TECHNICAL REPORT OF STUDENT

INDUSTRIAL WORK EXPERIENCE SCHEME

(SIWES)

DONE AT

DEGRILLS INTEGRATED SERVICES

20, Adeleke Street, Gbagada, Lagos State

BY

NWABACHILI OKECHUKWU MICHAEL

MATRICULATION NO: 20181085797

DEPARTMENT OF BIOTECHNOLOGY

SCHOOL OF BIOLOGICAL SCIENCES

FEDERAL UNIVERSITY OF TECHNOLOGY, P.M.B. 1526,

OWERRI, IMO STATE

IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR

THE AWARD OF BACHELOR OF TECHNOLOGY (B.Tech.)
DEGREE IN BIOTECHNOLOGY

AUGUST, 2021

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 DEDICATION
This work is dedicated to almighty God for his guidance and protection towards
me and my family.

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 ACKNOWLEDGEMENT
With a deep sense of appreciation, respect and gratitude, I want to say a big
thank you to my parents, brother, sisters, and other relatives and non-relative for
their caring attitude and support from the beginning of my pursuit for B. Tech in
Bioctechnology to this point. I also want to express my appreciation to the
management and workers of Degrills Integrated Services for their intellectual
support during our work together. Not forgetting my other IT colleagues. My
sincere appreciation also goes to everyone that has been by me all this while. A
Big thanks to you all.

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 TABLE OF CONTENTS
DEDICATION ................................................................................................................................ 2
ACKNOWLEDGEMENT .............................................................................................................. 3
CHAPTER ONE ............................................................................................................................. 6
1.0 INTRODUCTION .............................................................................................................................. 6
1.1 HISTORY OF STUDENT INDUSTRIAL WORK EXPERIENCE SCHEME ................................. 6
1.2 SCOPE OF SIWES ............................................................................................................................. 7
1.3 AIMS AND OBJECTIVES OF SIWES ............................................................................................. 7
1.4 OBJECTIVES OF THE SIWES REPORT ......................................................................................... 8
CHAPTER TWO ............................................................................................................................ 9
DESCRIPTION OF THE ESTABLISHMENT .............................................................................. 9
2.1 BRIEF HISTORY AND LOCATION OF THE ESTABLISHMENT ............................................... 9
2.2 MISSION AND VISION .................................................................................................................... 9
2.3 ORGANIZATIONAL STRUCTURE AND CHART ...................................................................... 10
CHAPTER THREE ...................................................................................................................... 11
LABORATORY TERMINONOGIES AND PRACTICES ......................................................... 11
3.1 GENERAL EQUIPMENTS IN THE LABORATORY AND THEIR USES .................. 11
3.2 GENERAL SAFETY PRECAUTIONS IN THE LABORATORY ................................. 15
3.3 COLLECTION AND PROCESSING OF CLINICAL SAMPLES .................................... 16
3.4 LABORATORY EXAMINATION OF BLOOD SAMPLES .......................................... 17
3.4.1 MALARIA PARASITE ............................................................................................... 17
3.4.2 BLOOD GROUPING ................................................................................................. 19
3.4.3 FASTING BLOOD SUGAR/GLUCOSE ................................................................. 20
3.4.4 PACKED CELL VOLUME ...................................................................................... 22
CHAPTER FOUR ......................................................................................................................... 23
4.1 LABORATORY EXAMINATION OF URINE SAMPLES ........................................... 23
4.1.1 URINE MICROSCOPY, CULTURING AND SENSITIVITY ................................. 23
4.1.2 URINE ANALYSIS ................................................................................................... 26
4.2 LABORATORY EXAMINATION OF SWABS ............................................................. 27
4.2.1 WOUND SWAB ........................................................................................................ 28
4.2.2 EAR SWAB................................................................................................................ 28
4.2.3 HIGH VAGINA SWAB ............................................................................................. 29
4.2.4 ANTIBIOTICS SENSITIVITY AND SUSCEPTIBILITY TESTING ........................ 29
4.2.5 SUSCEPTIBILITY TESTING TECHNIQUES ......................................................... 30

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CHAPTER FIVE .......................................................................................................................... 34
5.0 SUMMARY, CONCLUSION, AND RECOMMENDATION ........................................ 34
5.1 SUMMARY AND CONCLUSION .................................................................................. 34

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 CHAPTER ONE

1.0 INTRODUCTION

1.1 HISTORY OF STUDENT INDUSTRIAL WORK EXPERIENCE

SCHEME
SIWES, which is the acronym for the Students’ Industrial Work Experience Scheme is a

skilled training scheme and integral part of the fulfillment of award of Bachelor of Technology

(B.Tech). It provides each student the opportunity for practical and work experience outside the

university through attachment to establishments / companies. It helps to compliment knowledge

gained by students during their stay in school.

The Student Industrial Work Experience Scheme (SIWES) was established by the Industrial

Training Fund (I.T.F) in 1973 (following a promulgation of decree No 47 of 8Th October 1971)

to solve the problem of lack of adequate practical skills preparatory for employment in industries

by Nigerian graduate of tertiary institutions, sponsored by the Federal Government of Nigeria.

