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UNIVERSITI PUTRA MALAYSIA
BIOREMEDIATION OF VEGETABLE OILY BALLAST WASTEWATER UNDER TEMPERATE CONDITION USING ANTARCTIC BACTERIA
MARYAM ABUBAKAR
FBSB 2018 9
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BIOREMEDIATION OF VEGETABLE OILY BALLAST WASTEWATER
UNDER TEMPERATE CONDITION USING ANTARCTIC BACTERIA
By
MARYAM ABUBAKAR
Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia,
in Fulfillment of the Requirements for the Degree of Master of Science
December 2017
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COPYRIGHT
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icons, photographs, and all other artwork, is copyright material of Universiti Putra
Malaysia unless otherwise stated. Use may be made of any material contained within
the thesis for non-commercial purposes from the copyright holder. Commercial use
of material may only be made with the express, prior, written permission of
Universiti Putra Malaysia.
Copyright © Universiti Putra Malaysia
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DEDICATION
This thesis is dedicated to my parents Engr Abubakar Magaji Gusau and Hajiya
Fatima Abubakar Magaji for their love, prayers and assistance towards the success of
thesis.
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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment
of the requirement for the degree of Master of Science
BIOREMEDIATION OF VEGETABLE OILY BALLAST WASTEWATER
UNDER TEMPRATE CONDITION USING ANTARCTIC BACTERIA
By
MARYAM ABUBAKAR
December 2017
Chairman : Associate Professor Mohd Yunus Abd Shukor, PhD
Faculty : Biotechnology and Biomolecular Sciences
Spills of vegetable oily waste especially palm oil as a result of ballast water
discharge from vegetable oil tankers in temperate waters are of environmental
concern because they cause serious effects on marine life and coastal environments.
The ongoing reclassification of oil palm ballast waste as a hazardous substance by
the European Union will seriously affect the Malaysian economy. Biodegradation by
indigenous cold-tolerant microorganisms is an important and potentially remediating
process solving this current problem. This study aims to investigate the
biodegradability of vegetable oil (palm oil) under the influence of a cold-tolerant
bacteria (Rhodococcus erythropolis ADL36) previously isolated from Antarctica.
The strain was cultured at different oil concentrations, temperature, pH, salinity, and
inoculum size under simulated conditions of oily ballast waste water. Furthermore,
the influences of the independent variables were optimised using response surface
methodology (RSM). A Plackett-Burman screening was carried out prior to RSM.
Three factors namely temperature, oil concentration and inoculum size appears to be
the most significant factors among the five while two factors; pH and salinity show
non-significant effect on the degradation of palm oil. The results of the research have
shown that maximum growth and biodegradation occurred at 1% (v/v) of the oil, at
25oC, pH 6.8, 2% of NaCl and an inoculum size of 5% (v/v) after OFAT. The
difference in the peaks of the oil component was seen in the GCMS result.
Moreover, the results of RSM showed that oil concentration, temperature and
inoculum size showed significant effects on the biodegradation of the oil. Out of the
eight primary models utilize, the modified Gompertz model was the best in modeling
the bacterial growth. Based on the growth rate constants obtained from the primary
modelling, a secondary modelling was carried out using various models such as
Luong, Yano, Tessier-Edward, Aiba, Haldane, Monod, and Han and Levenspiel. The
best model was Haldane giving the calculated value for the Haldane’s constants such
as maximal growth rate ( max), half saturation constant for maximal growth (Ks) and
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growth inhibition constant (Ki) tolerated were 0.74±0.12 day-1, 1.23±0.14 palm oil
(% v/v), and 3.12±0.16 palm oil (% v/v), respectively. In conclusion, the results
indicated the efficiency of such a system as a potential treatment for oily ballast
wastewater (vegetable oil).
