Bioresource Technology 100 (2009) 4969–4975

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

Biological kinetics evaluation of anaerobic stabilization pond treatment

of palm oil mill effluent
Yee Shian Wong a,*, Mohd Omar A.B. Kadir b, Tjoon Tow Teng b
a
School of Environmental Engineering, Universiti Malaysia Perlis, Kompleks Pusat Pengajian Jejawi 3, 02600 Arau, Perlis, Malaysia
b
School of Industrial Technology, Universiti Sains Malaysia, 11600 Gelugor, Pulau Pinang, Malaysia

a r t i c l e i n f o a b s t r a c t

Article history: Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill efflu-
Received 18 December 2008 ent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was oper-
Received in revised form 28 April 2009 ated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8 l of raw POME for a day. Chemical
Accepted 29 April 2009
oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed
Available online 26 June 2009
that the growth yield (YG), specific biomass decay (b), maximum specific biomass growth rate (lmax), sat-
uration constant (Ks) and critical retention time (Hc) were in the range of 0.990 g VSS/g CODremoved day,
Keywords:
0.024 day1, 0.524 day1, 203.433 g COD l1 and 1.908 day, respectively.
Biological kinetic
Palm oil mill effluent
Ó 2009 Elsevier Ltd. All rights reserved.
Monod equation
Chemical oxygen demand
Anaerobic stabilization pond

1. Introduction
Malaysia is the largest producer and exporter of crude palm oil
(CPO). The production of CPO reached 15 million tones in the year
2005 from 14 million tones in the previous year (MPOB, 2006). This
amount will continuously increase in proportion to the world de-
mand of edible oils seeing as palm oil already is bio-diesel product.
Although the palm oil industry is the major revenue earner for our
country but it has also been identified as the single largest source
of water pollution source due to the palm oil mill effluent (POME)
characteristic with high organic content and acidic nature.
In palm oil mills, liquid effluent is mainly generated from ster-
ilization and clarification processes in which large amounts of
steam and hot water are used (Zinatizadeh et al., 2006). For every
ton of palm oil fresh fruit bunch, it was estimated that 0.5–0.75
tones of POME will be discharged (Yacob et al., 2006). In general
appearance, palm oil mill effluent (POME) is a yellowish acidic
wastewater with fairly high polluting properties, with average of
25,000 mg/l biochemical oxygen demand (BOD), 55,250 mg/l
chemical oxygen demand (COD) and 19,610 mg/l suspended solid
(SS). This highly polluting wastewater can cause several pollution
problems and also create odor problems to the neighborhoods of
the mills such as a nuisance to the passers-by or local residents
and river pollution.
* Corresponding author. Tel.: +60 4 979 8971; fax: +60 4 979 8636.
E-mail address: yswong@unimap.edu.my (Y.S. Wong).
Over the past 20 years, the technique available for the treat-
ment of POME in Malaysia has been basically biological treatment,
consisting of anaerobic, facultative and aerobic pond systems
(Chooi, 1984; Ma, 1999). The pond system has been applied in
our country for POME treatment since 1982 (Ashhuby et al.,
1996). Most of the pond system that has been applied for the treat-
ment of POME in Malaysia was classified as waste stabilization
pond. The configuration of this system consists of essentially a
number of ponds of different functions such as anaerobic, faculta-
tive and aerobic ponds. Thus, anaerobic ponds are one of the most
effective treatments that are being applied in Malaysia either in
pond system or close digesting tank systems to treat highly con-
centrated POME. This is because the anaerobic process has consid-
erable advantages such as (a) it demands less energy, (b) sludge
formation is minimal, (c) unpleasant odors are avoided and (d)
anaerobic bacteria efficiently break down the organic substances
to methane (Rincon et al., 2006).
Anaerobic digestion may be defined as the engineered metha-
nogenic anaerobic decomposition of organic matter. It involves dif-
ferent species of anaerobic microorganisms that degrade organic
matter (Cote et al., 2006). In the anaerobic process, the decomposi-
tion of organic and inorganic substrate is carried out in absence of
molecular oxygen. The biological conversion of the organic sub-
strate occur in the mixtures of primary settled and biological
sludge under anaerobic condition followed by hydrolysis, acido-
genesis and methanogenesis to convert the intermediate com-
pounds into simpler end products as methane (CH4) and carbon

0960-8524/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2009.04.074
4970 Y.S. Wong et al. / Bioresource Technology 100 (2009) 4969–4975

