ABSTRACT
The isolation and identification of the microorganisms associated with wood degradation in Umuariaga Village in Ikwuano Local Government Area was examined. The wood was collected from building sites in Umuariaga, district of Umuahia. All the materials and reagents used in this research work was gotten from the laboratories of Microbiology Department of Michael Okpara University of Agriculture, Umudike, (MOUAU) except the wood samples. The study found out the mean microbial counts from five (5) wood degradation sample. The Total heterotrophic plate count range from 3.8 x 105 to 8.7 x 105 cfu/ml and The Total coliform plate count were range from 2.8 x 105 to 5.3 x 105cfu/ml, Total fungal plate count 2.7 x 107 to 7.4 x 107 cfu/ml while Total staphylococcus plate count range from 1.3 x 105 to 4.1 x 105 cfu/ml respectively. The isolates from wood degradation samples were identified by morphological characteristics, pigmentation on media, microscopy, biochemical and sugar fermentation methods. This reveals the major bacterial isolates to belong to Bacillus species, Staphylococci aurues, Eschericial coli, Klebsiella species, streptococcus species and Serrieta species respectively. Fungal isolated from wood degradation samples were identify by their cultural characteristic and microscopic morphology namely; Aspergillus Niger, Aspergillus flavus, Rhodotorula species and Mucor Alternaria respectively. Distribution of bacterial isolates where a total of 27 bacterial and fungal strains were isolated from five (5) wood degradation samples collected at Umuariaga village of Umuahia namely; Sample A, B, C, D and E. The samples A, C and D has highest number of isolate with (6) and the lowest recorded at sample (F) with (4) isolate respectively. The percentage occurrence of bacterial and fungal species isolated from five (5) wood degradation samples presented in Table 4.4 The details of these isolates comprising; Serrieta species 5(18.5%), Staphylococci aurues 3(11.1%), Escherichia coli 4(14.8%), Streptococcus species 1(3.7%), Klebsiella species 2(7.4%), Bacillus species 3(14.8%) and Aspergillus Niger 1(3.7%), Aspergillus flavus 3(11.1%), Mucor Alternaria 3(11.1%) and Rhodotorula species 2(7.4%) respectively. The highest number of isolate found with Serrieta species 5(18.5%) follow by Escherichia coli 12(23.5%) and the lowest recorded with Aspergillus Niger 1(3.7%) and Streptococcus species 1(3.7%) respectively.
TABLE OF CONTENTS
Cover Page
Title Page i
Certification
ii
Dedication iii
Acknowledgement iv
Table of Contents v
List of Tables vii
Abstracts viii
CHAPTER ONE
INTRODUCTION
1.1 Background of the Study 1
1.2 Problem Statement 2
1.3 Aim of Study 3
1.4 Specific Objective 3
CHAPTER
TWO
LITERATURE REVIEW
2.1 Wood Decay 4
2.2 Bacterial Colonization of Wood 8
2.2.1 Edaphic and Atmospheric Sources of Bacteria 8
2.2.2 Bacterial Endophytes 9
2.2.3 Bacteria Co-Colonisation with Other
Organisms 9
2.3 Wet Wood 10
2.4 Bacterial Metabolism in Wood 11
2.4.1 Bacterial Nitrogen Fixation in Wood 11
2.4.2 Bacterial Wood Decomposition 13
2.5 Bacterial–Fungal Community Interactions 15
2.5.1 Community Competition and Co-Operation 15
2.5.2 Bacterial Endosymbiosis and Intimate Hyphal
Associations 18
2.5.3 Mycophagy and Predation 19
CHAPTER
THREE
MATERIALS
AND METHODS
3.1 Study Area 22
3.2 Materials, Reagents and Media to be Used 22
3.3 Sample Collection 22
3.4 Processing of Samples 23
3.5 Microbial
Analysis 23
3.5.1 Sterilization
Method 23
3.6 Isolation
of Micro Organisms 23
3.6.1 Sample
Inoculation 23
3.6.2 Isolation of Fungi 24
3.7 Microbial
Character Ization and Identification 24
3.7.1 Identification
of Bacterial Isolates 24
3.8 Biochemical
Tests 24
3.8.1 Identification
of Fungal Isolates 26
3.9 Screening
of Wood Isolates for Antibiotic Production 27
3.9.1 Preparation of Paper Disc 27
3.9.2
determination of Antimicrobial Activity 27
CHAPTER FOUR
RESULTS 29
CHAPTER FIVE
SUMMARY, CONCLUSION AND RECOMMENDATION
5.1 Summary 36
5.2 Conclusion 37
5.3 Recommendations 37
REFERENCES
LIST OF TABLES
Table 4. 1: Mean
count of microorganism isolates from wood degradation
sample 31
Table 4.2 Identification and characterization of Bacterial Isolates from wood degradation
sample. 32
Table
4.3. Identification and
Characterization of Fungal Isolates from wood degradation Samples 33
Table 4.4 Distribution of isolates from wood degradation samples 34
Table 4.5. Percentages
occurrence of isolates from wood degradation samples 35
CHAPTER ONE
INTRODUCTION
1.1 Background of the Study
Wood is an important renewable and
biodegradable natural resource with a multitude of uses. Wood is used
extensively as a structural material for buildings, wharves, telephone poles,
and furniture due to its high strength per unit weight, its versatility, and
its variety. Wood also serves as the industrial raw material for the