The reason behind the establishment of SIWES was due to observed changes in skills

requirement, need for competence building among students, a need to give students of Science,

Engineering and Technology hands-on training and work experience pertaining to their field.

Also, expectations from employers of labor – working experience, contribute to the need to

establish the SIWES program.

The SIWES program has objectives that can be summarized in a statement as follows: to

facilitate industrial skill and work experience acquisition by students of higher learning, thus

exposing students to work methods and techniques which afford them opportunity to apply into

practice their theoretical knowledge and at the same time affording the employers participation

in the educational process of the indigenous work force.The SIWES program started in 1974

with 748 students from 11 institutions, the scheme has evolve overtime and as at 2008 alone 204

institutions and 210, 390 students participated in the scheme, and this number is bound to
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increase over time. Training is a key factor in enhancing the efficiency and expertise of the

workforce. The SIWES program prepares students for labor markets. It has become an

innovative phenomenon in human resources development and training in Nigeria.

1.2 SCOPE OF SIWES

Practical knowledge is learning without which mastery of an area of knowledge may be too

difficult to achieve. Practical knowledge involves developing skills through the use of tools or

equipment to perform tasks that are related to a field of study. No society can achieve

meaningful progress without encouraging its youth to acquire necessary practical skills. Such

skills enable them to harness available resources to meet the needs of society. It was against this

background that SIWES, otherwise referred to as Industrial Training (IT), was introduced in

Nigerian tertiary institutions. SIWES is a cooperative industrial internship program that involves

institutions of Higher Learning, Industries, The Federal Government of Nigeria, Industrial

Training Fund (ITF), Nigerian Universities Commission (NUC) and NBTE/NCCE in Nigeria.

SIWES forms part of the approved minimum academic standards in these institutions.

1.3 AIMS AND OBJECTIVES OF SIWES

The specific objectives of SIWES were summarized by the Federal Government in its Gazette of

April, 1978 as follows:

 It exposes students to the use of machinery and equipment that are not available in the

Universities.

 To equip students with valuable skills which will give them a competitive edge in today’s

job market.

 It serves as a forum for preparing students for industrial working conditions, methods and

environment.

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  It provides students with the opportunity to apply their theoretical knowledge in real work

situation, thereby bridging the gap between University’s theoretical work and actual

practical.

 It enhances students’ contact for job placement after graduation

Sequel to the mandate lay down by SIWES, students from the Federal University of Technology

(FUTO) are harnessed during their second and fourth year for I.T experience to broaden their

horizon, in respect to this, I had my I.T experience at Degrills Integrated Services Healthcare

centre

1.4 OBJECTIVES OF THE SIWES REPORT

 To provide a detailed account of the knowledge and experience gained during the training

period by the student.

 To provide a technical report on the area covered by the student to the authority

concerned in the fulfilment of the IT semester program.

 To express the advantages of the scheme relevant to the field of study.

 To recommend ways by which the scheme can be improved.

My two weeks SIWES was undertaken at Degrills Integrated Services located at No 20, Adeleke

street Gbagada, Lagos state at Laboratory Department of the firm.

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 CHAPTER TWO
DESCRIPTION OF THE ESTABLISHMENT

2.1 BRIEF HISTORY AND LOCATION OF THE ESTABLISHMENT

Degrills Integrated Services Healthcare Center situated at Gbagada, Lagos commenced

operation in the year 2008 with the founder Dr. Okeke, two nurses and a Laboratory technician.

This Health Centre has now grown having its own permanent site. The rented space with the

initial one doctor, two nurses and a laboratory technical has now increased its employment

capacity by additional one doctors , 12 nurses, laboratory scientist, laboratory technician, student

nurses and a number of IT students from various school.

2.2 MISSION AND VISION

Mission

 To offer comprehensive and affordable health care services to all categories of health

care consumers

Vision

 To be the leading hospital that offers clinical and medical services

 To provide quality health care delivery to all categories of health care consumers most

efficiently.

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2.3 ORGANIZATIONAL STRUCTURE AND CHART
Organogram of Degrills Integrated Services Healthcare Center has a Medical Director. The

details is represented below;

Organogram Degrills Integrated Services Healthcare Center

MEDICAL
DIRECTOR

ADINISTRATION PHARMACIST MEDICAL DOCTOR

DEPT

LABORATORY NURSES
SCIENTIST STUDENT
NURSES

IT STUDENT

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 CHAPTER THREE
LABORATORY TERMINONOGIES AND PRACTICES

3.1 GENERAL EQUIPMENTS IN THE LABORATORY AND THEIR

USES
Different equipments are known to be used in the laboratory to carry out different clinical

purposes. Some are used as preservatives, storage purpose, and transportation purpose.

Some of these equipments include:

 Microscope: It is used for viewing small objects that cannot be seen with naked eyes for

example, parasite, Red Blood Cells, White Blood Cells, micro-organism, and so on.

