A TECHNICAL REPORT

ON

STUDENT INDUSTRIAL WORK EXPERIENCE SCHEME (SIWES)

UNDERTAKING AT

UNIVERSITY OF BENIN TEACHING HOSPITAL (UBTH)

BENIN CITY, EDO STATE

BY

PAUL SHARON OJONOKA

FPI/ND/SLT/23/215

SUBMITTED TO THE DEPARTMENT OF SCIENCE LABORATORY

TECHNOLOGY , SCHOOL OF TECHNOLOGY, IDAH, KOGI STATE.

IN PARTIAL FULFILMENT TO THE REQUIREMENT TO THE AWARD OF

NATIONAL DIPLOMA (ND) IN SCIENCE LABORATORY TECHNOLOGY.

JULY 2024 –NOVEMBER 2024

 DECLARATION

I, PAUL SHARON OJONOKA hereby declare that this SIWES Report has
been carried out by me.

____________________ _________________

Signature Date

ii
 CERTIFICATION

This is to certify that this report of the student industrial working experience Scheme
(SIWES) program was carried out by PAUL SHARON OJONOKA , a student
from the department of Science Laboratory Technology, School Of Technology,
Federal Polytechnic Idah, Kogi State. Matric number: FPI/ND/SLT/23/215, at
University of Benin Teaching Hospital (UBTH) Ugbowo, Benin City.

HEAD OF DEPARTMENT SIGN. DATE

SIWES COORDINATOR SIGN. DATE

iii
 DEDICATION

I dedicate this report to my family for their support and encouragement during my industrial

training.

iv
 ACKNOWLEDGEMENTS

I would love to acknowledge God almighty for his love and grace upon my life. I am grateful
for the staffs at University of Benin Teaching Hospital, who did everything to make sure that
my IT was a success. I am grateful to my director at UBTH, Dr. Igunma, for his contribution
he showed to me during my industrial training. I can’t neglect my supervisor, Dr. Ebeigbe,
Sir. Kelvin Osakpolor Nosa for their efforts and teachings during my industrial training. I
also want to appreciate the doctors, who also make out time to lecture me during my
industrial training. To my fellow IT students, thanks for the support. Lastly, I want to thank
my family for supporting me financially and emotionally throughout my academic pursuit,
may God bless you richly.

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 TABLE OF CONTENT

Page
COVER PAGE……………………………………………………………………… i
CERTIFICATION…………….………………………………………………………ii
DEDICATION………….……………………………………………………………..iii
ACKNOWLEDGEMENT…..…………………………………………………………iv
TABLE OF CONTENT………………………………..……………………………….v

CHAPTER ONE: ABOUT INDUSTRIAL TRAINING FUNDS

1.0 What is SIWES………………………………..………………………… 1
1.1 The History of SIWES ………………………………..…………… 1
1.2 Vision Statement………………………………..………………………… 2
1.3 Mission Statement………………………………..……………………… 2
1.4 Aims of SIWES………………………………..………………………… 2
1.5 Objectives of SIWES………………………………..…………………… 2
1.6 Scope of SIWES………………………………..………………………… 3
1.7 Importance of SIWES to AEB students………………………………….. 3
1.8 About the Company………………………………..…………………….. 4
1.8.1 Brief History of University of Benin Teaching Hospital…………………. 4
1.8.2 Mission Statement of the Hospital………………………………………… 5
1.8.3 Vision Statement of the Hospital………………………………………… 5
1.8.4 Departments in the Hospital…………………………………………..… 6
1.9 About Medical Microbiology Laboratory………………………………… 7
1.9.1 A Laboratory Department………………………………………………… 7
1.9.2 Key Services……………………………………………………………… 7

CHAPTER TWO: INTRODUCTION TO LABORATORY EQUIPMENT

2.1 Laboratory Equipment…………………………………………………… 10
2.2 Laboratory Safety and Precautions……………………………………… 11
2.3 Laboratory Waste Management…………………………………………… 12

CHAPTER THREE: BACTERIA STRUCTURE AND FUNCTION

3.1 External Structures of Bacteria…………………………………………..… 13
3.2 Internal Structures of Bacteria……………………………………………...13
3.3 Morphology of Bacteria…………………………………………..……….. 14
3.4 Preparation of Smears…………………………………………………..… 15
3.5 Fixing of Bacterial Smears……………………………………………….…15
3.6 Staining of Bacteria……………………………………………………..… 16
3.7 Types of Staining………………………………………………………… 17

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CHAPTER FOUR
4.0 Culture Media…………………………………………..………………… 20
4.1 Classification of Culture Media………………………………………….… 20
4.1.1 Based on Consistency…………………………………………..………… 20
4.1.2 Based on Nutrient…………………………………………..……………… 21
4.1.3 Based on Application………………………………………………………. 22
4.2 Components of Culture Media…………………………………………..… 22
4.3 Choice of Culture Media…………………………………………..……… 22
4.4 Methods of Culturing…………………………………………..………… 23
4.5 Biochemical Tests…………………………………………..…………… 23
4.5.1 Catalase Test…………………………………………..………………… 23
4.5.2 Coagulase Test …………………………………………..…………. 24
4.5.3 Citrate Utilization Test…………………………………………..………… 26
4.5.4 Urease Test…………………………………………..…………………… 27
4.5.5 Indole Test…………………………………………..…………………… 28
4.6 Anti-microbial Sensitivity Test ………………………………………30
4.7 Mode of Action of Anti-microbial Agents………………………………… 30
4.8 Anti-microbial Drugs of Clinical Importance……………………………. 31
4.9 Determination of Inhibitory Activity of Anti-microbial Agents…………… 32
4.10 Urine Microscopy, Culture and Sensitivity……………………………… 33
4.11 Collection and Transport of Urine Sample………………………………… 33
4.12 Urine Processing in the Laboratory for Urinary Tract Infection……………34

CHAPTER FIVE
5.0 Conclusion…………………………………………..………………….. 36
5.1 Experience Gained During my Industrial Training………………………. 36
5.2 Challenges Encountered During my Industrial Training………………….. 36
5.3 Recommendation…………………………………………..……………… 36
REFERENCE…………………………………………..………………………. 37
APPENDIX…………………………………………..………………………… 38

vii
 CHAPTER ONE

ABOUT INDUSTRIAL TRAINING FUNDS

1.0 WHAT IS SIWES?

The Student Industrial Work Experience Scheme (SIWES), also known as

INDUSTRIAL TRAINING is a compulsory skills training program designed to expose and
prepare students of Nigerian universities and other institutions for industrial work experience,
he/she is likely to meet after graduation.

The scheme provides students the eventuality of introducing and showing up

themselves to the needed experience in handling the best equipment and machinery that are
usually not available in their institution. It supports students and makes them independent.

Before the foundation of scheme, there was a growing thought among industrialists,
that graduates of institution of higher learning had inadequate practical background studies
preparatory for employment in industries. Thus, employers were of opinion that the
theoretical education in higher institutions wasn't liable to the needs of the employers of
labor.

