HRT de-aromatized and converted into methane
and CO2. The maximum specific degradation
rates of DMP, MMP, and phthalate were 415, 88,
and 36 mg (g VSS day)−1, respectively
Anaerobic/anoxic/oxic treatment system (AAO) DnBP The optimal HRT and SRT for DnBP removal were Increasing SRT from 10 to 15–25 days resulted in [64]
18 h and 15 days, respectively. In the men- increasing removal efficiency from 90 to 95%.
tioned conditions, about 72.66% of DnBP was Higher SRT improved DnBP biodegradation
degraded by the selected process, 24.44% was efficiency. The removal efficiencies of anaerobic,
accumulated in the system, 2.44% was released anoxic and oxic reactors were 17.14, 15.02, and
in the effluent, and 0.5% remained in the waste 63.46% of the total DnBP removal, respectively
sludge
Submerged membrane bioreactor (aerobic DEHP The removal efficiency of DEHP under HRTs of 4 The removal of DEHP was closely dependent on [65]

condition) and 6 h, SRT of 140 days, and sludge concentra- the membrane pore size and about 74% of inlet
tion of 11.5 and 15.8 g VS L−1 ranged between DEHP was biodegraded
91 and 98%
DMP dimethyl phthalate, DEHP bis(2-ethylhexyl) phthalate, DnBP di-n-butyl phthalate, BBP butyl benzyl phthalate, DOP di-n-octyl phthalate, DAP diallyl phthalate
Page 8 of 15
about 0.27 to 0.36 L C H4/g COD [68, 69]. Moreover, the particularly short-chains phthalates [e.g., DEP and dime-
methane production rate is usually expected to be placed thyl phthalate (DMP)], can be removed and mineralized
around 0.35 L C H4/g COD in STP condition (or 0.382 L in significant amounts. However, some other phthalates
CH4/g COD at 1 bar and 298.15 °K), which is the theo- with long chains like di(2-ethylhexyl) phthalate (DEHP)
retical methane yield [34, 69, 70]. The obtained higher may show resistance to digestion [6]. Although the pro-
methane yields in this study might be the result of further duced sludge in the AnFFBR had less adsorbed DEP, the
conversion of biosolids produced from dead biofilm cells UAnFFFBR had sludge with less VSS/TSS ratio (effluent
and their lysis into methane and other biogases. It can volatile suspended solids/effluent total suspended solids),
be confirmed that the effluent volatile suspended solids which can make it more feasible for digestion and further
( VSSe) and effluent total suspended solids (TSSe)/VSSe uses. The minimum and maximum VSS/TSS ratios were
ratios were considerably lower for both of the bioreactors observed as 0.537 and 0.671 for the AnFFBR, and 0.427
in study phase (A-5), as compared to other study phases. and 0.562 for the UAnFFFBR [both the minimum ratios
As an example, for the AnFFBR, the V SSe and V SSe/ were observed in study phase (A-5)]. The minimum VSS/
TSSe ratios in study phase (A-5) were 48.4 mg day−1 and TSS ratio for the UAnFFFBR (0.427) was considerably
0.537, respectively, which were considerably lower than less than the one reported for the aerobic moving bed
the VSSe and VSSe/TSSe ratios of 224.4 mg day−1 and biofilm reactor which ranged from about 0.52 to 0.68 [8]
0.671, respectively, obtained in study phase (A-1). This and can be considered as an important further sludge
point illustrates that the remained tissue of dead bacteria management advantage for this anaerobic reactor.
was converted into other inert solids and final products
including methane. Metabolic pathways of DEP biodegradation
However, it should be noted that there are many stud- According to the observed by-products, DEP in both bio-
ies which have reported the methane production rates reactors was primarily and mainly decomposed to mono-
between 0.4 and 0.47 L C H4/g COD [71–73]. ethyl phthalate (MEP) by the de-alkylation of its first side
chain and, then, the bioreaction continued to produce
Sludge quality phthalic acid (PA) by removing the other side chain of
Another essential factor which is particularly important MEP. This enzymatic reaction is known as de-esterifi-
for land application is the concentration of DEP adsorbed cation and commonly reported as the main phthalates’
by biomass, especially as it is known that the anaerobic biodegradation route [75]. In addition, the trace amount
digestion of biomass can produce residues that are rich in of other center metabolites including dimethyl phthalate
nutrients and have the potential to be used as the fertilizer (DMP) and mono-methyl phthalate (MMP) can be the
[74]. According to the results, DEP can be considerably result of de-methylation which is a less common and pre-
adsorbed by biofilm and, consequently, observed in efflu- dominant biodegradation pathway [75, 76]. Amir et al.
ent TSS. Moreover, the adsorbed concentration of DEP in [76] reported similar observation and stated that during
sloughed biofilm depends on influent DEP concentration sludge composting of di(2-ethylhexyl) phthalate (DEHP),
and organic loading rate. Minimum DEP concentrations DMP can be produced as the by-products of diethyl
in TSS for both the bioreactors were observed in study phthalate biodegradation due to de-methylation pathway.
phase (A-5) which had the minimum organic loading Similar observed metabolites for both bioreactors have
rate and influent DEP concentration and were as 4.1 and indicated that the biodegradation pathway is not different
5.2 mg DEP g TSS−1 for the AnFFBR and UAnFFFBR, and microbial activity in both bioreactors’ condition can
respectively. It should be noted that, for most of the lead to the same by-products.
study phases, the DEP concentrations in TSS were lower The other important degradation metabolites pro-
in the AnFFBR than UAnFFFBR, demonstrating that the duced before ring cleavage included protocatechuic
sludge of the AnFFBR can be used more safely. This fact acid, 4-hydroxyphthalic acid, benzoic acid, 4,5-dihy-
becomes more important when the produced sludge is droxyphthalic acid, catechol, and 3,4-dihydroxy benzoic.
used for land application and especially for agriculture Then, the biodegradation of the remained benzene ring
purposes, because the previous research has shown that of the mentioned by-products can result in the produc-
some phthalates can reduce the plant growth and cause tion of observed volatile fatty acids (VFAs), which have
chloroplast disintegrating [9]; others have stated that commonly been reported in the anaerobic biodegrada-
they can be introduced into the food chain and, conse- tion of organic compounds [77]. The final products of
quently, produce human exposure. However, both the biodegradation were methane, carbon dioxide, dihy-
bioreactors’ sludge should further be treated like other drogen oxide, and hydrogen as mentioned before in the
sludge. The performed research for evaluating digestion biogas. It should be noted that the VFAs and some of the
of phthalates in sludge has indicated that such phthalates, other by-products can be produced in each of the other

