TECHNICAL REPORT ON STUDENT INDUSTRIAL WORK EXPERIENCE SCHEME (SIWES) BY EZE GLORIA TOBECHUKWU REG.

NO: FPI/SLT/ND/10/081

DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY FEDERAL POLYTECHNIC IDAH KOGI STATE IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF NATIONAL DIPLOMA (ND) IN SCIENCE LABORATORY TECHNOLOGY, FEDERAL POLYTECHNIC IDAH

DEDICATION I dedicate this report, first to the Almighty God, who saw me through the four(4) months students industrial work experience scheme (SIWES) programme. Also to my parents for their love and support, also to all those who have in one way or the other contributed to my academic career.

ACKNOWLEDGMENT I want to use this opportunity to highly appreciate the Almighty God for his care, protection, good health and provision during the course of my programme. I want to acknowledge the efforts and contributions of so many individuals that brought this work or programme to a successful end. First, I thank the proprietor as well as the management of Mount Arafat Hospital for accepting and attaching me in their clinic for my industrial training. Secondly, I thank and acknowledge the efforts and training contributions of the medical laboratory technologist Miss Eze Philomena. Thirdly , I thank my dear parents Mr. and Mrs Eze Cyprail for their prayer, love and care and financial contribution rendered in the course of this programme in order to bring it to a successful end. Finally, I thank all and sundry, who have contributed to the success of this programme whose name cannot be mentioned in this paper.

ABSTRACT This report contains all experience and knowledge acquired during the four months programme (SIWES) training as undergon in the Laboratory department of Mount Arafat Hospital, 59 Enugu Road, Nsukka, Enugu State. As a matter of fact, science technology play relevant role in all aspect of human life reformed which modern machines and to adequate health standard which maintains the continuity of life world wide. This has gone along way in reducing the prevalence and intensity of the general society suffering from various kinds of disease.

FOREWORD Student Industrial Work Experience Scheme (SIWES) programme that must be undertaken as partial fulfilment for the award of National Diploma to science students in Nigerian polytechnic helps students in acquiring experience in their respective discipline.

TABLE OF CONTENT Title page Dedication Acknowledgement Abstract Forward Chapter one 1.0 Introduction 1.1 1.2 1.3 Chapter two 2.0 2.1 2.2 2.3 2.4 2.5 2.6 2.7 2.8 Hematology Pack cell volume Blood Grouping Haemolobin Genotype HIV Screening Erythrocyte sedimentation Rate (Esr) Malaria Parasite (MP) test Widal (Typhoid) test Hepatitis Test Aim and Objectives Introduction of equipments Collection of Sample

Chapter three 3.0 3.1 3.2 3.3 Microbiology Concept of culture Preparation of agar (media) Culture and sensitivity test

3.4 3.5 3.6 3.7 3.8 3.9

Sterilization High Vagina Swab (HVS) Pregnancy Test Urinalysis Urine Microscopy Urine culture and sensitivity

3.10 Stool Culture and sensitivity Chapter four 4.0 Chemical pathology 4.1 Chapter five 5.0 5.1 5.2 Conclusion Recommendation References Fasting and Random blood sugar

CHAPTER ONE 1.0 INTRODUCTION The student work experience scheme (SIWES) is to expose student to machine and equipment, professional working method and ways of safeguarding students in an organization. 1.1 AIM AND OBJECTIVES The main of technical report is to document the practical work experience acquired during successful four (4) months industrial training. This SIWES report covers the comprehensive work experience, findings and attachment. This objective are to provide an avenue for students to acquire industrial skills and experience in their course of study. Also to provide student an opportunity to apply their theoretical knowledge in practical work. 1.2 INTRODUCTION OF EQUIPMENT Firstly, I was introduced to different benches, told of the kind of test carried out on the different benches, the equipment found in each of them. The equipment is as follows: Centrifuge:- This is used to spin blood, urine centrifuging. It is ensured that the test-tubes are balanced (fixed) in position i.e (opposite test-tubes containing the same volume of fluid) to avoid damage of the machine and is covered to avoid splash of blood out of the test-tube. It is timed for about 5 minutes. Microscope:- This is used to view minutes organisms based on their higher or lower objectives lenses. Capillary Tube: This is a small narrow capillary tube, which is manufactured with an anti-cougulant called AEPARINE. It is used in P.C.V test. Micro-Heamatocrit Reader: This is an equipment used in taking PCV reading of the blood sample from a patient. It is measured in percentage (%).

