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The effect of hydraulic retention time on granular sludge biomass in treating textile wastewater
Khalida Muda a,*, Azmi Aris a,**, Mohd Razman Salim a, Zaharah Ibrahim b, Mark C.M. van Loosdrecht c, Azlan Ahmad a, Mohd Zaini Nawahwi b
a

Department of Environmental Engineering, Faculty of Civil Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia Department of Biological Sciences, Faculty of Biosciences and Bioengineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia c Department of Biotechnology, Delft University of Technology, Julianalaan 67,2628 BC Delft, The Netherlands
b

article info
Article history: Received 12 February 2011 Received in revised form 7 May 2011 Accepted 12 May 2011 Available online 20 May 2011 Keywords: Textile wastewater SBR Anaerobic/aerobic reaction Biokinetics Color

abstract
The physical characteristics, microbial activities and kinetic properties of the granular sludge biomass were investigated under the inuence of different hydraulic retention times (HRT) along with the performance of the system in removal of color and COD of synthetic textile wastewater. The study was conducted in a column reactor operated according to a sequential batch reactor with a sequence of anaerobic and aerobic reaction phases. Six stages of different HRTs and different anaerobic and aerobic reaction time were evaluated. It was observed that the increase in HRT resulted in the reduction of organic loading rate (OLR). This has caused a decrease in biomass concentration (MLSS), reduction in mean size of the granules, lowered the settling ability of the granules and reduction of oxygen uptake rate (OUR), overall specic biomass growth rate ( `overall), endogeneous decay rate (kd) and biomass yield (Yobs, Y). When the OLR was increased by adding carbon sources (glucose, sodium acetate and ethanol), there was a slight increase in the MLSS, the granules mean size, `overall, and biomass yield. Under high HRT, increasing the anaerobic to aerobic reaction time ratio caused an increase in the concentration of MLSS, mean size of granules and lowered the SVI value and biomass yield. The ` overall and biomass yield increased with the reduction in anaerobic/aerobic time ratio. The HRT of 24 h with anaerobic and aerobic reaction time of 17.8 and 5.8 h respectively appear to be the best cycle operation of SBR. Under these conditions, not only the physical properties of the biogranules have improved, the highest removal of color (i.e. 94.1 0.6%) and organics (i.e. 86.5 0.5%) of the synthetic textile dyeing wastewater have been achieved. 2011 Elsevier Ltd. All rights reserved.

1.

Introduction

Granulation techniques for dye degradation processes have been reported by many researchers (van der Zee, 2003; Isik and Sponza, 2008). Most of the studies focused on the application of anaerobic process since major decolorization process occurs under this condition (Isik and Sponza, 2005; Somasiri et al., 2008).

Nevertheless, complete mineralization of dye containing wastewater requires both anaerobic and aerobic conditions. Several studies have therefore been conducted to investigate the removal efciency under both operating conditions with series of anaerobic and aerobic reactor system (Isik and Sponza, 2004a, 2004b). Isik and Sponza (2008) reported that more than 91% and 84% removal efciency of COD and color

* Corresponding author. Tel.: 60 7 5531522; 60 7 5531581; fax: 60 7 5566157. ** Corresponding author. E-mail addresses: khalida@utm.my (K. Muda), azmi.aris@utm.my (A. Aris). 0043-1354/$ e see front matter 2011 Elsevier Ltd. All rights reserved. doi:10.1016/j.watres.2011.05.012

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respectively, were observed when a mixed dye compounds in synthetic textile wastewater were treated using partially anaerobic granule/activated sludge in a sequential anaerobic/ aerobic reactor system. Previous study also shows that the overall percentage of color removal in textile wastewater treatment is very much affected by the hydraulic retention time (HRT) particularly during anaerobic phase as the cleavage of the eNNe bond reported to occur more under anaerobic condition (Pasukphun and Vinitnantharat, 2003; Buitron et al., 2004; Ong et al., 2008). Color removal has been found to increase as the HRT of anaerobic phase increases (Buitron et al., 2004; Li and Xi, 2004; Van der Zee and Villaverde, 2005). However, different types of biomass (i.e. suspended cells, biolm or granules) used in the treatment system may have different HRT requirements and will affect the design of the system. Successful cultivation of microbial granular sludge using synthetic and raw textile dyeing wastewater in a single reactor column operating under intermittent anaerobic and aerobic reaction phases has been reported earlier (Muda et al., 2010; Ibrahim et al., 2010). While the developed granules appear to be capable in treating the textile wastewater, their performance under different range of HRT is still unknown. The paper discusses how the granules properties, reactor performance, and biokinetics can be changed due to the variation in the HRT of the reactor.

