CHAPTER ONE

INTRODUCTION

BACKGROUND OF SIWES

The students industrial work Experience Scheme (SIWES) was initiated in 1973 by the
industrial training fund (ITF). The initiation helps in promoting and encouraging the acquisition
of skills in industries commerce with the view to generating a trained indigenous manpower
sufficient to meet the needs of economy.

Since the introduction of SWIES by the ITF, the scheme has gone through series of
reforms restructuring. For instance, its management has changed hands from the ITF to the
various regular agencies, National University Commission (NUC), National Board for Technical
Education (NBTE) National Commission for Colleges of Education (NCCE) and back to the
ITT. Similarly, the structure operational framework have been reviewed, streamlined and made
more fictional at the various levels of operations.

OBJECTIVES OF SWIES

The industrial Training Funds policy Document No. 1 of 1973 which establishment
SWIES outlined the objectives of the scheme. They are:

 Provides an avenue for students institution of higher learning to acquire industrial skills
and experience during their course study

 Prepare students for industrial work situation that they are likely to meet after graduation.

 Expose students to work methods and techniques in handling equipment and machinery
that may not be available in their institution

 Make the transition from school to the world of work easier and enhance students contact
for later job placement

 Provide students with the opportunities to apply their educational knowledge in real work
situation thereby bridging the gap between theory and practice

1
 Enlist and strengthen employers involvement in the entire educational process through
SWIES

BENEFITS OF INDUSTRIAL TRANING

There are several benefits accrue from industrial training, some of which includes.

 Opportunity for students to blend theoretical knowledge acquired in the classroom with
practical hands on application of knowledge required to perform work in industry.

 Enhancing students contacts with potential employers while on training.

 Enabling students appreciate the connection between their courses of student and other
related disciplines in the production of goods and services.

 Enabling students bridge the gap between the knowledge acquired in institution and the
relevant production skills (RPSs) required in work organizations.

 Provision of an enabling environment where students can develop and enhance personal
attributes such as critical thinking, creativity, initiative, resourcefulness, leadership, time
management, presentation skills and interpersonal skills amongst others.

2
 CHAPTER TWO

2.0 DESCRIPTION OF THE ESTABLISHMENT OF ATTACHMENT

2.1 The Establishment Profile/Brief History

Olabisi Onabanjo university Teaching Hospital (OOUTH) (Formerly called, Ogun

State University Teaching Hospital, OSUTH) is situated at Sagamu, Ogun State, South West

Nigeria. The teaching hospital was established in the year 1986 with primary aim of teaching

medical students from Olabisi Onabanjo University and provision of healthcare service to the

indigene of Ogun State. OOUTH was founded on the 1 st of January 1986 in partnership of

Obafemi Awolowo college health sciences to provide medical training for medical student of

Olabisi Onabanjo University. The teaching hospital was located at the former state hospital,

Sagamu, Ogun State, Nigeria. The pioneer chief medical director (CMD) of the hospital was

Prof. L.A. Salako. The hospital is managed by governing council appointed by the state

government. The current 2016 chairman of the governing council is Prof. Emmanuel O. Otolorin

3
 ORGANIZATION CHART(ORGANOGRAM)

4
 CHAPTER THREE

GENERAL INSTRUCTIONS IN THE LABORATORY

 No unauthorized person is allowed to come into the microbiology laboratory. All staff or
personnel who come into the laboratory must be adequately dressed in clean laboratory
coat.

 Tuck in your long hair and beard apprpriately to avoid inflammation.

 Hands must be properly washed and disinfected before and after every days work.

 Note that sneezing, coughing, spitting or picking of nose, and nails are highly prohibited
in the microbiology laboratory.

 Do not walk barefooted in the laboratory in the laboratory; Always put on shoes.

 Do not eat,drink,share confectioneries or apply lipstick in the laboratory.

 Fighting and noise making are not allowed.

 Always wash your hands with soap and disinfectant accordingly after visiting the toilet.

 Benches and tables must be cleaned before and after every days work.

 Glass wares should be kept away from bench edges to avoid or reduce accidents.

 Floors must be mopped with detergent and water before commencement of work.

 Spill over water or samples must be cleaned immediately to avoid or reduce accident.

 The benches must be cleaned and swabbed with methylated spirit and cotton wool before
and after every work.