biodegradation steps. Simplified biodegradation pathway respectively. In this regard, the Grau model can be used
is presented in Fig. 1. for predicting the performance of both of the bioreac-
tors. Evaluating the KG values demonstrated this model
Modeling and analysis of kinetic coefficients to be in compliance with the obtained results, including
As shown in Fig. 2, the first-order model for both of the COD, DEP, and TOC removal, because the KG values for
bioreactors had low coefficients of determination, which the AnFFBR were higher than those for the UAnFFFBR.
were 0.377 and 0.305 for the AnFFBR and UAnFFFBR, In study step (A), as HRT increased, the KG values in the
respectively. Consequently, this model cannot be used for reactors were significantly raised, which may be due to
predicting performance of both of the bioreactors. the bioavailability of microorganisms to the substrate. In
For computing saturation value constant (KB) and max- study step (B), the KG values did not considerably change.
imum substrate removal rate (Umax), Eq. (4) was plotted With regard to the increasing DEP concentration and
for both bioreactors in Fig. 3. Moreover, semi-strong val- organic loading rate in study step (B) and stability of the
ues of coefficient of determination were obtained as 0.929 calculated KG, it can be concluded that both of the bio-
and 0.858 for the AnFFBR and UAnFFFBR, respectively, reactors can tolerate high organic loading rate and their
indicating that this model can be applied for designing performance was not affected by DEP concentration.
and predicting the selected bioreactors. Higher Umax and This point can be the advantage of these anaerobic sys-
KB of 4.04 (mg COD L−1 day−1) and 4.296 (mg L−1 day−1) tems over aerobic or other treatment processes for indus-
for the AnFFBR compared to 2.404 (mg COD L−1 day−1) trial wastewater treatment [30].
and 2.507 (mg L−1 day−1) for the UAnFFFBR, respec- According to Table 5 [correlation of methane yields
tively, demonstrated that the microbial community had (L CH4/g CODrem) and Grau coefficients], although in
better conditions for biodegrading DEP and stabilizing study step (B) for both bioreactors, strong correlation
COD in the AnFFBR than UAnFFFBR. (R2 > 0.7) between methane production rates and KG val-
Equation (6) was plotted in Fig. 4 for computing KG ues was not observed and, therefore, KG values could not
coefficient. After calculating (m) and (n) coefficients, KG be used for predicting methane yield with a high degree
was determined using Eq. (7). Table 4 presents the Grau of precision, strong coefficients of determination (R2) of
coefficient (KG) values. This model clearly had a high study step (A) as 0.791 and 0.937 for the UAnFFFBR and
degree of precision with the coefficients of determina- AnFFBR, respectively, can be used for the mentioned pre-
tion of 0.987 and 0.976 for the AnFFBR and UAnFFFBR, diction. Positive slopes of the obtained equations in study
Fig. 1 Biological degradation pathway of diethyl phthalate