Nomenclature

ABSR anaerobic bench scale reactor COD chemical oxygen demand (mg/l)
Alk total alkalinity TN total nitrogen (mg/l)
BOD biochemical oxygen demand (mg/l) TS total solid (mg/l)
D dilution rate, 1/HRT (day1) TVS total volatile solid (mg/l)
CPO crude palm oil UASFF up-flow anaerobic sludge fixed film
HRT hydraulic retention time (day) V reactor volume (l)
l liter VFA volatile fatty acid (mg/l)
MABR modified anaerobic baffled reactor VSS volatile suspended solid (mg/l)
NH3-N ammonia nitrogen (mg/l) X biomass concentration in the reactor, (mg/l)
O&G oil and grease (mg/l)
OLR organic loading rate (kg/m3/d) Greek letter
POME palm oil mill effluent YG growth yield (g VSS/g CODremoved day)
Q flow rate (l/day) b specific biomass decay (day1)
S substrate concentration in the reactor (mg/l) rX specific substrate utilization rate (day1)
S1 influent substrate concentration (mg/l) rv substrate utilization rate per volume (l1 day1)
S2 effluent substrate concentration, (mg/l) rx,max maximum specific substrate utilization (day1)
SCOD soluble chemical oxygen demand (mg/l) Ks saturation constant for substrate (g COD/l)
SRT solid retention time lmax maximum specific biomass growth rate (day1)
SS suspended solid (mg/l) Hc critical retention time (day)
t time (day) l specific biomass growth rate (day1)

dioxide (CO2) (Gee and Chua,1994; Guerrero et al., 1999). There- 2.2. Experimental set-up
fore, the anaerobic digestion process offers great potential for rapid
disintegration of organic matter to produce biogas that can be used The laboratory scale experimental set-up consisted of a feed
to generate electricity and save fossil energy (Linke, 2006). tank, a feed pump, an anaerobic reactor and an effluent tank. The
Nowadays, the anaerobic pond systems are designed depending schematic configuration of the experimental set-up is shown in
on a few common parameters such as hydraulic retention time Fig. 1. The feed tank, anaerobic reactor and effluent tank were fab-
(HRT), solids retention time (SRT), influent and effluent concentra- ricated of flexi glass. The total volume of the anaerobic reactor was
tions, sludge age and others; however the behavior or bio-kinetic 54 l and the actual working volume was 38 l. The anaerobic reactor
factor is not taken into consideration. Additionally, the literature was operated as an open top reactor and exposed to the ambient
is abound with results of research on advanced anaerobic treat- outside condition which has same functions as waste stabilization
ment such as high rate up-flow anaerobic sludge fixed film (Zinati- pond. Two panes of flexi glass were fixed at an angle of 45o at the
zadeh et al., 2006), modified anaerobic baffled reactor (Faisal and bottom of the anaerobic reactor to have good settling zone
Unno, 2001), membrane anaerobic system (Fakhrul Razi and Noor, designed to minimize the active biomass losses. Feeding was
1999) and anaerobic hybrid digester (Borja et al., 1996) for the carried out using peristaltic pump (Model 7520-35). HRT was
treatment of POME but there is scarcity of information in the liter- adjusted volumetrically through controlling the flow rate of the
ature about the anaerobic stabilization pond system. influent feed. The effluent from anaerobic reactor was streamed
Bio-kinetic coefficients are useful tools to obtain information on to the collection tank and overflowed to the effluent tank. No pH
the rate of microbial growth and consumption of substrate, which is control was exerted on the system. Mixed samples were taken
essential to determine the volume of the reactor and understanding from the effluent tank after 24 h for analysis at each batch of
well the process control through system simulation. Meanwhile, HRT. The anaerobic reactor was monitored constantly to ensure
bio-kinetic coefficients also play an important role to illustrate smooth operation.
the development of microorganism and substrate balances, the pre-
diction of effluent concentration, the development of process de- 2.3. ABSR operation
sign factors and the effects of kinetic coefficients on the process
of design, performance and stability (Metcalf and Eddy, 2003). The seeding that required for starting the ABSR was taken from
The aim of this work is focused on the anaerobic stabilization anaerobic pond of MALPOM Industries Bhd wastewater treatment
pond system because anaerobic digestion process was the first plant. About 38 l of the anaerobic digested POME were used to
treatment of waste stabilization pond system for POME. Therefore,
an anaerobic bench scale reactor (ABSR) was operated continu- Table 1
Characteristics of palm oil mill effluent (POME).
ously at different hydraulic retention time in order to evaluate
the bio-kinetic coefficients for POME. Parameter Concentration range (mg/l) Mean ± Standard deviation
pH 4.15–4.45 4.3 ± 0.11
2. Methods COD 45,500–65,000 55,250 ± 7534
SCOD 20,500–24,500 22,500 ± 1436
BOD 21,500–28,500 25,000 ± 2607
2.1. Wastewater preparation TS 33,790–37,230 35,515 ± 767
TVS 27,300–30,150 28,697 ± 1054
The POME was obtained from MALPOM Industries Bhd, in Nibong SS 15,660–23,560 19,610 ± 2908
Tebal, Penang. The characteristic of POME that been used in the O&G 1077–7582 3822 ± 2483
TN 300–410 355 ± 37
experiment is shown in Table 1. The wastewater was stored in the
NH3-N 8.3–8.5 8.4 ± 0.08
refrigerator at 4 °C until further detailed analysis could be con-
ducted. This storage had no observable effect on the composition. Unit for all parameter is mg/l except pH.
 Y.S. Wong et al. / Bioresource Technology 100 (2009) 4969–4975 4971

