Accepted Manuscript

Performance of integrated anaerobic/aerobic sequencing batch reactor treating

Ahmed Rahomi Rajab, Mohd Razman Salim, Johan Sohaili, Aznah Nur Anuar,
Salmiati, Sivarama Krishna Lakkaboyana

PII: S1385-8947(16)31561-3
DOI: http://dx.doi.org/10.1016/j.cej.2016.10.144
Reference: CEJ 16000

To appear in: Chemical Engineering Journal

Received Date: 16 September 2016

Please cite this article as: A. Rahomi Rajab, M. Razman Salim, J. Sohaili, A. Nur Anuar, Salmiati, S. Krishna
Lakkaboyana, Performance of integrated anaerobic/aerobic sequencing batch reactor treating poultry slaughterhouse
wastewater., Chemical Engineering Journal (2016), doi: http://dx.doi.org/10.1016/j.cej.2016.10.144

This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers
we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and
review of the resulting proof before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
Performance of integrated anaerobic/aerobic sequencing batch reactor treating poultry

slaughterhouse wastewater.

Ahmed Rahomi Rajaba,b* Mohd Razman Salima,c*, Johan Sohailia*, Aznah Nur Anuara,

Salmiatia,c and Sivarama Krishna Lakkaboyanac

a
Department of Environmental Engi1neering, Faculty of Civil Engineering, Universiti

Teknologi Malaysia, 81310, Johor Bahru, Malaysia.

b
Department of Civil Engineering, Faculty of Engineering, University of Anbar, 31001, Al-

Anbar governorate, Iraq.

c
Center of Environmental Sustainability and Water Security (IPASA), RISE, Universiti

Teknologi Malaysia, 81310, Johor Bahru, Malaysia.

Abstract

The present study investigates the performance of a new configuration laboratory-scale

bioreactor comprising of two regimes (anaerobic & aerobic) in one reactor with physical

separation and it is known as Integrated Anaerobic/Aerobic Sequencing Batch Reactor

(IAASBR). The IAASBR is designed for treating high-strength wastewater such as poultry

slaughterhouse wastewater (PSW) along with the simultaneous removal of organic carbon and

Abbreviations: A. Eff., aerobic effluent; An. Eff., anaerobic effluent; An. Inf., anaerobic influent; ASBR,
anaerobic sequencing batch reactor; DO, dissolved oxygen; EC, electrical conductivity; FOG, fat, oil &
grease; HRT, hydraulic retention time; IAASBR, integrated anaerobic/aerobic sequencing batch reactor;
MLSS, mixed liquor suspended solids; MLVSS, mixed liquor volatile suspended solids; NH3-N,
ammoniacal nitrogen; OLR, organic loading rate; ORP, oxidation reduction potential; PO4-3-P, total
phosphate; PSW, poultry slaughterhouse wastewater; Q, flow rate; SBBR, sequencing batch biofilm
reactor; SBR, aerobic sequencing batch reactor; SCOD, soluble chemical oxygen demand; SLR, sludge
loading rate; SVI, sludge volume index; TCOD, total chemical oxygen demand; TDS, total dissolved
solids; TKN, total Kjeldahl nitrogen; TOC, total organic carbon; TSS, total suspended solids; TVSS,
total volatile suspended; solids Van/Va, anaerobic-aerobic SBR volumetric ratio; Vf/VT, volumetric
exchange ratio; VSS, volatile suspended solids.

1
ammoniacal nitrogen (NH3-N). The IAASBR exhibits that the average removal efficiency of

total COD (TCOD), soluble COD (SCOD), NH3-N, fat, oil & grease (FOG), and total suspended

solids (TSS) were (97% ± 2%), (95% ± 3%), (98% ± 1.3%), (90% ± 11%), and (96% ± 3%)

respectively. The laboratory comparison test revealed that IAASBR configuration has enhanced

the sludge settle-ability for aerobic SBR more than the conventional SBR or settling tank.

Furthermore, IAASBR could tolerate the shock loading occurrence, handle organic loading rate

(OLR) up to 4.5 kg (TCOD) m-3 d-1 and produce a high-quality effluent complying with

Malaysian standards of industrial’s effluents.

Keywords: Integrated Anaerobic/Aerobic SBR, Poultry slaughterhouse wastewater, Settling

sludge enhancement, Shock loading tolerance, Fat, oil & grease removal.

∗
Corresponding authors E-mail addresses: R.R. Ahmed (ahmed.rahomi@gmail.com), S. Mohd.

Razman (mohdrazman@utm.my), and S. Johan (johansohaili@utm.my).

1. Introduction

The continuous increase in the meat production for the protein needs of the ever growing

world population has some pollution problems also attached to the environment. Poultry

slaughterhouse and chicken processing plants are one of the widest spread plants in Malaysia.

These plants produce relatively high amount of wastewater between 8 to 15 L per bird

slaughtered (200 – 700 m3/day) [1] depending on the production capacity and water

management policy that is followed by each plant. Furthermore, the characteristics of the

wastewater generated from these plants are classified as a high-strength wastewater [2],

where the contaminants’ concentrations have relatively high COD (3102±688 mg L-1),

suspended solids (SS) (872±178 mg L-1), oil & grease (O&G) (375±151 mg L-1), nitrogen

2
measured as total Kjeldahl nitrogen (TKN) (186±27 mg (N) L-1), and total phosphate (PO4-3-P)

(76±36 mg L-1). These contaminants are there due to the presence of high organic materials

such as blood, fat from skin, protein, oil from boiling birds for feather removal. Besides,

urine and faeces are also one of the main sources of nitrogen [1,3-7]. The discharge of this

wastewater without a proper treatment will have realistic damage on the environment and

municipal wastewater treatment plants.