 Autoclave: This is an indispensible tool for sterilization in the laboratory. Pressure is

used in producing high temperature steam to achieve sterilization. The autoclave is

usually operated at 121ºc for 15 minutes. It is used for sterilizing media, used plates or

slopes and used bottles such as MacCartney bottle, Bijou bottle and so on.

 Test tube: It is used for collection of blood or urine sample for centrifugation

 Universal bottle: It is used for collection of urine, stool, cerebro spinal fluid, semen

sample, and so on.

 Centrifuge: This is a machine that sediments particles (cells, parasites, casts, bacteria

and blood components) suspended in fluid (urine, blood) by exerting a force (centrifugal

force) greater than that of gravity. The centrifugal force increases with the speed of

rotation. Its unit is revolution per minute (rpm), e.g. 5000/rpm for 5 minutes. The

centrifuge is used to obtain plasma and serum from blood and can be used to perform

parasite concentration techniques.

 Swab stick: It is used to collect swabs from wound, eye, ear or vagina.

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  Glass or Microscopic slide: It is used for Malaria Parasite test, preparation of stool and

other substances for microscopy.

Eye piece

Objective lens

Stage

Coarse adjustment knob

Light source

Plate1: light binocular microscope

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 Pressure gauge

Cover

Knob

Figure 2: Autoclave

 Cover slip: It is used to cover a prepared stool and urine sample for microscopy view.

 Needle and syringe: It is used for blood collection from a patient.

 Glove: it is used for protection from infection.

 Pyrex bottle: It is used in media preparation.

 Refrigerator and freezer: This is used for storage of specimen and reagents.

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 Tourniquet: Used to tie patient’s hand to locate the prominent vein during phlebotomy.

 Measuring Cylinder: It is used for measuring chemicals and water.

 Petri dish: It is used for preparation of media, culturing and isolation of clinical isolates.

 Inoculating or wire loop: It is used for picking isolates from plates already cultivated. It

can also be used to cultivate microbes on plates by transferring inoculums for streaki

 Thomas pipette: It is used to withdraw and dispense samples and reagents.

 Reagent bottle: It is used to collect or store reagents such as stains, alcohol, lugol’s

iodine.

 Whatmann filter paper: It is used for sterilization process, blotting and oxidase test.

 Immersion oil: It is used when a stained slide is prepared to raise the refractive index of

the stained slide.

 Forceps: It is used to hold materials such as antibiotics disc.

 Blood culture bottle: It is used to culture organisms in blood samples.

 Anaerobic jar: It is used for the cultivation of anaerobic organisms in anaerobic

condition.

 Staining rack: This is used for the placement of slides that contains smear which is to be

stained.

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 3.2 GENERAL SAFETY PRECAUTIONS IN THE LABORATORY
 Read manuals and instructions before carrying out any practical.

 Work area/ surfaces must be disinfected before and after use.

 Hand gloves should be worn when collecting blood samples from patient and while

working in the laboratory.

 Laboratory coat should be worn and button up while working in the laboratory. It should

not be worn outside the laboratory.

 Application of cosmetics is prohibited.

 Care must be taken when removing needle from syringe.

 Only close toes shoes are to be worn, sandals are not permitted.

 Always wash your hand before and after carrying out each test or while leaving the

laboratory

 No food/drink is permitted in the lab at any time.

 Open cuts or wounds should be covered with plaster while working in the laboratory

 Sharp object like lancet and needle should be disposed into the safety box after usage to

avoid direct inoculation.

 Flame transfer loops, wires or needles immediately after use to transfer biological.

 Always label reagent bottle.

 Return all chemicals, reagent, cultures and glass wares to appropriate place.

 Glass wares should be washed with soap and water then rinsed with distilled water.

 Dispose wastes in proper containers.

 Don’t pour chemicals down the sink.

 Avoid pipetting with your mouth.

 Use a nose mask to prevent inhalation of aerosols and gaseous suspension of chemicals.

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  Any chemical or biological fluid spills must be reported immediately to the medical

laboratory scientist.

3.3 COLLECTION AND PROCESSING OF CLINICAL SAMPLES

In the laboratory, specimen collection is very important and is the first step in obtaining an

accurate laboratory diagnosis of an infection which depends upon timing, selection and method

used. Several specimens are collected with different methods and specimen bottles. The

specimen is:

 Collected before administration of any antimicrobial agent.

 Prevented from contamination by external microorganisms present.

 Immediately transferred into the appropriate specimen bottle which is sterile and free

from any contaminant, properly labelled and dated as the case may be.

 Handled aseptically during the necessary procedures of diagnosis

 Sampled at the appropriate time to prevent any form of alteration of the results.

Sample collection: Samples to be collected here must come in the right container (sterile

universal bottle, swab sticks and blood culture bottles) and must have been collected in the right

(that is, according to the procedure for the collection of sample) way before being registered.

