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BIODEGRADATION OF VARIOUS OIL PALM FIBRE WASTE VIA VERMICOMPOSTING

NAHRUL HAYAWIN BT ZAINAL

UNIVERSITI SAINS MALAYSIA

2011

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2

BIODEGRADATION OF VARIOUS OIL PALM FIBRE WASTE VIA VERMICOMPOSTING

by

NAHRUL HAYAWIN BT ZAINAL

Thesis submitte d in fulfillme nt of the re quire ments for the de gree o f

Maste r of Scie nce

FEBRUARY 2011

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3 iii ACKNOWLEDGEMENTS

Assalamualaikum and Alhamdulillah.

First and foremost to my field-supervisor, Dr Astimar Abdul Aziz from Malaysian Palm Oil Board (MPOB) and to MPOB itself for providing financial assistance to conduct this work in the form of a Graduate System Academic Scheme (GSAS) that made the research more smooth running. Praise and thanks only to Allah for giving me opportunity and strength to complete my master project within the time given.

My supervisors, Prof. Dr. Abdul Khalil Shawkataly and Assoc. Prof. Dr. Mahamad Hakimi Ibrahim who have given me a lot of ideas, guidance and criticism that has strengthened my thesis. Besides that, with this help, I could do my research smoothly.

My deep appreciation to my beloved parents Zainal b Katan and Mahanom bt Mohamad for giving me valuable support and encouragement during the research. Besides that, my brother and sister for their support and encouragement that has made me more confident and happy while doing the research.

To other staff from MPOB especially En Abdul Hafiz Shukor, En Wan Hasamuddin Wan Hassan, Nik Fadzillah Nik Mat, Sri Rahayu, Pn Hajah Masnon and Amir Hamzah for their help while doing the research. Without them, I would never be able to complete this research for my master project.

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4 TAB LE OF CONTENTS

ACKNOWLEDGEMENT iii

TABLE OF CONTENTS v

LIST OF FIGURES viii

LIST OF TABLES x

LIST OF ABBREVIATIONS AND SYMBO LS xii

ABSTRAK ABSTRACT

xiv xvi CHAPTER 1 : INTRODUCTION

1.1General 1.2Objectives

1 4 CHAPTER 2 : LITERATURE REVIEW

2.1 Biomass Introduction And Uses In The Oil Palm Industry 2.2 Distribution Of Oil P alm Planted Area

2.3 Availability Of Oil P alm Biomass 2.4 Cell Wall Components

2.5.1 Celluloses 2.5.2 Hemicelluloses 2.5.3 Lignin

2.5.4 Extractives

2.5 The R ole of Earthworm In Organic Wastes 2.6 Earthworm As Biodegraders Of Wastes Biomass 2.7 Breakdown Of Sewage Wastes By Earthworm

2.8 Breakdown Of Animal, Vegetable And Urban Industrial Organic Wastes B y Earthworms

2.9 Characteristics Of Different Earthworm Species 2.10.1 Eisen ia Fetida

2.10.2 Eu drilu s Eugenia e 2.10.3 Perio nyx Exca va tes 2.10.4 Dend robena Ven eta 2.10.5 Po lyph eretima Elonga ta 2.10.6 Lu mbricu s Rub ellus

2.10 Vermicomposts As P lant Growth Media And S oil A dditives 2.11 Growth Of Plants In Vermicomposts

2.12 Element Cycling

5 6 8 11 13 14 16 16 17 20 20 21 22 23 23 25 25 26 26 28 29 30

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5 CHAPTER 3 : THE DEGRADATION O F LIGNOCELLULOSIC

COMPO NENTS DURING THE VERMICOMPOSTING OF O IL PALM B IOMASS FIBRES

3.1 Introduction

3.2 Materials and Methods 3.2.1 Preparation details

3.2.2 Determination of moisture content 3.2.3 Determination of pH

3.2.4 Determination of extractive content 3.2.5 Determination of holocellulose content 3.2.6 Determination of α-cellulose content 3.2.7 Determination of lignin content 3.2.8 Determination of ash content 3.2.9 Determination of nutrient content 3.2.10 E lemental analysis and microscopy 3.2.11 C ompost and vermicompost preparation 3.3 Results And Discussion

3.3.1 Loss of lignocellulosic components during vermicomposting 3.3.2 pH , nutrient quality and heavy metals concentration

3.3.3 Scanning Electron Microscopy 3.4 Conclusion

33 35 37 37 37 38 40 41 43 43 45 45 46 52 62 64 CHAPTER 4 : THE GRO WTH AND REPRODUCTION OF

EISENIA FETIDA AND EUDRILUS EUGENIAE IN EMPTY FRUIT BUNCH, MIXED CULTURES WITH SOLID PALM OIL MILL EFFLUENT

4.1 Introduction

4.2 Materials and Methods 4.2.1 EFB and solid POME 4.2.2 Vermicomposter preparation 4.2.3 Statistical Analysis

4.3 Results And Discussion

4.3.1 Growth and cocoon production of E. fetida and E.eu genia e in different vermicomposters

4.4 Conclusion

65 67 67 70 70 74

CHAPTER 5: VERMICOMPO STING OF EMPTY FRUIT BUNCH WITH ADDITION OF SOLID PALM OIL MILL EFFLUENT

5.1 Introduction

5.2 Materials and Methods 5.2.1 E.fetida

5.2.2 Empty fruit bunch (EFB) and Palm Oil Mill Effluent (P OME) 5.2.3 Experimental Set-Up

5.2.4 Physico-chemical analysis of vermicompost 5.2.5 Statistical analysis

75 78 78 79 80 81

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6 5.3 Results And Discussion