Researchers have spent efforts for treating slaughterhouses wastewater using different

kinds of processes or for making a combination between them. Such work includes aerobic,

anaerobic [8,9], attached growth [10,11], suspended growth systems [12], mixed anaerobic

attached growth and aerobic suspended growth bioreactors in sequence [13,14],

anaerobic/anoxic/aerobic (suspended growth system)-biological contact oxidation (attached

growth system) integrated separated sludge bioreactors (A2/O–BCO) [15], and fixed bed

granular sludge with/without static activated sludge [16]. The combination of anaerobic-

aerobic fixed-film bioreactors connected in series (attached growth system in two reactors)

for treating the organic matter of poultry wastewater have achieved high removal efficiency

[17]. In another work, Del Pozo and Diez [18] put effort on treating PSW with the integrated

anaerobic-aerobic fixed-film reactor (attached growth system in one reactor). Their study

shows improvement with an overall high organics and nitrogen removal efficiency, when the

anaerobic:aerobic volume ratio was 2:3. Integrated anaerobic-aerobic bioreactor in one

reactor (mixed attached growth and suspended growth system) is used for treating high-

strength palm oil mill effluent (POME) wastewater [19]. The result shows that the system

had achieved a high constituent’s removal efficiency of BOD5, COD, and TSS. This result

was previously supported by using anaerobically digested POME as an influent to the

3
aerobic SBR for getting effluent within the discharge limits by changing OLR, sludge

loading rate (SLR) and mixed liquor volatile suspended solids (MLVSS) for attaining

optimum SBR performance [20].

A combination of anaerobic-aerobic SBR system in two reactors (suspended growth

system) was conducted by Bernet et al. [12] for treating piggery wastewater. The work

concentrates on the effect of the recycle-to-influent ratio for biological nitrogen and carbon

removal. The removal of total organic carbon (TOC) and TKN were fairly high. However,

the high hydraulic retention time (HRT = 22.5 d) in this study made it less suitable for

practical purposes.

Del Pozo and Diez [17] described the significance of using the anaerobic stage for

treating high-strength and high variance of wastewater’s characteristics before the aerobic

stage. They suggested conducting characterization for the wastewater before designing the

treatment plant. Anaerobic pre-fermentation stage is essential in practical slaughterhouse

wastewater treatment for high COD, NH3-N and total phosphate (PO4-3-P) removal efficiency

before entering SBR [21,22]. Del Nery et al. [23] further deduced that the effluent of an

individual anaerobic process fails to meet discharge requirements, and the aerobic process

alone requires more area and relatively high operating costs. The above statements are

important with respect to the applicability of the present study by using the raw wastewater

generated from poultry slaughterhouse as influent for the IAASBR.

The current investigation on the reactor configuration is motivated by and parallel to the

work [24]. The new reactor configurations are not only a necessity as an alternative to

already established systems but are of prime importance to develop optimized configurations

4
 that provide maximum performance, operational safety, and minimum cost [24,25].

Therefore, this study focuses on developing a novel configuration for existing technique

(SBR) to contribute and achieve the objectives stated afterward in an integrated manner

(anaerobic/aerobic processes in one reactor with physical separation using suspended growth

system only) and its practical use.

An attempt has been made to explore the performance efficacy of the proposed reactor

configuration that is IAASBR for simultaneous removal of carbonaceous organic matter and

ammoniacal nitrogen from PSW. This is, in conjugation with offering a suitable practical

solution for achieving the balance between the broiler chicken producer’s requirements

(needs for cheap, efficient, flexible, small footprint, less maintenance, and less sludge

disposal treatment technique) and the restricted regulations that are imposed by the

environmental protection agencies [26]. These requirements were necessary due to the

several critical issues which were found in the existing conventional treatment plants. These

challenges include high-cost of chemicals usage, non-compliance of effluents to standard

specifications, especially for NH3-N, and the defect of handling shock-loading.

2. Methodology and materials

2.1. Poultry slaughterhouse wastewater (PSW) characterization

PSW was collected from selected poultry slaughterhouse located in Johor (Southern state of

Malaysia Peninsular) called AYAMAS Food corporation Sdn. Bhd. This corporation is one of

the biggest poultry slaughterhouses in the state and has a production rate of 30,000 – 45,000

birds/day with average wastewater flow rate of 800 m-3 d-1. All samples used in this study were

collected from the flow coming from the fixed strainer instrument which is located after feathers

removal equipment. The characterizations of PSW are based on six visits; each visit lasts for one
5
working day. Equal volumes of samples were taken every 2 hours and gathered in a container.

Composite PSW had been analysed in a triplicate manner and presented in Table 1.

2.2. Reactor setup

Lab-scale IAASBR was made from acrylic, and it consists of two compartments. The first

compartment is for the anaerobic process with a maximum water volume of 12 L. This

compartment was supplied with a central mixer and has monitoring apparatuses for pH,

dissolved oxygen (DO), and oxidation reduction potential (ORP). Moreover, it has many ports

such as for anaerobic influent (An. Inf.) using water distributor at the bottom of the reactor,

aerobic sludge recycling, anaerobic effluent (An. Eff.), anaerobic sludge wasting, and gas-meter.

