BULE HORA UNIVERSITY

COLLEGE OF NATURAL AND COMPURTAIONAL SCIENCE

DEPARTMENT OF BIOTECHNOLOGY

SUBMITED BY:-BINIYAM MENGESHA. ... IDNO .

RU4566/13

ISOLATION,IDENTFICATION AND CHARACTERIZATION OF

BACTERIAL COMMUNITY FROM MUNICIPAL SOLID
WASTE,BULE HORA TOWN,NORTHEN ETHIOPIA

ADVISOR:-NIGUSE HOTESA

BULE HORA, ETHIOPIA

JANUARY, 202
 Catalog
LIST OF ABBRIVETION......................................................................................................iii
CHAPTER ONE.....................................................................................................................iv
1.INTRODUCTION................................................................................................................4
1.1.BACK GRAOUND OF THE STUDY.......................................................................4
1.2. STATEMENTOF PROBLEM...................................................................................3
1.3. Objectives..................................................................................................................4
1.3.1.General Objective.............................................................................................4
1.3.2.specific objectives.............................................................................................4
CHAPTER-TWO.....................................................................................................................4
2. Literature review.................................................................................................................4
2.1 Municipal Solid Waste (MSW)..................................................................................4
2.2. Types of wastes..........................................................................................................5
Table 3.1 summary of types of municipal solid wastes and their sources(Daniel and Laura,
1999)........................................................................................................................................5
2.3. Characterization of Bacteria......................................................................................6
2.3.1 Microbiological characterization of bacteria....................................................7
2.3.2 biochemical characterization of bacteria..........................................................7
2.3.3 Molecular Characterization of Bacterial Strains...............................................7
2.4. Municipal solid waste management...........................................................................8
2.4.1. composting.......................................................................................................8
2.4. 2 Incineration....................................................................................................10
2.4.3 Landfills.......................................................................................................10
CHAPTER-THREE...............................................................................................................11
3. Material and methods.........................................................................................................11
3.1.Study area..................................................................................................................11
3.2 Types of research design..........................................................................................11
3.3.Sample collection......................................................................................................11
3.4 Data collection instrument........................................................................................11
3.5 Culture media............................................................................................................11
3.6 Isolation of bacteria from waste Samples.................................................................12
3.7 characterizations of bacterial isolates.......................................................................12
3.7.1 Microscopic characteristics of bacterial isolates.............................................12
3.7.2 biochemical characteristics of bacterial isolates.............................................12
4.Expeted OutCome...............................................................................................................13
5. WORK PLAN AND BUDGET BREAK DOWN.............................................................14
5.1 Work plan and activity..............................................................................................14
Table 1 WORK PLAN AND ACTIVITY.............................................................................14
5.2.BUDGET BREAKDOWN.......................................................................................14
Table 2 Budget Breakdown...................................................................................................14
Referance...............................................................................................................................15

I
LIST OF ABBRIVETION

MSW: MUNCIPLAL SOLID WASTE

EPA: ENVIRONMENTAL PROTECTION AGENCY
MSWM: MUNICIPAL SOLID WASTE MANEGEMENT
USEPA: UNITED STATE ENIVIRONMENTAL PROTECTION AGENC

II
CHAPTER ONE

1.INTRODUCTION

1.1.BACK GRAOUND OF THE STUDY

Municipal solid waste (MSW) generation is an issue of worldwide concern. The
generators of municipal solid waste are broadly classified as residential, industrial,
commercial, institutional, construction, demolition, municipal and agricultural types
(Sehker ,1998). Municipal solid waste is also generated by human and animal activities
that are discarded as useless or unwanted waste. Municipal solid waste commonly
known as trash or garbage (US), refuse or rubbish (UK) is a waste type consisting of
everyday items we consume and discard. It predominantly includes food wastes, yard
wastes, containers and product packaging, and other miscellaneous inorganic wastes
from residential, commercial, institutional, and industrial sources (Ahsan, 1999). Examples
of inorganic wastes are appliances, newspapers, clothing, food scrapes, boxes, disposable
tableware, office and classroom paper, furniture, wood pallets, rubber tires, and
cafeteria wastes. Municipal solid waste does not include industrial wastes, agricultural
wastes, and sewage sludge.Economic development, urbanization and improving living
standard in cities of developing have lead to increase in the quantity and complex
composition of municipal solid waste. Management of municipal solid waste resulting
from rapid urbanization has become a serious concern for government departments,
pollution controlling agencies, regulatory bodies and public in most of the developing
countries (Erdogan et al.,2016).
Several other factors like education standard and infrastructure of the country have significant
effect on municipal solid waste generation. The estimation and prediction of municipal
solid waste generation play an important role in municipal solid waste management. The
quantity of municipal solid waste in developing countries has been consistently rising
over the years (Kansal et al., 2016). The municipal solid waste composition varies from
place to place and also bears a rather consistent correlation with the average standard
of living (Visvanathan et al., ( 2004). The waste generated in the developing countries
is similar in composition, the variation between regions being dictated by the climatic,