Influent Feed Open Top Reactor

Scum Layer
Peristaltic
Pump Effluent

Feeding Tank
Effluent Tank
Anaerobic Reactor

Fig. 1. Scheme diagram of ABSR experimental set-up.

acclimatize the laboratory ABSR. The start-up of the ABSR involved tration, V = reactor volume, rv = substrate utilization rate per vol-
step increases in influent organic volumetric loading rates from ume, and t = time. As only when the reactor became steady
0.89–0.92 kg COD/m3/day to 1.34–1.37 kg COD/m3/day. This accli- condition, dS
dt
¼ 0. Therefore,
matization phase was continued for about 50 days to allow the
rv ¼ ðS1  S2 ÞD ð2Þ
microorganisms present in the mixed liquor perfectly acclimatized
to the new environment and reached a steady state condition be- where D = dilution rate = QV = HRT
1

fore preceded to the next phase of research studies. Specific substrate utilization rate, rX is defined as follows:
This preliminary period was followed by a series of continuous
rv
experiments using feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and rX ¼ ð3Þ
X
3.8 l of raw POME per day, which correspond to HRT of 60, 50, 40,
30, 20 and 10 days. The rate of influent substrate concentration of where X = biomass concentration in the reactor. Therefore, Eq. (2)
COD was controlled in the range of 52,000–55,000 mg/l, respec- can be rewritten as:
tively. For each batch of HRT, 500 ml of samples from ABSR were ðS1  S2 ÞD
collected after 24 h of input raw POME. The samples were sub- rX ¼ ð4Þ
X
jected to the analysis of the following parameters such as feed
and effluent of COD, SCOD and VFA, ABSR pH, alkalinity, suspended On the other hand, the biomass balance gives the following
solid and volatile suspended solids based on American Public equation:
Health and Association (APHA) standard methods for water and dX QX
wastewater analysis (APHA, 1992). ¼ þ lX ð5Þ
dt V
For each batch of HRT, the ABSR was continuously operated un-
where l = specific biomass growth rate. At steady state conditions,
til steady state condition was achieved. This process was indicated dX
¼ 0, hence, Eq. (5) yields
by the concentration of mixed liquor volatile suspended solid dt

(MLVSS) inside the ABSR and effluent concentration of COD be- 1

came constant. Besides, the samples were collected and analyzed
l¼D¼ ð6Þ
HRT
for five consecutive days. The steady state value of tested parame-
The growth of microorganism was proportional to the substrate
ter was taken as average value of these consecutive analysis
utilization, and biomass decay or endogenous metabolism should
amounts for each batch of HRT. The variation of the actual results
also be taken into account to describe the specific growth rate as
from the mean values was <3%.
follows (Timur and Ozturk, 1999):