1.1 THE HISTORY OF SIWES

The early phase of science and technology in Nigeria was characterized by the
theoretical lectures in polytechnics and universities which have been proven to be an ill
method of teaching. Students in universities and polytechnics graduate with little or no
technical experience in their course of study. In the same vein, students’ inability to
contribute to the society is hampering the growth and development of our country. It was in
this view that SIWES was introduced to the industrial and educational sector.

SIWES was established in the year 1973 in order to improve the standard of education
in Nigeria so as to achieve the needed technological advancement. SIWES was solely funded
by ITF (Industrial Training Funds) during its early stage until it was difficult to continue due
to economic stress then the responsibility was shared between the ITF and the Federal
Government. The federal government took over the funding of the scheme and ITF took over
the managerial position by managing the funds given to them by the federal government in
order to sustain the scheme. The effective management of SIWES has been as a result of the
cooperation and well played role of the federal government, ITF and supervising agencies.

1
1.2 VISION STATEMENT

To be the prime skills training development organization in Nigeria and one of the
best in the world.

1.3 MISSION STATEMENT

To set and regulate standards and offer direct training intervention in industrial and
commercial skills training and development, using a corps of highly competent professional
staff, modern techniques and technology.

1.4 AIMS OF SIWES

The aim of SIWES is to promote industrialization in Nigeria and an avenue between

the world of teaching, learning, and work with reference to a field of study such as
engineering, science, technology, dietetics and other professional education programs.

1.5 OBJECTIVES OF SIWES

SIWES provides avenue for students to acquire industrial skills and experience in
their approved course of study. It also prepares students for their industrial work situation
after graduation.

The objectives of SIWES are:

 The program provides an avenue for students to acquire industrial skills for
experience during their course of study.

 It exposes students to work methods and techniques that may not be available during
their course of study.

 Bridges the gap between theory and practice by providing a platform to apply
knowledge, relate school to real life work situations.

 Enables the easier and mother transition from school by equipping students with
better contact for future work placement.

 Introduce students to real work atmosphere so that they can know what they will most
likely meet when they graduate.

2
1.6 SCOPE OF SIWES

The SIWES is a skill training program designed to expose and prepare students for the
industrial work situation they are likely to meet after graduation. It is also a planned and
structured program based on stated and specific career objectives charged towards developing
the occupational competencies of participants [MAFE, 2009]. Consequently, the SIWES
program is a compulsory graduation requirement for Nigerian university student offering
certain courses.

1.7 IMPORTANCE OF SIWES TO ANIMAL AND ENVIRONMENTAL

BIOLOGY STUDENTS

 It enables students in animal and environmental biology to gain first-hand experience

working as a biologist.

 It helps students in animal and environmental biology apply their theoretical

knowledge to real life situations.

 It enables students with microscopic work with other biologists.

 It enables students in animal and environmental biology to witness the functioning

and organization of hospitals, laboratory and institutes.

3
1.8 ABOUT THE COMPANY

1.8.1 BRIEF HISTORY OF UNIVERSITY OF BENIN TEACHING HOSPITAL

From very humble beginnings as Midwest medical center in 1971, the University of
Benin Teaching Hospital has risen to become the second largest teaching hospital in the
country. The Midwest medical center was the brain child of hen Military governor of
Midwestern state (later Bendel and now Edo and Delta states), Late Brigadier General
Samuel Ogbemudia who established the center with Edict no.12 of 1971 within a year, it was
renamed University of Benin Teaching Hospital and officially opened to the public on the
12th of May 1973 by Her Excellency, Mrs. Victoria Gowon.

The University of Benin Teaching Hospital was taken over by the federal government
on April 1st, 1975 as the fifth teaching hospital coming after Ibadan teaching hospital and
Lagos teaching hospital.

From the inaugural board headed by late Dr. [Mrs.] Irene E. B. Ighodaro to the current
board headed by Bashorun Adedoja Adewuolu (MFR), the hospital has consistently
witnessed expansion in every area of health care service delivery, education and research,
The institution has been decisively responding to challenges as they arise, to the extent that
UBTH could boastfully say that it has effectively discharged its mandate.

For over forty years now, University of Benin Teaching Hospital, widely
acknowledges as the center of excellence has remarkably and effectively served as the last
port call for expert management for diverse and varied disease condition in Edo, Delta, part
of Kogi and Ondo state which largely form its catchment area and sometimes further away,
With an initial bed capacity of 360 when it officially opened may 12 th of 1973, the UBTH of
today boasts of a bed capacity of over 850 as at August 2017 and still increasing with its
multiplicity of departments offering a wide range of services of which combined to make the
institution the Center of Excellence as its stands today.

Today UBTH boasts of a dialysis center, CT scanning service, a fully integrated

accident center and a fully functional oxygen production plant to effectively meet the needs
of the hospital. In areas of training and research, UBTH is at the fore front in West Africa,
after 35 years of remarkable health care advances, UBTH will continue to spear head
innovation in research, clinical services and compassionate care.

4
1.8.2 MISSION STATEMENT OF THE HOSPITAL

"Working as a team to improve our clients’ health by the integration of consistent

quality care, education and research in a compassionate environment".

1.8.3 VISION STATEMENT OF THE HOSPITAL

"To be the leader in the providing quality health care solutions in West Africa"

1.8.4 DEPARTMENTS IN THE HOSPITAL

The hospital consists of the following clinical and diagnostics departments:

 Anesthesiology
 Cancer Registry
 Chemical Pathology
 Child Health
 Community Health
 Dental-Oral and Maxilla facial Surgery
 Dental-Oral Diagnosis
 Dental-Oral Medicine and Pathology
 Dental-Orthodontics Dentistry
 Dental-Periodontics Dentistry
 Dental-Preventive Dentistry
 Dental-Restorative Dentistry
 Dietetics and Nutrition
 Ear, Nose and Throat
 Family Medicine
 Hematology
 Histopathology
 Internal Medicine
 Medical Microbiology
 Medical Social Service
 Mental Health
 Morbid Anatomy
 Nursing Service
 Obstetrics and Gynecology

5
  Occupational therapy
 Ophthalmology
 Orthopedics and Traumatology
 Pharmacy and Pharmaceutical Services
 Physiotherapy
 Public Health Nursing
 Radiology
 Radiotherapy and Clinical Oncology
 Radiotherapy and Nuclear Medicine
 Surgery

OUT-PATIENTS CLINICS AND HEALTH CENTERS OF THE HOSPITAL

 Accident and Emergency Centre

 Achieving Nigerian Health Initiative (ANHI) Clinic
 Assisted Reproduction / In Vitro Fertilization[IVF]Centre
 Children Emergency Clinic
 Comprehensive Health Centre, Ogbonna
 Comprehensive Health Centre, Ugo
 Consultant Out-patient Clinic
 Dental Out-patient Clinic
 Female Maternity Clinic
 General Practice clinic
 National Health Insurance Scheme [Clinic]
 Presidents Emergency Plan for AIDS Relief [PEPFAR] Clinic
 South-South Zonal Pharmacovigilance Centre
 Stem Cell Transplant Centre
 Stroke Centre

1.9 ABOUT MEDICAL MICROBIOLOGY LABORATORY

1.9.1 A LABORATORY DEPARTMENT

6
 Medical Microbiology is a branch of medical science concerned with the diagnosis,
prevention, and treatment of infectious disease. In addition, this field of science studies
various clinical applications of microbes for the improvement of health. A medical
microbiologist studies the characteristics of pathogens, their modes of transmission,
mechanism of infection and growth. Using this information, a treatment can be devised.
Medical microbiologists often serve as consultants for consultants providing identification of
pathogens and suggesting treatment options.

