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Acclimatization Process of Microorganisms from Activated Sludge in Kenaf-

Retting Wastewater

Chapter · November 2013

DOI: 10.1007/978-1-4614-6208-8_8

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Acclimatization Process
of Microorganisms from Activated Sludge
in Kenaf-Retting Wastewater

Z. Zawani, L. Chuah-Abdullah, F.-R. Ahmadun

and K. Abdan

Abstract Acclimatization process of microorganisms in Kenaf-retting wastewater

was studied using return activated sludge (RAS). RAS was collected from near
sewage treatment plant. Initial concentration of chemical oxygen demand (COD)
in the wastewater and mixed liquor suspended solids (MLSS) of activated sludge
were 2,300 and 1,000 mg/l, respectively. The MLSS and COD concentrations
were stabilized at day-25th. The highest substrate removal was recorded at 85 %,
and it is found that activated sludge bacteria are well adapted in the new envi-
ronment. At the end of the experiment, microorganisms, namely Achromobacter
sp, Bacillus sp and Acinetobacter sp, were found dominated in the wastewater.

Introduction

Kenaf is known as a renewable source and often used in developing new envi-
ronmental-friendly products. It consists of multiple useful components (e.g., stalks,
leaves and seeds), and within each of these plant components, there are various
usable portions (e.g., fibres and fibre strands, proteins, oils and allelopathic
chemicals). The most important part in kenaf is bast, which is a long fibre strands

Z. Zawani  L. Chuah-Abdullah  F.-R. Ahmadun

Chemical and Environmental Engineering, Department of Engineering,
Universiti Putra Malaysia, 43300 UPM Serdang, Selangor, Malaysia
L. Chuah-Abdullah (&)
Institute of Tropical Forestry and Forest Products (INTROP),
Universiti Putra Malaysia, 43300 UPM Serdang, Selangor, Malaysia
e-mail: chuah@eng.upm.edu.my
K. Abdan
Biological and Agricultural Engineering, Department of Engineering,
Universiti Putra Malaysia, 43300 UPM Serdang, Selangor, Malaysia
e-mail: khalina@eng.upm.edu.my

R. Pogaku et al. (eds.), Developments in Sustainable Chemical 59

and Bioprocess Technology, DOI: 10.1007/978-1-4614-6208-8_8,
 Springer Science+Business Media New York 2013
60 Z. Zawani et al.

that are composed of many individual smaller fibres, and it is contained in the bark
of kenaf stalk. The bast can be obtained from retting process. Retting is a fibre
separation process, and it can be done in several ways like dew, chemical or water-
retting. The process is a slow degradation process that can take up a few days to
complete. Water-retting generally is applied in industrial sector due to its effec-
tiveness and simplicity of the process itself. However, this procedure utilizes huge
amount of water. Wastewater that generated from the retting process produced
greenish colour, high in suspended solids and odour. The condition of wastewater
is unacceptable from an aesthetic value. Generally, the wastewater will be released
to watercourse without any proper treatment. Stringent regulations of water
quality, public awareness on environmental issues and a call for water reclamation
encourage an immediate action to resolve this issue.
Aerobic technology has been applied in wastewater treatment plant for a very
long time. Being one of the biological treatment processes, aerobic process
involved decompositions of organic and inorganic materials with the presence of
oxygen molecule and aerobic microorganism. Activated sludge (AS) from aerobic
process in wastewater treatment plant consists of various types of microorganisms
(MOs). Therefore, AS is often selected as a candidate for adaptation process in
new wastewater environment. The studies on acclimatization process are impor-
tant in introducing the MOs to unknown or foreign pollutants, which at some point
MOs can enhance degradation of pollutants compound effectively. Furthermore,
research on the acclimatization process is also necessary to improve WW tech-
nologies and the acclimatized MOs will help to increase the efficiency of bio-
logical treatment process.
Some researchers have studied the acclimatization process. Most of them exam-
ined the capability of MOs from activated sludge process (ASP) to adapt either
with new environment or under extreme condition. In 2007, Özbelge et al. were
studied about an exposure of heavy metal (HM) on MOs from activated sludge
process. They found that high level of HM exposure to the acclimatized MOs can
affect the ASP performance and it is difficult to predict the response of microor-
ganisms (MOs) in the AS as different MOs have different sensitivity to the factor
imposed to them. In contrast, other researchers studied on the capability of MOs
and mechanisms that may occur during acclimatization period. Wiggins et al
(1987) stated that several investigators have proposed that acclimatization process
is a result of the time needed for enzymes to be induced (Stephenson et al. 1984;
Torstensson et al. 1975), while others suggested time needed by MOs for mutation
or genetic exchange to occur (Schmidt et al. 1983; Walker and Newman 1956).
Mutation or appearance of new genotypes by genetic exchange may take place in
acclimatization process (Wiggins et al 1987).
The main purpose of this study is to investigate acclimatization process of
activated sludge in the kenaf-retting wastewater (KRW).
Acclimatization Process of Microorganisms 61