Table 5 Correlation of methane yields (L CH4/g CODrem)

AnFFBR → y = 2.7508x + 205.59 (R² = 0.3773)
and Grau coefficients
UAnFFFBR → y = 1.765x + 227.07 (R² = 0.3056)
900
800 AnFFBR UAnFFFBR AnFFBR
700 UAnFFFBR
600
Study phase A Y = 0.117x − 0.041 Y = 0.156x − 0.05
(S0-S)/HRT
500
(R2 = 0.937) (R2 = 0.791)
400 Study phase B Y = −0.39x + 1.892 Y = −0.560x + 1.917
300 Bioreactor K1(d-1) (R2 = 0.571) (R2 = 0.390)
200 AnFFBR 2.75
100 UAnFFFBR 1.765
0 step (A) (0.156 and 0.117 for the UAnFFFBR and AnFFBR,
0 50 100 150 200 250
respectively) can demonstrate that HRT had a positive
S
impact on both KG and methane yield values and these
Fig. 2 First-order model for the AnFFBR and UAnFFFBR
dependent variables changed with a similar direction.
Higher calculated slope for the UAnFFFBR than AnFFBR
can demonstrate that HRT had a more impact on meth-
AnFFBR → y = 1.0634x + 0.2475 (R² = 0.9298) ane production of the UAnFFFBR and, considering lower
3.5
UAnFFFBR → y = 1.0432x + 0.416 (R² = 0.8588) obtained methane yields for the UAnFFFBR, the AnFFBR
3 AnFFBR had more advantages due to its less dependence on HRT.
2.5 UAnFFFBR Furthermore, with regard to the insignificant variations
of the calculated KG and negative slopes of study step
V/Q.(S0-S)
2
(B) as 0.56 and 0.39 for the UAnFFFBR and AnFFBR,
1.5 respectively, it can be concluded that methanogens activi-
1
ties were affected more than other non-methanogens by
Bioreactor Umax (mgCOD/L.d) KB (mg/L.d)
higher DEP loading rates. In addition, higher negative
0.5 AnFFBR 4.04 4.296
UAnFFFBR 2.404 2.507 slope for the UAnFFFBR can demonstrate that this biore-
0 actor was more sensitive to higher organic and DEP load-
0 0.5 1 1.5 2 2.5 3
ing rates and its methane yield was more affected.
V/(Q.S0)
The kinetic coefficients including (KS) and (K) were
Fig. 3 Stover–Kincannon model for the AnFFBR and UAnFFFBR
computed using Eq. (10), which is plotted in Fig. 5. Half
saturation constant and overall reaction rate were com-
puted as 31.34 mg L−1 and 1.13 day−1 for the AnFFBR,
and 24.87 mg L−1 and 1.03 day−1 for the UAnFFBR,
2 AnFFBR → y = 0.8913x + 0.363 (R² = 0.9879) respectively. It should be noted that the anaerobic waste-
1.8 UAnFFFBR → y = 0.8211x + 0.5362 (R² = 0.9764) water treatment has slower substrate utilization rate as
1.6 AnFFBR one of their disadvantages [31], which can be confirmed
(S0*HRT)/(S0-S)
1.4
1.2
UAnFFFBR by the obtained overall reaction rates. As an example,
1 for an aerated submerged fixed film reactor with glu-
0.8 cose as the substrate, the overall reaction rate has been
0.6
Bioreactor m (d-1) n (dimentionless) computed about 2.7 day−1, while for other conventional
0.4 AnFFBR 0.3636 0.8913
0.2 UAnFFFBR 0.5362 0.8211
aerobic treatments, it is reported up to 12 day−1 [30,
0 49]. Half saturation constant presents the substrate con-
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6
centration at half of the maximum substrate utilization
HRT (day)
rate. The previous studies have declared that a substrate
Fig. 4 Second-order (Grau) model for the AnFFBR and UAnFFFBR
with higher biodegradability has more KS [8]. Under low
Table 4 Grau second-order kinetic coefficients (KG) for AnFFBR and UAnFFFBR
Study phase A-1 A-2 A-3 A-4 A-5 B-1 B-2 B-3 B-4
KG for AnFFBR 2.038 2.779 3.253 3.64 3.822 3.92 3.92 4.023 4.01
KG for UAnFFFBR 1.522 2.112 2.300 2.524 2.723 2.875 2.828 2.875 2.909