Furthermore, anaerobic SBR (ASBR) was provided with solenoid valves connected to a balloon

for balancing the internal pressure generated during the withdrawing and feeding time. The

anaerobic reactor was partially submerged inside the aerobic reactor (the second compartment)

depending on the aerobic water volume of each volumetric ratio that is subject to investigation in

this study. The aerobic compartment was also supplied with pH and DO meters, as well as with

air diffusers, mechanical mixers, and magnetic stirrer for providing air and mixing. Aerobic SBR

has many ports such as; aerobic influent (i.e. An. Eff.) with water distributor at the bottom,

aerobic effluent (A. Eff.), aerobic sludge recycling, and sampling ports. All experimental works

were carried out at a room temperature of 26-28 ºC. Figure 1 illustrates the reactor configuration,

and it works as one unit (i.e. flow rate (Q) is constant), where An. Inf. and A. eff. are equal. All

IAASBR’s operation phases concerning filling, reacting, settling, withdrawing, idle, and

anoxic/oxic phases and other complementary devices such as solenoid valves, air pump, and

mixers are controlled by timers. Excess aerobic sludge was transfer into the ASBR (i.e. ASBR is

6
the sole source for sludge disposal) for reducing the amount of sludge disposal and for achieving

one and more previously stated requirements of the abattoir’s industries.

2.3. Sludge developing and acclimatization process

Sludge for aerobic bioreactor (SBR) had been developed in the lab by adding 25% of

reactor’s total volume with raw wastewater (PSW). The reaction lasted for 23 hrs aerobically (5

L (air) min-1), in order to provide enough time for the aerobic biodegradation of the high-polluted

wastewater which is essential for the aerobes growth [2]. Then the same volume of supernatant

was discarded at the end of the settling phase. This procedure was repeated every day for

approximately three months. Excess aerobic sludge produced was used as inoculum sludge for

the ASBR, which was assigned for developing anaerobic sludge, a similar procedure used for

developing the aerobic sludge. This stage then continued for an extra three months. While

developing the anaerobic sludge, the acclimatization stage for aerobic sludge was initiated by

feeding the aerobic SBR with anaerobic effluent instead of raw wastewater (PSW). In other

words, this stage was carried out for adapting the developed aerobic sludge with the new feed

regime proposed for the IAASBR. Acclimatization stage lasted for one month, before starting up

for the first part of the current study.

2.4. Changing of anaerobic/aerobic volumetric ratio (Part I)

After the acclimatization period, the IAASBR started up by changing the anaerobic-aerobic

SBR volumetric ratio (Van/Va). To obtain the optimum ratio, the Van/Va ratios of 1, 1.5 and 2

were investigated. Other operating parameters for this configuration were listed in Table 2. The

water volume of ASBR (Van) was constant (12 L) and the water volume of aerobic SBR (Va) has

been changed accordingly with the designed Van/Va ratios. The flow rate (Q) for the system (6 L

7
d-1) and the volumetric exchange ratio for aerobic SBR (Vf/VT = 0.25) were constant. Hence, the

numbers of cycles for the system were 2, 3, and 4 cycles/d for the Van/Va ratios 1, 1.5, and 2

respectively.

2.5. Changing of organic loading rate (OLR) (Part II)

The optimum Van/Va ratio was selected from part one to be applied in the second part of this

study (i.e. Changes in OLR). This part investigated four ranges of OLR regimes i.e. 0.5-1.5, 2.0-

3.0, 3.5-4.5 and 5.0-6.0 kg (TCOD) m-3 d-1 by using real PSW. The real-time application of the

PSW discharge into the reactor is the sole reason for the OLR range utilized in this study instead

of a specific number. These OLR ranges were obtained by changing the concentration of

wastewater for keeping the Q and the HRT constants. The objective of this part was to quantify

the maximum OLR range that could be tolerated by the optimum anaerobic/aerobic volumetric

ratio (Van/Va) of the proposed IAASBR. This range is required for treating PSW with efficient

contaminants removal, producing a high-quality effluent that complies with the Malaysian

standards of discharged industrial effluents and to achieve most of the broiler’s chicken industry

requirements. Each OLR range was examined for at least three weeks, followed by a minimum

of one-week transitional period.

2.6. Analytical methods

The performance of the IAASBR has been observed by collecting samples from each process

such as An. Inf., An. Eff., and A. Eff. The parameters BOD5, TSS, volatile suspended solids

(VSS), mixed liquor suspended solids (MLSS), MLVSS, alkalinity, and FOG were determined

using standards methods [27]. Whereas the parameters of TCOD, SCOD (after filtration using

0.45µm filter paper), and NH3-N were measured using HACH system (DR6000)

8
Spectrophotometer (Method 8000-reactor digestion method for COD and Method 8038-Nessler

method for NH3-N). The pH, DO, and temperature (T) were monitored using Orion pH & DO

meter (Orion 4 star). Moreover, the sludge volume index (SVI) for both processes were

monitored, and the SVI parameter was measured according to the method stated in Metcalf &

Eddy [2]. All parameters mentioned earlier have been measured in a duplicate mode.

3. Results and discussion

3.1. Reactor performance and selection of the best volumetric ratio

IAASBR was operated more than four months for investigating its optimum performance by

changing the anaerobic-aerobic volumetric ratio (Van/Va) parameter. The results revealed that the

volumetric ratio of 2 has a maximum average removal efficiency of TCOD (97% ± 2%), SCOD

(95% ± 3%), NH3-N (98% ± 1.3%), FOG (90% ± 11%), TSS (96% ± 3%) and TVSS (98% ±

1.3%) as shown in Figure 2. Previously, researchers reported that the overall efficiency achieved

was 81-91% (measured as TOC) and 94.7% for NH3-N for the test period III [12]. This

configuration (IAASBR) as compared to the aforementioned study, where HRT was more than

22 days, exhibits higher performance and add to more practical operational mode as per results.