That is, samples should be collected into a wide mouth container. The container should be leak

proof, transparent and must have a screw cap. In-patients’ samples are collected by the Doctor or

Nurse on duty. Different samples have different registers which contains the full vital and basic

information like date, laboratory number, name of patient, age, sex, hospital number,

ward/department, amount paid, specimen required, nature of test, receipt number and remarks.

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Procedure involved in sample collection

A patient comes to the laboratory with a request form which is billed by the laboratory scientist

on duty depending on the type of test required. The patient pays and brings back the receipt

which is checked before collection of samples in the case of an outpatient or issuing of sample

container depending on the test required. For an in-patient, after payment, the appropriate sample

container is given out for the samples to be collected in the ward and returned to the laboratory.

On return of the sample and request form, they are both cross checked for any mistake which

when seen disqualifies the sample for analysis. In the absence of any mistake, the sample is

registered and given laboratory number, which is the serial number on the register by which the

sample will be identified henceforth. The patient is then asked to come at an appropriate time for

the result. The specimen or samples with their corresponding laboratory request form are then

taken to the designated bench in the laboratory for processing.

3.4 LABORATORY EXAMINATION OF BLOOD SAMPLES

3.4.1 Malaria parasite

Malaria is a major public health disease which is a cause of suffering and premature

deaths in tropical and subtropical countries. Malaria is becoming so difficult to control in some

endemic areas because of the resistance of the parasite to anti – malaria drugs and the failure of

vector control measures

Malaria is caused by a protozoan which invades human red blood cells. The organism is

the Plasmodium spp. Plasmodium falciparum is the predominant specie in Nigeria. Other species

of malaria parasite include: Plasmodium malariae, Plasmodium ovale, Plasmodium vivax

Malaria parasites are transmitted by the bite of an infected female anopheles mosquito.

Sporozoites contained in the saliva of the mosquito are inoculated into the blood of a human host

17
when the mosquito takes blood meal. Infection can also occur by transfusion of infected donor,

and occasionally, congenitally.

For the detection of malaria parasites (Plasmodium species) in blood, thick and thin blood films

was prepared. Blood for malaria parasite test was collected from the capillaries however, venous

blood could be used. Thick blood film was prepared to determine the intensity of the parasite

while thin film was prepared to identify the species of malaria parasite causing the infection in

the patient’s blood.

Procedures for thin blood film are:

 the patient’s finger was cleaned with an alcohol swab after which was pricked with a

lancet

 a drop of the patient’s blood was placed on a clean microscopic slide

 the lancet was discarded immediately into a bin

 the edge of a cover slip was used to spread the blood and a tail was formed

 the slide was left to air dry and was fixed with methanol

 after fixation, the film was covered with Giemsa stain for 15-30 minutes after which the

stain was rinsed off

 the slide was left to air dry, was covered with immersion oil and was viewed under 100x

objective lens

 the ring form of the cytoplasm with a chromatin dot was observed

Procedures for thick blood film are:

 the patient’s finger was cleaned with an alcohol swab after which was pricked with a

lancet;

 a drop of the patient’s blood was placed on a clean microscopic slide;

18
  the lancet was discarded immediately into a bin;

 the edge of another slide was used to spread out the drop of blood;

 the slide was left to air dry after which was fixed with methanol;

 the slide was stained with Leishman stain for 20-30 minutes after which was rinsed off;

 the slide was left to air dry, was covered with immersion oil and was viewed under the

microscope at 100x objective.

Expected Result

Trophozoites of Plasmodium spp will be seen for positive result

Observation are reported as +, ++ and +++ when seen and Negative if not seen.

Few (1 to 10) number of Plasmodiumspp (+)

Moderate (10 to 100) number of Plasmodiumspp (++)

Numerous (more than 100) (+++)

3.4.2 Blood grouping

This was done to determine the group (A, B, AB & O) of a patient.

Materials: Tourniquet, syringe or lancet, EDTA tube, alcohol prep pad, blood plate or ceramic

tile, antisera (A, B, D).

Procedures:

 Tourniquet is tied around the upper arm to force the vein to rise with blood.

 Used an alcohol prep pad to moisture the surface the skin and insert the syringe to collect

blood sample.

 Removed the needle and released the blood into the EDTA tube.

 Placed the tube into the tube rack.

 Placed a drop of the blood sample on the blood plate

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  Added a drop of anti-sera on each of the blood on the blood plate and mix well.

 Rock for 5 minutes and observe if there is any agglutination.

Result:

ANTISERM ANTISERUM ANTISERUM RESULT

A B D

+ - + A+

+ - - A-

- + + B+

- + - B-

+ + + AB+

+ + - AB-

- - + O+

- - - O-

KEY

+ indicates the presence of agglutination

- indicates the absence of agglutination

3.4.3 Fasting blood sugar/glucose

This is a method of learning or determining how much of glucose(sugar) in a blood sample taken

after an overnight fast. The fasting blood sugar test is commonly used in the detection of diabetes

mellitus. Blood glucose test measures the amount of sugar in blood sample. Glucose is the major

source of energy for most cells of the body, including those in the brain. Carbohydrates are found

in fruits, cereal, bread pasta, and rice. They are quickly turned into glucose in the body and this

raise your blood sugar level. Glucose level can also be tested using urine sample.