2.4. Bio-kinetic model development l ¼ Y G rX  b ð7Þ

where YG = growth yield coefficient and b = specific biomass decay
In present study, the mass balance, rate equation and Monod of
coefficient.
mathematical models were applied to the ABSR for determination
Combining Eqs. (6) and (7) gives the following (Timur and Oz-
of bio-kinetic coefficients. The ABSR was developed based on the
turk, 1999):
following assumption (Truax et al., 1995; Al-Malack, 2006):
D ¼ Y G rX  b ð8Þ
(i) The reactor was completely mixed and fed continuously or
without solid recycle.
(ii) Influent substrate concentration remains constant (this was D b
rX ¼ þ ð9Þ
achieved by using POME as the influent substrate). YG YG
(iii) No microbial solids were contained in the influent substrate. The substrate utilization rate depends on the substrate concen-
(iv) The volume of the reactor must constant. tration and several types of equations have been proposed to de-
(v) Steady state conditions prevail throughout the system. scribe the relationship. However, the Monod model is the
simplest and the most widely applied in practice (Kesavan and
The equations describing the performance of the system are the Law, 2005). The Monod equation is as follows:
mass balance equations of the both the substrate and biomass. The
substrate balance can be expressed by Timur and Ozturk (1999): S
rX ¼ rX;max ð10Þ
dS Q ðS1  S2 Þ KS þ S
¼  rv ð1Þ
dt V where, rX,max = maximum specific substrate utilization rate, KS = sat-
where S = substrate concentration in the reactor, Q = flow rate, uration constant for substrate, and S = substrate concentration in
S1 = influent substrate concentration, S2 = effluent substrate concen- the reactor.
4972 Y.S. Wong et al. / Bioresource Technology 100 (2009) 4969–4975

As stated in Eq. (10), the value of rX is half of rX,max when the 100

Removal efficiency (%)

substrate concentration is KS. This equation was quite similar to
the equation proposed by Monod, which described the relationship
between the growth rate and the substrate concentration (de Lucas
et al., 2005). One of the following three types of linear plot can be
used to evaluate rX,max, and KS by using the experimental data of rX
and S.
  
1 KS 1 1
¼ þ
rX r X;max S r X;max
r 
X
r X ¼ r X;max  K S
S
S S KS
¼ þ
r X rX;max r X;max
The above mentioned linear plots are called Lineweaver–Burk
plot, Eadie plot and Hofstee plot. Maximum specific biomass
growth rate, lmax, is obtained by substituting rX,max for rX in Eq.
(7) as follows:
lmax ¼ Y G rX;max  b
A critical retention time, HC is defined as the retention time be-
low which substrate utilization does not occur because of the
washout of the microorganisms happen in the reactor (Borja
et al., 2003). By using Eq. (6), HC is expressed as:
1 the VFA concentration steadily increased from 200 mg CH3COOH/
HC ¼
lmax
3. Results and discussion
3.1. Operation parameter and biodegradability
3.1.1. Anaerobic digestion process efficiency of ABSR
Table 2 summarized the steady state performance of ABSR at six
different HRT of 10, 20, 30, 40, 50 and 60 days. The variation of the
percentages of COD and SCOD removed at respective HRT for ABSR
is shown in Fig. 2. The COD and SCOD removal efficiency increased
from 10 until 60 days HRT. Hence, the COD and SCOD removal effi-
ciency were in the range of 66.38–85.41% and 26.8–73.19%. As can
be seen, the COD removal efficiency was not much significance dif-
ference with 85.41% and 84.77% at the higher HRT of 50 and
60 days. Meanwhile, the SCOD removal efficiency also does not
show significant difference at the level of 73.19% and 71.67% dur-
ing the HRT of 60 and 50 days. On the other hand, the COD and
SCOD removal efficiency became poorer at the shorter HRT. At
HRT of 10 days, the COD and SCOD were reduced to 66.38% and
26.8%. This was largely due to the wash-out phase of the reactor
since the biomass concentration increased in the system as shown
in Table 2. Moreover, Schiener et al. (1998) established that soluble
biodegradable material in the effluent increased with the rise of or-
80
60
40 COD
20 SCOD
0
ð11Þ 0 10 20 30 40 50 60
Hydraulic Retention Time (days)
ð12Þ
Fig. 2. COD and SCOD removal efficiency of ABSR under steady state condition on
various hydraulic retention time.
ð13Þ
ganic loading rate. Therefore, the SCOD in the effluent of ABSR in-
creased as the HRT decreased as shown in the Table 2. In addition,
the layer of scum on the surface area of ABSR are getting thicker
though the decreasing of HRT.
ð14Þ
3.1.2. Volatile fatty acid as acetic acid and pH
Fig. 3 showed the VFA as acetic acid and pH level of ABSR at
steady state condition between HRT of 10 and 60 days. The VFA
concentration was fairly high at HRT of 10 days and gradually re-
duced as the HRT increased. During the HRT of 60 until 30 days,
ð15Þ l to 529.14 mg CH3COOH/l. Thus, the VFA concentration increased
sharply to 1244.57 mg CH3COOH/l and 2102.86 mg CH3COOH/l be-
tween the HRT of 20 and 10 days. This was largely due to the accu-
mulation of intermediate products such as VFA at the shorter HRT
(Faisal and Unno, 2001). Besides, the low level concentration of
VFA at higher HRT could be recognized to low organic loading rate
(Faisal and Unno, 2001). In addition, the high concentration of
effluent VFA at shorter HRT also reflected in lower COD removal
efficiency as shown in Table 2. Meanwhile, the pH level of ABSR
was increased from 10 to 60 days of HRT. According to Grady
et al. (1999), the sensitivity of methanogenic bacterial to pH were
coupled with the VFA, which are the intermediates of the anaerobic
stabilization of organic matter, bring about the negative response
by the anaerobic system. From Fig. 3, it can be seen that the pH le-
vel of ABSR was in the range of 7.64–8.55, although no chemicals
were added for adjusting the pH level inside the reactor.
3.1.3. Alkalinity and VFA:Alk ratio fraction in the ABSR
The alkalinity and ratio fraction between VFA and alkalinity of
ABSR under steady state condition at various HRT is outlined in
Fig. 4 and Table 2. The alkalinity concentration of ABSR increased
from 12,772 mg CaCO3/l until 18,320 mg CaCO3/l by the HRT of
10 until 60 days. Thus, the VFA:Alk fraction ratio was on conversed
trends as the VFA:Alk decreased with the increase of HRT. Accord-
ing to Zhao and Viraraghavan (2004), the proper ratio for volatile