Whereas, comparison regarding the nutrients removal will be the future research scope (i.e.

results are not discussed in this paper). The high performance of IAASBR with a volumetric ratio

of 2 is attributed to the increasing number of cycles per day. Which means, that the ASBR had

been getting on a continuous feeding regime as it is recommended by Appels et al. [28] for the

high-rate anaerobic digester. Although the HRT for ASBR was constant (2 days) for all ratios

investigated, the HRT for aerobic SBR (SBR) keep on changing for each Van/Va parameter. The

change in HRT for aerobic SBR is because of using the same volumetric exchange ratio (Vf/VT)

with different aerobic SBR’s water volume, and the HRT is equal to 24 hr. for the volumetric

9
ratio of 2. This HRT value assisted the SBR to achieve a high performance [2]. By considering

the more detailed data during the timeline of this part, the high fluctuations of An. Inf.

contaminants concentrations can be seen in Figure I(a-e) of the supplementary data. These

fluctuations are common characteristics of such wastewater. As depicted, the An. Eff. have same

oscillation but less in their magnitude. The final products of IAASBR (i.e. A. Eff.) are more

stable and mostly less than the standard specification limits of effluent discharge, especially for

the high volumetric ratios 1.5 and 2. These results reveal that it is necessary for such kind of

wastewater (PSW) to use two processes (anaerobic and aerobic) in sequential order to overcome

the problems of highly polluted wastewater and its high fluctuation as well [29]. However, the

results support what is addressed as a future work by Mane and Munavalli [30], regarding using

the optimal design for SBR systems with different kinetics regimes.

3.1.1. Bearing of shock-loading

To be assured of the ability of IAASBR tolerance with the shock-loading condition, the

bioreactor configuration of volumetric ratio equal to 2 had been loaded with a high

contaminated PSW (high blood content) for two weeks. Then influent contamination level was

gradually reduced until it reaches the normal PSW within extra two weeks. This step proves that

the IAASBR could be handling the shock-loading occurrence, even though the final product (A.

Eff.) was not complying with standard specifications. However, the system could revert rapidly

to its previous performance depending on its resilient. This effect is observable from Figure 3.

3.1.2. Enhancement of sludge settling

In this configuration of IAASBR, the cylinder of the ASBR submerged into the cylinder of

aerobic SBR enhanced the sludge settle-ability within the aerobic SBR. The wash-out of the

10
aerobic sludge is not noticeable until the MLSS concentration becomes closer to 6000 mg L-1.

This phenomenon was not obvious although the sludge settling velocity was very slow and SVI

values were mostly more than 150 ml g-1 due to the fatty consistency of the sludge. Figure 4(a-c)

illustrates the comparison of sludge blanket level between the conventional SBR (normal 1 L

cylinder) and the simulated IAASBR configuration (blocked 0.5 L cylinder inverted into 1 L

normal cylinder) after 30 minutes settling time by using different MLSS concentrations. When

MLSS is equal to 7400 mg L-1, there is no difference in sludge blanket level between

conventional and new configuration SBR as shown in Figure 4a. Whereas this difference clearly

appears after reducing the MLSS concentration to 6000 mg L-1. The sludge volumes of the

conventional and the new configuration SBR were 950 ml and 730 ml respectively as depicted

in Figure 4b. The reduction in the MLSS concentration to 4600 mg L-1 also produces similar

results. Figure 4c shows that the sludge levels for conventional and new configuration SBR

were 595 ml and 375 ml respectively. The sludge of high concentration (more than 1000 mg L-1)

follows the hindered sludge settling theorem [2]. Therefore, this new configuration offers more

space or volume for developing compressed sludge faster at the lower/circle cross-section zone

of the reactor, while leaving the hindered sludge to be settled in the upper/ring cross-section

zone with more free space. Sludge blanket levels for the two cylinders as presented in Figure 4

are moving downward at a constant rate for the first 15 minutes of settling time. However, after

15 minutes elapsed, the difference of sludge levels between them is noticeable. This

phenomenon supports the above statement, regarding the development of compressed sludge in

the circle cross-section zone and making a reasonable explanation for why putting the all

aerobic effluent ports at the ring cross-section zone of the aerobic SBR.

3.2. Organic loading rate (OLR) examination (Part II)

11
 After selection of the best anaerobic/aerobic volumetric ratio from the part I, this ratio has

been adopted for the second part of this study, where the IAASBR was undergone to examine its

capability on handling the maximum OLR while maintaining its performance. Figure 5 presents

the overall results of part II by measuring the average concentrations of selected parameters

(TCOD, SCOD, NH3-N, FOG, and TSS) for each OLR range. The study reveals that the

IAASBR could withstand high OLR up to 4.5 kg (TCOD) m-3 d-1 and produces high-quality

effluents which are still within the Malaysia’s standard (B) for industrial effluents except for

FOG parameter. The average A. Eff. concentrations and the average removal efficiencies for

each OLR range, and the Malaysia’s standard (B) for industrial effluents are listed in Table 3.

The details of data for part II according to the timeline for the selected parameters are presented

in the Figure II(a-e) of the supplementary data. This result is compatible with the

recommendation made by Ruiz work [31] for the maximum OLR that should be applied to

ASBR which treats slaughterhouse wastewater is 4.5 kg (TCOD) m-3 d-1, However, it is

observed that only anaerobic process is not enough for achieving the aim of this study without

followed by an aerobic process. A comparison of IAASBR configuration has been made with the

most related bioreactor configuration in the literature [32]. The comparison shows that IAASBR

exhibit high performance in contamination’s removal efficiency of 97.9 %, while overall

removal efficiency of former research was only 94 % at the same OLR (0.9 kg (COD) m-3 d-1),

same temperature (28 ºC) but different in cycle time (12 hr.) and wastewater type (synthetic

wastewater, 800 mg (COD) L-1).

