ABSTRACT
The study was to isolate and characterize microorganisms involved in wood degradation. A total of six bacterial and four fungal isolates were found to be responsible for the deterioration of wood samples. These microorganisms were isolated using the pour plate technique. The bacteria species isolated and identified using morphological and biochemical characterization included; Bacillus species, Staphylococcus aureus, Escherichia coli, Klebsiella species, Streptococcus species and Serratia species, while the fungi species are Aspergillus niger, Aspergillus flavus, Rhodotorula species and Mucor alternaria. Amongst the five (5) wood samples investigated for degradation by microorganisms, Fibreboard had the highest bacteria count at 8.7x105, while the least was recorded for Beach wood (1.3x105). The fungi count showed that Beach wood had the highest counts at 7.4 x107 while Veneer wood recorded the least fungi counts at 2.7 x107. It was also observed in this study that Serratia species is the most frequently occurring isolates with a high percentage occurrence of 5(27.7, while amongst the various woods accessed for its degradation potentials, Veneer wood had the highest number of bacteria species 5(27.7%. This suggests that Veneer wood can easily be degraded by microorganisms. Among the fungal isolates, Aspergillus flavus and Mucor alternaria 3(33.3) was the most predominant and frequently occurring isolates on the wood types. The extracellular enzymes activities for isolate in this study revealed that Staphylococcus aureus, Bacillus spp and Streptococus species were found to exhibit the highest enzymes production. Conclusively, the major bacteria and fungi species generally associated with the deterioration of wood types as revealed in this study are Serratia species, Aspergillus flavus and Mucor alternaria. Therefore, man can depend on microorganisms in making the world a better place to live as a result of their degradation potential of recycling wood wastes thereby maintaining environmental quality
TABLE OF
CONTENTS
Title
Page i
Certification iii
Dedication iv
Acknowledgements v
Table
of Contents vi
List
of Tables ix
List
of Figures x
Abstract xi
CHAPTER
ONE
1.1 Introduction 1
1.1 Aim
and Objectives 2
CHAPTER
TWO
2.0 Literature
Review 4
2.1 Wood
and Its Component 4
2.1.1 Cellulose 4
2.1.2 Hemicellulose 5
2.1.3 Lignin 5
2.2 Types
of Wood 6
2.2.1 Beech Wood 6
2.2.2 Ash Wood 7
2.2.3 Fibreboard 7
2.2.4 Plywood 7
2.2.3 Veneer 7
2.3 Classification
of Wood 8
2.3.1 Hardwoods 8
2.3.1.1 Maple 8
2.3.1.2 Mahogany 9
2.3.1.3 Cherry 9
2.3.1.4 Walnut 10
2.3.2 Softwoods 10
2.3.2.1 Pine 10
2.3.2.2 Ash 10
2.3.2.3 Birch 11
2.3.2.3 Cedar 11
2.3.2.4 Redwood 11
2.3.2.4 Hemlock 12
2.4 Where
Woods Can Be Found in Nigeria 12
2.4.1 The Makoko-Oko Baba Wood Market 12
2.4.2 The Umuahia Modern Timber Market 13
2.5 Degradation
of Wood by Fungi 13
2.5.1 White-rot Fungi 14
2.5.2 Brown-Rot Fungi 14
2.6 Microbial
Colonisation of Wood 15
2.7 Diversity
of Bacterial Communities in Wood 16
2.7.1 Edaphic and Atmospheric Sources of Bacteria 17
2.7.2 Bacterial Endophytes 18
2.8 Bacterial
Nitrogen Fixation in Wood 18
2.9 Bacterial
Wood Decomposition 20
2.10 Isolation
and Characterization of Microorganisms Involved in
Degradation
of Wood Wastes 20
CHAPTER
THREE
3.0 Materials
and Methods 22
3.1 Collection
of Samples 22
3.2 Sterilization
of Materials 22
3.3 Preparation
of Culture Media 22
3.4 Isolation
of Hydrolytic Microorganisms from Wood Chips 22
3.5 Screening
of Hydrolytic Enzyme-Producing Microorganisms from Wood Chips 23
3.6 Identification
of the Isolates 23
3.7 Gram
Staining 24
3.8 Biochemical
Tests 24
3.8.1 Catalase Test 24
3.8.3 Citrate Utilization Test 24
3.8.4 Hydrogen Sulphide (H2S) Production Test 25
3.8.5 Starch Hydrolysis 25
3.8.6 Motility, Indole, Urease (MIU) 25
3.8.7 Coagulase Test 26
3.8.8 Oxidase Test 26
3.9 Identification
of Fungal Isolates 26
3.9.1 Wet Preparation 26
3.9.2 Colonial Morphology 26
3.10 Qualitative
Screening for Extracellular Enzyme Producing Isolate by Plate Assay 27
3.10.1 Purification of Isolate 27
3.10.2 Production
of Xylanase Enzyme 27
3.10.3 Production of Amylases Enzyme 27
3.10.4 Production of Cellulases Enzyme 28
CHAPTER
FOUR
4.0 Results 30
CHAPTER
FIVE
5.0 Discussion,
Conclusion and Recommendation 38
5.1 Discussion 38
5.2 Conclusion 40
5.3 Recommendation 41
References
LIST OF
TABLES
|
S/N
|
TITLES
|
PAGE NO
|
|
4.1
|
Total Viable Microbial Mean Counts of Isolates from Wood Samples
|
32
|
|
4.2
|
Identification and Characterization of Fungi Isolates from the
Wood Samples
|
33
|
|
4.3
|
Identification and Characterization of Fungi Isolates from the
Wood Samples
|
34
|
|
4.4
|
Distribution and Percentage Occurrence of
Bacterial Isolates from the Wood Samples