1.9.2 KEY SERVICES

 Training of students from University of Benin Medical School

 Training of Resident doctors for National and West African Post graduate Medical
Colleges

 Training of Medical Lab Scientist interns.

 Training of Rotating Residents from other Pathology Departments, Community

Health, Internal Medicine, Ophthalmology and Family medicine

 Conducting antibiotic sensitivity and resistant test to drug companies.

SWAB

 Wound swab
 Ear swab
 Eye, vagina swab
 Endocervical swab

URINE

 MCS
 Microscopy
 Culture
 Sensitivity

CSF
 Microbiology
 Culture

7
  Sensitivity

STOOL
 Oval and parasites
 Stool Antigen Studies

BLOODCULTURE:
 Microscopy
 Culture
 Sensitivity

PARASITIES
 MP
 Skin smear for filarial
 Blood for Micro Filarial
 Vagina swab for T. vaginolis

TB LAB SERVICES:
 Sputum
 Aspirate
 Skin smear for AFB

GASTRIC AND ENDOSCOPIC BIOPSY FOR HELICOBACTER PYLORI

HIV
 Screening—Elisa
 Confirmation—PCR
 Monitoring—ViralloadandCD4Count.

INFECTIOUS DISEASE CLINIC

 Infectious Disease
 Infectious control service

8
 HEAD OF
DEPARTMENT

CONSULTANTS MOST SENIOR MOST SENIOR ADMINISTRATIVE CONFIDENTIAL

INFECTION MLS DLS STAFF SECRETARY
CONTROL NURSE

CHIEF
RESIDENT
ADLS
INFECTION CLERICAL
CONTROL OFFICERS
NURSE
CMLS
SSENIOR
REGISTRARS
ATTENDANTS
ACMLS / CLEANERS

JUNIOR
PMLS
REGISTRARS

SMLS

MLS

INTERN
MLS

Fig 1: Organogram of UBTH Medical Microbiology Department

CHAPTER TWO

INTRODUCTION TO LABORATORY EQUIPMENTS

9
2.1 LABORATORY EQUIPMENTS

Microbiology labs stand as pivotal spaces for unraveling the mysteries of

microorganisms, and the significance of appropriate equipment cannot be over stated in
achieving precise and efficient research outcomes.

Examples of some laboratory equipment used in a microbiology laboratory include:

 The Microscope: The microscope takes center stage, serving as the fundamental tool
for examining microorganisms at the cellular level. Its capabilities span from basic
observations using light microscopy to more intricate imaging through electron
microscopy.

 The Incubator: The incubator emerges as a critical asset, furnishing controlled

temperature and humidity settings necessary for cultivating and nurturing
microorganisms under specific growth conditions.

 Autoclave: The autoclave is an apparatus that harnesses high- pressure steam to

sterilize equipment, media, and waste, ensuring the maintenance of a septic conditions
and the safety of researchers.

 Centrifuge: It is used to rapidly sediment particles such as cells which may be

suspended in a fluid. The principle is that the centrifuge exerts a centrifugal force
which is greater than that of gravity and causes particles in the fluid to sediment. The
greater the centrifugal force, the faster and more effective is the dissemination.

 Refrigerator and Freezers: In the microbiology lab, culture media and bacteria
cultures are refrigerated to preserve the culture and prevent contamination. Some
samples and reagents are kept frozen to ensure that the antigenic properties of the
samples and the potency of the reagents are retained.

 Glass and Plastic wares: A number of glass wares and plastic wares are required in
the microbiology laboratory. The use of clean, unbroken glass ware is one of the many
factors that ensure accurate results in laboratory investigations. Some examples of
glass ware include test tubes, centrifuge tubes, measuring cylinders, volumetric
pipettes, and Pasteur pipettes, amongst others. Some examples of plastic ware include
tube racks, specific containers, wash bottles, translucent and amber screw cap reagent
bottles.

10
2.2 LABORATORY SAFETY AND PRECAUTIONS

A laboratory is a building where all scientific works are carried out. However,
accidents cannot be totally ruled out during such activity but can be minimized if the
laboratory rules and safety precautions are strictly adhered to. The following are salient rules
that govern all laboratory processes:

 Appropriate laboratory outfit (Personal Protective Equipment) should be worn

before performing any laboratory work.
 Ensure to locate the first aid box, fire extinguisher, and power supply in case of
emergency when entering any laboratory.
 Chemicals kept on shelves or bench tops should be well labeled for easy
identification.
 Always read the specifications written on any reagent bottle before taking any part
of it for preparation.
 Always wash your hands before and after any laboratory work.
 Always ask your supervisor about anything in the laboratory that you do not
understand.
 Eating and drinking are prohibited in the laboratory.
 Do not attempt an unauthorized experiment.
 Clean laboratory surfaces and benches before and after work
 Avoid pipetting any solution with your mouth. Use the dropping pipette or pipette
sucker method.
 Always switch off all laboratory electrical appliances after use.
 Shut all water taps after use.
 Remember to check all sections of the laboratory before closing for the day.
 Ensure to understand and take note of all laboratory safety symbols.

2.3 LABORATORY WASTE MANAGEMENT

Laboratory waste management refers to the proper handling, storage, treatment, and
disposal of waste generated in laboratory settings.

Some recommended laboratory waste management practices are highlighted as follows:

11
 Source segregation: This involves separating waste at the point of generation. E.g.,
separate bins can be used for recyclables (paper, plastic), organic waste (food scraps,
garden waste), and non-recyclable / non-biodegradable waste, etc.

 Containment and Storage: Laboratory waste should be well placed in leak-proof,

puncture resistant, and appropriately labeled containers. The containers, afterward,
should be stored in secure, designated areas to prevent accidental spills or access by
unauthorized individuals.

 Autoclaving: Prior to disposal, bio hazardous waste, such as cultures, stocks, or

contaminated equipment should be autoclaved at appropriate temperature and
pressure settings to ensure microbial inactivation.

 Incineration: It involves the controlled combustion of waste materials at high

temperatures, in specialized incinerators designed for this purpose.

