ISOLATION AND CHARACTERIZATION OF INDIGENOUS MICRO ORGANISMS FROM WASTE DUMP SITES IN UMUAHIA METROPOLIS

ABSTRACT

This study evaluated the isolation and characterization of indigenous microorganisms from waste dump sites in Umuahia metropolis. Ten (10) waste samples were collected from five (5) different dumpsites located within Umudike area of Umuahia, out of which a total of 28 bacteria and fungi were obtained and were identified as six (6) different bacteria genera which includes; Staphylococcus aureus, E. coli, Bacillus spp, Klebsiella spp, Micrococcus spp and Serretia spp, and three (3) different fungi species., which includes; Rhizopus stolonifer, Aspergillus niger, and Rhodotorula spp, using a pour plate technique. The total bacterial mean count was found to be higher in dump site located at Government College (1) 13.6 x105 cfu/g followed by Umudike Junction 11.1 x105cfu/g while the least bacterial count was recorded at Government College (2) 4.5 x105 cfu/g. The high level of bacteria growth recorded at Government College (1) could be as a result of higher waste generation from the increased population and residential areas in the urban areas. From the findings in this study, it was observed that Escherichia coli and Staphylococcus aureus are the most frequently occurring isolates with a percentage occurrence of 25.0% each, while among the fungi species isolated, Rhizopus stolonifer had the highest occurrence at 14.3%, indicating that human and animal faecal wastes were dispersed with the refuse. The degree of acidity (pH), reported in this study for all the waste dump sites ranged from pH 4.5 to 5.7. In the waste dump sample collected from Government College (2), the total nitrogen present was 0.11% indicating that it is low while values recorded for available Phosphorus under Government College were higher than the initial value observed. From this study, it can be concluded that the open dump system of waste disposal is indeed a potential environmental quality problem which takes the form of unsightliness, land and water pollution, it reduces the quality of air by the emission of foul odours and different gases derived from the anaerobic decomposition as well as occasional burning.

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

1.0 Introduction 1

1.1 Aim and Objectives 4

CHAPTER TWO

2.0 Literature Review 6

2.1 Municipal Solid Waste 6

2.2 Municipal Solid Waste Management 7

2.3 Reduce, Reuse and Recycle of Solid Wastes 8

2.3.1 Waste Reduction and Reuse 8

2.3.2 Recycling 9

2.4 Treatment and Disposal of Solid Wastes 9

2.4.1 Thermal Treatment 10

2.4.2 Biological Waste Treatment 12

2.5 Health Hazard Associated With Solid Waste Dump Sites 13

2.5.1 Occupational Health Hazards 13

2.5.2 Hazards to the General Public 13

2.6 Micro Organisms in Waste Dump Sites 14

2.7 Isolation and Characterization of Bacteria from Solid Waste 15

2.8 Isolation and Characterization of Bacteria Isolated from Municipal Solid

Waste for Production of Industrial Enzymes and Waste Degradation 16

CHAPTER THREE

3.0 Materials and Methods 18

3.1 Sample Collection 18

3.2 Initial Soil Sampling 18

3.3 Physiochemical Analyses of Waste Sample 18

3.3.1 Determination of Physiochemical Parameters of the Waste Sample 18

3.3.1.2 Determination of Available Phosphorus (Bray Ii Method) 18

3.3.1.3 pH Determination Using pH Meter (Electrode Method) 19

3.3.1.4 Determination of Total Nitrogen by Modified Kjeldahl Method 19

3.3.1.5 Determination of exchangeable potassium Capacity of the Waste Sample  (Ammonium Acetate Method) 19

3.4 Microbial Analysis 20

3.5 Sterilization Method 20

3.6 Sample Preparation and Isolation of Micro Organisms 20

3.6.1 Sample Inoculation 20

3.6.1.1 Isolation of Fungi 20

3.7.2 Quantitative Estimation of Bacteria and Fungi Isolates 21

3.8 Microbial Characterization and Identification 21

3.8.1 Identification of Bacterial Isolates 21

3.8.1.1 Gram Staining 21

3.8.2 Biochemical Tests 22

3.8.2.1 Indole Test 22

3.8.2.2 Carbohydrate Utilization Analysis 22

3.8.2.3 Catalase Test 22

3.8.2.4 Coagulase Test 22

3.8.2.5 Citrate Utilization Test 23

3.8.2.6 Motility Test 23

3.8.2.7 Voges-Proskauer Test  23

3.8.3 Identification of Fungal Isolates 24

3.8.3.1 Wet Preparation 24

3.8.3.2 Colonial Morphology 24

CHAPTER FOUR

4.0 Results 25

CHAPTER FIVE

5.0 Discussion and Conclusion 35

5.1 Discussion 35

5.2 Conclusion 37

References

LIST OF TABLES

LIST OF FIGURES

CHAPTER ONE

1.0 INTRODUCTION

Waste (also referred to as rubbish, trash, refuse, garbage, or junk) can be described as unwanted or unusable materials (Khupe, 2006). 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. With population increase, there is increase in solid waste production making garbage pollution a serious problem (Khupe, 2006). Waste management in developing countries is usually equated with land disposal or discharge into water bodies (Cilinskis and Zaloksnis, 2006).