1
cultural and industrial, infrastructural and legal factors. Inefficient management and disposal
of municipal solid waste is an obvious causefor degradation of environment in the
developing countries. Ecological impacts such as land degradation, water and air
pollution are related with improper management of municipal solid waste (Khajuriaet
al.,2010).
Three primary sources of waste are classified as municipal solid waste. These are residential,
institutional and commercial waste, and municipal services wastes (street sweeping).
Residential wastes are high in quantity and vary with time and season. Over time,
municipal solid waste is not only increasing but the composition is also changing. The
organic wastes are decreasing and the paper and plastics are increasing in the waste stream,
indicating the growing preference for consumption of packaged food.
Bacteria are the most abundant group and the most important microbe for decomposing
waste. Bacteria use wastes for their own metabolism and finally they produce some simple
and useful compounds which are important for soil health, plant growth and over all to keep
well balance of natural ecosystem (Alexander, 1961).
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. 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 acids, enzymes, perfumes,
microbial mining .

1.2. STATEMENTOF PROBLEM

Now a days the population is extremely increasing throughout the world, that utilize and
consume greater amount of raw materials for surviving their living. This leads the generation
of high amount of municipal solid wastes. The problem is especially more prominent in urban
areas than rural areas due to the increased population number and the presence of different
industries and manufactories that release by-products in to the environment. The more

2
accumulation of municipal solid wastes which leads to environmental pollution.This is useful
to determine what management will be implemented to resolve the problem of municipal
solid waste in Bule Hora town. Municipal solid waste management costs are expensive and
require other alternative management for example incineration, sanitary landfill and their
technologies. But like incineration method is burning of municipal solid waste and it
increases carbondioxide green house gas, which is one cause of global warming. Dueto this
reason this research aims to isolate and characterize bacterial isolates which will use to
decomposed municipal solid wastes. Composting is one of solid waste management that
is economical, convenient and can produce useful organic fertilisers for crops.

1.3. Objectives
1.3.1.General Objective
The main objective of this project is to isolate and characterization of bacterial
community from municipal solid wastes to assess their capability to decompose solid
wastes.

1.3.2.specific objectives
 To isolate bacterial colonies from municipal solid wasteby microbiological methods
 To identify and characterize bacterial strains from municipal solid wastes by different
culture and biochemical methods
 To evaluate the decomposing potential of bacterial isolates, those can degrade solid
wastes

3
CHAPTER-TWO

2. Literature review
2.1 Municipal Solid Waste (MSW)

Municipal solid waste (MSW) is a term often used for the solid heterogeneous byproduct of
different human activities in the municipal area of the city. The waste generally contains
discarded material like papers, plastic, glass, metal fine earth particles, ash, sewage
sludge, dead animals, etc. (Thitame et al., 2010).
These wastescome from homes; institutions such as schools and hospitals; and commercial
sources such asrestaurants and small businesses. EPA’s definition of MSW does not
includemunicipal wastewater treatment sludges, industrial process wastes, automobile bodies,
combustionash, or construction and demolition debris. Once generated, MSW must be
collected and managed.Common management methods include recovery for recycling or
composting, combustion (with theresulting energy used to generate electricity or steam in
some cases), and landfill disposal. Manywastes that are disposed of in landfills represent a
loss of materials that could be reused, recycled, or converted to energy to displace the use of
virgin materials. Beyond the environmental impacts of disposal, patterns in MSW generation
can help reveal acomponent of the total materials a society creates and uses, which is an
important aspect ofsustainability. Usually society creates and consumes more materials, it
demands more resources (e.g., water, energy, minerals, land) and generates greater quantities
of pollutants andwaste. (Fiksel,2006).