 Waste disposal: Licensed waste management companies should be contacted for

transportation and proper disposal of bio hazardous, chemical or other regulated
waste.

CHAPTER THREE

12
3.0 BACTERIA STRUCTURE AND FUNCTION

Bacteria are microscopic unicellular organisms that exist in millions in every

environment both inside and outside other organisms. The structure of bacterial is known for
its simple body design. Bacteria are single- celled microorganisms with the absence of the
nucleus and other cell organelles; hence, they are classified as prokaryotic organisms. They
are also very versatile organisms, surviving in extremely in hospitable conditions.

3.1 EXTERNAL STRUCTURES OF BACTERIA

These structures protrude from the cell into the surrounding environment, these include:

 The Flagella: they are slender filaments that originate from the cytoplasm. They
function as organs of motility. They are too thin to be seen in ordinary stained
preparations but becomes visible only in silver impregnation preparation or in
electron microscopy

 The Pili: extruding from the cytoplasmic membrane, are shorter and finer filaments
than the flagella, these are the Pili. Majority of them are referred to as common Pili or
Fimbriae. They are organs of adhesion that help bacteria attach to host cells.

 Capsule: This is a layer of loose sliming material which surrounds some bacteria cell.
They are composed mainly of polymers or polysaccharides. They are identified by
negative staining due to their low affinity for simple stain.

 Cell wall: This is a prominent distinguishing feature of the bacteria cell, with a
rigidity and strength that protects the cellular content. The chemical constituents of
the cell wall (peptidoglycan) which gives rigidity and shape to the organism.

 Cytoplasmic membrane: This is a double layered structure within the cell wall. It is
composed of lipid and protein and acts as a semi- permeable membrane through
which nutrient pass into and out of the cell wall.

3.2 INTERNAL STRUCTURES OF BACTERIA

 Mesosomes: They are convoluted in variations within the cytoplasmic membrane.

They play an important role during cell division and the secretion of certain enzymes.

13
  Nuclear materials: within the cytoplasmic membrane, the cell itself has the nucleus
which has no nuclear membrane and lacks a definite shape. It is a single circular gland
of the deoxyribonucleic acid (DNA) which acts as a nucleus (chromosome).

 Ribosome: They are located throughout the cytoplasm and acts as the site of protein
synthesis. They are important for conveying genetic code of the nucleus into
instructions in the manufacture of cellular components. They are composed of
ribonucleic acid and protein.

 Cytoplasmic inclusions: These are seen in some bacteria and appear to be sources of
reserved food for energy.

 Spores: These are dense structures produced by the bacteria that enable them to
survive adverse conditions. They develop within and at the expense of the vegetative
cell. They are resistant to heat, stains, desiccation, and disinfectants.

3.3 MORPHOLOGY OF BACTERIA

The morphological classification of bacteria is based on the following types of shapes of

cells:

 The Spheroid or Coccus: They are round or oval bacteria. The bacteria multiply by
binary fission. During multiplication, the daughter cell is attached to the parent cell
but it gets detached before fission occurs again. Cocci in pairs are called diplococcic
e.g., meningococci and gonococci. If fission continues while they remain attached,
chains of cocci are formed, and are referred to as streptococci. If the division is not in
one plain and clusters of cocci are formed randomly, they are known as
staphylococci. Tetrads (ortetracocci) are formed when the cocci remain in pairs for
two consecutive divisions and form regular aggregates off our cocci.

 Bacillus: They are stick–like or rod-like bacteria. They have rounded or swollen ends.
Short rods with rounded ends are called coccobacilli. If the bacilli remain attached
after division, they are known as streptobacilli. Lactobacilli are bacilli that form
branching chains.

 Vibrio: They are small slightly curved rods. They are motile with single flagellum at
one end.

14
  Spirilla: They are small, regularly coiled rigid organisms. They are motile with
groups of flagella at both ends.

 Spirochaete: They are flexible, coiled, motile organisms. They have rapid body
movement. They are not easily stained by gram technique.

3.4 PREPARATION OF SMEARS

Smears can be made from liquid or solid cultures or from clinical specimens.

Preparation of smears from solid media

 Sterilize the wire loop in Bunsen flame.

 Place one drop of sterile saline on a grease-free clean slide with the sterilized loop.

 Re-sterilize the loop.

 With the wire loop, pick a small portion of bacterial growth and emulsify it in the
drop of saline and spread it to give a thin homogenous film or smear on the slide.
Sterilize the loop.

 Allow the smear to air dry.

Preparation of smears from liquid media

 Sterilize the wire loop in the Bunsen flame.

 Using a septic technique, remove a loopful of the culture.

 Place the culture on a clean slide and spread it with the loop to give a fairly thick film
of culture. Sterilize the loop.

 Allow the film to air dry.

3.5 FIXING OF BACTERIAL SMEARS

The smear having been made and allowed to dry is fixed quickly by passing it two or
three times over a Bunsen flame to a temperature of about 55 to 60°c. This is to coagulate the
bacteria proteins which make them adhere to the slide. Fixation should be such that the
bacterial morphology remains intact, preventing the smear from being washed off during
staining. Overheating will lead to distortion of morphology.

15
3.6 STAINING OF BACTERIA

Stains are mostly salts, comprising a base and an acid. They are classified in to acid,
basic and neutral stains. Basic stains have coloring substance contained in the basic radical
with the acid radical being colorless while those stains in which coloring substance are in the
acidic radical and base components being colorless are acid stains. When coloring substance
is contained in both acid and base components it is neutral stains.

The bacteria cell, which is rich in nucleic acid, has affinity for basic stains and so is stained
by basic stain.

3.7 TYPES OF STAINING

 Indirect Staining: this is when an organism is stained only in the presence of a

mordant e.g., Grams stain.

 Direct Staining: this is a simple one- step staining procedure in which the presence
and morphology of bacteria are demonstrated.

 Negative Staining: this is when the organism remains unstained against a stained
background. This is one of the few methods where acid stains such as nigrosin or
Indian ink are used.

 Metachromatic Staining: in this staining method the organism or part of the

organism is stained a different shade of color from that of the stain.

STAINING METHODS

 Simple Stains: this makes use of the direct type of staining. It used to demonstrate the
presence and the morphology of bacteria.

 Differential Stains: this staining method divides bacteria into two groups.

 Special Stains: these are specialized staining methods to demonstrate certain bacterial
components e.g., spore.

DIFFERENTIAL STAINING METHODS

16
 1. Gram Staining: In 1884, Christian Gram, a Danish bacteriologist, described
this staining method. It divides bacteria into two groups: Gram-positive and
Gram-negative bacteria. The gram-positive has an intact cell wall which the
negative does not. Their action is based on the ability of the organism to resist
decolorization with acetone alcohol aniline oil after the initial staining and
treatment with a mordant.

Gram staining method involves four steps:

 Primary staining: a primary stain such as crystal violet, methyl violet blue
and gentian violet stains all the bacteria in the smear.