5.3.1 C/N ratio of the vermifertilizer 5.3.2 Fertilizer quality of the vermicompost

5.3.3 Heavy metal concentrations in final vermicomposts 5.4 Conclusion

81 82 86 89

CHAPTER 6 : THE ANALYSIS OF LEACHATE FROM THE VERMICOMPOSTING O F EFB FIBRES AMENDED WITH POME SLUDGE

6.1 Introduction

6.2 Materials and Methods 6.2.1 Earthworm (E.fetida)

6.2.2 Empty fruit bunch (EFB) and Palm Oil Mill Effluent (P OME) 6.2.3 Experimental Set-Up

6.2.4 Physico-chemical analysis of vermicompost 6.2.5 Statistical analysis

6.3 Results And Discussion

6.3.1 Total nitrogen (TN), total phosporus (TP) and total potassium (TK) in vermicompost leachate.

6.3.2 Heavy metals (Zn, Cu, M n, Mg, Ca, and Fe) in vermicompost leachate.

6.4 Conclusion

91 92 93 93 95 96 96 99 103 CHAPTER 7 : CONCLUSIONS AND RECOMMENDATIONS

7.1 Conclusions 7.2 Recommendations

105 106

REFERENCES 108

APPENDICES 122

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7 LIST OF FIGURES

Figure 2.1 (a, b and c)

Sources of oil palm biomass 9

Figure 2.2 ß-D-gluco-pyranose 12

Figure 2.3 Cellobiose residue 13

Figure 2.4 The nitrogen cycle 33

Figure 3.1 (a, b and c)

Sources of oil palm biomass after refined 37 Figure 3.2 The trends of EFB cellulose degradation (with and without

earthworms)

51

Figure 3.3 The trends of OP F cellulose degradation (with and without earthworms)

51

Figure 3.4 The trends of OP T cellulose degradation (with and without earthworms)

52

Figure 3.5 The trends of EFB lignin degradation during the vermicomposting (w ith and without earthworms)

52

Figure 3.6 The trends of OPF lignin degradation during the vermicomposting (w ith and without earthworms)

53

Figure 3.7 The trends of OPT lignin degradation during the vermicomposting (w ith and without earthworms)

53

Figure 3.8 The patterns of TOC content during vermicomposting of the oil palm biomass fibres

58

Figure 3.9 Patterns of TKN content during the vermicomposting of the oil palm biomass fibres

59

Figure 3.10 Relationship between EFB WORM, OP F WORM and OPT WORM in C/N ratio during vermicompost process

60

Figure 3.11 Morphological structures of oil palm biomass. 65

Figure 4.1 Experimental set up 71

Figure 4.2 Earthworms and cocoons were separated, counted and weighted

71

Figure 4.3 Mean worm biomass of E.fetida and E.eug eniae (g) in different feed mixtures after 12 weeks.

73

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8 Figure 4.4 Total number of cocoons produced in different feed

mixtures after 12 weeks

75

Figure 5.1 Fiber sample of EFB before and after shredded and refined into loose fibrous material

61

Figure 6.1 Solid palm oil mill effluent (P OME) 95 Figure 6.2 The leachate collected from each vermicomposter

(vermileachate)

96

Figure 6.3 Nutrient composition of vermileachate at days 84 101 Figure 6.4 Heavy metals content in different vermicomposter of

vermicompost leachate at days 84

104

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9 LIST OF TABLES

Table 2.1 Oil palm planted area in Malaysia 8

Table 2.2 Area under oil palm (mature and immature) by states: 2008 (Hectares)

8

Table 2.3 The amounts of oil palm biomass (wet weight) 11 Table 2.4 Availability of oil palm biomass by states (tones/year; dry

weight)

11

Table 2.5 Productivity of earthworms in animal and vegetable waste- length of life cycle

25

Table 2.6 Major plant nutrient elements in earthworm-processed animal wastes

29

Table 3.1 Chemical composition of the oil palm biomass (%, dry weight basis)

48

Table 3.2 Chemical analysis of the compost and vermicompost prepared from different oil palm wastes at different days

57

Table 3.3 Heavy metal content (mg/kg) in the initial feed substrates and vermicompost obtained in different vermicomposters (mean ± SE, n= 3) (dry basis).

63

Table 5.1 Composition of empty fruit bunch (EFB) 78 Table 5.2 Characteristics of Palm O il Mill Effluet (POME) 78

Table 5.3 Composition of treatments 82

Table 5.4 Variation in C/N ratio during vermicomposting of the vermifertilizer (mean ± SE, n= 3) (dry basis).

84

Table 5.5 Physico-chemical characteristics of initial feeds and vermicomposts obtained from different ratio of EFB + POME feed mixtures.

88

Table 5.6 Heavy metal content (mg/kg) in initial feed substrates and vermicomposts obtained from EFB + POME

vermicomposters.

91

Table 6.1 Composition of empty fruit bunch (EFB) 97 Table 6.2 Nutrient content of the oil palm biomass waste, 100

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10 vermileachate at the initial feed

Table 6.3 Heavy metal content of the oil palm biomass waste, vermileachate at the initial feed

103

Table 6.4 Initial values of metal concentrations of vermileachate in different vermicomposter

105

Table 6.5 Final values (84 days) of metal concentrations of vermileachate in different vermicomposter

106

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11 LIST OF ABBREVIATIONS AND SYMBO LS