3.2 EQUIPMENTS IN THE LABORATORY AND THEIR USES

 REFRIGERATOR

 INCUBATOR

 AUTOCLAVE

5
 MICROSCOPE

 CENTRIFUGE

 ELECTRONIC COMPACT SCALE

 COLONY COUNTER

 OVEN

 MEASURING CYLINDER

 TEST TUBE

 TEST TUBE RACK

 CALIBRATED PIPETTE

 PETRI DISH

 INOCULATING LOOP

 SWAB STICK

 WATER BATH

 SYRINGE / NEEDLE

 SAMPLE BOTTLES

 ALUMINUM FOIL

REFRIGERATOR

For preserving specimens and stock cultures at low temperature.

INCUBATOR

It is an instrument used for providing optimal temperature for bacteria growth.

6
AUTOCLAVE

This is a device for boiling water under pressure. It is the most practicable agent of sterilization

for materials that cannot be damaged by high temperature e.g. glasswares, surgical instruments

e.t.c.

7
MICROSCOPE

To view and examine specimens that cannot be seen with the unaided eye and to magnify object

for easy visibility.

CENTRIFUGE

It is an instrument used for spinning liquid, microbial culture, biochemical preparation to

separate a subnatant from supernatant.

8
ELECTRONIC COMPACT SCALE

For measuring weight or calculating masses of substances.

OVEN

For sterilization of the items that can withstand thermal heat.

9
MEASURING CYLINDER

For measuring accurate amount of liquid molecules.

TEST TUBE

For mixing, heating and holding small quantities of liquids during experimental processes.

TEST TUBE RACK

For holding test tube

CALIBRATED PIPETTE

For measuring and transferring small amount of liquids

PETRI DISH

For preparation of media and culturing microorganisms

INOCULATING LOOP

For collection of specimen, making smear and inoculating microbes

SWAB STICK

For collection of samples such s urine and stool

WATER BATH

It is an instrument used for boiling, homogenizing and upraising the temperature of culture and
biochemical preparations.

SYRINGE / NEEDLE

For collection and storage of samples such as serum.

SAMPLE BOTTLES

For collection of samples.

10
ALUMINUM FOIL

It is used for wrapping tubes and for shielding objects that are prone to contamination.

CANISTER

It is an instrument used for keeping petri dishes and pipette to shield them breakages.

11
WORKDONE IN DEPARTMENT OF ATTACHMENT

CHARACTERIZATION AND IDENTIFICATION TECHNIQUES

GRAM STAINING

Gram staining or Gram stain, also called Gram's method, is a method of staining used to

differentiate bacterial species into two large groups (gram-positive and gram-negative). The

Gram stain was introduced by Hans Christian Gram, a Danish physician in 1884.Gram staining is

a bacteriological laboratory technique used to differentiate bacterial species into two large groups

(gram-positive and gram-negative) based on the physical properties of their cell walls. Gram

staining reaction depends on whether or not the organism resists decolourisation with acetone or

alcohol after staining with dye and further treatment with mordant ( Gram's iodine). Bacteria

which resist decolorisation and remain dark purple or deep blue after counter staining are called

Gram-positive bacteria while bacteria which gets decolorized and take the light pink or red

colour after counterstaining with safranin, or dilute carbon fushin are called Gram-negative

bacteria.

APPARATUS: Bunsen burner, glass slide, inoculating loop, microscope, forceps, culture

organism.

Reagents: 95% alcohol/acetone, crystal violet, iodine, safranin.

Methodology:

 Grease or oil from the fingers on the slide was removed by washing the slides with soap and

water and wiping with spirit or alcohol or by passing the slide through flame 3-5 times. After

cleaning, the dry slides are placed them on an already disinfected staining tray, ready for use.

12
 A loopfull of the broth culture was transferred to the surface of a clean glass slide, and spread

over a small area (smear). The film was allowed to air dry and fixed by passing it briefly through

the Bunsen flame two or three times without exposing the dried film directly to the flame.

NT: The slide should not be so hot as to be uncomfortable to the touch.

The slide was flooded with crystal violet solution for up to one minute, washed off briefly with

tap water (not over 5 seconds), and drained.