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To prepare for the test, the test may be conducted while fasting or at random.

 If a fasting glucose (sugar) test is to be conducted, the patient is not to eat or drink for

eight hours before the test.

 If a random sugar test is to be conducted, it could be done at any time of the day, but the

result depends on what the patient eat or drink before the test as well as the activity.

Materials nedded: flouride Bottle, glucometer, syringe or lancet, and blood

How the test is carried out

 Tourniquet is tied around the upper arm to force the vein to rise with blood or the use of

lancet to pick the thumb.

 Used an alcohol prep pad to moisture the surface the skin and insert the syringe to collect

blood sample.

 Removed the needle and released the blood into the flouride tube.

 Picked a drop of blood sample and placed on the glucometer

 Observed the reading taken on the glucometer

Result:

Up to 100(mg/dl) is considered to be normal for fasting blood sugar test.

For level between 100 and 125(mg/dl) have an impaired fasting glucose.

For level of 126(mg/dl) and above, the patient is diagnosed of diabetes.

NOTE: Patient with blood sugar level between 100 and 125mg/dl is known to be paradiabetic

and this level is considered risk factor of typetwo diabetes and its complications.Normal value

ranges may vary slightly among different laboratories. The example above shows the common

measurements for results for these tests. Some laboratories use different measurement or may

test different specimen.

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3.4.4 Packed Cell volume
Sample of packed cell volume (PCV) can be collected by cleaning the thumb of a patient with

spirit swab then prick the patient thumb by lancet. After which capillary tube are used to collect

the sample blood, then the sample taken to the laboratory. We now take the put sealant at the

edge to avoid blood splitting from the tube. The tube is placed in the haematocrit centrifridge and

balance with other tube to avoid noise, covered and spin for 5 minutes. Three segment of the

following result will be obtained;

I. Plasma

II. Buffy coat

III. Red blood cell

Packed cell Volume normal range

Male 40-52%
Female 37-47%
Neonate 40-60%
Children 30-42%

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 CHAPTER FOUR

4.1 Laboratory examination of urine samples

The analysis of urine is called urinalysis. Urinalysis (routine and microscopy, R&M) is an array

of test performed on urine. This can be used via dipstrips or test strips or through the use of

microscope to view wet preparation, culturing and sensitivity (MCS).

Macroscopic examination

Once the urine is collected for analysis, the macroscopic or appearance of the urine is noted.

However, if a urine sample is not to be analyzed immediately, it may be preserved under 4-6ºc in

a refrigerator or by adding boric acid to prevent changes in the urine when left unattended to.

Normal urine is supposed to be amber in colour and should be aromatic enough when perceived.

However, a cloudy urine with an unpleasant odour suggest a bacterial urinary infection and one

with a fruity odour may suggest diabetes; a red and cloudy urine due to the presence of

erythrocytes suggest urinary schistosomiasis; yellow-brown urine due to bilirubin suggest viral

hepatitis or jaundice; yellow urine may be as result of an intake of a sulphurnamide drug; a

greenish urine may be as a result of an intake of neurobium and may suggest pyelonephritis or

glomerunephritis; a black colouration may be as a result of intake of too much iron containing

drug.

4.1.1 Urine microscopy, culturing and sensitivity

 Wet preparation: This is done to detect the presence of excess leucocyte in the urine, red

blood cells in cases of haematuria (blood in urine), casts (hyaline cast, waxy cast,

cellular cast and granular cast), crystals (such as cholesterol crystals, sulphurnamide

crystals, cysteine crystals) epithelial cells and parasites such as Trichomonasvaginalis,

eggs of Schistosomahaematobium. Procedure for wet preparation of urine is as follows:

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  2 urine centrifuge tube, the urine in a universal bottle and a Thomas pipette was

provided;

 3ml of the urine was placed in each centrifuge tube and was covered;

 The tubes were placed in the centrifuge opposite each other to balance the

centrifuge and to avoid vibration;

 the urine was spun at 3000 revolution per minute (rpm) for 5 minutes;

 after spinning, the supernatant was discarded into the disinfectant jar;

 the sediment left in the tube was tapped to remix;

 a drop of the sediment was placed on the microscope slide and was covered with a

cover slip;

 the preparation was viewed under the microscope at 40x objective lens and the

observations were noted.

When bacteria and / or white blood cells were observed, direct graming and culturing

was done.

 Culturing and sensitivity: When a bacterial urinary infection is suspected or when

bacterial cells are present in the wet preparation, the urine was cultured on differential

media such as MacConkey or Cysteine Lactose Electrolyte Deficient medium after

which was incubated overnight at 37ºc.