EFB Empty Fruit B unch

OPT Oil Palm Trunk

OPF Oil Palm Frond

POME P alm Oil Mill Effluent TKN Total Kjeldahl N itrogen

TOC Total Organic Carbon

TP Total Phosporus

TK TCa

Total Potassium Total Calcium

Mg Magnesium

Mn Manganese

Zn Zinc

Ca Calcium

Fe Ferum

Cu Cuprum

N Nitrogen

P P hosporus

C Carbon

K P otassium

S Sulfur

H Hydrogen

O2 Oksigen

C/N ratio Carbon over Nitrogen ratio

BOD Biochemical Oxygen Demand

COD Chemical O xygen Demand

CO2 Carbon Dioxide

NH+4 Ammonium

NO-3 Nitrate

NaOH Sodium hydroxide

H2SO4 Sulfuric acid HClO4 Hydrochloric acid HNO3 Nitric acid

NO-2 Nitrite

Norg Organic nitrogen

MPOB Malaysian Palm Oil Board

AAS Atomic Absorption S pectrophotometer E.fetida Eisenia fetida

E.eugeniae Eud rilus eugeniae E.andrei Eisenia Andrei L.Rubellus Lumbricuc Rubellu s

SEM Scanning E lectron Microscopy

SE Standard error

i.e which is

e.g º

Example Degree

ºC Degree celcius

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12

α Alpha

ß Beta

% percentage

< Less than

> More than

± P lus minus

≈ Almost equal to

* Asterisk

² Superscript tw o

³ Superscript three

g Gram

cm Centimeter

mL Milliliter

µm Micrometer

mg/kg Milligram per kilogram

meq Milliequilibrium

nm Nanometer

mm Millimeter

ppm P art per million

kV Kilovolt

Pa P ascal

e Eksponen

R2 Regression line

wt Weight

v/v Volume over volume

w/v Weight over volume

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13 BIODEGRADASI PELBAGAI J ENIS SERAT K ELAPA SAWIT DENGAN

MENGGUNAKAN CACING SEBAGAI AGEN PEREPUTAN ABSTRAK

Satu kajian tentang biodegradasi pelbagai jenis sisa kelapa sawit, iaitu tandan kosong kelapa sawit, pelepah kelapa sawit dan batang kelapa sawit telah dijalankan dengan menggunakan cacing Eudrilus Eugeniae (cacing Africa) dan E isenia Fetida (cacing merah). Berat cacing tanah dan jumlah kokon yang dihasilkan ditimbang dan dikaji setiap minggu. Serat sisa kelapa saw it yang direputkan dengan menggunakan cacing telah dilakukan selama 84 hari dengan suhu kawalan 25 ± 1.0ºC. 5 gram cacing tanah spesies E udrilus Eugeniae dan Eisenia Fetida telah ditambah ke dalam setiap bekas yang mengandungi 150 gram serat sisa kelapa saw it. Dalam kajian tindak balas cacing tanah terhadap komponen selulosa dan lign in menunjukkan peratusan pereputan selulosa sangat tinggi di dalam tandan kosong kelapa sawit berbanding pelepah kelapa saw it dan batang kelapa sawit. Manakala, biodegradasi serat kelapa sawit tanpa menggunakan cacing tidak menunjukkan pereputan selulosa dan lignin yang tinggi. Di sebalik itu, hanya sedikit sahaja berlakunya pereputan terhadap lignin dengan menggunakan cacing di dalam ketiga-tiga serat kelapa sawit. P ereputan menggunakan cacing menunjukkan kadar nitrogen dan fosforus yang tinggi di dalam tandan kosong kelapa sawit malah menurunkan kadar kepekatan logam berat.

Mikroskop electron juga menunjukkan berlakunya pereputan pada struktur morfologi sebelum dan selepas biodegradasi dengan menggunakan cacing terhadap tandan kosong kelapa sawit dan pelepah kelapa saw it. Batang kelapa saw it tidak menunjukkan pereputan yang jelas. Data menunjukkan tandan kosong kelapa sawit paling sesuai dijadikan sebagai bahan makanan (subtrat) bagi cacing tanah berbanding pelepah kelapa saw it dan batang kelapa sawit. Kajian yang seterusnya

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14 telah dilakukan dengan menggunakan tandan kosong kelapa sawit dicampur dengan sisa pepejal kelapa sawit dalam pelbagai nisbah dengan membuat perbandingan antara dua spesies cacing tanah, Eudrilus Eugeniae dan Eisenia Fetida. Pertumbuhan dan pembiakan kedua-dua spesies cacing tanah telah dikaji setiap minggu apabila diletakkan di dalam campuran tandan kosong kelapa sawit dan sisa pepejal kelapa sawit. Sifat fizik dan kimia telah dijalankan selama 12 minggu terhadap najis cacing dan sisa air yang telah dihasilkan oleh kedua-dua spesies cacing tersebut. N ilai tertinggi nitrogen telah direkodkan di dalam campuran 50% tandan kosong kelapa sawit + 50% sisa pepejal kelapa saw it. P enurunan C/N dan peningkatan jumlah fosforus, jumpah kalium, jumlah kalsium dan jumlah magnesium juga telah direkodkan. Jumlah kandungan logam berat kelihatan tinggi tetapi masih lagi mengikut ketetapan yang dibenarkan. Secara keseluruhan menunjukkan campuran 50% tandan kosong kelapa sawit + 50% sisa pepejal kelapa sawit sangat sesuai dalam pereputan sisa kelapa sawit dengan menggunakan cacing tanah. Hasil daripada pereputan sisa-sisa kelapa sawit ini akan menghasilkan baja organik yang berkualiti tinggi di samping mengurangkan kos pemprosesan sisa.

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15 BIODEGRADATION OF VARIOUS O IL PALM FIBRE WASTE VIA

VERMICOMPOSTING.