Table 1 Characteristics of Parameter Value (mg/l)

wastewater used in the
experiment pH 4.0
Total suspended solids (TSS) 550
Total solids (TS) 2,400
Colour 2,390
Turbidity 500
Phosphorus 7.7
Nitrogen 13.5
Chemical oxygen demand (COD) 2,300.0
Biological oxygen demand (BOD) 20.0

Methodology

Wastewater

Wastewater used in this experiment was synthetically prepared. Kenaf bast fibres
were retted in water for ten (10) days with the ratio of fibre to water 1 g: 40 ml as
suggested by Hongqin and Chongwen (2010). On day 10, the fibres were removed
from the container and the wastewater was kept in the container prior to the use.
The characteristics of sample are tabulated in Table 1.

Inoculums

For this experiment, activated sludge was obtained from University Putra Malaysia
(UPM) sewerage treatment plant. The sludge was collected from return activated
sludge (RAS) point and preserved in the cold room.

Experimental Set-Up

Sequencing batch reactor (SBR) system is adapted in order to acclimatize the MOs
collected from the treatment plant. The working volume of the system is 1.5 L
with the total volume of 2 L. Peristaltic pump (Watson-Marlow Bredel 323 Dz)
equipped with silicon tubing was used for feeding and withdrawing the sample.
Aeration from air pump and mixing were provided in the reactor. The concen-
tration of biomass in this experiment was set to 1,000 mg/L. Temperature and pH
of the system were kept constant at 28.0 ± 0.2 C and pH 7.0 ± 0, respectively.
The pH of medium and samples were adjusted using 0.1 N sulphuric acid (H2SO4)
and 0.1 N sodium hydroxide (NaOH).
62 Z. Zawani et al.

The SBR was operated in 24-h cycle period in four stages; feed and decant time
is 1 h (which total hours is 2 and there was no air provided during this period—
anoxic condition), reaction time is 20 h (with air supply) and settle/idle time is 2 h
(without air supply). The experiment was continuous running until the concen-
tration of COD in treated sample did not show further significant reduction.

Sample Analysis

The treated wastewater was analysed for concentrations COD and mixed liquor
suspended solids (MLSS). All parameters followed standard methods (D. Eaton,
H. Franson, & American Public Health Association 2005).
The acclimatized sludge samples were cultured and isolated for identification of
microorganism. Then, the samples were sent to accredited laboratory for identi-
fication process.