2.2. Types of wastes

Municipal solid waste is generated from households, offices,hotels, shops, schools and other
institutions. The major components are food waste,paper, plastic rags,metal and glass,also
demolition and construction debris is often included in collected waste as are small quantities
of hazardous waste. Such as electric light bulbs, batteries,automotive parts and discarded
medicines and chemicals(keen et al., 1997).

4
 Table 3.1 summary of types of municipal solid wastes and their
sources(Daniel and Laura, 1999).
Source Typical waste generators Types of solid wastes

Residential Single and multifamily dwellings Food wastes, paper, cardboard, plastics,
textiles, leather, yard wastes, wood, glass,
metals, ashes, and household hazardous wastes
Industrial Light and heavy manufacturing, Housekeeping wastes, packaging, food
fabrication, construction sites, wastes,
power and chemical plants construction and demolition materials,
hazardous wastes, ashes, special wastes
Commercial Stores, hotels, restaurants, Paper, cardboard, plastics, wood, food
markets, office buildings, etc wastes, glass, metals, special wastes,
hazardous wastes
Construction New construction sites, road Wood, steel, concrete, dirt, etc
and repair,
demolition renovation sites, demolition of
buildings
Agricultural Crops, orchards, vineyards, Spoiled food wastes, agricultural wastes,
dairies, hazardous wastes (e.g. pesticides)
feedlots, farms
municipal Street cleaning, landscaping, Street sweepings, landscape and tree
services parks, trimmings, general wastes from parks,
beaches, other recreational areas, beaches; and other recreational area,
water Sludge
and wastewater treatment plants

2.3. Characterization of Bacteria

Bacteria can be described and classified in three major ways, namely; microscopic
examination, cultural characteristics, and molecular phylogenetics. The study of microbial
diversity represents a major opportunity for advances in biology and biotechnology. There is
a massive capacity for genetic diversity of bacteria in the biosphere (Rondon et al., 1999).
One of the major problems that hamper studying the bacterial diversity in the environment is

5
the inability to obtain many of bacteria in culture . Over the years, diverse natural
microorganisms have yielded important biological materials useful to humans in
medical, industrial and agricultural fields . Recent progress in molecular microbial ecology
shows that the extent of microbial diversity in nature is far greater than previously thought
(Torsvic et al., 1996; Dees and Ghiorse, 2001).

2.3.1 Microbiological characterization of bacteria

Gram staining is an old and reliable method for observing the bacteria. Gram negative
bacteria were decolourized by alcohol, losing the purple colour of crystal violet. Gram
positive bacteria did not decolourized and remained purple .
Pure culture techniques have been the core of standard microbiology. Nonetheless, the
traditional culture-dependent methods are strongly biased and limiting, as only less than
five per cent of bacteria can be cultured with the rest of them being unamenable to this
approach (Amann et al.,1995). Janssen and group revisited the question of culturability
of soil bacteria using simple methodologies, and have found that there is considerable
scope for improving both the apparent culturability and the phylogenetic range of bacteria
that can be cultivated . Parallel study of widespread but previously uncultivated groups
of organisms strongly complement molecular ecological investigations and enhance
research into the roles of environmental bacteria and their biotechnological potentials .
Though limiting in terms of the number of cultivable bacteria, the culturing techniques
still provide non-overlapping information as compared to molecular screening
(Brambilla et al., 2001; Dees and Ghiorse, 2001).

2.3.2 biochemical characterization of bacteria

This type of characterization is characterization of bacteria by using different biochemical

testes such as catalase test, urease test, motility test and citrase test methods.Catalase test
used to test weather the bacteria can produce catalase enzyme or not.Citrase test is used to
test weather the bacteria can produce citrase enzyme that hydrolyse citrate in to oxaloacetic
acid and acetic acid.Urease test is used to determine the ability of microorganisms to degrade
urea by means of enzyme. Motility used to determine weather an organism is equipped with
flagella and thus capable of swimming.