AB STRACT

A study of the biodegradation of various palm oil mill wastes, viz. empty fruit bunch (EFB), oil palm frond (OPF), oil palm trunk (OPT) and palm oil mill effluent (P OME) using earthw orm species Eu drilu s Eugenia e (E.eugeniae) and Eisen ia fetida (E.fetid a) was conducted under laboratory conditions. Earthw orms were weighed weekly and the numbers of cocoons produced per week were assessed. The oil palm biomass was vermicomposted for 84 days under controlled conditions (25 ± 1ºC). 5 g of clitellated adult worms E.eug en ia e and E.fetid a were added to vermicomposter. In the study of the effects of the earthworms’ digestability towards the major lignocellulotic component (cellulose and lignin) of the EFB in the vermireactors (EFB WORM), results showed significant degradation rate of the cellulose. A similar trend was also observed for the lignin degradation. Meanwhile, lower percentages of the lignocellulosic degradations were observed for the control reactor (w ithout earthworms). The vermicomposts were rich in nitrogen and phosphorus and had low levels of heavy metals. Scanning E lectron Microscopy revealed the morphological structure of the oil palm fibre wastes before and after vermicomposting. The data revealed that vermicomposting (using E.eug en ia e) is a suitable method for the decomposition of oil palm wastes into value added material, especially for EFB rather than OPT and OPF. The next stage was to determine the growth and reproduction of E.fetida and E.eugeniae to evaluate their suitability for vermicomposting of EFB when amended w ith POME. A total of six vermicomposters filled with different ratios of EFB and POME were maintained for this study under laboratory conditions. Physico-chemical properties of both solid

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16 (vermicast) and liquid (vermileachate) were monitored for 12 weeks. E.fetid a appeared as more efficient than E.eu geniae in terms of palm oil mill mineralization rate. The growth and cocoon production were also better in E.fetid a for EFB wastes.

Highest value of total kjeldahl nitrogen (TKN) was recorded in 50% EFB + 50%

POME (V6) feed mixture containing vermicomposter. A significant decrease in C/N ratio and increase in total phosporus (TP), total potassium (TK), calcium and magnesium content were recorded. The heavy metal content was higher in vermicompost obtained in all the reactors than in the initial feed substrates. Overall, V6 vermibed appeared as an ideal substrate to manage oil palm industrial waste effectively. Vermicomposting can be proposed as a low-input basis technology to convert industrial waste into value-added biofertilizer.

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17 CHAPTER 1

INTRODUCTION

1.1 Ge ne ral

Oil palm is the main agricultural crop in Malaysia with the total planted area recorded in 2009 at 4.69 million hectares, and based on the distribution rate of biomass per hectare that has been formulated by Gurmit, (1994) and widely accepted; the total oil palm biomass availability (empty fruit bunch, EFB; oil palm trunk, OP T and oil palm fronds, OPF) was estimated about 65.46 million tonnes (dry weight). Hereby, agro-wastes from the oil palm industry (EFB, OPT and OPF) have attracted attention as potential sources for new value-added materials. Oil palm biomass (EFB, OP T and OP F) is an agricultural by-product periodically left in the field during the replanting, pruning and milling processes. O il palm biomass are largely unutilized after a 25-year economic life of the tree and causes severe environmental pollution problems (Abdul Khalil et al., 2001; Sreekala et a l., 2002).

Oil palm wastes is a suitable raw material for recycling because it is produced in large quantities in mill areas especially EFB.

A palm oil mill generates large volumes of palm oil mill effluent (POME), estimated around 0.7 tonne per tonne fresh fruit bunch (FFB) processed. In general, the palm oil milling process can be categorized into a dry and a wet (standard) process. The wet process of palm oil milling is the most common and typical way of extracting palm oil, especially in Malaysia. For each tonne of crude palm oil produced, it is estimated that 5.0– 7.5 tonnes of water are required, and more than 50% of this water

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18 ends up as palm oil mill effluent (POME) (Ahmad et al., 2003). Some environmental problems are considered too caused by high biochemical oxygen demand (BOD) (25,000 mg/L), chemical oxygen demand (COD) (53,630mg/L), oil and grease (8370 mg/L), total solids (43,635 mg/L) and suspended solids (19,020mg/L) if the untreated effluent is discharged into watercourses (Ma, 2009). Besides, POME also contains appreciable amounts of nitrogen (N), phosporus (P ), potassium (K), magnesium (Mg) and calcium (Ca) which are essential nutrient elements for plant growth.

Lignocellulosic materials are the most abundant polymer in nature and constitutes to a large carbon pool for microorganisms, the main agents responsible for soil organic matter decomposition. Cellulolysis occurs as the result of the combined action of fungi and bacteria towards the lignocellulosic materials during composting. There are several factors that limit the degradation of cellulose in soils, which make the process slow. These factors also influence the activities of the cellulases, such as concentration, location, and the mobility of the enzymes (Sinsabaugh & Linkins, 1988). Moreover, the production of cellulases is regulated by the speed of accumulation of enzyme products during the process. According to Lynd et al., (2002), hemicellulose and lignin content, as well as the degree of the cellulose crystalline can also determine the rate at which the cellulose is metabolized.

Some problems and challenges are faced by oil palm industry in exploiting the waste products, but in the recent development, this oil palm biomass have been exploited for the wood based products (Nor Y uziah & Paridah, 2008) ; bioenergy (Sansubari,

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19 2008); and for bio-fertilizer (Khalid & Tarmizi, 2008). At controlled rates, the land applications have been shown to give positive responses to oil palm yields and negligible pollution to the environment (Tam et a l., 1982; Lim et al., 1991; Mohd Hashim, 1991; L im & Chan, 1993). T he traditional disposal methods such as open dumping and land filling practices of these materials are not only increasingly expensive, but impractical as open space becomes limited. Therefore, there is a need of such ecologically sound technologies which are not only cost effective, but also sustainable in terms of possible recovery of recyclable constituents from effluents as they are rich in nutrients and have higher organic matter content.