Table 2
Experimental results obtained under steady state condition at six different HRT.

Parameter HRT (day)

60 50 40 30 20 10
Effluent COD (mg/l) 7850 ± 2 8180 ± 1 9750 ± 2 11150 ± 3 14060 ± 2 18310 ± 2
Effluent SCOD (mg/l) 6065 ± 2 6358 ± 2 7943 ± 2 9350 ± 2 12255 ± 1 16520 ± 3
Effluent VFA (mgCH3COOH/l) 200.00 ± 2.57 251.43 ± 1.48 325.71 ± 2.63 529.14 ± 1.52 1244.57 ± 1.57 2102.86 ± 1.55
Total alkalinity in ABSR (mgCaCO3/l) 18320 ± 2 17252 ± 1 16538 ± 2 15290 ± 4 14516 ± 3 12772 ± 2
Total VFA in ABSR (mgCH3COOH/l) 1916.57 ± 1.15 1957.71 ± 1.10 2067.43 ± 2.09 2253.71 ± 1.13 2958.86 ± 2.18 3821.72 ± 2.15
VFA:Alk 0.11 ± 0.01 0.11 ± 0.01 0.13 ± 0.01 0.15 ± 0.01 0.20 ± 0.01 0.30 ± 0.01
MLVSS in ABSR (mg/l) 18418 ± 2 20582 ± 1 22378 ± 2 25646 ± 1 27236 ± 2 28694 ± 3
pH in ABSR 8.55 ± 0.01 8.47 ± 0.01 8.28 ± 0.01 8.16 ± 0.01 7.78 ± 0.01 7.64 ± 0.01
COD removal efficiency (%) 85.41 ± 0.08 84.77 ± 0.06 82.12 ± 0.07 79.55 ± 0.07 74.20 ± 0.09 66.38 ± 0.08
SCOD removal efficiency (%) 73.19 ± 0.08 71.67 ± 0.06 64.79 ± 0.07 57.78 ± 0.06 45.09 ± 0.08 26.80 ± 0.09

Results are mean of 10 values ± standard error of the mean (SEM).

 Y.S. Wong et al. / Bioresource Technology 100 (2009) 4969–4975 4973

2500 8.6

utilization, rx(day-1)
0.14

Specifc substrate
8.4 0.12
2000 0.1
Effluent VFA (mg

0.08
CH3COOH/l)

8.2
1500 0.06
eff. VFA
0.04

pH
8
1000 pH 0.02
7.8 0
500
0 10 20 30 40 50 60
7.6 HRT (day)
0 7.4
10 20 30 40 50 60 Fig. 5. Specific substrate utilization rate, rx of COD under steady state at various
Hydraulic Retention Time (days) HRT.