3.2.1. Alkalinity demand and ammoniacal nitrogen removal

Alkalinity concentration provided by raw PSW is sufficient for the anaerobic process for

all conditions applied on the proposed system (IAASBR), where the pH values for anaerobic

12
effluents were neutral (i.e. in the range of 6.5-7). Whereas for the aerobic process, alkalinity

concentrations were insufficient even though its concentrations had been elevated by anaerobic

and denitrification processes (anoxic phase for 1 hour during aerobic reactor’s feeding time with

An. Eff.). Hence, sodium bicarbonate (NaHCO3) has been added to the An. Eff. for increasing

the alkalinity buffer and producing A. Effs. within the normal pH values (in the range 6.5-8.5).

The quantity of the alkalinity required for accomplishing the nitrification process was varied

(750-1250 mg L-1 as HCO3) depending on the OLR applied.

From Figure II(c) at supplementary data, it can be observed that the level of NH3-N in An.

Eff. is much higher than An. Inf., especially in high OLR values (more than 4.5 kg (TCOD) m-3

d-1) as in alkalinity concentrations (data not presented here). This increase in NH3-N and

alkalinity is attributed to the anaerobically degradation of proteins of the raw wastewater (high

blood content). In the first step, proteins are hydrolysed to smaller molecules and produced α-

amino acids to be able to pass through the cell wall. Alternatively, the process of deamination of

these acids is held by anaerobic microbes causing the increase in the quantity of NH3-N and

alkalinity by producing ammonium bicarbonate (NH4HCO3) [33]. These concentrations of

alkalinity are not enough for completion of nitrification process and remove most of the

ammoniacal nitrogen generated from anaerobic-process without the addition of NaHCO3 to the

An. Eff. in a concentration that must be enough to maintain the alkalinity’s concentration of A.

Eff. more than 100 mg L-1 as CaCO3 at the end of aeration phase. Otherwise, the pH for A. Eff.

would be acidic (pH < 4) and the final NH3-N concentrations higher than the standard level as

illustrated in Figure 6. Consequently, the alkalinity is a crucial parameter for nitrification and

denitrification processes as revealed in this research work and as stated by other researchers

[34,35].

13
3.2.2. Fat, oil and grease (FOG) removal

Biochemistry of FOG’s degradation in its initial phases is similar for both aerobic and

anaerobic processes, where FOG is hydrolysed firstly with production of glycerol and fatty

acids. These fatty acids then undergo for further breakdown to smaller molecules consisting of

acetic acids in accordance with beta oxidation theory [33]. It is well known that the most

influential parameter on any biological treatment for PSW is FOG [22]. This parameter requires

for pre-treatment, such as commonly used of dissolved air flotation (DAF) system for this

purpose, before proceeding for any secondary treatment using any biological technique because

the lipids will be acting as a barrier between the microorganisms and its substrate causing

reduction of microorganisms activity and sludge wash-out [23,36]. On the contrary, in this study

PSW was added to the IAASBR without any pre-treatment in order to investigate its endurance

of added FOG concentrations and the potential of dispensing the DAF unit (economic aspect) on

one side, and to exploit the FOG content for producing high quantity of biogas on the other side

after the adaptation of the anaerobes microorganisms with this kind of wastewater [36,37] as

stated in the acclimatization section. From Figure II(d) at supplementary data, it could be said

that the average aerobic effluent FOG concentration of the OLR range (0.5-3.0 kg (TCOD) m-3

d-1) could be maintained below 20 mg L-1. However, this is only possible, if the anaerobic

influent FOG concentration is between 75 mg L-1 and 370 mg L-1 for the OLR range earlier

stated, while the average of A. Eff. FOG concentration was increasing more than 20 mg L-1 for

OLR higher than 3.0 kg (TCOD) m-3 d-1. The main effect of increasing OLR is the increasing of

FOG concentration in the influents of IAASBR which leads to; firstly, reduction of sludge’s

settling characteristics because of being lighter and secondly, forming a floating layer of sludge

at the water surface inside the anaerobic reactor. Moreover, entrapped gas bubbles by the sludge

14
flocs play a significant role in increasing the thickness of this layer. In other words, the

supernatant of anaerobically treated wastewater would be in between two layers of sludge. This

problem can be solved by increasing the settling time from 1 to 2 hrs and putting the

withdrawing point in a suitable place along the height of ASBR. However, the concentration of

FOG did not affect the performance of IAASBR unless the OLR does not exceed 4.5 kg

(TCOD) m-3 d-1. This result supports to the use of recurrent pulse feeding strategy (nature of the

SBR’s operation mode) for reducing of long-chain fatty acids (LCFA) inhibition effect on the

anaerobic treatment of slaughterhouse wastewater [38].

4. Conclusion

The operation/examination of the proposed configuration (IAASBR) exhibits high

organic and ammoniacal nitrogen removal efficiency accompanied with steady performance

and shock-loading tolerance. This system also has a high capability of handling OLR up to

4.5 kg (COD) m-3 d-1 with producing high-quality final effluents. Furthermore, the sludge

settling has been enhanced in the aerobic SBR due to this system's configuration in

conjunction with the overall decline in sludge production, because the ASBR is the sole

source of sludge wasting (0.37 kg (dry solids) m-3 (treated wastewater)). In addition, the pre-

treatment of raw wastewater is not required for removing FOG constituent. Hence, IAASBR

successfully achieved the target and it can be also tested for treating other kinds of high-

strength wastewaters in the future.