 Mordanting: application of a mordant such as Lugols iodine results in the

formation of a complex with the primary stain.

 Decolorization: some bacteria resist the removal of the dye-iodine complex

with decolorizing agent and retain the primary stain due to the presence of
teichoic acid in their cell wall which appears blue or deep purple in coloration
after staining. These are called Gram-positive bacteria. On the other, Gram-
negative bacteria get decolorized with the decolorizing agent due to the
presence of high lipid content, giving are decoloration after staining.
Decolorizing agents include acetone alcohol, 95% ethanol, and methylated
spirit.

 Counter staining: a counter stain, contrasting in color with the primary stain,
is used to stain Gram negative bacteria which are decolorized in the third step.
Examples of counter staining include neutral red, safranin and dilute carbol
fuschin.

PROCEDURES

 Degrease the slide by passing through flame.

 Sterilize wire loop by passing through the flame.
 Label the slide appropriately.
 Make a smear; allow drying and then fixing with gentle heat by passing the slide
two or three times over a Bunsen flame.
 Stain with crystal violet for one minute. Wash with tap water.
 Flood the slide with iodine for one minute then wash with tap water

17
  Decolourise with acetone oral cohol until no more colour appears to ooze out of
the smear (about 2 seconds for acetone and 1- 2 minutes for alcohol). Wash
immediately with tap water.
 Counter stain with neutral red or safranin or dilute carbol fuschin for one minute.
Wash with tap water.
 Place the slide in a draining rack to air dry.
 Examine microscopically with oil immersion.

RESULT

Gram-positive bacteria: Blue-black/Violet

Gram-negative bacteria: Red

2. Acid-Fast Stain: Acid- fast staining divides bacteria into two groups: acid-fast
and non-acid fast. Members of the genus Mycobacterium are acid-fast in
nature. Mycobacteria have high lipid content, especially mycolic acid, in their
cell wall. Ordinary aniline dye solutions cannot penetrate the mycobacterial
cell wall. Strong staining solutions containing phenol preferably with the
application of heat are used for staining them. Once stained, they resist
decolorization with mineral acid. Therefore, they are called Acid-Fast Bacilli
(AFB).

Acid-fast staining is performed in 3 steps:

 Primary staining: a strong primary stain containing phenol is applied, usually with
heat, to penetrate the cell wall. Carbol fuchsin is commonly used.

 Decolorization: 20% sulphuric acid is used to decolorise non acid-fast bacteria and
other cells. The acid-fast bacteria uses 3% alcohol which resist decolorization and
retain the color of the primary stain.

 Counter staining: decolorized non acid-fast bacteria are counter stained in a contrast
color (blue or green) to differentiate them from the acid-fast bacteria.

PROCEDURES

18
  Degrease the slide by passing through the flame.

 Sterilize the wire loop by passing it through the flame.

 Label the slide appropriately.

 Make a smear, allow it to dry, and then fix with gentle heat by passing the slide two or
three times over a Bunsen flame.

 Flood the slide with the carbol fuchsin solution and heat gently until steam rises. Do
not allow stain to boil. Stain for 5-10 minutes. Wash off stain with tap water.

 Decolorize with 3% alcohol or 20% sulphuric acid for 3minutes. Wash with tap water.

 Counter stain with 0.5% methylene blue or malachite green for 1 minute. Wash off
stain with tap water.

 Place the slide in a draining rack to air dry.

 Examine microscopically with oil immersion.

RESULT

 Acid-alcohol fast bacteria: Red

 Background and other bacteria: Blue or Green.

CHAPTER FOUR

4.0 CULTURE MEDIA

19
 To study and identify microorganisms, they have to be cultivated and isolated in pure
culture. Most bacteria can be cultured in artificial media if the media meet the nutritional and
physical growth requirement of the bacteria. Culture media are designed to support the
growth, identification, transport, and isolation of microorganisms. There are two categories of
culture media, chemically defined or simple medium, and chemically undefined or complex
medium. A defined media is referred to as a medium having a known concentration of
ingredients like sugar and nitrogen.

4.1 CLASSIFICATION OF CULTURE MEDIA

The classification of culture media is based on consistency, nutritional component,

and application or chemical composition.

4.1.1 Based on Consistency

This classification depends on the amount of solidified materials (agar or gelatin)

present in the media.

 Solid media: These are used to grow microorganisms in their full physical form.
They are used to prepare bacteria pure cultures and also to isolate bacteria to study
colony characteristics. They are free from growth-inhibiting substances. Bacteria
growth can appear as mucoid, round, smooth, rough, filamentous, irregular and
punctiform. Examples of these media include blood agar, nutrient agar, MacConkey
agar, chocolate agar, etc.

 Semi-solid media: It is mainly used to study the motility of microorganisms. It

distinguishes motile and non-motile organisms through U-tube and Craigie’s tube. It
has 0.2-0.5% of agar concentration. It appears like a soft jelly-like substance due to
reduced agar concentration. They are used mainly as transport media and biochemical
tests. Examples of these media include Hugh and Leifosons oxidation fermentation
medium, Stuarts and Amies media, Mannitol motility media.

 Liquid media: They are called broth. They allow uniform and turbid growth of
bacteria strain. They are used for biochemical testing, blood culture, and motility
testing and enrichment media. They lack solidifying agents. The large growth of
bacteria colonies can be observed. Examples of these media include Alkaline peptone
water, Nutrient broth, Tryptic Soy broth, and Cary Blair medium.

20
4.1.2 Based on Nutrient

This classification is divided into three:

 Simple or Basic media: This is a general-purpose media that support the growth of
non- fastidious microbes; it is primarily used for the isolation of microorganisms.
Examples include nutrient agar, nutrient broth, and peptone water.

 Complex media: they are media containing nutrients in unknown quantities that are
added to bring about a particular characteristic of a microbial strain. Examples
include: Tryptic soy broth, Blood agar, Nutrient broth.

 Synthetic agar: They are chemically defined media produced from pure chemical
substance. It is generally used in scientific research. An example is Czapek Dox
medium.

4.1.3 Based on Application

 Basic media: They are simple media with carbon and nitrogen source. They boost the
growth of microorganisms. They are also known as general purpose media. They are
non- selective media suite for growing Staphylococcus and Enterobacteriaceae.
Examples include nutrient broth, nutrient agar, and peptone water.

 Enriched media: They are used to grow fastidious microorganisms. They require
additional substances like blood, serum, or egg yolk in the basic medium. It is used
for specimens collected from sites that are normally sterile to ensure the rapid
multiplication of a pathogen. Examples include Chocolate agar, Blood agar, Loeffler’s
serum slope.

 Selective media: These are solid media that allow the growth of selected microbes
and inhibit the growth of others. They are used to isolate microorganisms. Selective
growth of microbes is determined by adding substances like antibiotics, dyes, bile
salts or by pH adjustments. They are used on sites having normal flora to prevent
unwanted contaminants. Examples include Salmonella-Shigella agar, Wilson and
Blair’s agar, MacConkey agar, and Mannitol salt agar.

