ABSTRACT
A total of 10 wastes samples were collected from different waste dump sites and analyzed for microbial contamination also checking their extracelluar enzymatic activities. 0.1ml volumes of each samples were homogenate separately, 10 fold serial dilution was made and was cultured on MacConkey agar, Nutrient agar, Saboraud Dextrose agar and Blood agar respectively and incubated at 370C for 24hours. The isolated bacteria species after microbial analysis were Escherichia coli, Klebsiella sp, Bacillus sp Staphylococcus aureus, Micrococcus sp, Aspergillus niger, Aspergillus flavus, Rhizopus stolonifer, and Rhodotorula spp. the Total Viable Counts (TVC) of Isolate from Waste Dump Samples, Total Viable Counts (TVC) of Isolate from Waste Dump Samples, The Total heterotrophic plate count range from 3.9 x 10-9 to 6.2 x 10-9 and The Total coliform plate count were range from 2.0 x 10-9 to 5.1 x 10-9 while Total staphylococcus plate count range from 3.2 x 10-9 to 7.1 x 10-9 TFPC and Total fungal plate count range from 2.2 x 10-9to 5.0 x 10-9 respectively. The percentage occurrence of the study showed that the bacterial isolates, Staphylococcus species and Escherichia coli were found to have highest percentage of occurrence with 5(27.78%) each, follow by Bacillus and Klebsiella spp with 3(16.66%) and the least was found in Micrococcus spp with 2(11.11%) respectively while the fungal species Aspergillus Flavus and Rhizopus Stolonizer has the highest of percentage with 4(33.33%) each, follow by Aspergillus Niger with 3(25.01%) and Rhodotorula spp the least with 1(8.33%) respectively. The extracellular enzymatic activities showed that Staphylococcus aureus, Bacillus spp and Micrococus sspp found to be highest enzymes production follow by Escherichia coli, Rhizopus stolonizer, Aspergillus niger and Aspergillus flavus and Klebsilla spp has the list enzymatic production respectively. In conclusion, the microbial qualities of the evaluated samples from different waste dump sites were averagely poor, and are certainly not fit for human consumption if by chance any crop grows around it as they are of low quality threshold. This may be due to direct contamination by animal and human excreta and other anthropogenic activities such as swimming, washing of clothes, farming etc., and thus, require further purification to ensure their suitability for human utility therefore they are considered highly valuable as they are used in fermentation processes, much as brewing, baking, cheese and butter manufacturing, chemical manufacturing such as ethanol, acetone, organic acid, enzymes, perfumes etc., microbial mining and they produce various antibiotics, vaccines, steroids as well as other therapeutically useful compounds with diverse biological activities.
TABLE OF CONTENTS
Title
Page i
Certification
ii
Dedication
iii
Acknowledgements
iv
Table
of Contents v
Lists
of Tables viii
Abstract
ix
CHAPTER ONE
1.0 Introduction
1
1.1
Aims and Objectives 3
1.2 Objectives 3
CHAPTER TWO
2.0 Literature Review 4
2.1
Municipal Solid Waste 4
2.2
Municipal Solid Waste Management 5
2.3
Composting Process 6
2.4
Composting Microorganisms 8
2.5
Factors Affecting Composting 10
2.6 Municipal Solid Waste for
Production of Industrial Enzymes 11
and Waste
Degradation
CHAPTER THREE
3.0
Materials and Method 13
3.1 Collection of Sample 13
3.3 Sterilization 14
3.4 Isolation of Microorganisms 14
3.5
Characterization and Identification of The Bacterial isolates 14
3.5.1 Grams Staining 14
3.5.2 Motility Test by
Hanging Drop Method 15
3.6 Biochemical Characteristics of
the Isolates 15
3.6.1
Catalase
Test 15
3.6.2
Coagulase Test 16
3.6.3
Citrate Test 16
3.6.4
Oxidase Test 16
3.6.5
Indole Test 17
3.6.6
Urease Test 17
3.6.7
Methyl Red Test 18
3.6.8
Voges-proskaeur Test 18
3.6.9
Sugar Fermentation Test 18
3.8 Qualitative Screening for Extracellular Enzyme Producing 19
Isolate by Plate Assay
3.8.1 Purification of isolate 19
3.8.2 Production of protease
enzyme 19
3.8.3 Production of Amylases enzyme 19
3.8.4 Production of Cellulases enzyme 20
3.9
Characterization and Identification of the Fungal Isolates 20
3.9.2
Lactophenol Cotton Blue Staining 20
CHAPTER
FOUR
4.1 Results 21
CHAPTER FIVE
5.0 Discussion, Conclusion and
Recommendation 30
5.1
Discussion 31
5.2
Conclusion 31
5.3 Recommendation
32 References 33
LIST OF TABLES
Table Title
Page
1: Total Mean microbial counts from
waste dump site sample 23
2: Cultural Characteristics of
Bacterial Isolates 24
3.