2.3.

Reactor set-up

The schematic illustration of the reactor set-up is given in Fig.1. The design of the reactor system was based on Wang et al. (2004) and Zheng et al. (2005) with several modications. The height of the reactor column was 100 cm with 8 cm internal diameter. The column was designed to be operated with 4-L working volume. The wastewater enters the reactor from the bottom of the column. A ne air bubble diffuser was placed at the bottom of the column for aerobic phase. Air bubbles was introduced into the reactor intermittently and controlled by a timer. The withdrawal of the wastewater took place via an outlet port located at 40 cm height from the bottom of the reactor which gave 50% of the volumetric exchange rate.

2.4.

Analytical methods

2.
2.1.

Materials and methods

Granular biomass

The effect of HRT was investigated with respect to the changes in the physical characteristics, microbial activity and removal performance of the granular biomass in the reactor system. Physical characteristics include particle size distribution, settling velocity (SV) and sludge volume index (SVI). The particle size distribution was measured throughout the experiment using wet sieves analysis where the volume of each different granular size was expressed as a fraction of the total volume of the granule. The SV was determined by recording the average time taken for an individual granule to settle at a certain height in a glass column lled with tap water. The SVI was determined based on Beun et al. (1999). The bed volume was obtained by measuring the bed height of the sludge biomass that settle within 5 min in the reactor

The development of the microbial granules has been explained in detail in Muda et al. (2010). The size of the microbial granules selected for this experiment was in the range of 0.3e2.5 mm.

2.2.

Wastewater composition

Synthetic textile dyeing wastewater which was applied in this study was prepared according to Tan et al. (2005) and Sirianuntapiboon and Srisornsak (2007) with several modications. The wastewater contains a mixture of an equal ratio (based on the COD concentration) of glucose (0.5 g L1), ethanol (0.125 g L1) and sodium acetate (0.5 g L1) as the carbon sources. Other components including NH4Cl 0.16 g L1, KH2PO4 0.23 g L1, K2HPO4 0.58 g L1, CaCl22H2O 0.07 g L1, MgSO47H2O 0.09 g L1, EDTA 0.02 g L1 and trace solution 1 mL L1 were also added in the media solution. Several trace elements recommended by Smolders et al. (1995) were used as the essential mineral content. A total concentration of 50 mg L1 of Sumix Black EXA, Sumix Navy Blue EXF and Synozol Red K-4B were used as mixed dyes in this study. These azo dyes represent some of the dyestuffs that are commonly used in dyeing cotton, polyester and polyacrylic fabrics (Correia et al., 1994). The COD and BOD of the synthetic wastewater were 1270 mg/L and 312 mg/L, respectively with COD over BOD ratio of about 4.1 indicating the relatively non-biodegradable nature of the wastewater.

Fig. 1 e Schematic layout of the reactor system (Wang et al., 2004; Zheng et al., 2005).

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Table 1 e Detail of experimental condition of the reactor system. Stage Days covered Anaerobic
I II III IV V VI 49 43 51 43 46 46 1.42 2.92 5.92 5.92 8.92 2.92

Phase (h) 1st Aerobic

1.42 2.92 5.92 5.92 2.92 8.92

HRT (h) 2nd

OLR (kgCOD m3 day1)

Anaerobic
1.42 2.92 5.92 5.92 8.92 2.92

Aerobic
1.42 2.92 5.92 5.92 2.92 8.92 6 12 24 24 24 24 2.5 1.3 0.6 0.8 0.8 0.8

OLR X/VtotalVadd/t, where X COD concentration of the inuent (mg L1); Vadd Volume of inuent added in each cycle operation (mL); Vtotal Total working volume of the experiment (mL); t Hydraulic retention time (h).

after the aeration phase stopped and was multiplied with the surface area of the inner reactor column. Then, the obtained bed volume was divided with the dry weight of the biomass in the reactor. The microbial activity of the microbial granules was conducted by measuring the oxygen utilization rate (OUR) at different HRT. The OUR (mg DO L1 h1) measurements were performed by following the Standard Methods (APHA, 2005). Sludge retention time (SRT) was determined according to Beun et al. (1999). Other parameters such as mixed liquor suspended solid (MLSS), mixed liquor volatile suspended solid (MLVSS), COD and color were analyzed according to Standard Methods (APHA, 2005).

in reduction of the organic loading rate (OLR) supplemented into the reactor from 2.5 to 0.6 kg COD m3 day1. The HRT for Stage III to VI was kept constant, i.e. 24 h, but the durations of anaerobic and aerobic reaction phases were varied. From stage III onwards, the OLR was increased to 0.8 kg COD m3 day1 by increasing the concentration of the carbon sources in the synthetic textile dyeing wastewater. The temperature of the treatment system was kept constant at 30.0 2.0  C while the pH throughout the experiment was between 6.3 and 8.0.