 Enrichment media: These are liquid media used to increase the relative of certain
microbes before culturing them on a solid medium plate. They inhibit the growth of
commensal species of microorganisms. They are used in isolating fecal and soil

21
 microorganisms. Examples include Selenite F broth (for isolating Salmonella typhi
from fecal sample), Tetrathionate broth, and alkaline peptone water.

 Differential media: These media contain indicators like dyes (neutral red, phenol red,
methylene blue), or metabolic substrates that give different colors to colonies of
different microbial species. It allows the growth of more than one microorganism.
Examples include blood agar, Mannitol salt agar, MacConkey agar, TCBS agar.

 Transport media: They are useful for clinical specimens that are required to be
transferred immediately to labs in order to maintain the viability of potential
pathogens and to prevent over growth of commensals or contaminating
microorganisms. They are semi solid in consistency. Transport media ensure the
survival of both aerobic and anaerobic pathogens. Examples include Pikes medium,
Cary Blair’s transport and Venkahalan Ramakrishana media, and Amies transport
medium.

4.2 COMPONENTS OF CULTURE MEDIA

 Water

 Peptone

 Meat extract

 Yeast extract

 Mineral salt, and

 Carbohydrate

4.3 CHOICE OF CULTURE MEDIA

The choice of culture media use in a medical microbiology laboratory depends on:

 Major pathogens to be isolated, their growth requirements, and features by which they
are recognized

 Whether the specimen being cultured are from sterile sites or from sites having a
normal microbial flora.

4.4 METHODS OF CULTURING

 Streak method

22
  Lawn method

 Pour plate method

4.5 BIOCHEMICAL TESTS

Biochemical tests are used to identify bacteria species by differentiating them on the
basis of biochemical activities. These biochemical tests include:

4.5.1 CATALASE TEST

This test is used to differentiate those bacteria that produce the enzyme catalase, such
as Staphylococci from non-catalase-producing bacteria such as Streptococci.

PRINCIPLES OF CATALASE TEST

Catalase acts as a catalyst in the breakdown of hydrogen peroxide to oxygen and

water. Organism is tested for catalase production by bringing it in contact with hydrogen
peroxide. The production of catalase is indicated by the presence of oxygen bubble.

PROCEDURES

 Pour 2-3 ml of the hydrogen peroxide solution into a test tube

 Using a sterile wooden stick, or a glass rod, remove several colonies of the test
organisms and immense in the hydrogen peroxide solution

 Look out for immediate bubbling.

RESULT

Active bubbling= Positive catalase test

No bubbling= Negative catalase test

Organisms in Positive Catalase Test

 Micrococcus sp.
 Norcardia sp.
 Pseudomonas
 E.coli
 Staphylococcus sp.
 Heplicobacter pylori

23
Organisms in Negative Catalase Test

 Enterococcus sp.

 Streptococcous sp.

 Kingella kingae

4.5.2 COAGULASE TEST

The test to identify Staphylococcus aureus which produces the enzyme Coagulase

PRINCIPLES OF COAGULASE TEST

Coagulase causes the plasma to clot by converting fibrinogen to fibrin. Two types of
coagulase are produced by most strains of S. aureus; Free coagulase and Bound coagulase.

Free coagulase: It converts fibrinogen to fibrin by activating a coagulase reacting factor

present in plasma.

Bound coagulase: It converts fibrinogen to fibrin directly without a coagulase reacting

factor. It is detected by the clumping of bacterial cells in the rapid slide tests.

PROCEDURES OF COAGULASE TEST

1. Slide Test Method (Detects Bound Coagulase)

 Place a drop of distilled water on each end of a slide or on two separate slides.

 Emulsify a colony of the test organism in each of the drop to make thick suspensions.

 Add a loopful of plasma to one of the suspensions and mix gently.

 Look out for clumping of the organisms within 10seconds.

RESULT

Clumping within 10secs= S.aureus

No clumping within 10secs= No bound coagulase

2. Tube Test Method (Detects Free Coagulase)

 Take three small test tubes and label in this order: T= Testorganism (18-24hrs.broth
culture), Pos= positive control (18-24hrs.S.aureus broth culture) and Neg= Negative
control (Sterile broth).

24
  Pipette 0.2ml of plasma into each tube.

 Add 0.8ml of the test broth culture to tube T.

 Add 0.8ml of S.aureus culture to tube Pos.

 Add 0.8ml of sterile broth to tube Neg.

 After mixing gently, incubate the three tubes at35-37oc.

 Examine for clothing after 1hr, if no clotting has occurred, examine after 3hrs. If the
test is still negative, leave the tube at room temperature overnight and examine again.

RESULT

Clotting of tube content= Coagulase Positive test

No clotting of tube content= Coagulase Negative test

Organisms in Coagulase Positive Test

 Staphylococcus hyicus

 Staphylococcus delphini

 Staphylococcus schleiferi

 Staphylococcus intermedius

 Staphylococcus aureus

Organisms in Coagulase Negative Test

 Staphylococcus epidermidis

 Staphylococcus saprophyticus

 Staphylococcus wameri

 Staphylococcus hominis

 Staphylococcus caprae

4.5.3 CITRATE UTILIZATION TEST

It is used in the identification of citrate utilization species from those that cannot. It is
based on the ability of an organism to use citrate as its only source of carbon. It is based on

25
the ability of certain bacteria to produce the enzyme citrate permease which enables the
uptake of citrate from the environment.

PRINCIPLES OF CITRATE UTILIZATION TEST

Citrate permease is capable of converting citrate to pyruvate. Citrate agar is used to

test an organism's ability to utilize citrate as a source (NH 4H2PO4) as the sole source of
nitrogen. Bacteria metabolize citrate; ammonium salts are broken down to ammonia. This
increase alkalinity which turns the bromothymol blue indicator in the medium from green to
blue. Christensen citrate test medium does not require organism to use citrate as sole carbon
source. It contains both peptone and cysteine.

CITRATE USING SIMMONS CITRATE AGAR

PROCEDURES

 Prepare slope of the medium in bijou bottles and store at 2-8°c.

 Using sterile straight wire, first streak the slope with a saline suspension of the test
organism and stab the butt.

 Incubate at 35°c for 48hrs.

RESULT

Bright blue= Positive Citrate test

No change in color= Negative Citrate test

Positive Citrate Test Organisms

 Salmonella sp.

 Proteus mirabilis

 Klebsiella sp.

 Enterobacter aerogenes

 Citrobacter freundii

 Providencia sp.

Negative Citrate Test Organism

26
  Escherichia coli

 Shigella sp.

 Yersinia pestis

 Morganella morganii

 Staphylococcus aureus

 Streptococcus pyogene

4.5.4 UREASE TEST

It is used to identify organisms capable of producing the enzyme urease which catalysis the
hydrolysis of urea into ammonia and carbon dioxide. It is used in differentiating between
organisms within the family of Enterobacteria. Proteus strains are strong urease producers,
Yersinia enterocolitica are weakly urease producers and Salmonella-Shigella do not produce
urease.