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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai
memenuhi keperluan untuk ijazah Master Sains
BIOREMEDIASI MINYAK SAYURAN AIR SISA BUANGAN DIBAWAH
KEADAAN SUHU SEJUK DENGAN MENGGUNAKAN BAKTERIA
ANTARTIKA
Oleh
MARYAM ABUBAKAR
Disember 2017
Pengerusi : : Profesor Madya Mohd Yunus Abd Shukor, PhD
Fakulti : Bioteknologi dan Sains Biomolekul
Pencemaran dari tumpahan sayur-sayuran berminyak terutamanya kelapa sawit
akibat pelepasan air balast dari kapal pembawa minyak sayuran di perairan yang
sejuk menjadi kebimbangan kepada alam sekitar kerana ia menyebabkan kesan yang
serius terhadap kehidupan laut dan persekitaran pantai. Pengkelasan semula bahan
buangan kelapa sawit yang berterusan sebagai bahan berbahaya oleh Kesatuan
Eropah akan memberi kesan serius kepada ekonomi Malaysia. Biodegradasi oleh
mikroorganisma yang toleran kepada suhu sejuk adalah proses pemulihan yang
penting dan berpotensi untuk menyelesaikan masalah semasa ini. Kajian ini
bertujuan untuk mengkaji kebolehpenguraian minyak sayuran (minyak kelapa sawit)
di bawah pengaruh bakteria yang toleran pada suhu sejuk (Rhodococcus sp. strain
ADL36) yang sebelum ini dipencilkan dari Antartika. Strain ini ditumbuhkan pada
kepekatan minyak, suhu, pH, saliniti dan saiz inokulum yang berbeza di bawah
keadaan simulasi air sisa balast berminyak. Selain itu, pengaruh pembolehubah
bebas dioptimumkan menggunakan kaedah permukaan respon (RSM). Penyaringan
Plackett-Burman telah dijalankan sebelum RSM. Tiga faktor iaitu suhu, kepekatan
minyak dan saiz inokulum menjadi faktor yang paling penting di antara lima
manakala dua faktor; pH dan kemasinan menunjukkan kesan yang tidak signifikan
terhadap penguraian minyak kelapa sawit. Keputusan menunjukkan bahawa
pertumbuhan maksimum dan biodegradasi berlaku pada 1% (v /v) minyak, pada
suhu 25oC, pH 6.8, 2% NaCl dan saiz inokulum 5% (v /v) selepas RSM. Perbezaan
di puncak-puncak komponen minyak dilihat dalam pemerhatian berasaskan GCMS.
Selain itu, keputusan RSM menunjukkan kepekatan minyak, suhu dan saiz inokulum
menunjukkan kesan yang ketara terhadap biodegradasi minyak. Daripada lapan
model utama yang digunakan, model Gompertz yang diubahsuai adalah yang terbaik
dalam memodelkan pertumbuhan bakteria. Berdasarkan pemalar kadar pertumbuhan
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yang diperoleh dari pemodelan primer, pemodelan sekunder dilakukan dengan
menggunakan pelbagai model seperti Luong, Yano, Tessier-Edward, Aiba, Haldane,
Monod, dan Han dan Levenspiel. Model terbaik adalah Haldane yang memberikan
nilai-nilai pemalar Haldane seperti kadar pertumbuhan maksima (max), pemalar
ketepuan tepu untuk pertumbuhan maksimum (Ks) dan kepekatan yang merencat
pertumbuhan (Ki) pada 0.74 ± 0.12 hari-1, 1.23 ± 0.14 minyak sawit (% v /v), dan
3.12 ± 0.16 minyak sawit (% v /v), masing-masing. Kesimpulannya, hasil kajian
telah menunjukkan kecekapan bakteria ini yang berpotensi dalam merawat air
kumbahan balast berminyak (minyak sayuran).
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ACKNOWLEDGEMENTS
In the name of Allah, the Most Gracious and Most Merciful. All thanks and praises
are due to Allah (SWT) for giving me the opportunity to successfully conduct this
research. May all blessings be upon His Prophet and Messenger, Muhammad
(SAW).