3.
3.1.

Results and discussions

Physical prole of the reactor System

2.5.

Experimental procedures

The reactor system was operated according to a sequential batch reactor (SBR). A complete cycle of the operation comprised of ll (15 min), react (6e24 h), settle (5 min), decant (5 min) and idle (5 min) steps. The react step consisted of alternate anaerobic and aerobic reaction phase. During the anaerobic reaction phase, circulation of wastewater was carried out using peristaltic pump (ColeeParmer System Model, 6e600 rpm) at a ow rate of 18 L h1. Air was diffused into the reactor using an air pump at a supercial air velocity of 2.6 cm s1 throughout the aerobic react phase. The details of the experimental conditions are shown in Table 1. The HRT of the experiment was increased from 6 to 24 h in Stage I to Stage III. The increase in the HRT resulted

The changes in the biomass concentration at different HRT (Stage I to III) are shown in Table 2 and Fig. 2. It is apparent that the biomass concentration (MLSS) in the reactor decreases from about 35 g L1 to 25 g L1 as the HRT increases from 6 to 24 h. The reduction of the biomass concentration in the reactor is expected to be due to the reduction of OLR applied in the reactor system (i.e. from 2.5 to 0.6 kg COD m3 day1) as a result of the increase in HRT. Reduction in the OLR means less carbon was supplied to the microorganisms in the reactor and hence, less growth is taking place. When the OLR was increased to 0.8 kg COD m3 day1, the MLSS started to increase again to 30.5 3.4 g L1 and 31.6 3.7 g L1 in Stage IV and V. Similarly, the ratio of the

Table 2 e Biomass concentrations at different stages of the experiment. Reaction Phase I

Anaerobic (h) Aerobic (h) MLSS (g L1) MLVSS (g L1) MLVSS/MLSS Efuent (VSS g L1) SRT (day) 2.8 2.8 35.3 31.9 0.90 0.34 27.6 5.8 5.8 28.7 24.5 0.85 0.31 42.4

Stage II III
11.8 11.8 25.2 1.8 18.5 2.2 0.73 0.26 0.19 78.9 23.9

IV
11.8 11.8 30.5 3.4 26.0 3.4 0.85 0.34 0.11 70.1 23.9

V
17.8 5.8 31.6 3.7 22.4 2.0 0.71 0.33 0.10 72.5 23.3

VI
5.8 17.8 23.3 0.8 20.2 0.8 0.87 0.55 0.22 41.6 18.4

1.6 1.8 0.16 13.4

0.6 2.2 0.11 10.2

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Since the system was operated with no sludge wastage at any of the operating time, Eq. (1) can be simplied as Eq. (2) (Tchobanoglous et al., 2004; Liu and Tay, 2007). q Xvss Vr tc Xe Ve (2)

Fig. 2 e Prole of biomass concentration at different stages of the experiment. (C) MLSS, (,) MLVSS. Stage I: anaerobic (2.8 h): aerobic (2.8 h); Stage II: anaerobic (5.8 h): aerobic (5.8 h); Stage III and Stage IV: anaerobic (11.8 h): aerobic (11.8 h); Stage V: anaerobic (17.8 h): aerobic (5.8 h); Stage V: anaerobic (5.8 h): aerobic (17.8 h).

Based on Eq. (2), the SRT of the reactor system increases from 27.6 13.4 to 78.9 30.8 d when the period of the HRT increases from 6 to 24 h (Stage I to III). This is mainly due to the decrease of MLSS and also MLVSS caused by lower OLR as previously explained. With HRT of 24 h, increase of anaerobic reaction phase up to 18 h (Stage IV to V) has slightly increased the SRT from 70.1 23.9 to 72.5 23.3 d. The SRT value changes in each stage of the experiment. According to Wijffels and Tramper (1995), the favorable sludge age for high removal efciency for COD and nitrication process is more than 4 days. Based on the SRT obtained, this granular system is capable of simultaneous nitrication process and COD removal. Since the treatment goal is to remove recalcitrant dyeing compound, the SRT value of all stages evaluated in this experiment was in the acceptable range for the degradation of xenobiotic compounds (Grady et al., 1999).