In recent years, the industrial waste management is using vermicomposting (Elvira et al., 1997). Vermicomposting is stabilization of organic material involving the joint action of earthworms and microorganisms. According to Vinceslas-Akpa & Loquest, (1997) and S uthar (2007a), earthworm accelerates the transformation of organic waste material into more stabilized forms by aeration and bioturvation, either by their excreta and qualitative or quantitative influence upon the telluric microflora.

Vermicomposting with earthworms often produces a product with a low er processing time, lower mass, phytotoxicity is less likely, an additional product (earthworms) produced, greater fertilizer value, and more N released (Lorimor et al., 2001).

The transformation of wastewater sludge into vermicompost is of double interest: on the one hand a waste is converted into value added product, and on the other, it minimizes a pollutant that is a result of increasing industrialization. E lvira et a l.

(1998) have reported that vermicomposting of paper mill sludge was using E isenia

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20 And rei (E.and rei) under laboratory as well as field conditions. Nogales et a l. (2005) then reported that the vermicomposting of winery waste was using E.andrei under laboratory conditions. Studies have revealed that vermicomposting could be an appropriate technology to transfer organic wastes to value-added products, which is vermicompost (Kale, 1998).

1.2 Obje ctive s o f the study:

1) T o determine the degradability of lignocellulosic components of oil palm biomass fibres during vermicomposting.

2) T o determine the suitability of EFB amended with P OME sludge cake for vermiculture of Eudrilus Eug en iae and Eisenia Fetida .

3) To evaluate nutrients quality of various mixtures of EFB and POME sludge cake.

4) To analyse leachate from vermicomposting of EFB amended with POME sludge cake.

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21 CHAPTER 2

LITERATURE REVIEW

2.1 Bio mass pro duction and its use s in the o il palm industry

Oil palm is one of the most versatile crops where nearly every part of the palm, from oil to the entire biomass can be utilized. There is a need to look beyond crude palm oil (CP O) as the sole source of revenue, especially utilization of biomass, in order to optimize total returns. Through intensive research and development efforts, large quantities of oil palm biomass from the Malaysian oil palm industry have now found applications in several commercially viable bio-based products in country. The most important is the utilization of the ligno-cellulosic materials from the biomass for a number of value-added products through physical, biolog ical, and chemical innovations (C han, 2009).

Since the establishment of the oil palm, cocoa and rubber plantations, the biomass from these crops was used as a mulching material when practicing ‘green’

agriculture (Chan, 1999). This would mean that their management practices would include planting of tree crops, ground legume covers, using of trunk and frond biomass as chips and mulch during replanting, planting on conservation terraces and using of silt pits to conserve soil, nutrient and moisture. Such green practices have since been expanded to include the utilization and recycling of mill biomass residues on land as well as their use for manufacturing of non-wood products and for steam power generation (Yusof & Chan, 2003). Through the practice of zero burning of plant residues of the former stand at replanting, the plantation industry has

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22 consciously been using the biomass in a balanced manner in the field to maintain organic matter and soil fertility and also to provide fibre for substitution of timber trees.

Oil palm biomass of empty fruit bunches, fronds, trunk, fibre, shell and effluent are obtained under two situations in the field and in the mill. Firstly, during productive life of the palm covering about 25 years, the two common co-products are fresh fruit bunches (FFB) and pruned fronds from oil palm growing in the plantations and also the additional trunk biomass during replanting. Secondly, during processing of FFB in the mill, the four common co-products biomass materials are the empty fruit bunches fibre, shell and effluent from the mills, despite of the palm oil, kernel oil and meal (Chan, 2009).

2.2 Distribution of O il Palm Plante d Are a

Malaysia is the top producer of palm oil in the world and at present, the total area under oil palm cultivation is about 4.48 million hectares, with the total palm oil production at 26.47 million tonnes (MPOB, 2008). Table 2.1 shows the distribution of oil palm planted area in Malaysia in year 2008. The total area under oil palm cultivation in Peninsular Malaysia was 2.4 million hectares or 53% of the total area, while Sabah and Sarawak was about 2.08 million hectares or 46.4% of the total area.

However, Sabah shows the largest planted area at 29.7%, followed by Johor and Pahang at 15.3% and 14.4% respectively. Distribution of the planted area for the mature and immature palms by state is shown in Table 2.2. From the table, it shows

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23 that in Peninsular Malaysia, about 2.1 million hectares of plantation are under mature palm (fruit production) which means that it is ready for replanting while about 260 575 hectares are still immature (less than 25 years old) (MP OB, 2008).

Planted area from the private estates is about 59% while for the Government Schemes are about 21%. In the Government Scheme, FELDA contributes the highest planted areas as compared to FELCRA and RISDA. P rivate estates contributed about 60% of the total planted area in Malaysia while from the Government Scheme;

FELDA contributed the highest planted area which is about 16%. Planted area by FELCRA and RISDA is only about 4% and it may be increased in year 2009 (MP OB, 2008).