The slide was then flooded with Gram's Iodine solution, and allowed to act (as a mordant) for

about one minute. It was washed off with tap water and drained.

 Excess water was removed from the slide and blotted. The slide was then flooded with 95%

alcohol for 10 seconds and washed off with tap water. The slide was drained.

 After the slide had been completely drained, it was flooded with safranin solution and allowed

to counter stain for 30 seconds, after which it was washed off with tap water. The slide was

drained and blot dried gently with bibulous paper.

 The stained bacterial slide was examined under oil immersion lens.

Result and Discussion

Gram positive bacteria - Purple or deep blue colour

Gram negative bacteria - Pink or red colour

Gram staining differentiates bacteria by the chemical and physical properties of their cell walls

by detecting peptidoglycan, which is present in the cell wall of gram-positive bacteria. Gram-

positive bacteria retain the crystal violet dye, and thus are stained violet, while the gram-negative

13
bacteria do not; after washing, a counter stain is added (commonly safranin) that will stain these

gram-negative bacteria a pink color.

Some organisms are gram-variable (meaning they may stain either negative or positive); some
are not stained with either dye used in the Gram technique and are not seen.

CATALASE TEST

Aim: To differentiate strains of Escherichia coli.

Materials: Glass slide, inoculating loop, hydrogen peroxide solution, broth culture of test

organism, forceps, staining tray

Method:

The Bunsen burner is lit.

The glass slide is fixed in flamed 3 to 5 time to eradicate any form of grease.

The glass slide is placed on the staining tray.

Few drops of hydrogen peroxide solution is dropped on the slide.

The inoculating loop was flamed and allowed to cool. A loop full of the test organism was taken,

and a smear was made with the hydrogen peroxide solution on the slide.

The slide was allowed to stand for a while, and the slide was observed and the result recorded.

Result and Discussion

After a short while, about 5 minutes, bubbles were seen all over the smear indicating gas

production and a positive reaction, therefore proving that Escherichia coli is catalase positive.

14
ASSESSMENT OF ANTIMICROBIAL ACTIVITY

An antimicrobial is an agent that kills microorganisms or inhibits their growth. Antimicrobial

medicines can be grouped according to the microorganisms they act primarily against. For

example, antibiotics are used against bacteria and anti fungals are used against fungi.

Minimum Inhibitory Concentration (M.I.C) Wide Range

This is a measure of the minimum concentration of a compound, which will inhibit growth. Such

a concentration may not be lethal to the cells. The method consists of including the compound

(bacteriostatic agent) under test in suitable nutrient medium in graded concentrations. To this

medium, a fixed inoculum of a suspension of test culture is added and after suitable incubation

the medium is examined for growth.

DETERMINTION OF MINIMUM BACTERICIDAL CONCENTRATION (MBC)

EVALUATION OF THE BACTERIOSTATIC ACTIVITY OF 0.5% PHENOL

Aim: To evaluate the minimum bacteriostatic concentration of the test compound

against bacteria

Materials: Test tubes, nutrient broth medium, sterile pipette, 0.5% phenol solution, 4 hours

broth culture of test organism, bunsen burner.

Method:

 A solution of the compound under investigation was diluted serially with the nutrient

broth medium, such that the concentration was halved in each tube in the series of 8 tubes

excluding the controls.

15
 Using a pipette, 5ml of solution of the test compound was aseptically added to 5ml of

double strength medium in tube 1 and mixed by shaking. Using a fresh pipette, 5ml of the

mixture was transferred to tube 2 (containing 5ml single strength medium) from tube 1 and

mixed also.

 This procedure was repeated to the 6th tube. Also, 5ml from tube 6 was discharged into
tube 01(positive control) whereas, no antimicrobial agent was discharged into tube 02, left alone
5ml of nutrient broth being the control for the viability of test organism(negative control).

 Finally, to each test tube (except tube 01), 0.1ml inoculum of test organism was added.

 Afterwards, a loop full of the control tube of the test organism was transferred to a
nutrient agar (seeding) and then poured into petri dish and allowed to set and cool.

 The tubes and the plate were then incubated at 37°C overnight before the results were

recorded.

 The lowest concentration inhibiting the growth of the test organism was recorded as

minimum inhibiting concentration (M.I.C) .

 All tubes not showing visible growth was sub-cultured on a nutrient agar plate.