volatile biomass (MLVSS) to total biomass (MLSS) reduced from Stage I to III mainly due to decrease in the OLR as the HRT increased from 6 to 24 h. It was also observed that when the duration of the anaerobic phase was increased, the MLVSS/MLSS ratio was decreased (i.e. 0.71). The decrease in MLVSS/MLSS ratio may indicate an increase of inert particles within the granules. The same observation was reported by Panswad et al. (2001) that increase of inert solids in the biomass was observed when the system was exposed to high anoxic/anaerobic condition in the SBR cycle. When the duration of aeration phase was increased up to 18 h, the biomass started to reduce again (Stage VI) and increase of VSS (Table 2) in the efuent was observed. This indicates that too long of aerobic reaction phase is not suitable for granular biomass system as the turbulent caused by the aeration may rupture the granules causing them to leave the reactor during decant phase due to their small size and long settling time. The sludge retention time (SRT) of the biomass in the SBR system can be calculated by using Eq. (1) (Tchobanoglous et al., 2004; Liu and Tay, 2007). q Xvss Vr tc Xd Vd Xe Ve (1)

3.2.

Physical properties of the granular biomass

The effects of HRT variation (6e24 h) on the physical properties of the granules are given in Table 3. In general, the mean size of the granules reduces with the increase in the HRT which resulted in higher SVI and lower SV values. The mean granular size in the reactor was the largest (i.e. 843 44 `m) during Stage I which has the shortest HRT and highest OLR. When the HRT increases from 6 to 24 h, the OLR was reduced from 2.5 to 0.6 kg COD m3 day1. This condition may contribute to the smaller granules formation due to the reduction in the food supply. Furthermore, as the HRT increases from 6 to 24 h, the duration of the aeration phase also increases from about 3 to 12 h. Hence, the reduction in the mean granular size may also be due to the long exposure of the granules to the shear force imposed by aeration process. When the OLR was increased from 0.6 to 0.8 kg COD m3 day1from Stage III to IV, the mean granular size slightly increased due to increase in food supply. Fig. 3 shows the particle size distribution of granular biomass in the reactor at each stage of the experiment. The gure shows that the particle size distribution was clearly affected by the HRT and aeration time which imposed shear force to the granules.

Table 3 e Physical properties of the granular biomass at different stages of experiment. Reaction Phase I
Anaerobic (h) Aerobic (h) Mean size (mm) SVI (mL g1) SV (m h1) 2.8 2.8 843 44 13.1 0.4 41.3 3.1

Stage II
5.8 5.8 590 55 18.8 1.5 35.1 0.8

III
11.8 11.8 440 40 21.4 1.6 24.5 1.1

IV
11.8 11.8 567 79 16.8 1.3 28.4 1.3

V
17.8 5.8 575 46 15.5 1.3 33.4 2.5

VI
5.8 17.8 385 22 24.8 0.9 21.3 0.5

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In Stage V, although the HRT was similar to Stage IV (i.e. 24 h), the mean granular size was observed to slightly increase. This is apparently due to the shorter aerobic phase (i.e. 6 h) as the anaerobic phase was prolonged to 18 h. This shows that bigger granules could still be maintained in the reactor at high HRT provided that the aerobic phase is reduced by increasing the anaerobic phase. This condition seems to be suitable for treating textile wastewater that requires both anaerobic and aerobic phases. Increase in the size of the granular biomass in Stage V may be achieved by the equilibrium between the growth factor and detachment of the granules due to the shear force effect. The intermittent anaerobic and aerobic reaction phase will inuence the production and consumption rate of EPS that eventually causes the changes on the surface charges of the granular biomass (Foster, 1991). A reasonable amount of EPS production with suitable shear force imposed by the aeration rate may lead to the successful formation and growth of granular biomass (Li et al., 2006). The SVI and SV of the granular sludge were used to evaluate the granular settling ability. It is anticipated that bigger granules would have higher SV and hence, reduce the SVI value, indicating good settling ability. The SVI value changes with the same pattern as the granular biomass concentration as well as the mean particle size of the granules. As the particles of the granular biomass decreased in size, the SVI value increased. The SVI value improved when the anaerobic reaction phase was prolonged in Stage V indicating such reaction pattern would help to develop granules with better settling prole. The SVI value improved as the size of the granules become bigger. It could also be possible that the increased accumulation of inert particles within the granules have contributed to improvement of SVI properties of the granular biomass. Fig. 4 shows the prole of SVI throughout the experiments. The SVI value in Stage V was reduced from 16.8 1.3 mL g1 (in Stage IV) to 15.5 1.3 mL g1. This is expected to be due to the accumulation of more inert solids within the granules as shown with small ratio of MLVSS/MLSS in Stage V (0.71). Despite changes in HRT that caused decrease in the granular sizes, the SVI values of the whole experiments were good except for Stage VI. During Stage VI, the prolonged duration

of the aerobic phase (i.e. 17.8 h) which was operated at high supercial air velocity (2.5 cm s1), caused the granular biomass to rupture. At this stage, the size of the granular biomass became smaller causing the settleability of the particles to reduce and was demonstrated with the increase in the SVI value.