Table 2.1: Oil palm planted area in Malaysia (2008) (MPOB, 2008)

State Oil palm planted area (hectares)

Johor 687, 906

Kedah 77, 080

Perlis 251

Kelantan 103, 636

Melaka 48, 408

Negeri Sembilan 171, 647

Pahang 647, 879

Pulau P inang 13, 001

Perak 363, 022

Selangor 135, 529

Terengganu 161, 660

Peninsular Malaysia 2, 410, 019

Sabah 1, 333, 566

Sarawak 744, 372

Sabah & Sarawak 2, 077, 938

Total, Malaysia 4, 487, 957

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24 Table 2.2: Area under oil palm (mature and immature) by states: 2008 (Hectares)

(MP OB, 2008)

State Mature Immature Total

Johor 615, 834 72, 072 687, 906

Kedah 73, 057 4023 77, 080

Kelantan 82, 771 20, 865 103, 636

Melaka 45, 047 3, 361 48, 408

N.Sembilan 153, 940 17, 707 171, 647

Pahang 568, 937 78, 942 647, 879

P.Pinang 12, 711 290 13, 001

Perak 330, 938 32, 084 363, 022

Perlis 232 19 251

Selangor 126, 235 9, 294 135, 529

Terengganu 139, 742 21, 918 161, 660

P.Malaysia 2, 149, 444 260, 575 2, 410, 019

Sabah 1, 197, 284 136, 282 1, 333, 566

Sarawak 569, 196 175, 176 744, 372

Sabah/Sarawak 1, 766, 480 311, 458 2, 077, 938

MALAYSIA 3 , 915 , 924 572, 033 4 , 487 , 957

2.3 Av ailab ility of Oil Palm Biomass

Besides the oil, the oil palm industry also generates massive quantities of oil palm biomass such as empty fruit bunches (EFB), fronds (OP F), and trunks (OPT) as shown in F igure 2.1 (a, b and c). The biomass is available from the replanting and through routine field and mill operations. Generally, OPF are pruned from each matured oil palm tree per year during the harvesting of fresh fruit bunches (FFB) in the field (Anis et a l., 2008). F or processing, the FFB are then collected and transported to the palm oil mills. Next, at the palm oil mill, the sterilized fresh fruit bunches go through a threshing process to separate the fruit nuts from the empty fruit bunches.

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25 a) Empty fruit bunch (EFB)

b) Oil palm frond (OPF) c) Oil palm trunk (OPT) Figure 2.1(a, b and c): Sources of oil palm biomass

Studies showed that the empty fruit bunch consists of around 75-80% of spikelets and 20-25% of stalk (Kamarudin et a l., 1996). The oil palm fronds are collected during pruning and replanting activities. The collected fronds during the pruning activity currently give an average of 43.12 million tonnes (wet basis) in Peninsular Malaysia and 32.7 million tonnes (wet basis) in Sabah and Sarawak. It is estimated that more than 70 thousands hectares of oil palm will be due for replanting every year. This w ould involve the felling of approximately 9 million palms.

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26 About 53.9% (dry weight) of fibre bundle can be extracted from a trunk; with the remaining parts are followed by parenchyma tissues (31.7%) and the bark which contribute 14.5% (dry weight) of the trunk respectively. Hence the amount of oil palm biomass (wet weight) available annually is estimated to be from OP F, 75.90 million tonnes from the field, EFB about 16.05 million tonnes from the mills, and OPT about 13.92 million tonnes or 21.63 million cubic meters from replanting (Table 2.3) (Anis et a l., 2008). Thus the quantities at hand could make a very substantial contribution to the production of palm-based composites for a wide range of applications without depleting the nations’ fibre resources from the natural forest and forest plantations. T he amounts of oil palm biomass by states are shown in Table 2.4 where Sabah shows the highest amount.

Table 2.3: T he amounts of oil palm biomass (wet weight) (Anis et al., 2008) Unit

Sources of oil palm biomass

M illion tonnes Million cubic

per year (wet weight) meter per year Peninsular Malaysia

1. Oil palm trunk (OP T) 7.91 12.29 2. Oil palm frond (OP F) 43.12

3. Empty fruit bunch (EFB) 9.69 Sabah and Sarawak

1. Oil palm trunk (OP T) 6.01 9.34 2. Oil palm frond (OP F) 32.78

3. Empty fruit bunch (EFB) 6.36

TOTAL 105 .87 21 .63

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27 Table 2.4: Availability of oil palm biomass by states (tonnes/year; dry weight) (Anis

et al., 2008) State Empty fruit

Bunch

P runes oil palm fronds

Felled oil palm

trunks Total

Pahang 626 141 1 452 059 526 191 2 604 391

Terengganu 160 865 367 948 100 236 629 049

Kelantan 83 257 190 441 54 413 328 111

Johor 772 740 1 536 120 588 887 2 897 747

Selangor 237 647 442 027 98 585 778 259

N.Sembilan 181 040 336 780 100 919 618 739

Melaka 45 050 83 802 5841 134 693

Kedah 49 451 103 355 14 405 167 211

P.Pinang 19 153 39 980 8081 67 214

Perak 421 415 765 462 362 735 1 549 612

Sabah 943 401 2 017 747 548 386 3 509 534

Sarawak 154 845 350 873 129 580 635 298

TOTAL 3 695 005 7 686 594 2 538 259 13 919 858

2.4 Ce ll wall components

Plants are made up of individual cell which contain a cell wall, the morphology of the plant such as a structural support, and control the passage of water and nutrients (Schuerch, 1963). Plant cell wall contains of skeletal polysaccharides, hemicellu loses, polyuronides, lignin and proteins. The chief components of woody plant cell walls are cellulose, hemicelluloses, and lignin.

2.5.1 Cellulose

Cellulose is the most abundant organic material on earth, containing approximately 50% of all biomass for an annual production of about 100 billion tonnes. Preston, (1974) stated that it is a long chain polymer of ß -D-glucose in the pyranose form (F igure 2.2), linked together by 1,4 – glycosidic bonds to form cellobiose residues (F igure 2.3) which are the repeating units in the cellulose chain. The ß -linkage

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28 requires that the alternate glucose units must be rotated through 180º. An important implication of this structure is a marked tendency for the individual cellulose chains to come together to form bundles or by hydrogen bonds which held the crystalline order together. A major factor in the high stability of cellulose is the conformation of the pyranose rings, where the total energy lies close to the minimum.