Acknowledgements

The authors gratefully acknowledge the financial support provided by the Universiti Teknologi

Malaysia under Research University Grant: Vot. No. Q.J130000.2522.10H69. Ahmed Rahomi

15
expresses his appreciation to the Ministry of Higher Education and scientific research, University

of Anbar, Iraq for the personal financial support to perform this study.

Appendix A. Supplementary data

5. References

[1] E.M. Matsumura, J.C. Mierzwa, Water conservation and reuse in poultry processing plant—

A case study, Resources, Conservation and Recycling 52 (2008) 835-842.

[2] George Tchobanoglous, Franclin L. Burton, H.D. Stensel, Wastewater Engineering

Treatment and Reuse, 4th ed., McGraw-Hill New York, USA, 2003.

[3] W. Cao, M. Mehrvar, Slaughterhouse wastewater treatment by combined anaerobic baffled

reactor and UV/H 2 O 2 processes, Chemical Engineering Research and Design 89 (2011)

1136-1143.

[4] V. Del Nery, M.H. Damianovic, F.G. Barros, The use of upflow anaerobic sludge blanket

reactors in the treatment of poultry slaughterhouse wastewater, Water science and

technology 44 (2001) 83 - 88.

[5] A.K.B. Amorim, I.R. de Nardi, V. Del Nery, Water conservation and effluent minimization:

Case study of a poultry slaughterhouse, Resources, Conservation and Recycling 51 (2007)

93-100.

[6] V. Del Nery, I.R. de Nardi, M.H.R.Z. Damianovic, E. Pozzi, A.K.B. Amorim, M. Zaiat,

Long-term operating performance of a poultry slaughterhouse wastewater treatment plant,

Resources, Conservation and Recycling 50 (2007) 102-114.

[7] S. Bayar, Y.Ş. Yıldız, A.E. Yılmaz, Ş. İrdemez, The effect of stirring speed and current

density on removal efficiency of poultry slaughterhouse wastewater by electrocoagulation

method, Desalination 280 (2011) 103-107.

16
[8] M.R. Johns, Developments in wastewater treatment in the meat processing industry: A

review, Bioresource Technology 54 (1995) 203-216.

[9] D.I. Massé, L. Masse, The effect of temperature on slaughterhouse wastewater treatment in

anaerobic sequencing batch reactors, Bioresource Technology 76 (2001) 91-98.

[10] A. Saddoud, S. Sayadi, Application of acidogenic fixed-bed reactor prior to anaerobic

membrane bioreactor for sustainable slaughterhouse wastewater treatment, J Hazard Mater

149 (2007) 700-706.

[11] A. Jacome, J. Molina, R. Novoa, J. Suarez, S. Ferreiro, Simultaneous carbon and nitrogen

removal from municipal wastewater in full-scale unaerated/aerated submerged filters, Water

science and technology : a journal of the International Association on Water Pollution

Research 69 (2014) 217-221.

[12] N. Bernet, N. Delgenes, J.C. Akunna, J.P. Delgenes, R. Moletta, Combined anaerobic–

aerobic SBR for the treatment of piggery wastewater, Water Research 34 (2000) 611-619.

[13] A. López‐López, R. Vallejo‐Rodríguez, D. Méndez‐Romero, Evaluation of a combined

anaerobic and aerobic system for the treatment of slaughterhouse wastewater,

Environmental technology 31 (2010) 319-326.

[14] J.C. Leyva-Díaz, M.M. Muñío, J. González-López, J.M. Poyatos, Anaerobic/anoxic/oxic

configuration in hybrid moving bed biofilm reactor-membrane bioreactor for nutrient

removal from municipal wastewater, Ecological Engineering 91 (2016) 449-458.

[15] M. Zhang, C. Wang, Y. Peng, S. Wang, F. Jia, W. Zeng, Organic substrate transformation

and sludge characteristics in the integrated anaerobic anoxic oxic–biological contact

oxidation (A2/O–BCO) system treating wastewater with low carbon/nitrogen ratio,

Chemical Engineering Journal 283 (2016) 47-57.

17
[16] E. Debik, T. Coskun, Use of the Static Granular Bed Reactor (SGBR) with anaerobic sludge

to treat poultry slaughterhouse wastewater and kinetic modeling, Bioresource Technology

100 (2009) 2777-2782.

[17] R. Del Pozo, V. Diez, Organic matter removal in combined anaerobic–aerobic fixed-film

bioreactors, Water Research 37 (2003) 3561-3568.

[18] R. Del Pozo, V. Diez, Integrated anaerobic–aerobic fixed-film reactor for slaughterhouse

wastewater treatment, Water Research 39 (2005) 1114-1122.

[19] Y.J. Chan, M.F. Chong, C.L. Law, An integrated anaerobic–aerobic bioreactor (IAAB) for

the treatment of palm oil mill effluent (POME): Start-up and steady state performance,

Process Biochemistry 47 (2012) 485-495.

[20] Y.J. Chan, M.F. Chong, C.L. Law, Biological treatment of anaerobically digested palm oil

mill effluent (POME) using a Lab-Scale Sequencing Batch Reactor (SBR), Journal of

Environmental Management 91 (2010) 1738-1746.

[21] Y. Jia, C. Gao, L. Zhang, G. Jiang, Effects of Pre-fermentation and Influent Temperature on

the Removal Efficiency of COD, NH4+-N and PO43–-P in Slaughterhouse Wastewater by

Using SBR, Energy Procedia 16, Part C (2012) 1964-1971.