3.3. 3.3.1.

Microbial activity and biokinetics Oxygen uptake rate

Microbial activity was measured based on the oxygen uptake rate during the aerobic phase in a one complete cycle operation. The OUR was measured several times before each stage ended. Figs. 5 and 6 show the proles of the OUR during aerobic phase for Stage I to VI. The results of the OUR measurement show that most of the external substrate was consumed within the rst 30 min of each aerobic reaction phase. This is shown by the sharp decrease of the oxygen consumption at the initial stage of the rst aerobic phase for all Stage I to VI of the experiment. The OUR proles of Stage I to Stage III (Fig. 5) show that the OUR at the beginning of each aeration phase reduces as the HRT increases. This is possibly due to the reduction in the OLR as the HRT increases. As the organic loading is reduced, less oxygen is therefore required. After a sharp increase of OUR at the beginning of each aerobic phase in all stages (Stage I to VI), the OUR was constantly low until the end of the aerobic phase. The low value of OUR indicates that most of the external substrates have been consumed. These gures (5 and 6) show that no further degradation took place although the HRT was extended. During the aerobic phase, after all of the external substrate was consumed, the microorganisms in the reactor undergo starvation phase where endogenous respiration took place. At the beginning of the second aerobic reaction phase, a relatively smaller increase in the OUR could be observed. The increase in the OUR measurement was believed probably to be due to the mineralization of amines, the byproducts of dye degradation during the anaerobic reaction phase.

Fig. 3 e Distribution of size particles at different stages of the experiment. Stage I: anaerobic (2.8 h): aerobic (2.8 h); Stage II: anaerobic (5.8 h): aerobic (5.8 h); Stage III and Stage IV: anaerobic (11.8 h): aerobic (11.8 h); Stage V: anaerobic (17.8 h): aerobic (5.8 h); Stage V: anaerobic (5.8 h): aerobic (17.8 h).

Fig. 4 e Prole of sludge volume index throughout the experiment. Stage I: anaerobic (2.8 h): aerobic (2.8 h); Stage II: anaerobic (5.8 h): aerobic (5.8 h); Stage III and Stage IV: anaerobic (11.8 h): aerobic (11.8 h); Stage V: anaerobic (17.8 h): aerobic (5.8 h); Stage V: anaerobic (5.8 h): aerobic (17.8 h).

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Fig. 5 e OUR prole of (a) Stage I (Aerobic phase 2.84 h), (b) Stage II (Aerobic phase 5.84 h) and (c) Stage III (Aerobic phase 11.84 h).

Fig. 6 shows the Stage IV to VI which was operated with the same HRT and organic loading but different in the anaerobic and aerobic reaction phase HRT ratio. As the duration of the anaerobic reaction phase was increased, the short pulse of the OUR in the second aerobic phase has increased from 96 to 130 mg L1 h1 in Stage IV and V, respectively. However, the OUR in the second aerobic phase for Stage VI has reduced to less than 80 mg L1 h1. These changes were postulated to be related to the length of second stage of anaerobic reaction phase. Prolong of the anaerobic reaction phase has increased the production of amines which was further degraded under aerobic phase causing an increase in

the OUR. This condition is vice verse when the length of anaerobic stage was shorten (Stage VI).

3.3.2.

Biokinetic parameters

The total solid biomass concentration in a biological reactor system is governed by the rate of substrates utilization and biomass production by the microorganisms. The rates of such processes which are known as the biokinetic parameters would give prediction on the performance of the biological process in wastewater treatment. The understanding and information on the rate of biological reactions and basic principles governing the growth of microorganisms are

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Fig. 6 e OUR prole of (a) Stage IV (Aerobic phase 11.84 h), (b) Stage V (Aerobic phase 5.84 h), (c) Stage VI (Aerobic phase 17.84 h).

very important in developing an effective design and operation of the biological reactor system (Tchobanoglous et al., 2004). In this study, the biokinetic parameters of the granular sludge were also investigated in relation to the effect of different HRTs. The biokinetic parameters that were investigated are the overall specic biomass growth rate ( `overall), endogenous decay rate (kd), observed biomass yield (Yobs), and theoretical biomass yield (Y). The calculations for the biokinetic parameters are according to the equations listed in Table 4 and are based on Liu and Tay (2007) and Chen et al. (2008). The results of biokinetic parameters for all stages in this experiment are given in Table 5. When the experiment moved from Stage I to III, the SRT was increased from 27.6 13.4 to 78.9 30.8 d which have caused the `overall to reduce from 0.036 to 0.013/d. The results are in accordance with Li et al. (2006) that sludge biomass will lose their bioactivity when the SRT is increased. The reduction of the `overall as the HRT increased was also observed by Liu and