The successful completion of this research was made possible with the support and
assistance of many important people. First and foremost, I would like to express
profound gratitude to my supervisor Assoc Prof. Dr. Mohd Yunus Abd Shukor and
my co-supervisors; Dr Adeela Yasid and Dr. Siti Aqlima Ahmad Whom accepted
me as their student and offered me mentorship, moral support and care. This work
would not have been possible without their guidance and involvement. Equally, I
wish to express my appreciation to all my colleagues in the Bioremediation Lab for
the contributions they rendered during the conduct of this research.
Words cannot express my gratitude to my husband Dr. Abbas Sani Dahiru for the
support and encouragement towards the success of this research. He was always
beside me during the happy and hard moments to motivate me. I thought that it is
impossible to continue, you helped me to keep things in perspectives. I greatly value
his contributions and deeply appreciate his belief in me. Equally, my profound
gratitude goes to my brothers, sisters, friends and family members for their concern
and prayers which serves as a source of inspiration to me. My heart felt regard goes
to my father inlaw, mother in law for their love, prayers and support.
Last and not the least is my lovely daughter Ramlat I really appreciate you for
bearing with me and making the house lively despite my busy academic schedules.
Maryam Abubakar, 2018.
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This thesis was submitted to the Senate of the Universiti Putra Malaysia and has
been accepted as fulfilment of the requirement for the degree of Master of Science.
The members of the Supervisory Committee were as follows:
Mohd Yunus Shukor, PhD
Associate Professor
Faculty Biotechnology and Biomolecular Sciences
Universiti Putra Malaysia
(Chairperson)
Nur Adeela Yasid, PhD
Senior Lecturer
Faculty of Biotechnology
Universiti Putra Malaysia
(Member)
Siti Aqlima Ahmad, PhD
Senior Lecturer
Faculty of Biotechnology and Biomolecular Sciences
Universiti Putra Malaysia
(Member)
ROBIAH BINTI YUNUS, PhD
Professor and Dean
School of Graduate Studies
Universiti Putra Malaysia
Date :
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Declaration by graduate student
I hereby confirm that:
this thesis is my original work;
quotations, illustrations and citations have been duly referenced;
this thesis has not been submitted previously or concurrently for any other degree
at any institutions;
intellectual property from the thesis and copyright of thesis are fully-owned by
Universiti Putra Malaysia, as according to the Universiti Putra Malaysia
(Research) Rules 2012;
written permission must be obtained from supervisor and the office of Deputy
Vice-Chancellor (Research and innovation) before thesis is published (in the
form of written, printed or in electronic form) including books, journals,
modules, proceedings, popular writings, seminar papers, manuscripts, posters,
reports, lecture notes, learning modules or any other materials as stated in the
Universiti Putra Malaysia (Research) Rules 2012;
there is no plagiarism or data falsification/fabrication in the thesis, and scholarly
integrity is upheld as according to the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (Revision 2012-2013) and the Universiti Putra Malaysia
(Research) Rules 2012. The thesis has undergone plagiarism detection software
Signature: Date:
Name and Matric No : Maryam Abubakar, GS44411
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Declaration by Members of Supervisory Committee
This is to confirm that:
the research conducted and the writing of this thesis was under our supervision;
supervision responsibilities as stated in the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (Revision 2012-2013) were adhered to.