Fig. 3. Effluent VFA as acetic acid concentration and pH under steady state

utilization rate, rx (day-1)

condition on various hydraulic retention times.
0.14

Specific substrate
0.12
0.1
20000 0.35 0.08 y = 1.01x + 0.0244
TotalAlkalinity (mgCaCO 3 /l)

2
R = 0.99
0.3 0.06
VFA:Alk ratio fraction
15000 0.25 0.04
0.02
0.2
10000 0
0.15 0 0.05 0.1 0.15
5000 0.1 Dilution rate, D (day -1)
Alkalinity VFA:Alk
0.05
0 0 Fig. 6. Determination of growth yield, YG and specific biomass decay, b according to
10 20 30 40 50 60 Eq. (9).
Hydraulic Retention Time(days)

Fig. 4. Alkalinity and VFA:Alk fraction ratio of ABSR under steady state condition on 30
25
1/rx , (day)

various hydraulic retention times.

20
15 y = 367.74x + 1.8076
R2 = 0.96
10
acid to alkalinity is between 0.1 and 0.2 for anaerobic digestion 5
process. Besides, the VFA:Alk fraction ratio of ABSR was in the 0
range of 0.11–0.30 from 60 to 10 days of HRT. Therefore, the 0 0.02 0.04 0.06 0.08
VFA:Alk ratio at HRT 10 days was considered out of the proper 1/S (l/mg)
range as the acidification phenomenon may have occurred in the
Fig. 7. Determination of maximum specific substrate utilization, rx,max and satu-
system. However, the VFA:Alk ratio at 10 days of HRT operated
ration constant, KS according to Eq. (11).
without the risk of acidification because the requirement of
VFA:Alk ratio for the stability of anaerobic digestion process must
be <0.4 so that the digestion process was considered to be operat- Table 4
ing favorably without acidification risk (Raposo et al., 2004; Grady Summarize data for kinetic coefficients of ABSR.
and Lim, 1980). As a result, the performance of ABSR could
Kinetic coefficients Value R2
maintain active methanogenic activity by keeping the VFA concen-
Growth yield, YG (g VSS/g CODremoved day) 0.990 0.99
tration low, especially at a longer hydraulic retention time. Alter-
Specific biomass decay, b (day1) 0.024 0.99
natively, the adequate alkalinity or buffer capacity was essential Maximum specific substrate rate, rx,max (day1) 0.553 0.96
to maintain a stable pH in the digester for optimal biological activ- Saturation constant for substrate, KS (g COD/l) 203.433 0.96
ity (Agdag and Sponza, 2005). In addition, the bicarbonate alkalin- Maximum specific biomass growth rate, lmax (day1) 0.524
ity of anaerobic digestion must be above 12,500–14,000 mg CaCO3/ Critical retention time, HC (day) 1.908

l. The high alkalinity level indicates that the bacterial groups were
in equilibrium (Rubia et al., 2006). Hence, the alkalinity of ABSR 3.2. Determination of bio-kinetic coefficients
was enough to provide the optimal condition for methanogenic
activity within the range of 12,772–18,320 mg CaCO3/l from 10 un- The experimental data under steady state condition (Table 2)
til 60 days of HRT as shown in the Table 2 and Fig. 4. were analyzed and kinetic coefficients were evaluated by the

Table 3
Experimental results at six different HRT for kinetic analysis.

Steady state periods (days) Q (l/day) HRT (day) MLVSS (mg/l) S, (COD ABSR) (mg/l) COD, effluent (mg/l)
55–60 0.63 60 18,418 ± 2 16,500 ± 2 7850 ± 2
50–55 0.76 50 20,582 ± 1 17,560 ± 2 8180 ± 1
40–45 0.95 40 22,378 ± 2 20,800 ± 1 9750 ± 2
30–35 1.27 30 25,646 ± 1 25,540 ± 2 11150 ± 3
20–25 1.90 20 27,236 ± 2 33,500 ± 1 14060 ± 2
10–15 3.80 10 28,694 ± 3 47,520 ± 2 18310 ± 2

Results are mean of 10 values ± standard error of the mean (SEM).