Tay (2007). As mentioned earlier, the OLR was reduced when the HRT was increased from 6 to 24 h (from Stage I to III). The reduction in the OLR may also contribute to the reduction of `overall from Stage I to III. The `overall of Stage IV and V was the same when the SRT of these two stages slightly increased from 70.1 23.9 to 72.5 23.3 d, respectively. The `overall of Stage VI increased as the SRT was reduced to 41.6 18.4 d although Stage VI was operated with the same HRT as Stage IV and V. The reduction of the SRT in Stage VI may be contributed by the increase in the sludge washout that was shown by the increase in the suspended solids concentration in the efuent discharge. The rate of biomass lost due to endogenous respiration is represented by endogenous decay rate kd, as given in Eq. (4). The OUR that was measured during the last 10 min or before the second aeration phase stop of one cycle operation was used to calculate the kd. As the HRT increased from 6 to 24 h (Stage I to III), the kd values reduced. However, since the

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Table 4 e Coefcient of biokinetic parameters. Biokinetic coefcient

Overall specic biomass growth rate

Units
Per day

Formula 1 q

moverall
Per day

(3)

q sludge retention time Endogenous decay rate

kd

dO2 =dt OX M

(4)

dO2/dt oxygen uptake rate (mg/L h) Ox theoretical chemical oxygen demand which is assume as 1.42 mg O2/mg biomass M biomass concentration (mg VSS/L) Observed biomass yield mg VSS/mg COD

Yobs

Xe Ci Ce

(5)

Xe Efuent volatile solid concentration (g VSS/L) Ci COD concentration in the inuent (mg/L) Ce COD concentration in the efuent (mg/L) Theoretical biomass yield mg VSS/mg COD

Y Yobs 1 kd $q

(6)

reduction was also very small, the kd can be considered as constant when the HRT was increased. Furthermore, the kd value during 24 h HRT of Stage III to V can also be considered constant (i.e. 0.0075 to 0.0076/d). It can thus be concluded that the kd is considered constant throughout the experiment. The kd values calculated from this study were very small as compared to the kd values of aerobic granules (Chen et al., 2008) and of the activated sludge (Tchobanoglous et al., 2004). The observed biomass yield Yobs is the ratio of the biomass production rate to the substrate removal rate and is calculated according to Eq. (5). The Yobs is one of the most important parameter used in biological kinetic models. Eq. (5) is derived from the equation below (Liu and Tay, 2007): Yobs  Xvss2 Xvxss1 Vf Xe Ve tc Ci Ce Ve =tc (7)

Then Eq. (5) is simplied from Eq. (7) when the reactor system reached steady state and the biomass was maintained at a constant value (Chen et al., 2008). The Yobs can be used to describe the sludge productivity which relates to the net sludge production. The results in Table 5 show that the sludge production is inversely related to the value of SRT as shown in Stage I to

III. As SRT increased, the Yobs, value decreased. Since the biomass activity is reduced when the SRT increased, this has caused the reduction of the biomass yield. The results obtained from this experiment are in accordance with the ones reported by van Loosdrecht and Hence (1999). It is well known that the net sludge production in an activated sludge system decreases with increasing sludge age. The biokinetic parameters could give a good indication for the system performance. It can be used as a basis for the design and product optimization of a system reactor. The Yobs value of Stage IV to V, decreased from 0.269 to 0.217 mg VSS/mg COD as the SRT of those stages was increased from 70.1 23.9 to 72.5 23.3 d, respectively. Although Stage IV and V were operated with the same HRT, the ratio of anaerobic/aerobic reaction phase was different. It shows that when the ratio of anaerobic/aerobic time was increased, the Yobs decreased. The theoretical Y value is calculated using Eq. (6). It is expected that the theoretical Y value will be higher as compared to Yobs. The difference between Yobs and theoretical Y value is contributed by several factors including the endogenous metabolism of the microorganisms, predation, death and as well as lysis process. The theoretical Y value obtained in this study shows the same pattern as given by the Yobs. The results for the Yobs and theoretical Y value obtained

Table 5 e Kinetic coefcients of granular sludge at different stages of the experiment. Kinetic coefcients of granules sludge
Observed specic biomass growth rate (moverall) (per day) Endogenous decay rate kd (per day) Observed biomass yield (Yobs) (mg VSS/mg COD) Theoretical biomass yield Y (mg VSS/mg COD)

Stage I
0.036 0.0096 0.316 0.399

Stage II
0.024 0.0086 0.298 0.395

Stage III
0.013 0.0075 0.242 0.385

Stage IV
0.014 0.0075 0.269 0.410

Stage V
0.014 0.0076 0.217 0.338

Stage VI
0.024 0.0060 0.412 0.515

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in this experiment are within the typical reported range of conventional activated sludge system (Al-Malack, 2006; Tchobanoglous et al., 2004).