Signature:
Name of Chairman
of Supervisory
Committee:
Associate Professor
Dr. Mohd Yunus Shukor
Signature:
Name of Member
of Supervisory
Committee:
Dr. Nur Adeela Yasid
Signature:
Name of Member
of Supervisory
Committee:
Dr. Siti Aqlima Ahmad
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TABLE OF CONTENTS
Page
ABSTRACT i
ABSTRAK iii
ACKNOWLEDGEMENTS v
APPROVAL vi
DECLARATION viii
LIST OF TABLES xii
LIST OF FIGURES xiii
LIST OF ABBREVIATIONS xv
CHAPTER
1 INTRODUCTION 1
1.1 Research Background 1
1.2 Problem statement 3
1.3 Objectives 3
2 LITERATURE REVIEW 4
2.1 Biodegradation 4
2.2 Degradation of Fats and Oils by Lipases 4
2.3 Oil-Degrading Bacteria 5
2.4 Aerobic Biodegradation 5
2.5 Vegetable Oil 6
2.5.1 Properties of Vegetable Oil 7
2.5.2 Industrial Uses of Vegetable Oils 7
2.5.3 Biodegradation Characteristics of Vegetable Oils 8
2.6 Marine Pollution 9
2.6.1 Vegetable oil pollution 9
2.7 Remedies for Oil Spill in Marine Water 11
2.7.1 Physical Remediation Methods 11
2.7.2 Chemical Remediation Methods 13
2.7.3 Thermal Remediation Method 14
2.7.4 Bioremediation Method 14
2.8 Remedies for Oil Spill in Arctic Waters 15
2.8.1 Mechanical Recovery 15
2.8.2 Use of Dispersants 16
2.8.3 In-situ Burning 16
2.9 Methods of Optimisation of Vegetable Oil-Degrading Capacity
in Microorganisms 17
2.9.1 One Factor at a Time 17
2.9.2 Experimental Designs and Statistical Optimisations 17
2.10 Bioremediation of Ballast Water through Biodispersion 19
2.11 Growth Kinetics of Bacteria 20
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3 MATERIALS AND METHODS 28
3.1 Experimental Approach 28
3.2 Source and Maintenance of bacteria 28
3.3 Media Preparation 29
3.4 Growth Determination 29
3.5 Determination of Oil Degradation 30
3.6 Gas Chromatographic Analysis of Palm Oil Degradation 30
3.7 Optimization of Factors Affecting Oil Degradation Using OFAT 31
3.7.1 Optimization of Substrate Concentration 31
3.7.2 Optimization of Inoculum Size 31
3.7.3 Optimization of the Medium pH 31
3.7.4 Optimization of Temperature 31
3.7.5 Optimization of the Salinity 32
3.8 Optimization Using Statistical Approach 32
3.8.1 Plackett – Burman Factorial Design (PBFD) 32
3.8.2 Central Composite Design (CCD) and Response
Surface Methodology (RSM) 32
3.9 Modelling the Kinetics of Growth on palm oil 33
3.9.1 Fitting of the data 33
3.9.2 Statistical analysis 33
4 RESULTS AND DISCUSSIONS 35
4.1 One Factor at a Time (OFAT) 35
4.1.1 Effect of initial Substrate Concentration 35
4.1.2 Effect of Temperature 36
4.1.3 Effect of pH 37
4.1.4 Effect of Salt Concentration 38
4.1.5 Effect of Inoculum Size 39
4.2 GCMS Analysis 40
4.3 Statistical Optimizations 44
4.3.1 Selection of Significant Variables by Plackett–Burman
Design 44
4.3.2 Response Surface Methodology (RSM) 46
4.4 Kinetic Modelling of Growth 53
4.4.1 Growth on palm oil modelled using primary models 53
4.4.2 Growth on palm oil modelled using secondary models 60
5 SUMMARY, CONCLUSION AND RECOMMENDATIONS 65
66
84
87
REFERENCES
APPENDICES
BIODATA OF STUDENT
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LIST OF TABLES
Table Page
2.1 Bacterial growth models used in this study 22
2.2 Various mathematical models developed for degradation kinetics
involving substrate inhibition
24
4.1 GCMS analysis of 1% palm oil 42
4.2 GCMS analysis of control palm oil 43
4.3 Coded and actual values of significant factors used in plackett-
Burman factorial design
44
4.4 Experimental sdesign matrix of Plackett-Burman and the response 45
4.5 Analysis of ANOVA for the Model of Factors in Placket-Burman 45
4.6 Coded and actual values of significant factors used in central
composite design (CCD)
46
4.7 CDD experimental matrix generated by Design expert and
corresponded responses (actual and predicted)
46
4.8 Analysis of variance (ANOVA) for Response Surface Quadratic
Model
47
4.9 Comparison of the optimized condition between OFAT and RSM of
4% palm oil
53
4.10 Statistical analysis of the various fitted models 58
4.11 Bacterial growth coefficients at various palm oil as modelled using the
modified Gompertz model
59
4.12 Statistical analysis of kinetic models 64
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LIST OF FIGURES
Figure Page
2.1 Hydrolysis of TAG by lipase (Edmund, 2001) 4
2.2 Global vegetable oil production. Source: USDA, AMI 6
3.1 A schematic overview of the experimental stages 28
4.1 Effect of Palm oil Concentration (sole carbon source) on the growth
of Rhodococcus erythropolis ADL36. Error bars represent mean ±
standard deviation of triplicates.