[22] R. Lemaire, Z. Yuan, N. Bernet, M. Marcos, G. Yilmaz, J. Keller, A sequencing batch

reactor system for high-level biological nitrogen and phosphorus removal from abattoir

wastewater, Biodegradation 20 (2009) 339-350.

[23] V. Del Nery, M.H.Z. Damianovic, R.B. Moura, E. Pozzi, E.C. Pires, E. Foresti, Poultry

slaughterhouse wastewater treatment plant for high quality effluent, Water Science and

Technology 73 (2016) 309-316.

18
[24] D.S. de Oliveira, A.C. Prinholato, S.M. Ratusznei, J.A.D. Rodrigues, M. Zaiat, E. Foresti,

AnSBBR applied to the treatment of wastewater from a personal care industry: Effect of

organic load and fill time, Journal of Environmental Management 90 (2009) 3070-3081.

[25] A. Dutta, S. Sarkar, Sequencing Batch Reactor for Wastewater Treatment: Recent Advances,

Current Pollution Reports 1 (2015) 177-190.

[26] C.F. Bustillo-Lecompte, M. Mehrvar, Slaughterhouse wastewater characteristics, treatment,

and management in the meat processing industry: A review on trends and advances, Journal

of Environmental Management 161 (2015) 287-302.

[27] APHA, Standard Methods for the Examination of Water and Wastewater, 19th ed.,

American Public Health Association/American Water Works Association/Water

Environmental Federation, Washington DC., USA., 1995.

[28] L. Appels, J. Baeyens, J. Degrève, R. Dewil, Principles and potential of the anaerobic

digestion of waste-activated sludge, Progress in Energy and Combustion Science 34 (2008)

755-781.

[29] Y.J. Chan, M.F. Chong, C.L. Law, D.G. Hassell, A review on anaerobic–aerobic treatment

of industrial and municipal wastewater, Chemical Engineering Journal 155 (2009) 1-18.

[30] S.S. Mane, Dr.G.R.Munavalli, Sequential Batch Reactor- Application to Wastewater –A

Review, Proceeding of International Conference SWRDM-2012 (2012).

[31] C. Ruiz, M. Torrijos, P. Sousbie, J. Martinez, The anaerobic SBR process: basic principles

for design and automation, Water science and technology 43 (2001) 201 - 208.

[32] G. Kassab, M. Halalsheh, A. Klapwijk, M. Fayyad, J.B. van Lier, Sequential anaerobic-

aerobic treatment for domestic wastewater - a review, Bioresour Technol 101 (2010) 3299-

3310.

19
[33] Clair N. Sawyer, Perry L. McCarty, G.F. Parkin, Chemistry for environmental engineering

and science, 5th ed., McGraw-Hill, New York, USA, 2003.

[34] N. Callado, E. Foresti, Removal of organic carbon, nitrogen and phosphorus in sequential

batch reactors integrating the aerobic/anaerobic processes, Water Science and Technology

44 (2001) 263-270.

[35] P. Kundu, A. Debsarkar, S. Mukherjee, Treatment of Slaughter House Wastewater in a

Sequencing Batch Reactor: Performance Evaluation and Biodegradation Kinetics, BioMed

Research International 2013 (2013) 11.

[36] P.W. Harris, B.K. McCabe, Review of pre-treatments used in anaerobic digestion and their

potential application in high-fat cattle slaughterhouse wastewater, Applied Energy 155

(2015) 560-575.

[37] A.B.G. Valladão, A.G. Torres, D.M.G. Freire, M.C. Cammarota, Profiles of fatty acids and

triacylglycerols and their influence on the anaerobic biodegradability of effluents from

poultry slaughterhouse, Bioresource Technology 102 (2011) 7043-7050.

[38] J. Palatsi, M. Viñas, M. Guivernau, B. Fernandez, X. Flotats, Anaerobic digestion of

slaughterhouse waste: Main process limitations and microbial community interactions,

Bioresource Technology 102 (2011) 2219-2227.

20
Figure caption

Figure 1: Integrated anaerobic/aerobic sequencing batch reactor (IAASBR); (a) Schematic setup
(b) real setup.

Figure 2: Selection of the best volumetric ratio depending on the best average removal
efficiency of contaminations (error bars represent the standard deviation).

Figure 3: IAASBR performance during the shock-loading period.

Figure 4: Comparison of sludge blanket level after 30 min settling time between conventional
SBR (1) and simulated new configuration SBR for this study (2), where (a) for MLSS
= 7400 mg L-1, (b) for MLSS= 6000 mg L-1, and (c) for MLSS = 4600 mg L-1.

Figure 5: Average concentrations for selected parameters of aerobic effluents (A. Eff.) for each
OLR range (OLR measured as kg (TCOD) m-3 d-1).

Figure 6: illustrates the minimum concentration required of alkalinity in the A. Eff. for reducing
the ammoniacal-nitrogen and making its concentration within the standard level.

21
Legend of the Figure 1

1. Anaerobic SBR.
2. Mixers.
3. To the gas meter.
4. Aerobic SBR.
5. Magnetic stirrer.
6. Air diffusers.
7. Sludge disposal.
8. Anaerobic effluent.
9. Aerobic sludge recycling.
10. Aerobic effluent.
11. Aerobic effluent recycle (optional)
12. Influent
13. Air supplier
14. Anaerobic effluent sampling port

Solenoid
Air pump
valves
control Mixers

ASBR

Timers

Peristaltic
pumps
SBR
Final
effluent
tank

(a) (b)

Figure 1: Integrated anaerobic/aerobic sequencing batch reactor (IAASBR); (a) Schematic setup
(b) real setup.