Figure 2.2: ß-D-gluco-pyranose (Immergut, 1963)

Figure 2.3: Cellobiose residue (Immergut, 1963)

O

O

O

H H

H

H H O H

O H CH2O H

O

CH2OH H

H H

H OH

OH

H O O OH

H OH

H H

OH OH

H

CH2OH H

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29 Biodegradation of cellulose wastes by bacterial or fungal enzymatic activities represents a large area of research experiments concerning the influence of different biochemical and physical factors, where it interferes in cellular dynamics of such biotechnological processes (Verstraete, 1992). The cellulose biodegradation using bacterial and fungal cells immobilized in polymerized hydrogels is essentially based on the complex interactions between biotic factors, in first place, including: cell age, morphogenetic specificity of microorganisms, and cell wall composition. In addition, the cellulose composition, especially its complexity with hemicellu lose and lignin, as well as the abiotic ones, such as: physico-chemical surface properties of polymerized hydrogels which w ill be used as immobilization matrices, their pH value, porosity, and ionic strength of nutritive solutions added to substrata (Lowenfels & Lewis, 2006).

2.5.2 He mice lluloses

In nature, the major part of biomass is lignocellulose and, consequently, its degradation is essential for the operation of the global carbon cycle. Lignocellulose, such as wood, is mainly composed of a mixture of cellulose, hemicellulose, and lignin (Sjostrom, 1993). Closely associated with the skeletal polysaccharide cellulose in the cell wall are hemicellu loses, the other structural polysaccharides. The different between hemicelluloses w ith other cellulose is although hemicelluloses are water insoluble, but they can be dissolved in strong alkali. This characteristic of hemicellu lose may be used to separate the hemicelluloses from the total carbohydrate fraction called holocelluloses, leaving essentially pure or α–cellulose behind.

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30 Another property of hemicelluloses is they are more readily hydrolyzed by acid compared to cellulose (Schuerch, 1963).

Basically, there are only a few hemicellulose structures found in all plants, which shows small modifications within the same plant in different tissues, and so with the modification in different plants. In softwoods w hich contain about 25%

hemicellu loses, mannose, galactose, xylose, glucose and arabinose are the principal constituent sugars in decreasing abundance (Preston, 1974). Meanwhile, in hardwoods which contain about 30% hemicelluloses, xylose, galactose, and mannose are the principal constituents’ sugars in decreasing abundance, with minor amounts of rhamnose and arabinose. B oth types contain 4-O-methylglucuronic acid.

Generally, pentosans are rich in annual plants and hardwoods while in softwoods, the hexosans predominate.

2.5.3 Lig nin

The remaining 25% of the cell wall material in w ood plants is lignin, the third major of cell wall component. In fact, lignin which is woody is the necessary component for plant. L ignin serves as cement between the wood fibre, as a stiffening agent within the fibre, and as barrier to enzymatic degradation of the cell wall. According to Sarkanen & Ludwig (1971), most bacteria reach their limit when it comes to the noncarbohydrate lignin, another prevalent, molecularly complex plant material.

Lignin is a much more complex organic molecule than cellulose; these are resistant to the enzymes produced by most bacteria and are left for fungi to decay.

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31 In the production of chemical wood pulps, various chemical processes are techniques used for lignin to dissolve away. Preston (1974) reported that despite of their importance and tremendous natural abundance, second only to cell wall carbohydrates, lignin resisted structural characterization until quite recently and still poses a major problem for their utilization. The lignin from grasses, softwoods, and hardwoods differ in their composition, chiefly in methoxyl substitution and the degree of carbon-carbon linkage between phenyl groups. H owever, their common structural features predominate, and the conifer lignin schematic structure (Freudenberg & Harkin, 1964) shows the features important for conversion into chemicals.

The macromolecular properties and structural characteristics of lignin make biodegradation studies uneasy. Suitable model compounds of lignin are difficult to obtain and for biodegradation studies, there are only a few assays which are suitable (Buswell & Odier, 1987). The methods used for isolating lignin can be classified into two methods which are removed and recovered selective lignin from the final solution, and the other one is lignin is left as an insoluble residue follow ing dissolution of the carbohydrates. Determination of Kappa number and Klason lignin are the most common methods used to analyse lignin quantitatively. Kappa number is an important parameter in the pulp and paper industry and basically, it is determined by oxidized the lignin selectively from pulp using a potassium permanganate solution (Argyropoulos & Menachem, 1997). By using gravimetric technique, Klason lignin is determine after extracting the sample with sulphuric acid to dissolve out the other components (Dence, 1992).

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32 2.5.4 Extractive s

According to Buchanan (1963) and Hillis (1962), extractive is the term applied to the extraneous plant components that may be separated from the insoluble cell wall material by their solubility in water or organic solvents. They include many different kinds of chemicals and a large number of individual compounds. Traditional uses of chemicals from biomass have involved such extractives components as tall oil fatty acids, rosin, turpentine, tanning materials, rubber, volatile oils, gums and camphor.

Classification of extractives is made difficult by their great variety. Different structures contain different extractives even they are within the same plant (Preston, 1974). There are several major categories of extractives which are volatile oils, terpenes (turpentines, tropolones, cymene, resin acids, and sterols), fatty acids, unsaponifiables, polyhydric alcohols, nitrogen compounds, and aromatic compounds (acids, aldehydes, alcohols, phenylpropane dimmers, stilbenes, flavonoids, tannins, and quinones) (H illis, 1962).