36
4.2 Effect of temperature on the growth of Rhodococcus erythropolis
ADL36. Error bars represent mean ± standard deviation of triplicates
37
4.3 Effect of pH on the growth of Rhodococcus erythropolis ADL36.
Error bars represent mean ± standard deviation of triplicates
38
4.4 Effect of Salinity on the growth of Rhodococcus erythropolis ADL36.
Error bars represent mean ± standard deviation of triplicates
39
4.5 Effect of Inoculum size on the growth of Rhodococcus erythropolis
ADL3. Error bars represent mean ± standard deviation of triplicates
40
4.6 Inoculated Gas chromatographic analysis of fatty acids from
utilization of 1% (v/v) palm oil by the culture of Rhodococcus
erythropolis ADL36 after 7 days of inoculation with (a) uninoculated
(control) and (b) treated
41
4.7 Model diagnostic plots (a) predicted versus actual (b) studentized
residue versus predicted (c) normal plots of residue (d) outlier T
versus run
49
4.8 3D and 2D surface response view showing the interaction between oil
concentration and temperature
50
4.9 3D and 2D surface response view showing the interaction between
temperature and inoculum size
51
4.10 3D and 2D surface response view showing the interaction between oil
concentration and inoculum size
52
4.11 The bacterial growth curves of Rhodococcus sp. strain ADL36 at
various concentrations of palm oil over time after RSM optimization.
The error bars represent mean ± standard deviation of three replicates
54
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4.12 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the Huang model
54
4.13 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the Baranyi-Roberts model
55
4.14 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the modified Gompertz model
55
4.15 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the Buchanan-3-phase model
56
4.16 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the modified Richards model.
56
4.17 A. Growth profile of Rhodococcus sp. strain ADL36 on palm oil
fitted according to the modified Schnute model
57
4.18 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the modified Logistics model
57
4.19 Growth profile of Rhodococcus sp. strain ADL36 on palm oil fitted
according to the von Bertalanffy model
58
4.20 Fitting experimental data with the Luong model 60
4.21 Fitting experimental data with the Yano model 61
4.22 Fitting experimental data with the Teissier model 61
4.23 Fitting experimental data with the Aiba model 62
4.24 Fitting experimental data with the Haldane model 62
4.25 Fitting experimental data with the Monod model 63
4.26 Fitting experimental data with the Han-Levenspiel model 63
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LIST OF ABBREVIATIONS
% Percent
CCD Central composite design
cm Centimetre
dH2O Distilled water
et al and friends
G Gram
h Hour
kpa Kilopascal
L Liter
M Molar
min Minute
mL Mililiter
MSM Minimal salt medium
NA Nutrient agar
sec Second
ºC Degree Celcius
OD Optical density
RPM Rotation per minute
RSM Response surface method
µL Microlitre
µm Micrometer
mm Millimeter
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CHAPTER 1
1 INTRODUCTION
1.1 Research Background
More than 80% of commercial goods shipments are transported through Sea globally
(Ibrahim & El-Naggar 2012). In addition to the intentional shipment, over 12 billion
tons of ballast water is moved through massive coastal and sea areas annually
(Wattayakorn, 2012). Ships are made purposely to the transportation of goods such
as oil through aquatic. However, if the ship is travelling without the goods or has
discharged goods in one port on its way to other port of call, In other to achieve the
required safe, functional conditions that include keeping the ship sufficiently in the
water to make sure effective propeller and rudder procedure also to avoid the bow
developing from the water, mostly in heavy oceans, Ballast may be carried on board.