4974 Y.S. Wong et al. / Bioresource Technology 100 (2009) 4969–4975
Table 5
Kinetic coefficients in different type of reactor for the treatment of POME.
Type of reactor S0 (g/l) KS (g/l)
UASFF 34.75 0.982
MABR 16 0.313
ABSR 53.50 203.433
ABSR – present study.
model development mentioned in the previous Section of 2.4.
chemical oxygen demand (COD) as influent substrates was selected
for bio-kinetic study. The rate of influent substrate concentration
of COD was controlled in the range of 52,000–55,000 mg/l. Hence,
Table 3 summarized the parameters required for bio-kinetic
coefficients determination. By using these parameters, the value
of specific substrate utilization rate (rX) for the substrate of COD
was calculated according to Eq. (4). Fig. 5 showed the values of spe-
cific substrate utilization rate of COD at steady state condition be-
tween 10 and 60 days of HRT. The specific substrate utilization rate
of COD generally increased proportionally with the lowering of
HRT which indicated that the bacterial population in the ABSR
had been multiplied (Abdullah et al., 2005).
In order to determine the bio-kinetic coefficients, Figs. 6 and 7
were plotted using the data in Table 3. The bio-kinetic coefficients
of growth yield (YG), specific biomass decay (b), maximum specific
substrate utilization (rx,max) and saturation constant (Ks) were
calculated from the slope and intercept as shown in Figs. 6 and
7. Besides, the maximum specific biomass growth rate (lmax)
and critical retention time, HC were calculated from Eqs. (14)
and (15). All the kinetic coefficients calculated from the models
were summarized in Table 4.
The value of YG for ABSR was at 0.990 g VSS/g CODremoved day,
respectively. This value was considered higher as compared to
the anaerobic suspended and attached growth processes. This
could be due to the ABSR was a simulated reactor of the anaerobic
stabilization pond system. Moreover, the literature survey showed
that there was lack of information related to the bio-kinetic coeffi-
cients of anaerobic stabilization pond system for treating the
POME. As stated by Singh and Viraraghavan (1998), the high value
of growth yield in ABSR because the anaerobic bacteria were
immobilized by the process of spontaneous aggregation of bacteria
to dense compact granules with high activity and good settling
characteristics in the ABSR.
A comparison of the bio-kinetic coefficients obtained with the
ABSR in the present study with those obtained from literature such
as up flow anaerobic sludge fixed film (UASFF) and modified anaer-
obic baffled reactor (MABR) of treating POME was described in
Table 5. Throughout this research, the value of lmax for the sub-
strate of COD was closed to the reported value for the work done
by (Zinatizadeh et al., 2006) and (Faisal and Unno, 2001). This
was due to similar activity for microbial growth in the treatment
of POME. Moreover, the small value of lmax implied relatively with
high amount of biomass in the ABSR (Zinatizadeh et al., 2006).
According to Grady and Lim (1980), the values of the parame-
ters lmax and Ks are very dependent upon the organism and sub-
strate employed. If a given species of organism is grown on each
of several substrates under fixed environmental conditions, the
values of lmax and Ks observed will depend upon the substrates.
Likewise, if several pure cultures are fed the same substrate under
otherwise identical environmental conditions, the value of lmax
and Ks will depend upon the species of organisms. However, the
lmax and Ks were at 0.524 day1 and 203.433 g COD l1 for the
present study. Thus, the low value of lmax and high value of Ks
showed that the typical wastes were slowly biodegraded in the
comparison to more easily degraded substrate (Truax et al.,
1995). On the over hand, the critical retention time, HC at which
lmax (per day) Coefficient basis References
0.207 COD Zinatizadeh et al. (2006)
0.304 COD Faisal and Unno (2001)
0.524 COD This study
the washout of microorganisms occurred were on the HRT of
2 days for the substrate of COD. As a result, the ABSR was not al-
lowed to operate less than the critical retention time because the
washout of microorganisms will bring down the performance of
the anaerobic digestion process.
4. Conclusion
Throughout this research study, it can be concluded that the
anaerobic digestion process of anaerobic bench scale reactor
(ABSR) has provided useful information to analyze the bio-kinetic
coefficients for treating the high strength wastewater of POME.
The result of this study proved that the treatment of POME could
be treated effectively through ABSR at different HRT. Moreover,
the proposed kinetic equations were applicable for anaerobic
digestion process of POME with ABSR.
Acknowledgements
The authors would like to express their thanks for the support
by the Project for Environmental Technology of University Science
Malaysia, MALPOM Industries Sdn Bhd and School of Environmen-
tal Engineering of University Malaysia Perlis (UniMAP).
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