3.4. Chemical oxygen demand and color removal performance

The prole for COD concentration in the inuent, efuent and removal performance for all six stages of experiment is given in Fig. 7. The biogranular system shows consistent COD degradation performance with 84.2 0.9% removals after about 50 days of start-up period (acclimatization phase). The overall performance was almost consistent despite the decrease in biomass concentration and OLR as mentioned earlier. When the OLR was increased from 0.6 kg COD/m3day to 0.8 kg COD/m3day on the 194th day of the experiment (Stage III to IV), the COD removal efciency increased from about 84.4 0.4% at the end of Stage III (day 193) to 90.7 0.2% at the end Stage IV (day 236). Mohan et al. (2007) reported that the performance efciency of the SBR system was found to be affected by the operating OLR. The SBR system operating at higher OLR resulted with a high substrate uptake rate at the end of the cycle period. This was also observed by Ong et al. (2005). An increase in the percentage of COD removal efciency was also observed when the period of anaerobic phase was increased from 12 h to 18 h. The removal increased from 90.7 0.2% in Stage IV to 94.1 0.6% in Stage V. Pasukphun and Vinitnantharat (2003) claimed that the increase in the nonaeration phase in the SBR system would cause an alteration in the population of anaerobic microorganisms in the system which is expected to produce good COD and color removal for textile wastewater. However, according to Kapdan and Oztekin (2006), when the duration of anaerobic phase is too long, the contribution of aerobic reaction phase might be

Fig. 7 e Prole of COD removal performance of the reactor system at different stages of the experiment. (B) Inuent COD; (-) Efuent COD, (:) COD removal. Stage I: anaerobic (2.8 h): aerobic (2.8 h); Stage II: anaerobic (5.8 h): aerobic (5.8 h); Stage III and Stage IV: anaerobic (11.8 h): aerobic (11.8 h); Stage V: anaerobic (17.8 h): aerobic (5.8 h); Stage V: anaerobic (5.8 h): aerobic (17.8 h).

decreased. This is possibly due to the toxic effect of aromatic amines produced during dye degradation. Owing to the condition in the SBR system where different reaction phases occur in the same column, too long of anaerobic reaction periods will cause high accumulation of aromatic amine in the same compartment. High concentrations of aromatic amines may inhibit the activity of aerobic microorganisms during the aerobic phase. In this study, even though the anaerobic reaction phase was extended up to 18 h, there was no reduction in COD removal. This shows that there was no inhibition on the activity of aerobic microorganisms by the long accumulation of the byproduct produced from anaerobic degradation of the dye compound. It might be that the concentration of dye used during this experiment was not that high to produce enough concentration of the aromatic amines that may cause toxic effect toward the microorganisms within the biogranules. Furthermore, the biogranules might not be affected by the dyestuff degradation byproducts due to the structural form of the biogranules. The biogranules structure which consisted of EPS acts as a shield for microorganisms within the granules against any shock loading or toxic compound. At the nal stage (Stage VI) of the experiment, a surge drop of COD removal efciency was observed. As the aeration time was increased from 6 to 18 h, the COD removal reduced from 94.1 0.6% to 82.6 0.8%. The drop in the COD removal efciency was possibly due to the increase in biomass loss into the efuent. The MLSS in Stage VI was 23.3 0.8 g/L as compared to 31.6 3.7 g/L observed in the previous stages. Color removal was observed to increase from 66.7 1.6% to 76.5 0.8% as the HRT increased from Stage I to Stage III. Increase in the HRT allows longer contact time between the granules and the wastewater resulting in better color removal. Furthermore, when the OLR was increased from 0.6 kg COD/ m3day (Stage III) to 0.8 kg COD/m3day (Stage IV), improvement in color removal from 76.5 0.8% to 83.1 1.4% was observed. This may be caused by the increase in the microbial population. Ong et al. (2005) reported that the percentage of color removal efciency increased by 16% in anaerobic and 50% in aerobic SBR reactor systems when the OLR rate was increased from 2.66 to 5.32 g COD/Lday. An increase from 82% to 90% of color removal efciency was observed by Talarposhiti et al. (2001) when the COD loading was increased in a two-phase anaerobic packed bed reactor from 0.25 to 1 kg COD/m3day. Since more color removal took place in anaerobic condition (van der Zee et al., 2001; Dos Santos et al., 2007), the percentage of color removal was once again increased from Stage IV (83.1 1.4%) to Stage V (86.5 0.5%) when the anaerobic reaction phase was extended from 12 to 18 h of the 24 h reaction cycle. Improved decolorization process that occurs during the anaerobic stage enhances the overall wastewater biodegradation since more readily biodegradable substances could be degraded in the following aerobic treatment (Stolz, 2001). Fig. 8 shows the prole of the color removal performance. With respect to the mechanisms that are involved in color degradation, the addition of electronedonating substrate could considerably improve the decolorization reductive rate as reported by Dos Santos et al. (2005). Their studies using