A good weight-to-volume ratio is taken in a detached or empty cargo tanks which
are used only for ballast water and when a container is parting a port any residue that
is pumped into the ballast tanks that may be disturbed up, is released again in the
next port when takings on cargo. Hazardous nature of oil spill can effectively be
tackled through enforcing proper regulations in the best good working condition, but
can never be removed completely.
Vegetable or petroleum oil spills are considered as serious contaminants in marine
environments, due to their toxicity to marine organisms (El-Masry et al. 2004). Only
few oil spills in Arctic waters have been reported (Singsaas & Lewis 2011). Though,
with an increase in the activities of oils such as production, exploration, and
transport there is an increased risk for future occurrence. Various Arctic areas are
remote with inadequate infrastructure to support spill response. Additionally, oil spill
response in an extremely low temperature and darkness is much difficult. The
physical treatment of fats containing wastewaters is considered inadequate and hence
is costly while biological treatment provides the most efficient means that eliminate
fats and oil using lipases enzyme (Manan et al. 2014). These enzymes otherwise
known as triacylglycerol acylhydrolases are produced by many microorganisms
where they catalyse the synthesis or hydrolysis of fats (Shabtai et al. 1992).
According to the U.S. Environmental Protection Agency (USEPA), the release of
edible fats, oils, and greases (FOGs) into the environment does not constitute danger
to the environment or human health, However, it is required by regulation that
FOGs be treated by the same safety and spill response practices applied to petroleum
oils due to its definition under the general definition of ‘‘oil’’ as specified by 40
CFR 112.2 and Clean Water Act Section 311(a)(1), (USEPA 1997). It is evident
from the literature that discharges of FOGs into open seas, coastlines, or river bodies
can be as harmful as spills of petroleum oil, resulting in environmental and economic
damage (Bucas and Saliot 2002).
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Therefore, it is important to identify effective bacteria that degrade lipids through
culture-dependent technique. The lipid- degrading bacteria usually produce
triacylglycerol acylhydrolases which influences the structural arrangements of fats
and oil such as oleic acid and Tween 20, olive and palm oils (El-Masry et al. 2004).
El-Bestawy et al. (2005) reported many pathways through which triacylglycerol
acylhydrolases are produced and secreted by microorganisms (Nizam & Zuhan,
2008). Margesin and Schinner (2001) stated that hydrocarbon polluted environments
are categorized based on extreme temperatures, pH, and salinity. Despite such
extreme environmental conditions, microorganisms thrive and adapt to the
environment thereby utilising the oils as potential energy means.
The vegetable oil spill is the most common form of environmental pollution in the
United States which attracts many scientific attentions in the field of its toxicology
and the probable degradation response under aerobic and anaerobic conditions (Al-
Darbi et al. 2005). Palm oil pollution in the European seas has reached the limelight
as it has been proven to be the cause of deaths of dogs and animals consuming the
cold precipitated washed palm oil residues from oily ballast wastewater discharge
into the sea (Cudmore, 2017). The rise in the number of pollution cases in the
European waters is an alarming issue as the European Union has recently tried to ban
the use of palm oil in biodiesel production and pollution cases involving palm oil
will only make the ban a reality in the near future (Tornero et al., 2016).
