REMOVAL OF OIL AND GREASE FROM AGRO-FOOD INDUSTRIAL EFFLUENT USING Serratia ...

82 Pages·2017·1.72 MB·English

REMOVAL OF OIL AND GREASE FROM AGRO-FOOD INDUSTRIAL
EFFLUENT USING Serratia marcescens SA30 AND ITS KINETIC STUDY

SHAKILA BT ABDULLAH

UNIVERSITI TEKNOLOGI MALAYSIA
REMOVAL OF OIL AND GREASE FROM AGRO-FOOD INDUSTRIAL
EFFLUENT USING Serratia marcescens SA30 AND ITS KINETIC STUDY

SHAKILA BINTI ABDULLAH

A thesis submitted in fulfilment of the
requirements for the award of the degree of
Degree of Philosophy (Civil Engineering)

Faculty of Civil Engineering
Universiti Teknologi Malaysia

JUNE 2016
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This thesis is dedicated to:

Special dedication to my beloved father and mother, Abdullah bin Ibrahim
and Hasnah bt. Mohd Yatim, who have been with me throughout the hard time of my
study. Thanks for the du’a and prayer, guidance, continuous support and tremendous
of love throughout my course of study.

For my beloved auntie, Hajah Samsiah bt. Ibrahim, Norashikin bt.Mohd
Yatim and Sarina bt. Mohd Yatim, thanks for the du’a, advices and constant support
for every step of my PhD.

and lastly

Special thanks to my best friends Wan Haslinda bt. Wan Ahmad and Azila bt. Md
Sudin who always keep my spirit in many moments of crisis, her encouragement and
helping me to succeed and instilling in me the confidence to be a strong and capable
person.
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ACKNOWLEDGEMENTS

In the name of Allah S.W.T, the Most Gracious, the Most Merciful,
Alhamdulillah and thanks to Allah, He who has given me the strength and patience to
complete this research.

I would like to express my heartiest gratitude goes to my supervisors: Assoc.
Prof. Dr. Mohamad Ali Fulazzaky and Prof. Dr. Mohd Razman Salim for his
willingness to help, listen and assist in every way, in the midst of his heavy
responsibilities and duties and thank you for the advice, guidance, ideas, criticism
and encouragement throughout the research study.

A special note of thanks and appreciation to all staff in Centre for
Environmental Sustainability & Water Security (IPASA), Research Institute of
Sustainable Environment (RISE), Resources Sustainability Research Alliance,
Environmental Engineering, Faculty of Civil Engineering, Chemistry Department,
Faculty of Science, UTM, Chemistry Department, Faculty of Science, UM and
Institute Medical Research for allowing me to use their laboratory facilities.

Last but not least, I would like to acknowledge and thank to my research
colleagues: Hairul, Mimi, Irena, Budi, Ain, Syahrul, Shikin, Thana, Maria, Anis,
Hudai and Biotechnology lab members for their constructive comments and constant
support.
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ABSTRACT

Agro-food industrial effluent (AFIE) may contain high concentration of oil
and grease (O&G), which poses a major threat to aquatic environments, killing or
adversely affecting fish and other aquatic organisms. Even though biosorption
techniques are commonly used to remove inorganic and organic matters from
wastewater, the kinetics and mechanisms of O&G removal from AFIE by Serratia
marcescens SA30 immobilised in a packed-bed column reactor (PBCR) need to be
verified. The aims of this study were to perform characterisation of beneficial strain
of biosurfactant-producing bacteria in order to investigate their ability to remove
O&G from water, to develop kinetic models for predicting the efficiency of O&G
removal from AFIE and to apply modified mass transfer factor models for assessing
the mechanisms and mass transfer resistance for the biosorption of O&G from AFIE
by Serratia marcescens SA30. The performance of PBCR achieved 91% of
efficiency using Serratia marcescens SA30 as oil-degrading bacteria. The best
performance of nearly 100% efficiency can be achieved by experiments run at a
fixed volumetric flow rate of 0.18 L h-1, even during treatment using two different
concentrations of O&G at 26.9 and 33.5 g L-1 to feed the reactor. The results show
the applicability of linear and logarithmic equations with high validity. The
resistance to mass transfer could be dependent on intracellular accumulation at the
beginning and then on film mass transfer at the final stage of O&G biosorption by
Serratia marcescens SA30. The well verified experimental data of kinetic models
and mass transfer mechanisms give significant contributions to the development of
biosorption theory and an insight of using new approaches to improve environmental
quality. This study would provide a green and sustainable pathway for removing
O&G from water.
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ABSTRAK