PRINCIPLES OF UREASE TEST

Organism is cultured in a medium which contains urea and phenol red indicator. When strain
is urease producing, urease break down urea to give ammonia and carbon dioxide. Medium
becomes alkaline at the release of ammonia. Color changes to pink-red from light orange.

CHRISTENSEN MODIFIED UREA BROTH METHOD

Urea agar was developed by Christensen in 1946 for differentiation of enteric bacilli.

PROCEDURES

 Inoculate heavily the test organism in a bijou bottle containing 3ml sterile Christensen
modified urea broth.

 Incubate at 35-37°c for 3-12hrs

 Look out for a pink color in the medium.

RESULT

Pink colour= Urease Positive test

No pink colour= Urease Negative test

27
Urease Positive Organisms

 Proteus sp.

 Helicobacter pylori

 Yersinia sp.

 Klebsiella pneumoniae

 Morganella morganii

Urease Negative Organisms

 Escherichia coli

 Salmonella sp.

 Shigella sp.

 Pseudomonas aeruginosa

 Staphylococcus aureus

 Streptococcus pyogenes

4.5.5 INDOLE TEST

The indole test for indole production is used in differentiating Gram-negative bacilli
essentially the identification of enterobacteria.

PRINCIPLES OF INDOLE TEST

The test organism is cultured in a medium containing tryptophan. Indole production is

detected by Kovacs or Ehrlich’s reagent which contains 4-(dimethylamino) benzaldehyde.
This reacts with the Indole to produce a red-colored compound. Kovacs reagent is
recommended in preference to Ehrlich’s reagent for the detection of Indole from
enterobacteria.

PROCEDURES OF INDOLE TEST

 Grow the organism in peptone water overnight

 If using Ehrlich’s reagent, extract the indole by shaking the culture with equal volume
of xylene, ethyl ether or petroleum ether.

28
  Add about 1ml of Ehrlich’s reagent.

 Observe a red rose ring between the layer of ether and the peptone water.

 If using Kovacs reagent, add a few drops of the reagent to the overnight peptone water
culture.

 Observe a red ring above the peptone water. The red ring indicates positive indole
production.

RESULT

Presence of red ring= Indole Positive test

No changes= Indole Negative test

Indole Positive Organisms

 Aeromonas hydrophila

 Aeromonas punctata

 Bacillus alvei

 Edwardsiella sp.

 Escherichia coli

 Flavobacterium sp.

 Haemophilus influenzae

 Klebsiella oxytoca

 Proteus sp.

Indole Negative Organisms

 Actinobacillus sp.

 Aeromonas salmonicida

 Bacillus sp.

 Bordetella sp.

 Enterobacter sp.

29
  Haemophilus sp.

 Klebsiella sp.

 Neisseria sp.

 Proteus mirabilis

4.6 ANTI-MICROBIAL SENSITIVITY TEST

An antibiotic is an anti-microbial agent that is derived from a microorganism. The

term anti-microbial agent is used for a drug that acts primarily against infectious
microorganisms. It is derived from a natural source or may be a synthesized chemical.

Anti-microbial drugs differ in the range of bacteria they inhibit. Broad spectrum drugs
such as Ampicillin and Tetracycline are effective against both Gram-positive and Gram-
negative bacteria, whereas Flucloxacillin has an arrow spectrum and is active against Gram-
positive bacteria such as Staphylococcus aureus, but in effective against most Gram-negative
bacteria. It is preferable to use an arrow spectrum drug for a known pathogen because it does
not affect the composition of the normal flora at various sites. A broad-spectrum drug may
have an adverse effect on the normal flora.

4.7 MODE OF ACTION OF ANTI-MICROBIAL AGENTS

There are five main ways in which antimicrobial drugs act on bacteria. They include:

 Inhibition of cell wall synthesis: some drugs inhibit the cell wall synthesis of
bacteria by preventing the cross-linking of the polysaccharide chains in the
peptidoglycan layer of the cell wall, e.g., Penicillin, Cephalosporins and Vancomycin.

 Damage to cell membrane: some anti-microbial agents act as cationic detergents and
bind to the cell membrane. The binding result is the loss of semi-permeability of the
membrane, leading to the loss of cell contents and cell death, e.g., the Polmixins and
Amphotericin B.

 Inhibition of protein synthesis: such drugs arrest protein synthesis by affecting the
translation in the cell, e.g., Chloramphenicol, Aminoglycosides, Tetracycline,
Erythromycin, Lincomycin and Clindamycin.

30
  Inhibition of nuclei acid synthesis: these anti-microbial agents prevent
multiplication in bacteria, e.g., Quinolones and Nalidixic acid inhibit DNA replication
and Rifampicin inhibits RNA synthesis.

 Inhibition of folic acid synthesis

4.8 ANTI-MICROBIAL DRUGS OF CLINICAL IMPORTANCE

 Penicillin

 Cephalosporins

 Cotrimoxazole

 Aminoglycosides

 Tetracyclines

 Chloramphenicol

 Erythromycin

 Lincomycins

 Nalidixic acid

 Nitrofurantoin

 Vancomycin

 Polymixins

 Metronidazole

 Rifampicin

 Fusidic acid

 Anti-fungal agents (Amphotericin B, Nystain and Griseofulvin)

4.9 DETERMINATION OF INHIBITORY ACTIVITY OF ANTI-MICROBIAL

AGENTS

KIRBY-BAUER’S DISC DIFFUSION TEST

31
 In Kirby-Bauer method, the inhibition zone diameters of the test organisms are
compared with those of the control strains with reference to the corresponding minimum
inhibitory concentrations and interpretive guidelines published by the National Committee for
Clinical Laboratory Standards (NCCLS).

MATERIALS NEEDED FOR ANTI-MICROBIAL SUSCEPTIBILITY TESTING

 Muller Hilton Agar (MHA)

 Antibiotics disk

 Swab stick

 Test organism solution

 CLSI guideline (Clinical and Laboratory Standard guideline Institute which select
antibiotics for the infection).

Procedures for Anti-microbial Susceptibility Testing

 Prepare a bacteria solution which must be equal to 0.5% MacFarland standard.

 Select antibiotics according to CLSI guideline

 Dip a sterile cotton swab into the inoculum, squeeze off excess fluid against the side
of the bijou bottle, and make a lawn on the agar plate without touching the edge, then
go round the edge once and discard the swab stick.

 Allow 3-5 minutes, for the surface of the agar to dry before adding the anti-microbial
discs.

 Place the antibiotics discs on the surface of the agar, using either forceps or
dispensers. The discs should be placed at 22mm from each other and at 15mm from
the edge of the plate. 90mm diameter plate should contain no more than 6 antibiotics
discs while the one of 120mm not more than 9.