|
35
|
|
4.5
|
Distribution and Percentage Occurrence of Fungal Isolates from
the Wood Samples
|
36
|
|
4.6
|
Detection of Extracellular Enzymes Activities for Bacterial and
Fungal Isolates at 36°C
|
37
|
LIST OF
FIGURES
|
FIGURES
|
TITLES
|
PAGE NO
|
|
1
|
Used Wood obtained from residential buildings
|
31
|
|
2
|
Unused Wood obtained from timber market
|
31
|
CHAPTER
ONE
1.1 INTRODUCTION
Globally, fallen wood
stores more than 73 billion tonnes of carbon (Pan et al., 2011) and provides habitat for a wide range of saproxylic
(i.e. dead wood-inhabiting) organisms (Stokland et al., 2012). Understanding the rate, mechanisms and control of
wood decomposition is of major ecological and economic importance, and the key
to doing so lies in understanding the microbial communities that effect and
regulate decomposition. Fungi are the dominant agents of wood decomposition,
but it has long been known that bacteria also inhabit dead wood. There are
indications of great bacterial diversity within wood (Hoppe et al., 2015), but bacteria are very
poorly understood compared with fungi in the same environment. Wherever
bacteria and fungi co-occur, they must interact with and influence each other,
yet, although wood decay fungi are well known for being highly competitive,
relatively little attention has been paid to the fungus– bacteria relationship
(de Boer et al., 2005).
Fungal–bacterial interactions have already been studied in other contexts for
their importance in medicine, agriculture, and food and drink (Frey-Klett et al., 2011), but have been explored
far less with respect to decomposition. The suite of bacteria that surrounds
and interacts with a fungus effectively constitutes its microbiome, and as
such, they must be considered together.
Biodegradation is the natural
process of breaking down organic pollutants by microorganisms to harmless compound
or recycling wastes to nutrients, which can be used by other organisms.
Degradation is carried out by huge assortment of bacteria, fungi, insects,
worms and other organisms that eat materials and recycle them into new forms
(Singleton and Sambury, 2008). The end products of effective biodegradation are
non-toxic such carbon dioxide and water and can be accommodated without harm to
the environment and living organisms. The microorganisms also multiply in
numbers in the process (Okpokwasili, 2004). The economic uses of wood include
usage in ice houses to keep ice frozen during the summer, until the advent of
refrigerator. It is used as platforms in poultry houses, cow pens and horse
stalls and it is mixed with dirt and chicken manure for compositing. Wood is
also used for energy production in the United States of America (Rose, 2012).
The general recalcitrance of wood
components such as cellulose, lignin and hemicellulose and the importance of
their biodegradation in the environment have received much attention for
several years (Erikson et al., 2010).
In microbial ecology, cellulose, the most abundant as in naturally occurring
biopolymer is a vital component of the biospheric carbon cycle and its
bioconversion to fuel and chemicals is of great interest (Philips and Humphrey,
2003). Cellulose is totally insoluble in water and has about 2000–10,000
glucose subunits with molecular weight determination value that ranges from 200
000 to about 2.4 million. Cellulose fibrils have high tensile strength which is
used in the textile industry, paper and miscellaneous materials like vulcanized
fibre, plastic filters, filtering media and surgical cotton. Other uses include
adhesives, explosives, thickening agents, coated paper, cellophane, artificial
leather, films and foils (Hitchner and Leatherwood, 2002).
Wood has been reported to be
degradable by Lentinus squarrosolus (Mont)
singer, a basidiomycete also known as white rot fungi to form protein, glucose
and ethanol (Shide et al., 2004). Wu et al. (2008) reported the cultivation
of enzymes for the degradation of lignocellulosic materials such as wood. Fungi
of the classes hyphomycetes, zycomycetes, pyrenomycetes, hymenomycetes and the
actinomycetes and bacteria of the groups Cytophaga,
Erwinia, Pseudomonas, Sporoiytophaga,
Xanthomonas and Streptomonas degrade hemicelluloses content in wood (Wu et al., 2008).
1.1 AIM AND OBJECTIVES
To characterize and
identify microorganisms involved in wood degradation, while the specific
objectives are;
•
To isolate and identify various microorganisms
present in different wood samples
•
To determine the distribution and percentage
occurrence of isolates from different wood samples
•
To determine the extracellular enzymes activities for
bacterial and fungal isolates from the wood samples
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