 Afterwards the plate was incubated overnight before the result was recorded.

Result and Discussion

After incubation for 48 hours, there was no growth of the test organism recorded for tube

1(double strength broth with 5ml 0.5% phenol) and also, in tube 2(single strength). Every other

tube in the series excluding tube 01 (positive control) showed presence of growth of test

organism.

This indicates that at the dilution factor in tube 1(0.5) and (0.25) in tube 2, was potent enough to

16
carry out bacteriostatic action, with tube 2 at a concentration of 0.25 being the minimum

inhibitory concentration (M.I.C).

Further dilutions including the negative control all showed presence of growth of the test

organism, indicating no bacteriostatic action at the respective concentrations.

SOURCES OF MICROORGANISMS

Microorganisms may occur, live and multiply in a variety of environments, in the soil, in waters

of all types, in snow, in the air and in or on animals, plants and human. The following

experiments are designed to illustrate some of these sources.

ISOLATION OF MICROORGANISM FROM HUMAN FINGERS IN RELATION TO

SOURCES OF MICROORGANISMS

Aim: To identify sources of microorganisms in relation to human fingers.

Materials: Fingerprints, Petri dish, Soap and water.

Method:

 Nutrient agar was poured into a petri dish and allowed to set.

 A line was then drawn across the base of the plate to divide it into equal halves. A side

was labelled "washed" and the other "unwashed".

 Fingerprints was taken firmly and carefully from unwashed fingers on one section of the

plate.

 The hands was washed thoroughly with soap and water and the hands was allowed to dry
in air. The hands must not be dried with towel because it is also a source of microrganism.

17
 Fingerprints were then taken firmly from the washed fingers and placed on the second

section of the plate labelled 'washed'.

Result and Discussion

On examination of the plates, there was a large amount of growth on the part labelled

"unwashed" while the part labelled "washed" had just few colonies either as a result of the

microorganisms either from the water or air . No further test was carried out.

ISOLATION FROM THE AIR IN RELATION TO SOURCES OF MICROORGANISMS

Aim: To identify sources of microorganisms in relation to air.

Materials: Petri dish.

Method:

 Nutrient agar plate was poured and allowed to set firm.

 The plate was exposed to the atmosphere (by removing the lids) for 30 mins at different

points in the laboratory.

 It was the incubated inverted for 24 hours at 37°C

Result

On examination of the plates, there was a large amount of growth on the agar.No further test
was carried out.

18
ISOLATION FROM TAP WATER IN RELATION TO SOURCES OF
MICROORGANISMS

Aim: To identify sources of microorganisms

Materials: Sterile test tube

Method:

 5ml of water was collected in a sterile test tube from one of the laboratory taps.

 With the usual aseptic precautions, 1ml of water was transferred to the petri dish.

 Approximately 20ml of molten nutrient agar was added to the dish and mixed thoroughly.

 It was then allowed to set and incubated at 37°C for 24 hours.

Result

On examination of the plates, there was a large amount of growth on the agar.No further test

was carried out.

MICROBIOLOGICAL ANALYSIS OF RIVER WATER

Aim: To identify microorganisms present in river water.

Materials: Petri dish, test tubes, Syringe, Mannitol Salt Agar, Eosin Methylene Blue Agar,

Salmonella Shigella Agar, MaccConkey agar.

Method:

 10 bottles of water samples was collected from river water.

 10 test tubes of sterile water was prepared each containing 9mls each.

19
  The bottles and test tubes were labelled S1 to S10 respectively.

 1ml was transferred from bottle S1 to test tube S1 and shaken, after which 1ml was

discarded.

 1ml was also transferred from bottle S2 to test tube S2 containing 9ml of sterile water

and shaken after 1ml was discarded using syringes (labelled according to the bottles). The

same procedure was repeated for bottles and test tubes S3 to S10.

 20 bottles of nutrient agar approximately 20ml was prepared.

 20 petri dishes was also labelled.

 Pour plate by seeding (i.e. introducing 0.5 ml from each direct water and dilute water

sample into the nutrient agar and poured) was then carried out. It was then poured into

respective petri dishes and allowed to cool/solidify.

 All the plates were then incubated at 37°C for 24 hours for the determination of the

colony forming values.