AnFFBR → y = 0.0361x - 0.0319 (R² = 0.9203)

removal performances, the UAnFFFBR can be prefer-
0.016
UAnFFFBR → y = 0.0417x - 0.0402 (R² = 0.9686)
ably used when strong wastewater with high hydraulic
0.014
AnFFBR loading rate must be treated. Under this circumstance,
0.012
UAnFFFBR the insignificant difference observed for (Y) between the
0.01
selected bioreactors results in sludge production to dif-
1/S
0.008
fer significantly. Meanwhile, the biomass yields for both
0.006
Bioreactor KS (mg/L) K (d-1) bioreactors are very desirable when compared to those
0.004
AnFFBR 31.34 1.131 for aerobic based reactors. Experimental results for the
0.002 UAnFFFBR 24.87 1.035
biodegradation of similar compounds in aerobic biore-
0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 actors have shown that they have biomass yields more
Xatt/(Q.(S0-S) than 0.5 or even up to 0.78, which is more than five times
Fig. 5 Mathematical calculation of (KS) and (K) for the AnFFBR and greater than the calculated (Y) in this study [8, 48]. The
UAnFFFBR Kd coefficients, which present specific decay rate (and are
expressed as g VS of microbial mass loses/g VS of pre-
sents mass day or day−1) were computed as 0.107 and
0.100 day−1 for the AnFFBR and UAnFFFBR, respectively.
substrate availabilities and at the same maximum specific The difference of the values, however, was observed to
growth rate in the two microbial groups or even two dif- be insignificant between the bioreactors. These values
ferent bioreactors, smaller KS is obtained for microbial placed in the typical range of 0.06–0.15 day−1 for the
mass or a bioreactor with higher affinity to the substrate conventional treatment [30].
[78]. As the calculated maximum growth rate (µm) for the Consequently, maximum specific growth rate (day−1)
selected bioreactors is different, KS coefficients should values were calculated from Eq. (15) as 0.176 and
not be directly applied to compare the bioreactors’ per- 0.151 day−1 for the AnFFBR and UAnFFFBR, respec-
formances. By contrast, if the difference between two µm tively. This coefficient had good compliance with other
values is ignored, the UAnFFFBR can be stated to have experimental results (including COD removal) and with
better performance under low substrate concentrations the Stover–Kincannon and second-order (Grau) mod-
and, therefore, the microbial growth will be less influ- els, indicating that the AnFFBR had better performance.
enced by substrate concentration. It is a very important Considering the same inoculation used for both of the
point of view, especially for industries having different bioreactors, µm demonstrated that the microbial mass
work shifts, as it does not continuously generate waste- had more favorable conditions in the AnFFBR. Finally,
water or the substrate concentration is not stable in influ- (rsu) coefficients were computed for the study phases
ent wastewater. with maximum COD removal (phase B-4 for both of
The computed biomass yield coefficients (Y) plotted in the bioreactors). Higher rsu of 938.11 mg L−1 day−1 for
Fig. 6 were obtained as 0.156 and 0.146 (g VS produced/g the AnFFBR compared to 842.26 mg L−1 day−1 for the
COD utilized). The (Y) coefficient is an important param- UAnFFFBR confirmed other experimental results and
eter for sludge management and its subsequent disposal proved the AnFFBR to have better performance due to its
[77, 79]. From this point of view and without other faster utilization rate.
Conclusions
AnFFBR → y = 0.685x + 6.4058 (R² = 0.9884)
Both the bioreactors were capable of treating low-to-high
45 UAnFFFBR → y = 0.6877x + 6.8403 (R² = 0.9912)
strength DEP wastewater; however, the AnFFBR was
40 AnFFBR
preferred, since it could achieve 90.31% COD removal at
35 UAnFFFBR
36 h of HRT. This study suggested the Grau and Stover-
30
Kincannon models for predicting bioreactors due to
(S0-S)/Xe
25
their suitable coefficients of determination and good
20
conformity of their kinetic parameters to the obtained
15
Bioreactor Y Kd (d-1) results. Moreover, the experimental results and obtained
10 AnFFBR 0.156 0.107
5 UAnFFFBR 0.146 0.100
kinetic coefficients indicated that the AnFFBR had bet-
0
ter performance than the UAnFFFBR. Although both
0 10 20 30 40 50 60 these bioreactors can achieve nearly 90% of DEP removal,
Xatt/Q.Xe they are promising only as pre-treatment methods and,
Fig. 6 Mathematical calculation of (Y) and (Kd) for the AnFFBR and due to standard regulations, should be coupled with fur-
UAnFFFBR ther technology. Although from the sludge management