manufacture of paper and paper products, wood composites, and other products
made from cellulose, such as textiles and cellophane. In many parts of the
world wood is used as a fuel for heating and cooking. The primary biotic
decomposers of wood are basidiomycete decay fungi, which can attack and degrade
both wood in the forest and wood in service. In the forest ecosystem wood decay
fungi play an important role in carbon and nitrogen cycling and help to convert
organic debris into the humus layer of the soil. Some fungi attack living
trees; others invade downed timber and slash on the forest floor, lumber, and
wood in service. Wood decay basidiomycetes colonize and degrade wood using enzymatic
and nonenzymatic processes. Brown-rot fungi preferentially attack and rapidly
depolymerize the structural carbohydrates (cellulose and hemicellulose) in the
cell wall leaving the modified lignin behind. White-rot fungi can progressively
utilize all major cell wall components, including both the carbohydrates and
the lignin. As decay progresses the wood becomes discolored and loses strength,
weight, and density. Decay and discoloration caused by fungi are major sources
of loss in both timber production and wood use, with losses of 15 to 25%
marketable wood volume in standing timber and of 10 to 15% in wood products
during storage and conversion. Each year ca. 10% of the timber cut in the
United States is used to replace wood in service that has decayed, resulting in
the expenditure of hundreds of millions of dollars for raw materials, labor,
and liability.
Wood
decay fungi are considered the most destructive microbes that attack wood in
service, but the actions of bacteria are also important and are less studied in
wood degradation. Past studies have shown that bacteria are often the first
microorganisms to colonize wood as it decays. They initiate the decay process
by increasing permeability of the wood, hydrolyzing the waxes and pectin of
bordered pits, and breaking down wood extractives and preservatives. Bacterial
degradation of wood can be a concern (Blanchette 2011) but not typically in
residential lumber, because the process often takes decades. Past studies at
FPL (Clausen 2011) found bacteria capable of removing copper, chromium, and
arsenic from wood treated with chromated copper arsensate (CCA) and
demonstrated alternative routes for bioremediation of treated wood wastes,
typically confined to wood decay fungi. Bacteria have also been shown to
interact with decay fungi in the decay environment (Johnston et al. 2016). With
the increasing availability of next generation DNA sequencing, it is now easier
and faster to characterize bacterial species from environmental samples,
thereby overcoming the difficulties inherent in culturing methods of isolation.
The goal of this collaborative research is to identify bacterial communities in
wood treated with several preservative chemistries using high-throughput
sequencing to compare treatments and bacterial populations over time. The
outcomes of this study will provide much needed baseline data that will enable
us to better understand the contributions of bacteria to decay of
preservative-treated wood and eventually provide improved strategies for more targeted
inhibition of biodegradation.
1.2 PROBLEM
STATEMENT
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, Siitonen and Jonsson 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.
The
fungal community within dead wood has received considerable study, but far less
attention has been paid to bacteria in the same habitat. Bacteria have long
been known to inhabit decomposing wood, but much remains underexplored about
their identity and ecology. Bacteria within the dead wood environment must
interact with wood-decay fungi, but again, very little is known about the form
this takes; there are indications of both antagonistic and beneficial
interactions within this fungal microbiome. Fungi are hypothesised to play an
important role in shaping bacterial communities in wood, and conversely,
bacteria may affect wood-decay fungi in a variety of ways. This minireview
considers what is currently known about bacteria in wood and their interactions
with fungi, and proposes possible associations based on examples from other
habitats. It aims to identify key knowledge gaps and pressing questions for
future research.
1.3 AIM
OF STUDY
i.
To isolate and identify the microorganisms
associated with wood degradation in Umuariaga Village in Ikwuano Local
Government Area.
1.4 SPECIFIC
OBJECTIVE
ii.
To determine the microbial content of the
samples
iii.
To isolate the microorganisms associated
with wood degradation in Umuariaga village in Ikwuano Local Government Area.
iv.
To identify possible microorganisms
associated with wood degradation in Umuariaga Village in Ikwuano Local
Government Area.
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