In summary, seeding was carried out from both direct and diluted water samples.

S1 - S10 , S1Di - S10Di.

- 2 bottles (20ml each) of Mannitol Salt Agar, MacConkey Agar, and Eosin Methylene Blue agar

was also prepared and seeding from both diluted and direct water samples was carried out. Each

plates was labelled and incubated at 37°C for 24 hours.

20
Result and Discussion

After 24 hours, the result on the nutrient agar goes thus :

DILLUTION DIRECT

S1Di - 4 S1 - 1

S2Di - 2 S2 - No colony

S3Di - 170 S3 - 51

S4Di - 12 S4 - 5

S5Di - 3 S5 - No colony

S6Di - 35 S6 - 22

S7Di - 76 S7 - 14

S8Di - 70 S8 - 16

S9Di - 151 S9 - 32

S10Di - 175 S10 - 87

There was presence of yellow colonies on some mannitol salt agar which denotes fermentation

and positive to viable Staphylococcus aureus. On Eosin Methylene blue agar, green metallic

sheen was observed, positive to viable Escherichia coli while on MacConkey agar plate, mucoid

colonies was observed indicating Klebsiella pneumoniae. Pale colonies was also present on

MacConkey agar after when subcultured on Salmonella shigella agar, pale colonies appeared

indicating Shigella dysenteriae.

All these denotes that all microorganisms been identified are all present in the water samples.

21
 CULTURING TECHNIQUES

TRANSFERENCE OF BROTH CULTURE TO SOLID MEDIA:

Materials: Agar medium, broth culture, inoculating loop, bunsen burner, petri dish, McCartney

bottles, water bath, pipette, inoculating needle

STREAKING (INOCULATING WIRE LOOP)

Aim: To get discrete colonies of an organism when sub-culturing

Method:

Agar was prepared, sterilized, poured aseptically into dishes, and left to solidify.

The agar was oven dried at 50 degree C to prevent cracking.

The bunsen burner was lit.

An inoculating wire loop was flamed red hot and allowed to cool.

Aseptically, a loop full of inoculum from the broth culture (24 hour or less) was taken with the

wire loop.

The petri dish (agar) was opened in front of the flame, and a semi- circular smear was made at

the corner of the plate.

The wire loop was flamed again and allowed to cool.

The wire loop was then used to streak (make patterns) on the plates, and then incubated inverted.

22
SEEDING (PIPETTE)

Agar was prepared, sterilized and placed in a water bath (maintaining a temperature of 55 degree

C).

A sterile pipette was used to withdraw 0.2ml of the broth culture.

The McCartney bottle containing agar inoculated directly in its molten form by seeding. It was

rubbed between both palms gently for homogeneous distribution.

The plates was then poured aseptically, left to solidify and incubated.

SWABBING (SWAB STICK)

Agar was prepared, sterilized, poured aseptically into dishes, and left to solidify.

The agar was oven dried at 50 degree C to prevent cracking.

The bunsen burner was lit.

A sterile swab stick was soaked in a broth culture (not older than 24 hours).

The petri dish was held in the left hand, in front of the flame, and the surface was swabbed

evenly with the soaked swab stick.

The plate was left to stand for a while and incubated inverted.

23
 CHAPTER FOUR

PROBLEMS ENCOUNTERED DURING SIWES

Though the industrial; training program proved to be an eye opener, there were few

inconveniences encountered, just to mention a few:

Movement constraints during the last three months of my program due to some traditional and

cultural rights in the community where i undertook my program.

Also, and majorly, was the lack of funds during the training period. Allowances were not

allocated at any point in time whereas so much was spent on transportation and feeding.

SUGGESTION ON IMPROVEMENT OF SIWES

There should be payment to siwes student monthly.

They should collaborate with mega industries to improve working experience and training

facilities for the students.

SUMMARY AND CONCLUSION

In summary I have been able to learn how to use various microbiological equipments effectively,

learn various techniques and microbiological work ethics. I have learnt how to run quality

control of products, evaluate plant extracts, isolate microorganisms, as well as biochemical test.

In conclusion, the SIWES program has enlightened me to broader aspects of my discipline. I

have also been able interact with senior professionals who have embedded a lot of knowledge in

me. SIWES has been a tremendous success.

24