6
2.3.3 Molecular Characterization of Bacterial Strains
The use of molecular techniques in microbial ecology has made possible the discovery
of new microorganisms previously unknown. Identification of bacteria has been based
on the determination of 16S rRNA sequences of amplified and cloned genes derived
from the isolates’s DNA. The 16 S rDNA is the most widely accepted gene employed for
bacterial classification and identification(Goto et al.,2000) suggested that 5' end region (~275
bp) is the hypervariant (HV) region in the gene, highly specific for each type strain, and
considered as an useful index for identification or grouping of Bacillus sp. Gupta et al
emphasized that use of molecular markers like 16 S rDNA as species-specific
identification tool have provided with a truly "microscopic" sensitivity down to single-
cell detection. Bacteria which grow well in pH range from 9 to 11 on the pH scale
are of ecological, industrial and basic bioenergetics interests .

2.4. Municipal solid waste management

One of the characteristics of modern society is its ability to produce waste. Most
human activities generate large amounts of waste that are apparently useless and can cause
serious problems from both environmental and economic perspectives. Waste is generated in
the human activity, such as municipal solid waste or sewage sludge, while the other,
associated with certain economic activities, is recorded only in a few areas. Knowledge of
quantity and composition of municipal solid waste is fundamental for the planning of waste
management system. Most previous studies looked at the characteristics of municipal solid
waste at the final disposal sites. capacity of composting to produce a valuable product used
to improve soil fertility or as a growing medium in horticulture. Common waste was used
for composting are agricultural waste/plant residues, livestock/poultry waste, sewage
sludge (SS) and municipal solid waste (MSW) . There are common municipal solid waste
managements. Those are composting, landfill, and incineration.

2.4.1. composting
Composting process is an aerobic biological breakdown of unstable organic matter which
resultsin formation of a more stable end product (compost) (Liang et al., 2003). It occurs in
four main stages which include an initial mesophilic, then thermophilic, a second
mesophilic and finally maturation (stabilization) phases

7
During the process, microorganisms use the crude organic matter as source of food thereby
producing heat, carbon dioxide, water vapor and humus (Tiquia, 2005). This aerobic
bio-decomposition self-heating activity results in the reduced volume of 40-50% of the
original waste, which becomes safer for use in improving soil fertility, tilth and its water
holding capacity (Finstein, 1975, Paré, 1999). The compost matrix consists of the organic
material, which serves as a source of nutrients for microbial growth, a sink for metabolic
products, and a site for gas exchange and insulation of heat (Rodrigues and Lopez-Real,
1999). The end product should not contain pathogens or viable seeds to make it suitable
for use as a soil amendment (Finstein, 1975; Epstein, 1997).
Composting (from the Latin compositum, meaning mixture) refers to a biodegradation
process of a mixture of substrates carried out by the microbial community composed of
various populations in aerobic condition and in the solid states. Microbial transformation of
pure substrates goes under the name of fermentation or bio-oxidation, but not
composting. The main product is called compost, which may be defined as the stabilized
and sanitized product of composting that is compatible and beneficial to plant growth.
Compost has undergone: (1) an initial, rapid stage of decomposition; (2) a stage of
stabilization; and (3) an incomplete process of humification, (Bertoldi and Insam, 2007).

2.4.1.1. Application of Bacteria in Bio composting

Bacteria use wastes for their own metabolism and finally they produce some simple
and useful compounds which are important for soil health, plant growing and over all to
keep well balance of natural ecosystem. The municipal solid waste in the urban centers is
generated by domestic, commercial and industrial sources It contains mostly organic wastes
that can be decomposed by composting. The bacterial conversion of the organics present
in MSW in the presence of air under hot and moist conditions is called composting, and the
final product obtained after bacterial activity is called compost (humus), which has very
high agricultural value. It is used as fertilizer, and it is non odorous and free of pathogens
(Ahsan, 1999; Khan, 1994).
As a result of the composting process, the waste volume can be reduced to 50–85%. The
composting methods may use either manual or mechanical means and are accordingly
termed as a manual or mechanical process. Manual composting is carried out in smaller
urban centres and mechanical composting plants have been set up in big Indian cities.
(Bhide and Shekdar,1998; Chakrabarty et al.,1995). The main objectives of composting are to

8
reduce the solid volume, weight and moisture content, minimize odour, decrease pathogens
and the spread of disease and increase potential nutrients for agricultural applications.
Therefore, composting is emerging as a popular waste management alternative both in
developed and developing countries Implementation of composting technology has great
potential for mitigating several problems related to an ecological imbalance due to loss of
nutrients from ecosystems and the disposal of organic wastes that cause water, soil and
air pollution and corresponding health hazards.