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Fig. 8 e Prole of color removal performance of the reactor system at different stages of the experiment. (A) Inuent color, (-) Efuent color, (B) Color removal. (100 ADMI z 1 PteCo). Stage I: anaerobic (2.8 h): aerobic (2.8 h); Stage II: anaerobic (5.8 h): aerobic (5.8 h); Stage III and Stage IV: anaerobic (11.8 h): aerobic (11.8 h); Stage V: anaerobic (17.8 h): aerobic (5.8 h); Stage V: anaerobic (5.8 h): aerobic (17.8 h).

 The size and the SVI of the granular sludge reduced as the HRT of the system increased due to increase in the aeration time that resulted with the disintegration of the granules. Prolonged starvation condition may cause instability of the granular structure that lead to disruption of the granules. However the size and the SVI value were improved with the increase in the OLR and anaerobic reaction time and reduce in the aerobic phases.  Increase in the HRT resulted with an increase in the SRT. Since the SRT is inversely related to the moverall, increase in the HRT will cause a reduction in the moverall. Increase in the HRT has caused a reduction in the bioactivity of the granular sludge shown by the reduction of the moverall, Yobs and Y values. A slight increase in the SRT was observed with increase in the anaerobic/aerobic time ratio. This has caused a reduction in the Yobs and Y values but the moverall is considered constant. The kd is also considered constant throughout the experiment.  The percentage of COD removal in this study was not likely affected by the increase in the HRT which was mainly due to the decrease in the granular biomass and OLR. However, the COD and color removal was improved with prolong on the anaerobic reaction phase.

anaerobic and aerobic sequential wastewater treatment system indicated that the anaerobic stage was the main step for color degradation while the aerobic phase acted as the polishing step and enhancement in COD removal. Higher initial COD concentration did not improve color removal but caused deterioration in COD removal in the anaerobicaerobic SBR system (Kapdan and Oztekin, 2006). Pasukphun and Vinitnantharat (2003) reported that the duration of the anaerobic phase should be long enough to obtain better COD and color removal. Increase in the HRT would provide enough time for the degradation of the organics and inter-metabolites of textile wastewater in anaerobic or/and anaerobic/aerobic systems (Isik and Sponza, 2008). From this study, it shows that having longer anaerobic (18 h) and shorter aerobic (6 h) reaction phase resulted with the highest removal for color and slight improvement in the efciency of COD removal.

Acknowledgments
The authors wish to thank the Ministry of Science, Technology and Innovation (MOSTI), Ministry of High Education (MOHE) and Universiti Teknologi Malaysia for the nancial supports of this research (Grants No.: 79137, 78211 and 75221).

references

3.5.

Conclusions

Several conclusions can be derived from the study. They are as follows:  The granular biomass concentration in the reactor reduces as the HRT increases which is mainly due to the reduction in the OLR. However, with HRT of 24 h, the biomass concentration slightly improved when the anaerobic reaction phase was longer than aerobic reaction. The ratio of MLVSS/MLSS reduces when the anaerobic/aerobic reaction phase was set with 17.8/5.8 h which may be due to the increased accumulation of inert particles within the granules. Although with increase in the HRT, the concentration of granular biomass can be improved with the increase in the anaerobic reaction time and reduce in the aerobic reaction time.

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Abbreviations
q: Solid retention time (d) Xvss: Volatile solid concentration in the reactor system (g VSS/L) Vr: Working volume of the SBR system (L) Xd: Biomass concentration of manually discharged mixture (g VSS/L) Vd: Manually discharge mixture volume (L) Xe: Efuent volatile solid concentration (g VSS/L) Ve: Efuent volume of the SBR operating cycle (L) tc: Cycle time of the SBR operation (d) XVSS1: Volatile solid concentration at the beginning of cycle operation in SBR reactor (g VSS/L) XVSS2: Volatile solid concentration at the end of cycle operation in SBR reactor (g VSS/L)