Results and Discussion

From the experiment, it is found that MOs in the system required 25 days to
acclimatize in the wastewater (as shown in Fig. 1). From the graph, it is revealed
that on day 25th, the MOs start to stabilize and did not change significantly until
day 30th. At this stage, biomass concentration was recorded at 2,900 mg/L with
the increase in 65.5 %. Since day 1st, the biomass starts to increase gradually, and
this condition may occur due to diversity of MOs present in the sludge. The
variations in MOs in the wastewater have made adaptation process easy.
The highest removal of COD was recorded at 85 %, as shown in Fig. 2. As can
be seen from both graphs, degradation level of organic compound and the growth
of biomass were increased gradually since first day. However, after 25 days, the
degradation process became slow which shows no significant change in concen-
tration of COD. The same condition occurred in biomass growth. There is no

Fig. 1 The average biomass

concentration recorded in
acclimatization process
Acclimatization Process of Microorganisms 63

Fig. 2 The COD removal efficiency in the acclimatization process

significant change after 30 days. The main species of bacteria that survived and
responsible in degrading pollutant compounds, there are bacillus sp., achromo-
bacter sp. and acinetobactor sp.
Several events may occur in the acclimatization process as proposed by pre-
vious researchers. Frequently, it is suggested that adaptation process indicates the
time for induction of enzymes after exposure of the indigenous populations to the
compound of interest (Wiggins et al. 1987). The time required for enzyme
induction is usually minutes or hours (Richmond 1968). The exposure to unknown
and unfamiliar organic pollutant may give obstacle to MOs in degrading the
pollutant. However, a long period of exposure to the wastewater and the diversity
of microorganism present in the AS may alleviate acclimatization process.
Another factor that may account for the acclimatization process is mutation or
the appearance of new genotypes by genetic exchange. In 1983, Schmidt et al.
explained that in their studies, new genotype by plasmid transfer appeared during
acclimation of microbial communities to chlorophenols. However, the latter
suggestion shows that the possibilities of new genotype appearance remain
unidentified.

Conclusions

Activated sludge from sewage treatment plant has potential to be used as inocu-
lums for the biological treatment of wastewater due to its assortment. Wastewater
used in the experiment was prepared using the same procedure done by farmers
and industry. From the experiment, it is found that microorganisms took 25 days to
adapt in the kenaf-retting wastewater . The percentage removal of COD and the
number of biomass growth recorded in the experiment shows that AS is capable to
live and help degrading the organic pollutant that is present in the wastewater.
 64 Z. Zawani et al.

References

Eaton, A. D., Franson, M. A. H., & American Public Health Association (Eds.) (2005). Standard
Methods for the Examination of Water and Wastewater. Washington: Amer Public Health
Assn
Lewis, D. L., Kollig, H. P., & Hodson, R. E. (1986). Nutrient limitation and adaptation of
microbial populations to chemical transformations. Applied and Environment Microbiology,
51, 598–603.
Özbelge, T. A., Özblege, H. Ö., & Altinten, P. (2007). Effect of acclimatization of
microorganisms to heavy metals on the performance of activated sludge process. Journal
of Hazardous Materials, 142, 332–339.
Richmond, M. H. (1968). Enzymic adaptation in bacteria: Its biochemical and genetic basis.
Essays in Biochemistry, 4, 105–154.
Schmidt, E., Hellwig, M., & Knackmuss, H. J. (1983). Degradation of chlorophenols by a defined
mixed microbial community. Applied and Environmental Microbiology, 46, 1038–1044.
Stephenson, T., Lester, J. N., & Perry, R. (1984). Acclimatisation to nitrilotriacetic acid in
activated sludge process. Chemosphere, 13, 1033–1040.
Torstensson, N. T. L., & Stark, J. (1975). The effect of repeated applications of 2, 4-D and MCPA
on their breakdown in soil. Weed Research, 15, 159–164.
Walker, H. H., & Newman, A. S. (1956). Microbial decomposition of 2, 4-dichlorophenoxyacetic
acid. Applied Microbiology, 4, 201–206.
Wiggins, B. A., Jones, S. H., & Alexander, M. (1987). Explanations for the acclimation period
preceding the mineralization of organic chemicals in aquatic environment. Applied and
Environmental Microbiology, 53(4), 791–796.
Yu, H., & Yu, C. (2010). Influence of various retting methods on properties of kenaf fiber.
Journal of the Textile Institute, 101(5), 452–456.

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