Following incubation, the plates were harvested or read and the colonial morphology of

each plate was obtained. Gram staining, biochemical tests and antibiotic susceptibility were

also carried out. Pathogens that can be isolated include:

Escherichiacoli: Gram negative bacillus was observed under the microscope.On

MacConkey, the colonies was dry, raised and was pink in colour. It was positive to indole

24
test and negative to oxidase test. It is the major causative agent of urinary tract infection

(70-90%). Other biochemical test that can be done are VogesProskauer test (positive),

methyl red (positive) and sugar (sucrose, lactose, glucose) fermentation tests.

Proteusmirabilis: Gram negative bacillus was observed under the oil immersion lens of

the microscope. The colony on the chocolate agar plate was not distinct and was swarming,

on MacConkeyagar, pale coloured colonies were observed. The organism was urease

positive (pink colour) and indole negative. It is one of the causative agent nosocomial

infections, urinary tract infection following catheterization and occasionally, septicemia. It

belongs to the enterobacteriaceae family (coliforms). Other species is Proteus

bulgariswhichisureasepositive and indole positive, however, this specie is not common. The

swarming of Proteus species can be inhibited by adding salt to the culture medium.

Klebsiellaaerogenes: Gram negative bacillus was observed under the microscope’s oil

immersion lens. On MacConkey, wet, raised, mucoid and pink colonies was observed; on

CLED medium, the colonies was yellow, wet, mucoid and raised. The organism was

positive to citrate test and negative to indole test. It also belongs to the enterobacteriaceae

(coliforms). It is one of the causative agents of urinary tract infection and nosocomial

infections.

Viridans streptococci: Italso causes urinary tract infection. They are resistant to optochin

disc test and negative to bile solubility. Nitrofurantoin is an antibiotic of choice for urine

pathogen. Other antibiotics that were routinely used are chloramphenicol, co-trimoxazole,

ciprofloxacin, ofloxacin, tetracycline and so on.

25
 Colonies Escherichia coli on MacConkey

Proteus mirabilis on chocolate

4.1.2 Urine Analysis

 Urinalysis via test strips: This is a basic and rapid diagnostic tool used in determining

pathological changes in a patient’s urine. The test strip is referred to as “combi”, the

amount of parameters a strip can analyze determines its suffix. For example, a strip that

26
 can analyze 3 parameters is referred to as combi-3, 9 parameters combi-9. The

parameters that can be detected and analysed by a strip are glucose, protein, pH,

leucocytes, nitrite, urobilinogen, ketone, haemoglobin, specific gravity and so on.

Procedure for urinalysis using test strip include:

 Urine sample was provided in a universal container;

 The test strip was removed from its container and was dipped into the urine;

 The test strip was removed and was tapped to remove excess urine;

 The test strip was compared with the colour chart on the test strip pack and the

result was noted.

The variation in the result away from normal indicates a number of things. Presence of

excess protein may be as a result of an intake of too much proteinous meal which may lead to

odema in pregnant women or may indicate a urinary tract infection; an acidic pH maybe as

aresult of an intake of fruits or a urinary tract infection caused by Escherichia coli while an

alkaline pH may be as a result of urinary tract infection caused by Proteus mirabilis; glycosuria

(glucose in urine) may suggest diabetes, pyelonephritis or glomerunephritis.

In order to prevent the test strip from absorbing water (test strips are hygroscopic), the pack was

closed immediately. The cap of the pack has a drying agent in it that keeps the test strips dry.

4.2 LABORATORY EXAMINATION OF SWABS

Sterile swabs are often used in the collection of pus or exudates from different parts of the body

such as throat, ear, wound, vagina amongst others. Usually, the swabs were cultured before

direct gram staining so as to prevent the contamination of the swab which could lead to mixed

growth on the cultured plate.

27
4.2.1 WOUND SWAB
For culturing, nutrient agar, chocolate agar and blood agar plates were used after which direct

gram staining was done. The cultured plates were incubated at the required temperature and time.

The following day, the plate harvesting, gram staining, biochemical tests and antibiotic

susceptibility testing was done. Organisms that were isolated from the cultured plates include

Staphylococcus aureus: Gram positive cocci in clusters, non-haemolytic golden yellow, round,

entire colonies on blood agar and cream, round, distinct, entire colonies on nutrient agar. It was

positive to catalase and coagulase tests. It was also most susceptible to vancomycin when the

antibiotic susceptibility testing was done. It is an opportunistic organism that causes wound and

burn infection.

Pseudomonas aeruginosa: Gram negative bacilli, greenish pigmentation due to the production

of pyocyanin was observed on the nutrient agar plate and zone of haemolysis was observed

around the colonies on the chocolate agar plate. It was catalase positive and oxidase positive and

was most susceptible to anti-pseudomonal penicillin, imipenem, ceftazidime. It is also one of the

causative agents of wound and burn infection.

Other organisms that cause wound and burn infections are Streptococcus species, Proteus

species, and Staphylococcus aureus, Enterococcus and so on.