22
Figure 2: Selection of the best volumetric ratio depending on the best average removal
efficiency of contaminations (error bars represent the standard deviation).

23
 14,000 100

90
12,000
80
TCOD concentration (mg L-1)

10,000

Removal efficiency %
70

60
8,000
50
6,000
40

4,000 30

20
2,000
10

0 0
0 3 6 9 12 15 18 21 24 27 30 33 36
Time (d)

An. Inf. TCOD Conc. An. Eff. TCOD Conc.

Figure 3: IAASBR performance during the shock-loading period.

24
(a) (b) (c)

Ring
cross-
section
zone

Circle
cross-
section
zone

Figure 4: Comparison of sludge blanket level after 30 min settling time between conventional
SBR (1) and simulated new configuration SBR for this study (2), where (a) for MLSS
= 7400 mg L-1, (b) for MLSS= 6000 mg L-1, and (c) for MLSS = 4600 mg L-1.

25
Figure 5: Average concentrations for selected parameters of aerobic effluents (A. Eff.) for each
OLR range (OLR measured as kg (TCOD) m-3 d-1).

26
 NH3-N concentration (mg L-1) 50

40

30

20

10

0
0 50 100 150 200 250 300 350 400 450
Alkalinity (mg L-1 as CaCO3)

Figure 6: illustrates the minimum concentration required of alkalinity in the A. Eff. for reducing
the ammoniacal-nitrogen and making its concentration within the standard level.

27
Table 1: Characterization of PSW used in this study.

Parameter Unit Average ± SD

TCOD mg L-1 2711 ± 487

TBOD5 mg L-1 930 ± 96

TN-N mg L-1 153 ± 32

NH3-N mg L-1 85 ± 32

PO4-3-P mg L-1 51 ± 2

FOG mg L-1 281 ± 63

Alkalinity mg L-1 as CaCO3 160 ± 21

TSS mg L-1 835 ± 162

TVSS mg L-1 813 ± 168

TDS mg L-1 917 ± 135

pH 6.8 ± 0.2

Temperature ºC 29 ± 0.7

Turbidity NTU > 1000

EC µs cm-1 795 ± 109

28
Table 2: Operation mode parameters for IAASBR during part I investigation stage.
Volumetric ratio (Van/Va)
1.0 1.5 2.0
An. An. An. A.
Parameter Unit SBR A. SBR SBR A. SBR SBR SBR
No. of cycles cycle d-1 2 2 3 3 4 4
Total volume
L 12 12 12 8 12 6
(VT)
Raw Raw Raw An.
Influent type ---- An. Eff. An. Eff.
PSW PSW PSW Eff.
HRT hr. 48 48 48 32 48 24
Flow rate (Q) L d-1 6 6 6 6 6 6
SRT day 40 15 40 15 40 15
-1
MLSS mg L 6000 <= 4000 6000 <= 4000 6000 <= 4000
Temp. ºC 26 - 28 26 - 28 26 - 28 26 - 28 26 - 28 26 - 28
(Vf / VT)* 0.25** 0.25 0.167** 0.25 0.125** 0.25
Filling time hr. 0.5 0.5 0.5 0.5 0.5 0.5
Anoxic time hr. --- 1.0 --- 1.0 --- 1.0
React time hr. 10 9.5 6 5.5 4 3.5
Settling time hr. 1.0 0.5 1.0 0.5 1.0 0.5
Withdrawing
hr. 0.5 0.5 0.5 0.5 0.5 0.5
time
min. 5.0 5.0 5.0 5.0 5.0 5.0
Idle time
*
Volumetric exchange ratio of aerobic SBR which was fixed on 25%, where Vf = water volume of the
filling phase and VT = total volume of aerobic SBR.
**
Varied because the system works as one unit (i.e. constant Q).

29
Table 3: The average A. Eff. Concentrations and the average removal efficiencies for each OLR range

comparing with the Malaysia’s standard (B) for industrial effluents.

Organic loading rate (OLR) range (kg (TCOD) m-3 d-1) Malaysia’s
0.5 – 1.5 2.0 – 3.0 3.5 – 4.5 5.0 – 6.0 Standard
(B) for
A. Eff. A. Eff. A. Eff. A. Eff. industrial
Conc. Removal Conc. Removal Conc. Removal Conc. Removal effluents
(mg L- efficiency (mg L- efficiency (mg L- efficiency (mg.L- efficiency 2009 (mg L-
Parameter 1) % ± SD 1
) % ± SD 1
) % ± SD 1
) % ± SD 1)

TCOD 44.0 97.9±0.8 47.8 98.9±0.3 65.0 99.2±0.4 651 94.4±2.2 200

SCOD 24.3 96.1±1.3 26.5 97.8±0.8 45.5 98.0±0.9 276 96.0±1.3 ----

NH3-N 2.3 96.3±3.3 9.3 86.1±11 17.8 90.7±4.1 44.0 75.5±18.6 20

FOG 21.1 85.6±5.0 18.5 94.3±1.8 44.0 93.4±5.2 119 90.6±5.3 10

TSS 35.9 95.7±2.6 35.0 97.8±0.7 46.0 98.6±0.6 78.3 93.3±4.9 100

30
Highlights:

• A novel bioreactor configuration (IAASBR) is developed for treating PSW.

• The proposed bioreactor (IAASB) fulfilled most of poultry industries’ requirements.

• IAASBR configuration enhanced the sludge settle-ability for aerobic SBR.

• This system produces the high-quality effluents within Malaysian standards.

• In this system, the FOG inhibition effect has been successfully eliminated.

31