standpoint, the AnFFBR can be considered as a safer gratitude to Prof. Parvin Pasalar for her constant inspiration, encouragement,
and support throughout the whole period of this project.
treatment option than the UAnFFFBR due to its lower
DEP concentrations in sludge, the UAnFFFBR had less Competing interests
VSS/TSS ratio, which makes it more practical for diges- The authors declare that they have no competing interests.
tion. Moreover, the AnFFBR had more sludge yield which Availability of supporting data
should also be taken into consideration. Finally, both the All data generated or analyzed during this study are included in this published
bioreactors showed considerable methane yield; how- article with Additional file 1: Figure S1.
ever, compared to the UAnFFFBR, the AnFFBR had more Funding
potential for bioenergy production, which could result in This work was financially supported by grants from Occupational Health
saving more energy and costs. Research Center (OHRC), Iran University of Medical Sciences (Grant Number
92-03-132-23829).
Additional file
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in pub-
Additional file 1: Figure S1. Schematic diagram of the (A) UAnFFFBR;
lished maps and institutional affiliations.
and (B) AnFFBR.
Received: 8 January 2017 Accepted: 22 May 2017
Abbreviations
DEP: diethyl phthalate; AnFFBR: anaerobic fixed film baffled reactor; UAnFFFBR:
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Environmental Monitoring Center; USEPA: United State Environment Protec-
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Biotechnol Biofuels (2017) 10:139

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A comparative study of anaerobic fixed

*Correspondence: gholamim@iums.ac.ir; ehsanahmadi_eh@yahoo.com;

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Background organic matters in wastewater is an economic way for

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Moreover, (Y) and (Kd) as the biomass yield coefficient

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Biogas measurements were performed as described in Results and discussion

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lar weight was biodegraded with higher rate

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Fig. 1 Biological degradation pathway of diethyl phthalate

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Table 5 Correlation of methane yields (L CH4/g CODrem)

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AnFFBR → y = 0.0361x - 0.0319 (R² = 0.9203)

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