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. Umuahia city does not have a sanitary landfill (Moffat and Linder, 2006). As a matter of fact, waste materials are deposited in certain “open” dump sites in some areas in Umuahia and allowed to compile until they are taken away or incinerated. Improper disposal of untreated municipal solid waste is not only harmful to human health but also a threat to the environment (Ogbonna et al., 2006).

Pathogenic microorganisms and harmful chemicals in solid waste can be introduced into the environment when the waste is not properly managed (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).

In Nigeria as well as in most developing countries, the urban landscapes are littered with garbage, plastics, bottles, disposable cups, discarded tires and even human and live-stock faeces. These wastes are aesthetically unpleasant, constitute eyesores, produce unpleasant odour especially when their organic compositions are acted upon by putrefying bacteria. These refuse dumps thus constitute a habitat for vector and other nuisance organisms capable of transmitting or causing diseases such as typhoid, infantile diarrhoea and cholera in humans and animals (Onyido, et al., 2009). Refuse dumps refer to areas or land sites where material wastes from several sources and processes are deposited. Refuse dumps include both municipal solid wastes and industrial wastes including liquid effluents containing heavy metals (Olanrewaju, 2002). Refuse dumps provide a rich source of microorganisms most of which are pathogenic (Odeyemi et al., 2011).

This is usually as a result of the attraction of rodents and vector insects for which the dump serves as shelter and food source (Donderski et al., 2000). Although it is known that vector insects and rodents can transmit various pathogenic agents of diseases such as amoebic and bacillary dysentery, typhoid fever, salmonellosis, cholera, plague and so on. A good percentage of these infections are caused by bacteria which are suspended in air around these refuse dumps which may later settle and cause contamination. Activities involving the disposal of solid wastes even if properly controlled with proper precautionary measures adopted may have adverse impact on the environment especially air since most of the dumps are open (Odeyemi et al., 2011).

Microorganisms present in the refuse use the refuse as a food source. Under the anaerobic conditions typical in most dumps, these microorganisms convert the organic material in the refuse to methane and carbon dioxide. As the gas rises through the dump and escapes into the atmosphere, it some-times picks up other compounds. The presence of large amounts of methane in this uncontrolled environment may result in explosions and fires. Additionally, this untreated gas may contain other compounds that pose a substantial health risk to nearby communities (Lewis, and Gattie, 2002). Many microbes can remain viable even after extended periods of time aloft despite the challenges associated with surviving in the atmosphere, including extended UV expo-sure, low moisture levels and extremely oligotrophic condi-tions (Jones and Harrison, 2004). Atmospheric transport is a key mode of microbial dispersal (Stetzenbach et al., 2004) and the transmission of airborne plant and animal pathogens can have significant impacts on ecosystems, human health and agricultural productivity.

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. 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 molecule (Lewis, and Gattie, 2002). 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 (Womiloju, et al., 2003). Bacteria are present in diverse ecological habitats. 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 (Scarpino, and Quinn, 2010).

The ever-increasing waste generation in urban cities of developing countries resulting from escalating metropolitan populations has rendered the conventional waste management practices inadequate. The abundance of solid organic wastes in an ever decreasing land space in urban centers has become a nuisance and health hazard. Depletion of organic compounds in farmlands resulting in lower yields and eventual high costs of farm products are obvious. Consequently, there is a need for a technique that would proficiently reduce the volume of waste and convert it into agriculturally usable resource for the amendment of farm lands. It is therefore necessary to adopt an indigenous, innovative, cost-effective and sustainable microbial technology for rapid Municipal Solid Waste composting, given the role the microbes play in the process. In Nigeria, there are colossal volumes of biodegradable wastes in large dumps, which make this proposed microbial Municipal Solid Waste management technique ideal.

1.1 AIM AND OBJECTIVES

To isolate and characterize indigenous micro organisms from waste dump sites in Umuahia metropolis, while the specific objectives are;

· To isolate micro organisms from indigenous waste dump within Umuahia Metropolis

· To identify possible microorganisms associated with waste dump within Umuahia metropolis

· To characterize the waste dumps by the physiochemical parameters

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