Efluen perindustrian agro-makanan (AFIE) mungkin mengandungi kepekatan
minyak dan gris (O&G) yang tinggi dan boleh memberikan ancaman besar kepada
persekitaran akuatik, membunuh atau memberi kesan buruk kepada ikan dan
organisma akuatik yang lain. Walaupun teknik biopenjerapan sering digunakan untuk
menyingkirkan bahan tak organik dan organik daripada air sisa, kinetik dan
mekanisma penyingkiran O&G daripada AFIE oleh Serratia marcescens SA30 yang
dipegunkan dalam reaktor turus terpadat tunggal (PBCR) perlu disahkan. Tujuan
kajian ini adalah untuk melakukan pencirian ke atas bakteria pengeluar biosurfaktan
bagi menyiasat keupayaan bakteria tersebut untuk menyingkirkan O&G dari air,
untuk membina model kinetik bagi menganggarkan kecekapan penyingkiran O&G
daripada AFIE dan menggunakan model faktor pemindahan jisim yang telah
diubahsuai untuk menilai mekanisma dan rintangan pemindahan jisim bagi
biopenjerapan O&G daripada AFIE oleh Serratia marcescens SA30. Prestasi PBCR
boleh mencapai 91% kecekapan menggunakan Serratia marcescens SA30 sebagai
bakteria pendegradasi minyak. Prestasi terbaik dengan kecekapan hampir 100%
boleh dicapai apabila eksperimen dijalankan pada kadar alir isipadu tetap 0.18 L h-1,
walaupun pada rawatan menggunakan dua kepekatan O&G berbeza pada 26.9 dan
33.5 g L-1 untuk ditambah ke dalam reaktor. Keputusan menunjukkan kebolehgunaan
persamaan linear dan logaritma dengan kesahihan yang tinggi. Rintangan
pemindahan jisim boleh bergantung kepada pengumpulan intrasel pada permulaan
dan kemudian pada pemindahan jisim filem di peringkat akhir biopenjerapan O&G
oleh Serratia marcescens SA30. Data eksperimen model kinetik dan mekanisma
pemindahan jisim yang telah disahkan boleh memberikan sumbangan yang besar
kepada pembanggunan teori biopenjerapan dan memberi pandangan menggunakan
pendekatan baru untuk meningkatkan kualiti alam sekitar. Kajian ini akan
memberikan laluan hijau dan lestari untuk menyingkirkan O&G dari air.
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TABLE OF CONTENTS

CHAPTER TITLE PAGE

DECLARATION ii
DEDICATION iii
ACKNOWLEDGEMENT iv
ABSTRACT v
ABSTRAK vi
TABLES OF CONTENTS vii
LIST OF TABLES xiii
LIST OF FIGURES xv
LIST OF ABBREVIATIONS xx
LIST OF SYMBOLS xxi
LIST OF APPENDICES xxiv