2.4. 2 Incineration

Incineration is a waste treatment process that involves the combustion of organic substances
contained in waste materials (. Incinerators reduce the solid mass of the original waste by 80
85% and the volume (already compressed somewhat in garbage trucks) by 95-96%,depending
on composition and degree of recovery of materials such as metals from the ash for recycling.
It is a key process in the treatment of hazardous wastes and clinical wastes. It is mandatory in
many countries that the medical waste be subjected to the high temperatures of incineration to
destroy pathogens and toxic contamination it contains(IARI, 2013).
The actual plant design and configuration of incineration plant will differ
considerablybetween technology providers. However, an Incinerator with energy recovery
will comprise the following key elements: waste reception and handling,combustion
chamber, emissions clean-up for combustion gases, and energy recovery plant,bottom ash
handling and air pollution control residue handling(Edugreen ,2010).

2.4.3 Landfills

Landfilling is the ancient and most well established infrastructure for waste management in
all fifty states. Curb side collection for residential waste is prevalent in most counties. It is
very easy to spot waste collection trucks in any town and city. Incoming collection trucks are
weighed at the entrances of landfills, as recording of incoming wastes tonnages is required
for landfills. There are efforts to keep separate account of MSW-only tonnages, but many
landfills accept combinations of MSW and non-MSW materials. As a matter of fact, the
majority of MSW-designated landfills are not exclusively MSW landfills. These sanitary
landfills are allowed to accept combinations of few of the following This are MSW,
Household hazardous waste, Municipal sludge, Municipal waste combustion ash, Infectious
waste, Waste tires, industrial non hazardous, waste, Conditionally exempt small quantity
generator hazardous waste, Construction and demolition debris, Land clearing debris,

9
Agricultural wastes, Oil and gas wastes and Mining wastes(US Environmental Protection
Agency, 1991).

CHAPTER-THREE

3. Material and methods

In order to achieve the objectives outlined in above chapter, different experiments

Will be carried out following the methodologies mentioned below.

3.1.Study area

Municipal solid waste samples will be collected from waste disposal sites of Bule hora
university campuses and other municipal waste dumping sites in Bule hora town . Bule
hora town is located at 485 kmNorth from the capital city, Addis Ababa, Ethiopia.

3.2 Types of research design

The study design is experimental.

3.3.Sample collection
A total of ten waste samples will be collected six from waste disposal sites of Bule
hora university Tewodros, Maraki and Fasil campuses’ and the rest of the four from dumping
andwaste disposal sites of Bule hora town. Sample (soil mixed with waste) will be
collected in sterile zip-lock plastic maintaining aseptically condition , stored at 4 0C and
marked accordingly to their source and location.

3.4 Data collection instrument

The data collection is experimentally through basic laboratory techniques.

3.5 Culture media

Culture media will be prepared containing nutrient agar, beef extract, peptone, agar and
distilled water. Five culture media will be employed to isolate a vast variety of
bacteria with potential biodegradation properties of the common MSW constituents
and their secondary products, namely; cellulose, lipids, proteins and simple sugars.
Nutrient Broth (NB) enriched with meat extract will be for isolation of proteolytic
bacteria, Horikish-I Agar (HIA) for simple sugars (product of complex carbohydrate

10
 breakdown), Emulsified CeNAN-Olive Oil (ECO) and Carboxymethyl Cellulose
(CMC) for lipolytic and cellulolytic bacteria, respectively. The culture formulations
done according to Haruta et al., (2002) and Ruiz et al., (2005) will be : LB agar
medium: (10g peptone, 5g yeast extract, 10g NaCl, 15g agar, to 1L distilled water, pH
7); NB agar medium: (5g peptone, 2g yeast extract, 5g NaCl, 1g Meat extract, 15g agar to
1L distilled water, pH 7.4); HIA agar medium: (10g D(+)-glucose, 5g yeast extract, 5g
Bactotryptone, 1g K2PO4, 0.2g MgSO4.7H2O, Na2CO3, 20g agar, per 1L distilled
water, pH 10); ECO agar medium: (5.5g peptone, 3.5g yeast extract, 15g Agar, 1% v/v
olive oil, 0.0002% Rhodamin B per 1L distilled water, pH 7.5) and CMC
Medium: (5g yeast extract, 5g peptone, 5g NaCl, 20g Carboxymethyl cellulose, per 1L
distilled water, pH 8).