4.2.2 EAR SWAB

Culturing was done on nutrient agar and chocolate agar plate then the direct gram staning was

done. The cultured plate was incubated at 37ºc overnight. The following day, the colonial

morphology on each plate was read and noted, Gram staining was done, biochemical tests such

as catalase and oxidase test was done and antimicrobial susceptibility testing was also done. The

antibiotics used are gentamicin, polymixin, ceftazidime, imipenem, anti-pseudomonal penicillin

and ciprofloxacin. The organisms isolated were:

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Pseudomonas aeruginosa: Gram negative bacilli, greenish pigmentation (pyocyanin) was

observed on the nutrient agar plate and zone of haemolysis was observed around the colonies on

the chocolate agar plate. It was catalase positive and oxidase positive and was most susceptible

to anti-pseudomonal penicillin. It is one of the causative agents of otitis externa and otitis media

(inflammation of the external and internal ear).

4.2.3 High vagina swab

Culturing was done on Nutrient Agar and SabouraudDextroseAgar after which direct gram

staining was done. The plates were incubated at 37ºc for 24 hours. Following incubation, the

colonial morphology on the plates were read, gram staining was carried so also the biochemical

test-germ tube test. The organism isolated was:

Candida albicans: Gram positive oval which was much bigger than cocci was observed. On

the plates, cream coloured pasty colonies with a characteristic yeast smell were observed.

However, the colonies on the Sabouraud dextrose agar were more prominent. After the germ

tube test was incubated and viewed under the microscope, the budding form of the organism that

appeared tube like was observed. This organism is a majour cause of vaginitis i.e. inflammation

of the vagina. Mycotin is the drug of choice.

4.2.4 Antibiotics sensitivity and susceptibility testing

This is also referred to as antimicrobial susceptibility testing or antagonistic susceptibility

testing. Antibiotics are chemicals produced naturally by bacteria and fungi to act against other

microorganism. Antibiotics are also used in describing antimicrobial agents (usually

antibacterial) that can be used in treating infection. Antimicrobial agents include naturally

occurring antibiotics, synthetic derivatives of naturally occurring antibiotics (semi-synthetic

antibiotics) and chemical antimicrobial compounds (chemotherapeutic agents). Antibacterial

agents have various mode of action amongst which are:

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 i. Inhibition of bacterial cell wall synthesis, e.g. penicillins such as carbenicillin,

cloxacillin; cephalosporins such as ceftazidine, ceftriaxone; carbapenems such as

imipenem; glycopeptides such as vancomycin.

ii. Inhibition of protein synthesis, e.g. aminoglycosides such as gentamicin, streptomycin;

macrolides such as erythromycin, azithromycin; chloramphenicol; tetracycline.

iii. Inhibition of bacterial nucleic acid synthesis, e.g. quinolones such as ciprofloxacin,

norfloxacin; co-trimoxazole; metrinidazole.

Examples of antimycobacteria (bacteria with mycolic acid in their cell wall) agents are rifamprin,

ethambutol, isoniazid and so on. Examples of antiviral agents are acyclovir (against herpes group

viruses), gangiclovir. Some antimicrobials are bacteriostatic while some are bactericidal.

4.2.5 Susceptibility testing techniques

In the treatment and control of infectious diseases especially when caused by pathogens that are

often drug resistant, susceptibility testing is done to select most effective antimicrobial drug.

Factors that must be considered before prescribing an antibiotic includes age, weight, sex,

medical history of the patient, cost of antibiotics and the condition of the patient.

1.Dilution susceptibility testing technique: This was used to measure the minimum inhibitory

concentration (MIC) of an antibiotic on bacteria. Minimum inhibitory concentration is the lowest

concentration of an antimicrobial that will inhibit the visible growth of a microorganism after an

overnight incubation. The procedure used is as follows:

 an antibiotic (tetracycline) stock solution was prepared in a universal bottle by

dissolving 250g of tetracycline in 10ml of distilled water.

 250ml of nutrient agar media was prepared and was poured into 10 Petri dishes

which were left to cool and solidify.

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  2ml of distilled water was withdrawn with a needle and syringe and was

dispensed into test tubes labeled 1-10 which was placed on a test tube rack

 2ml of the tetracycline stock solution was withdrawn from with a needle and

syringe and was dispensed into the first test tube.

 2ml of the preparation in the first test tube was withdrawn and was place into the

second test tube. This procedure was repeated for the remaining test tubes.

 2ml of the preparation in the tenth test tube was withdrawn and was discarded into

a disinfectant jar since there was no other test tube.

 after the plates had solidified, they were flooded with a peptone broth culture of

Escherichia coli and were left for some minutes.

 with the aid of a cork burrower, holes were burrowed into each plate that had been

labelled 1-10.

 each hole of each plate was filled to the brim with corresponding antibiotic

dilution.

 the plates were incubated at 37ºc for 24 hours

On checking after incubation, plates 6,7,8,9 & 10 had no zone of inhibition; plate 5 had a very

little zone of inhibition; plate 4 has a zone of inhibition of 16mm in diameter (at thus

concentration [625цg/ml], tetracycline had the minimum inhibitory concentration); plate 3 had a

zone of inhibition of 22mm in diameter; plate 2 had a zone of inhibition of 24mm in diameter

and plate 1 had the highest zone of inhibition of 28mm in diameter.