Thus, ways to remediate oily ballast wastewater containing palm oil is in urgent
demand. In addition to bio treatment, many techniques used to remedy marine oil
spills involve mechanical and chemical methods. Boben and Yanting (1996)
recommended using in-situ burning as the primary means of response to the major
oil spill occurrence. However, these methods are expensive and not practical to treat
oily ballast wastewater that contains mainly palm oil and small amounts of minerals
in seawater. The treatment of oily ballast wastewater tends to focus on tropical and
subtropical regions, and concerns on the reduction of invasive microorganisms
chiefly phytoplankton and hence treatment usually involve filtration and UV
treatment (Tsolaki & Diamadopoulos, 2009). With the increasing problem in oily
ballast wastewater containing toxic hydrocarbon and lipid wastes, the use of
microorganism is being developed (Ganti et al., 2003) but is still limited. Further
aggravating this issue is that biological treatment for oily ballast wastewater in cold
seas, especially in the European waters centring on palm oil, is absence due to the
difficulty in finding suitable microorganisms to treat the waste before being
discharged. The ongoing reclassification of oil palm ballast waste as a hazardous
substance by the European Union will seriously affect Malaysian economy (Höfer &
Mez 2008) being the second world most palm oil production and exportation after
Indonesia.
Considering the facts discussed above, this research was conducted to address the
issue of oily ballast wastewater treatment using microorganisms as an alternative to
physicochemical methods. More specifically, this study aims to investigate the
aerobic biodegradability of palm oil under a simulated oily ballast wastewater
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conditions. Furthermore, refined commercial palm oil was selected for this study
which is one of the main sources of vegetable oil (Przybylski et al., 2005).
1.2 Problem statement
This is the first attempt to use an Antarctic diesel-degrader as a candidate for the
subtropical conditions remediation of oily ballast wastewater-containing palm oil as
a major pollutant under marine environment. Major issues that can hamper the
successfulness of this study include the ability of the bacterium to degrade high
concentration of palm oil in the simulated oily ballast wastewater. Another issue is
whether the degradation of oil palm can be effective under high salts and cold
conditions that are present in the Atlantic seas. The degradation will be carried out
under controlled conditions that can be found in the oily ballast wastewater.
As of now, there is no attempt in studying the possibility of using cold region
microbes in remediating oily ballast wastewater from palm oil tankers. This is the
first such study, and the effect of environmental factors in palm oil biodegradation
needs to be studied and optimized under cold conditions.
1.3 Objectives
The general objective of the study is to remediate oily ballast wastewater under
temperate condition using Antarctic bacteria.
The specific objectives of the study are;
1. To investigate the biodegradability of palm oil under simulated oily ballast
wastewater conditions by Rhodococcus erythropolis ADL36 using one factor
at a time (OFAT).
2. To optimize the degradation condition using response surface methodology
(RSM).
3. To determine the growth parameters of the bacteria in utilizing the oil using
kinetic models.
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6 REFERENCES
AbdEl-Mongy, Shukor, M., A., M.,S., Hussein, S., Ling A., P., K., Shamaan, N., A.
& Shukor, M.,Y. (2015) Isolation and characterization of a molybdenum-
reducing, phenol- and catechol-degrading Pseudomonas putida strain amr-12
in soils from Egypt. Scientific Study & Research Chemistry & Chemical
Engineering, Biotechnology, Food Industry 16, 353–369.
Abou-Shanab R., A., I., Khalafallah, M., A., Emam, N., F., Aly, M., A., Abou-Sdera
S., A., Matter, I., A. (2012) Characterisation and identification of carbofuran-
utilising bacteria isolated from agricultural soil. Chemistry and Ecology 28,
193–203.
Adhvaryu, A., Erhan, S., & Perez, J. (2002). Wax appearance temperatures of
vegetable oils determined by differential scanning calorimetry: effect of
triacylglycerol structure and its modification. Thermochimica acta, 395, 191-
200.
Agarry, S., Audu, T., & Solomon, B. (2009). Substrate inhibition kinetics of phenol
degradation by Pseudomonas fluorescence from steady state and wash-out
data. International Journal of Environmental Science and
Technology:(IJEST), 6, 443.
Ahmad, S., A., Ahamad, K., N., E., K., Johari, W., L.,W., Halmi, M., I., E., Shukor,
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