3.6 Isolation of bacteria from waste Samples

Bacteria will be isolated from municipal solid waste by using spread plate method .
Spread plate is useful for quantifying microorganisms.

3.7 characterizations of bacterial isolates

The bacterial isolates will be identified according to bergey’s manual specification by
examining its microscopic characters, microscopic morphology and biochemical
characters(bergey’s 1981).

3.7.1 Microscopic characteristics of bacterial isolates

Gram staining chemicals will be used for testing weather the bacteria is gram negative or
gram positive. This will be done according to the following procedure.
After the preparation of bacterial smears crystal violate will be added and left for one minute.
the smears will be then treated with iodine solution and left for one minute. After that acetone
will be added for decolorizing bacterial isolates. Finally saffranin will be added for staining
the decolorized bacteria. Between each steps the smears will be rinsed with distilled water.
After the slides are dried the smears will be observed under microscope.

3.7.2 biochemical characteristics of bacterial isolates

The selected isolates will be examined by using biochemical test

3.7.2.1 Catalase test

This test used to test weather the bacteria can produce catalase enzymeor not. The test
follows following procedure.

11
Some amount of bacterial colony will be transferred from the culture media to glass slide.
Immediately one drope of 3% of hydrogen peroxide will be placed on to organism on the
slide. Finally the slides will be observed to check weather the bubbles are formed or not.

3.7.2.2 . Citrate test

This test is used to test weather the bacteria can produce citrase enzyme that hydrolyse citrate
in to oxaloacetic acid and acetic acid.
Simmons media will be prepared the bacteria will be inoculated by inoculation loop. After
the incubation for 2-4 days at 20-24 0c the media will be observed to test colour change from
green to blue.

3.7.2.3. Urea’s test

This test is used to determine the ability of microorganisms to degrade urea by means of
enzyme.
Urea test broth will be prepared from 20 g of urea ,9.5 g of Na2HPO4 ,9.1 g of KH2PO4, 0.1
g of yeast extract and 0.01 g of phenol red.
Loopful organism will be taken out from the tryptic broth culture tube with a cooled loop.
the urea broth will be inoculated with inoculation loop containing bacteria. After the
incubation for 24-48 hours at 370c. finally the broths will be obtained from incubator and the
colour will be observed.

3.7.2.4 motility agar test

It is used to determine weather an organism is equipped with flagella and thus capable of
swimming.
Semisolid nutrient motility test medium with 0.3% agar will be prepared. One colony of
bacteria will be selected and inoculated to a medium. After the incubation at 35 0c for 18-24
hours, the growth around the line of innoculatin will be examined.

4.Expeted OutCome
The isolation and characterization of bacteria strains from waste dumoing of
Bule hora university campus and Bule hora town will be undertaken in this study. In the
result different bacterial species will be expected to isolate which are important for the
management of municipal solid wastes by producing Waste biodegrading enzymes. The

12
isolated bacteria will be Aerobic and an aerobic type,they will be mostly aerobic because
municipal solid wastes are exposed for air(oxygen).

5. WORK PLAN AND BUDGET BREAK DOWN

5.1 Work plan and activity

Table 1 WORK PLAN AND ACTIVITY

No. Activity Dec Jan Feb Mar Apr
1 Chemical ###
requirement and
Sample collection
2 Bacterial isolation ###
and cultural
characterization
of bacteria
3 Identification and ###
biochemical
characterization
of bacteria
4 Determination of ###
bacteria in to their
species
Over all report ###
writing

13
5.2.BUDGET BREAKDOWN

Table 2 Budget Breakdown

No. Phase Activity Duration Cost ( Birr)

1 Phase 1 Requirment of chemicals, 29 Dec/2017-25 Jan/ 17000
and sample collection 2018
2 Phase 2 Bacterial isolation and 26 Jan/2018- 16500
cultural characterization Feb25/2018
of bacteria
3 Phase 3 Identification and 01 Mar/2018- 15500
biochemical 20Apr/2018
characterization of
bacteria

14
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