2.5 The role o f e arth wo rms in organic waste .

Darwin (1881) highlighted the breakdown of organic debris of earthworms on the soil surface and in the soil turnover. Since then, it has taken almost a century to appreciate their important contribution in curbing organic pollution and providing topsoil in impoverished lands. T his realization, although late, has awakened the global population to serious thought on utilizing them for ecological benefits.

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33 By the turn of century, earthw orms potential as a biological tool should be much better understood to make organic farming and sustainable development a reality with the use of selected species of earthworms. By regulating the moisture levels and mixing the ingredients in proper ratios that can be accepted by the earthworms, coffee pulp (Arellano et a l., 1995), sugar factory waste (Kale et al., 1994), and pig solids (Dominguez & Edwards, 1994) could all be converted into good quality soil additives, along with the biomass production of earthworms.

The reports of Vinceslas-A pka & Loquest (1994) and Fredrickson et a l. (1994) have shown that the involvement of earthworms in the composting process decreases the time of stabilization of the waste and produces an efficient organic pool with energy reserves as vermicompost. Garcia et al. (1994) showed in their studies that the sludges from both agri based industries and domestics’ sewage plants not accepted as soil additives directly on fields can be a food source for composting earthworms, with suitable organic amendments such as plant litter or animal waste to produce the worm biomass and obtain better quality soil additives.

2.6 Earth worms as bio de grade rs o f waste biomass.

The decline in productivity status of these tropical soils is majorly caused by the loss of topsoil due to practices associated with the indiscriminate use of chemicals. This fact has indeed increase the use of organic wastes on the fields. The availability of cattle dung has declined and the current need is to get the required organic soil additives from unutilized available plant biomass residues in minimum time (Sinha et

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34 al., 2008). In addition to the old practice of pit composting, various methods have been practicing. Engineering skills are employed to minimize the time of composting and to provide good aeration of composting materials. For composting of lignocellulose-based materials, selected groups of fungi are used in different places (Chaoui et a l., 2003).

The diversity and richness of earthw orms species is not much less than in temperate regions. However, much of the importance given to earthw orms’ activity in temperate soils is lacking in tropical regions (Bano & Kale, 1991). T he help of earthworms in the breakdown of waste has been broaden by the help of food niches and strategies that have been developed among different species of earthw orms. A greater affinity for nitrogen and rich organic matter live in a more unstable environment has been showed by the E pigeic earthworms’ species. This Epigeic earthworms’ species resemble the spiral stages of ecological development, with a higher metabolic rate, higher fecundity, smaller body size, , and shorter life cycle (Doube & Brown, 1998). In natural conditions, their survival depends on the degree of biotic pressure from predators and the environment. T hese earthworms come from the components of the tropical forest floor community. They have entered the agricultural plantations with the loss of natural forests, (Edwards, 1998).

The farming community has been neglecting the importance of organic manures and additives due to the higher yields from the use of chemical fertilizers. Farmers have been unaware that in order to restore the cycle of events in the soil to keep it

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35 productive, the organic wastes are essential. In recent years, the chemicals applied to the soil has short-circuited this cycle of events and deprived the soil organisms of their energy source to store the health of the soil through their activity (Chaoui et al., 2003). Earthw orms are classified as shredders. As they search for food, the leaf litters in the garden and on the lawn are broken down, greatly spreading up the decomposition of plant material, directly and indirectly. Besides that, the bacteria and fungi have been given better access to the cellulose (and other carbohydrates) and lignin (a noncarbohydrate) in the organic matter as the earthworms opens up leaves and other organic matter. Thus, it is obvious that earthworm facilitate the recycling of nutrients back to the plants. At the same time, the composition of the food web community may change due to competition for nutrients between earthworm and fungi and bacteria. They also even eat the populations of fungi and bacteria (Lowenfels & Lewis, 2006).

The magnitude of the impact of earthworms is shown by the simple fact: leaves on the forest floor or in a garden or lawn only need three months to decay by the help of worm shredding. B ut, w ithout worm shredding, normally about one or tw o years are needed. In some parts of the United States and Canada, the earthw orms left by fishermen have invaded the forests (Doube & Brown, 1998). These have completely altered the floor habitat. Plus, the entire forests are affected as the litter layer is being decayed far faster than is healthy for the trees and the rest of the soil food web. The minute particles of organic litter that microorganisms can eat are the end results of worm shredding and digestion. Microbial populations in the soil are also enhanced.

This is caused by some microbes that are mixed into worm fecal pellets during their

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36 formation and elimination, creating protected enclaves of fungi and bacteria (Lowenfels & Lewis, 2006).

2.7 Bre akdown of se wage wastes by e arthwo rms

In the late 1970s at the State University of New York (SUNY), the research into the potential use of earthworms to break down and manage sewage sludge is begun (Hartenstein, 1978). It was demonstrated quite early on a laboratory scale that aerobic sewage sludge can be ingested by the earthworm E.fetida and egested as casts, and in the process the sludge is decomposed and stabilized (i.e., rendered innocuous) about three times as fast as non-ingested sludge, apparently because of the increases in rates of microbial decomposition in the casts (Edwards, 1995).

During this process, relative to non-earthworm ingested sludge, objectionable odors disappear quickly and there is a marked reduction in populations of the pathogenic microorganisms Esch erich ia coli, Sa lmon ella en triditis, and other Enteroba cteriaceae (Edwards, 1988). A lthough most of the sludge produced in sewage plants are anaerobic, and when fresh can be toxic to E.fetida, after becoming aerobic it becomes readily acceptable to this species. According to Edwards, (1998) mixing other materials, e.g., paper pulp sludge, garden wastes, or other lignin-rich wastes w ith sewage sludge and composting the mixture using earthworms can accelerate their decomposition. This is due to maceration and mixing of such materials during passage into earthworm casts and passage through the earthworm gut.

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