 Gently press each disc on the agar using forceps to provide uniform contact with the
surface.

 Incubate the plate at 37°c for 16-18 hours.

32
RESULT

At the end of 16-18 hours incubation, measure the inhibition zone. Measure the diameter of
the inhibition zone from one edge to the opposite edge. The zones are measured to the nearest
millimeter as the complete inhibition of growth appears to the unaided eye.

INTERPRETATION OF RESULT

Interpretative standards are provided by the CLSI guideline as Susceptible, Intermediate and
Resistant according to the inhibitory zone size for each antimicrobial.

4.10 URINE MICROSCOPY, CULTURE AND SENSITIVITY

The bladder and urinary tract are normally sterile. The urethra however may contain
some few commensals and also the perineum which can contaminate urine when it is being
collected. With female patients, the urine may become contaminated with organisms from the
vagina. Vaginal contamination is often indicated by the presence of epithelial cells and a
mixed bacteria flora.

4.11 COLLECTION AND TRANSPORT OF URINE SAMPLE

Give the patient a sterile, dry and clearly labeled container for collection of urine sample.

It depends on the type of process or target of the urine.

For Urinary Tract Infection (UTI): a clean catch-midstream urine sample is collected. To
collect sample from female patient, clean the area around the urethra opening with clean
water, dry the area and collect the urine with the labia held apart. For male patient, wash the
hands before collecting the specimen.

For a patient with terminal hematuria, terminal urine sample is sample is collected. For
gonococcal urethritis, first void urine sample is collected.

Once a sample is taken, immediate transfer to the laboratory is pivotal. When there is a delay
in transport acid, boric acid is added for preservation of the urine. Specimen with boric acid
need not be refrigerated.

33
4.12 URINE PROCESSING IN THE LABORATORY FOR URINARY TRACT
INFECTION

It follows a three-day microscopy, culture and sensitivity:

Day One

 Macroscopy of urine: In macroscopy,

1. Check for color, which can be greenish or reddish.

2. Also check if the urine is clear or cloudy.

 Urinalysis: In urinalysis, the urine test strip (dipstick) is inserted into the urine
and wait for few minutes for the color indicator to appear. The dipstick checks
for Nitrite, Leukocyte, Esterase, pH, Glucose, Protein, etc.
 Microscopy (Direct Wet Mount):

 Make a smear of the urine on a slide and cover with a coverslip.

 View under x10 or x40 magnification.

 Pus cells, red blood cells, yeast cells, epithelial cells, casts, sperm cells, oval of
Schistosomia hematobium are found in wet mount

 Direct Gram Staining: White blood cell and bacteria can be seen in direct gram
staining.
 Culturing: The type of culture media to be used is determined by the direct gram
staining.

Day two

 Plate reading
 Indirect gram staining
 Motility test if necessary
 Setup biochemical test using citrate, peptone water and urease
 Setup anti-microbial susceptibility test

Day three

 Read the biochemical test and the anti-microbial susceptibility test

34
  Report writing

CHAPTER FIVE

5.0 CONCLUSION

Industrial Training is a very important platform to introduce respective university

students to their various fields and it should be embraced and not taking for granted by using
the time met to get industrial experience to pursue various personal interests.

5.1 EXPERIENCE GAINED DURING MY INDUSTRIAL TRAINING

 I benefited a lot during my industrial training by been exposed to different work

methods.
 I learnt how to handle equipment in the laboratory that is not available in school.
 I visited the Intensive Care Unit (ICU), alongside my supervisors to collect samples
from patients.

35
  I took part in different seminar presentations given by the unit which enhanced my
public speaking skills.
 I was opportune to attended health conference and seminars where renowned health
personnel gave several insightful presentations and views in the health sector.

5.2 CHALLENGES ENCOUNTERED DURING MY INDUSTRIAL TRAINING

I didn’t experience much challenge other than trying to get acquainted to the
environment, medical terms and also different personalities like the doctors, my colleagues
and also some patients but aside that it was an amazing experience.

5.3 RECOMMENDATION

 It is recommended that the students partaking in the SIWES program must be

supervised by ITF and the school supervisors.
 A comprehensive report from the organizations or establishments where students do
their SIWES program be forwarded to both the school and ITF.
 The government should educate various establishments in the country about the need
of admitting students for industrial training.
 Orientation should be given to students on finding a placement in industries.
 All companies and industries should make it a priority to provide housing for their IT
students. This will reduce student’s idleness, reduce expenses and increase their
timeliness at work
REFERENCE

Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries (Part2).

Cambridge University Press.

Chiodini, P. L., Moody, A. H., & Manser, D. W. (2018). Atlas of Medical Helminthology and
Protozoology. CRC Press.

Eneanya, C. I., Okoli, N. J., Edeoga, H. O., & Otagburuagu, V. C. (2019). Significance of
industrial training in the microbiology curriculum. Journal of Advances in Medical
and Pharmaceutical Sciences, 21(4), 1-8.

Forbes, B. A., Sahm, D. F., & Weissfeld, A. S. (2007). Bailey & Scott's Diagnostic
Microbiology (12thed.). Mosby.

36
Garcia, L.S. (2009). Diagnostic Medical Parasitology (5 th ed.). American Society for
Microbiology.

Gerhardt, P., Murray, R. G. E., Costilow, R. N., Nester, E. W., Wood, W. A., Krieg, N. R., &
Phillips, G. B. (1981). Manual of Methods for General Bacteriology. Washington:
American Society for Microbiology.

Hugonnet, S., & Pittet, D. (2000). Sterilization methods: Radiation. Journal of Hospital
Infection, 46 (Supp l2), S29-S33.

Koneman, E. W., Allen, S. D., Janda, W. M., Schreckenberger, P. C., & Winn, W. C. (2015).
Color Atlas and Textbook of Diagnostic Microbiology (7 th ed.). Lippincott Williams &
Wilkins.

APPENDIX

LABORATORY EQUIPMENT

MICROSCOPE INCUBATOR AUTOCLAVE

37
CULTURE MEDIA

CULTURE MEDIA

Solid media Semi solid media

38
 BIOCHEMICAL TEST

Coagulase test
39

Urease test
 Catalase test

Citrate test

Indole test
URINE MICROSCOPY, CULTURE AND SENSITIVITY

40
Urine m/c/s

41
MORPHOLOGY OF BACTERIA

Bacteria morphology

42
 STAINING REACTION

1. GRAM STAINING PROCEDURE

2. ACID-FAST STAINING PROCEDURE

43
3. MICROSCPIC VIEW OF GRAM-POSITIVE AND GRAM-NEGATIVE
BACTERIA

ANTI-MICROBIAL SENSITIVITY TEST

Antimicrobial discs on inoculated MHA

plate

44
 PERSONAL PROTECTIVE EQUIPMENT

LAB COAT HAND GLOVES

SAFETY EYE GOGGLE

SAFETY BOOTS

45