Biochemical Characteristics of Bacterial
Isolates 25
4. Identification and characterize of
fungal isolates 26
5: Percentage Occurrence of Fungal
Isolates 27
6:
Percentage Occurrence of Bacterial Isolates 28
7:
Detection of Extracellular
Enzymes for Isolates at 36°C. 29
CHAPTER ONE
1.0 INTRODUCTION
Waste
(also referred to as rubbish, trash, refuse, garbage, or junk) can be described
as unwanted or unusable materials (Wikipedia, 2009). Wastes may be generated
during the extraction of raw materials, the processing of raw materials into
intermediate and final products, the consumption of final products, and other
human activities. Residuals recycled or reused at the
Place of generation are excluded
(Unsd, 1997). With population increase, there is increase in solid waste
production making garbage pollution a serious problem (Khupe, 1996; Yaliang,
1996).
Waste
management in developing countries is usually equated with land disposal or
discharge into water bodies (Cilinskis and Zaloksnis, 1996). This method of
waste management is unscientific; a nuisance to the public, hence, the
resultant effect is pollution. When waste is dumped on land, microorganisms
such as bacteria and fungi proliferate using the components of the waste
materials as source of nutrient for growth as well as degrading the organic
materials in the waste (Stainer et al.,
1989).
Pathogenic
microorganisms and harmful chemicals in solid waste can be introduced into the
environment when the waste is not properly managed (Wai-ogosu, 2004; Ogbonna et al., 2006). Waste can contaminate
surface water, ground water, soil and air which poses more
Problems for humans, other species,
and ecosystems (Obire et al., 2002).
Waste
generation and its control have taken an important role in our environment.
With the doubling of population and changing lifestyle pattern of the
inhabitants the quantity of municipal waste generated is increasing in an
alarming rate. Most of this waste is subjected to dumping in a specified
disposal yard (Chukwu et al., 2004).
The greatest challenge to the
environmentalists is the eco friendly management of this waste and application
of microorganisms in this context has got an age over other available
technologies.
Organic waste is consumed by the
bacteria, used as nutrients by the bacteria, and is no longer present to
produce odors, sludge, pollution or unsightly mess. When bacteria consume
waste, they convert the waste into safe by products and in due course of this
conversion they actually produce several metabolites to break down the complex
waste into simple compounds. Soil microorganisms are increasingly becoming an
important source in the search for industrially important molecules (Alexander, 2007).
Extent of microbial diversity in nature is
still largely unknown, thus there might be many more useful products yet to be
identified from soil microorganisms. In soil 80 to 99% of microorganisms remain
unidentified whereas these biological communities are known to play a dominant
role in maintaining a sustainable biosphere. Today both academic and industrial
interest in soil bacteria (due to their several advantages over other microorganisms)
is on the rise, in search of deriving these unique biologically active
metabolites and novel commercially important products from them. Bacteria are
present in diverse ecological habitats (Rajbanshi, 2008).
They
are considered highly valuable as they are used in fermentation processes, much
as brewing, baking, cheese and butter manufacturing, chemical manufacturing
such as ethanol, acetone, organic acid, enzymes, perfumes etc., microbial
mining and they produce various antibiotics, vaccines, steroids as well as
other therapeutically useful compounds with diverse biological activities.
Hence there is an immense possibility to screen effective bacterial strains
from waste dump sites with valuable applications. To cope up with the demand
for new organisms with properties of production of unique enzymes/ molecules
for industrial application and waste degradation there have been a constant
effort in isolating novel bacteria from diverse environment (Gupta et al., 2002).
1.1
Aims and Objectives
The aim of this study is to isolate the potential
degraders from waste dump sites based on their enzymatic activities.
The objectives are:
1. To isolate microorganisms that can
degrade wastes in Umuahia, metropolis.
2. To identify the isolated
microorganism from the waste dump sites in Umuahia, Abia State.
3. To determine the enzymatic
activities of microorganisms isolated from various waste dump sites in Umuahia metropolis.
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