2. Disc diffusion technique: This technique is mostly used in the laboratory to routinely test for

antibiotic susceptibility. The antibiotics used were either single discs (more effective) or multiple

disc that are made by impregnating filter or blotting paper with known concentration of

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antibiotics which is then placed at definite distance on the agar plate that has been inoculated

with the test organism. The antibiotic diffuses into the medium and the growth of the test

organism is inhibited at a distance from the disc that is related to the susceptibility of the

organism (zone of inhibition). Organisms that were resistant to the antibiotic had little or no zone

of inhibition. The choice of antibiotics used depended on the pathogen, specimen, and range of

locally available antimicrobials, toxicity, efficacy and local prescribing policies. The procedure

for carrying out disc susceptibility testing is as follows:

 The agar of choice (Mueller-Hinton or Nutrient agar) was prepared and was poured into

sterile Petri plates;

 The broth culture of the test organism which is of match to 0.5 McFarland standard of

turbidity was used to inoculate the plate using the pour plate or Lawn method. McFarland

standard was used as a reference to adjust the turbidity of bacterial suspensions so that

the number of bacteria will be within a given range to standardize microbial testing. If a

suspension used was too heavy or too dilute, an erroneous result (either falsely resistant

or falsely susceptible) for any given antimicrobial agent might occur.

 The plate was left to soak for about 5 minutes;

 With the aid of a sterile forceps, a multiple disc was introduced into the plate and was

incubated upside down for 24 hours. For single discs, a minimum of 6 and a maximum of

8 were used to prevent the zones of inhibition from entering into each other which could

make the diameter of each zone difficult to determine.

Several antibiotics were used routinely in the laboratory, some of which was specific for Gram

positive organisms, e.g. erythromycin, ampiclox, oxacillin, vancomycin and so on; some others

are specific for gram negative organisms, e.g. amikacin, chlormaphenicol, ofloxacin,

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nitrofurantin and so on; most can however be usedfor both, e.g. cefuroxime, gentamycin,

ceftriaxone, ciprofloxacin, pefloxacin and so on.

Some microbes are growing resistance to routinely used antibiotics as a result of extensive use

and misuse of antimicrobials thereby favoring the emergence and survival of resistant strains of

microorganism. Drugs resistance is common among Staphylococci (Meticillin Resistant

Staphylococcus aureus(MRSA), Gonococci, Meningococci, Pneumonococci, Enterococci,

Salmonella, Shigella, Klebsiella, Psuedomonas and Mycobacterium tuberculosis.

Antibiotics

Clear zone of inhibition

Plate5: Disc antibiotics susceptibility test

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 CHAPTER FIVE

5.0 SUMMARY, CONCLUSION, AND RECOMMENDATION

5.1 SUMMARY AND CONCLUSION

In the laboratory, different clinical samples such as swabs, (urine, high vaginal swab, wound

swab) and so on were analyzed using different culture media, staining techniques, biochemical

test and antibiotic susceptibility testing. Drugs of choice were deduced from the antibiotic

susceptibility testing depending on the organisms’ susceptibility to each antibiotic among other

factors such as age, sex, medsical history and condition of the patient and so on.

The industrial training program set up by the Student Industrial Work Experience Scheme

(SIWES) is aimed at equipping students with the practical training of the theoretical work taught

in schools, thereby giving students an opportunity to be exposed to the usage and operational

value of some of the facilities needed in their chosen field later in life.

The three-month industrial training has helped broaden my theoretical knowledge to practical

usefulness of Biotechnology. It has helped me to see from practical perspective most concepts

involved in Clinical Microbiology like microscopy, staining, collection of samples like blood,

urine and swab from wound, vagina and ear, culture, preparation of media, slide preparation and

operation of common laboratory apparatus like Autoclave incubator, centrifuge, and operational

procedure of equipments.

Most importantly, I have been able to see the various prospects available in the field and

also the various challenges. Indeed, the industrial training program has trained personally to be

more conscious and initiative with the limited resources available in Nigeria economy.

It has also changed my orientation about medical laboratory to be more relevant as I

initially thought and I now see it to be the backbone of the modern medicine

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5.2 Recommendation

With my experience during the three-month industrial training program, I hereby recommend

that

 Government should orientate establishment on the importance of the programme so as to

avoid refusal of applicants.

 The hospital management should pay more attention to staffs’ welfare and training in

order to improve working standard and efficiency.

 Establishments should make efforts to compensate student with little token to enable

them fund themselves during their period of attachment.

 The student should put into consideration the available modern equipment in the

establishment in which they intended to go for their Industrial Training before putting in

their application.

 Establishments should give students absolute access equipment and other sections for a

better understanding and knowledge of their discipline.

 The school authority should provide adequate arrangements for prospective I.T students

for easy accessibility into establishments/companies that is related to their field of study.

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