1 INTRODUCTION 1

1.1 Background 1
1.2 Problems Statement 2
1.3 Objectives 4
1.4 Scope of the study 4
1.5 Significance of the study 6
1.6 Thesis outline 7
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2 LITERATURE REVIEW 8
2.1 Industrial wastewater 8
2.1.1 Food processing industrial wastewater 9
2.1.2 Discharged regulations for oil and grease in
several countries 11
2.2 Oil and grease 13
2.2.1 Type and classification of oil-water mixture 15
2.2.2 Oily wastewater 15
2.2.3 Impacts of oily wastewater on the
environment 17
2.3 Oil-degrading bacteria 18
2.3.1 Biosurfactant 18
2.4 Supporting materials 27
2.5 Oily wastewater treatment processes 28
2.5.1 Physical treatment processes 28
2.5.1.1 Filtration 28
2.5.1.2 Flotation 31
2.5.1.3 Adsorption 32
2.5.2 Physico-chemical treatment processes 34
2.5.2.1 Coagulation 34
2.5.2.2 Ozonation 35
2.5.3 Biological treatment processes 36
2.5.3.1 Anaerobic Digestion 36
2.5.3.2 Aerobic digestion 37
2.5.3.3 Biosorption 44
2.5.3.3 Advanced biological process 47
2.6 Kinetic and mass transfer models 56
2.6.1 Adsorption kinetic models 56
2.6.2 Growth kinetic models 58
2.6.3 Mass transfer factor models 59

3 MATERIALS AND METHODS 63
3.1 Materials 63
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3.1.1 Glassware and apparatus 63
3.1.2 Growth media 63
3.1.2.1 Nutrient broth medium 63
3.1.2.2 Nutrient agar medium 64
3.1.2.3 Tween peptone agar medium 64
3.1.2.4 Blood agar medium 64
3.1.2.5 Basal salts medium 64
3.1.2.6 Oil agar medium 65
3.1.2.7 Luria bertani (LB)-glycerol stock
solution 65
3.1.2.8 Buffer solution 65
3.1.2.9 Active culture in Erlenmeyer
flask 66
3.1.2.10 Bacterial growth on nutrient agar
plate 66
3.2 Agro-food industrial effluent 67
3.2.1 Sampling locations and procedure 67
3.2.2 Characterisation of agro-food industrial
effluent 68
3.2.2.1 Determination of oil and grease 68
3.3 Isolation of oil-degrading bacteria 69
3.3.1 Isolation of single colony 69
3.3.2 Physico-chemical test for characterising
oil-degrading bacteria 69
3.3.2.1 Zeta potential test 71
3.4 Identification of bacterial gene using 16s rRNA
gene sequence analysis 71
3.5 Batch experiments for optimisation of
environmental conditions 71
3.5.1 Bacterial acclimatisation with synthetic
oily-wastewater 71
3.5.2 Test of oil and grease removed from
synthetic-oily wastewater 72
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3.5.3 Bacterial adaptation with agro-food
industrial effluent 73
3.5.4 Test of oil and grease removed from agro-
food industrial effluent 73
3.6 Microscopic identification for Serratia marcescens
SA30 74
3.6.1 Scanning electron microscopy 74
3.6.2 Transmission electron microscopy 74
3.7 Packed-bed column reactor 75
3.7.1 Supporting materials 75
3.7.2 Characterisation of supporting materials 75
3.7.3 Packed-bed column reactor and
experimental procedure 75
3.7.4 Experimental runs by packed-bed column
reactor 77

4 DEGRADATION OF OIL AND GREASE FROM
AGRO-FOOD INDUSTRIAL EFFLUENT 78
4.1 Overview 78
4.2 Characteristics of the agro-food industrial effluent 79
4.2.1 Composition of fatty acids in the agro-food
industrial effluent 81
4.3 Morphological features of biosurfactant-producing
bacteria 84
4.4 Results of screening the beneficial oil-degrading
bacteria 86
4.4.1 Growth and pH profiles of the isolated
bacteria 86
4.4.2 Bacterial growth in minimal medium 88
4.4.3 Opaque halo formation 89
4.4.4 Microbial adherence to hydrocarbon 90
4.5 Characteristics of biosurfactant-producing bacteria 93
4.5.1 Biosurfactant production and formation of