Autoclave: This is a machine used for moist heat sterilization at 1210C in 15 minutes. Beaker: This is used for collection of reagent from store-room or bottle to the lab. And also in working processes in the laboratory. Slide: It is an equipment used in smearing samples for microscopic viewing. Petri-Dish: this is used for cultuning samples and also storing agar. Weighing Balance: It is used for measuring agar and weighing sample (blood). Benches: It is a construction where laboratory equipment are kept or fixed for accurate use. TEST TUBE: This is equipment where bloods are dispensed PIPETTE: It is equipment used for transferring water, serum and reagents from beaker to test-tube to various strips. 2.1 COLLECTION OF SAMPLE

The collection of sample (blood) is done by collecting the blood directly from the vein. This method depends on the kind of test to diagnose. For instance, test that requires serum, venous method is required and test like malaria parasite (mp) the thumb or finger method is required. MATERIALS: Needles, souring, swab, tunicate and spirit. VENOUS COLLECTION: Tunicate is used to fasten the upper arm or the wrist of the patient depending on where the prominent vein is seen. The portion was cleaned with a swab (cotton wool) soak in spirit. They syringe is pierced into the vein and blood is collected depending on the ml required and transferred into a test-tube containing anti-coagulant agent to avoid clothing of the blood sample.

CHAPTER TWO 2.0 HAEMATOLOGY

Haematology is the science that deals with the nature, functions and diseases of blood. The study of blood and its analysis. 2.1 PACKED CELL VOLUME

It is a test to check the percentage of blood in the body PROCEDURE: Prick patient thumb and collect the blood in a capillary tube. Seal on end of the tube and spine it with haematocrite centrifuge for 10 minutes, then use haematocrite reader to read the result. Normal blood range for female is 35 45 and male39 56. 2.2 BLOOD GROUPING

This is done to ascertain the blood group a patient belong to. MATERIALS: The patients blood, grouping tile, anti sera A, B, O and mixer (stick). PROCEDURE Place three separate drops on the grouping tile. Add anti-sera A to the first drop, B to second drop and D to third drop. Mix each well with mixer. Rock for 2 3 minutes and observe for agglutinates. Result may be shown below:

A+

AB+

B+

O+

A-

= =

BO-

AB+

2.3

HAEMOGLOBIN GENOTYPES It is one of the test used to eliminate the problem of giving birth to stickler.

MATERIALS: Electrophoresis, cellulose paper, buffer water, patients blood, control (usually AS). PROCEDURE: Place a drop of sample on one slide of carbon paper and control (AS) on the opposite side. Add a drop of water to both blood and mix in order to lyse the blood. Put a pinch of the sample on the cellulose paper in a straight line with the sample one after the other three places. Pour buffer water into the tank of the electrophoreses. Place the cellulose paper inside the electrophoresis at a space provided and then cover with lid. Connect to electricity source and leave for 5 minutes and then read as follows:

___ ___ ___ AA Thick straight lines of the sample indicated AA. ___ ___ 2.4 ___ ___ ___ ___ AS Thick and thin scattered lines indicate AS.

HIV SCREENING Acquired Immune Deficiency syndrome (AIDS) is caused by a viral

transmitted sexual contact or by infected mother to her foetus or child during parental period, by exposure to blood also sharing of contaminated needles and syringe and by blood transfusion. MATERIALS REQUIRED: Blood, HIV strip or kit and buffer. PROCEDURE: Collect the HIV kit, pipette blood using pipette and dispense drop by drop in the blood space provided. Add two drops of the HIV buffer and the mixture continues to migrate through the solid phase. Allows for 10 minutes and then read. Single line shows negative while double line indicate positive as shown below.
C T T T C C

No line

Sample pad

Negative reaction T = Test at patient window C = Control bar 2.5

Positive reaction

Invalid reaction

ERYTHROCYTE SEDIMENTATION RATE (ESR)

An anti-coagulant is added in a blood sample and then allowed to stand n a tube, red cells slowly sediment to the bottom of the tube leaving clear plasma as the

supernatant. The rate of sedimentation estimate under standard condition is known as erythrocyte sedimentation rate (ESR). APPARATUS: Westergren tube, sedimentation rate rack, test tube, pipette, stop watch. PROCEDURE: About 0.5ml of sodium citrate solution is mixed thoroughly with 2ml of blood in a test tube. A Westergren tube is used to draw the blood from the tube. The tube is kept air tight so as to allow blood stay in the tube without falling and it should be without air bubbles. The tube is then place on the Westergren sedimentation rate rack and allowed to stand for one hour, after which the sedimentation rate is observed. The blood sediments by leaving a less dense solution on top (serum). Note: Sodium citrate acts as an anti-coagulant. PRECAUTIONS: 1. The tube must be clean 2. The tube must be filled properly without air bubbles 3. The tube must be vertical to ensure accuracy Normal value for Male 0.5 mm/hour Female 0.7 mm/hour This test is done to check whether one is having an internal hidden disease with no symptoms. 2.6 MALARIA PARASITE (MP) TEST

This is a test done to determine whether a malaria parasite is present in the blood stream of a patient. APPARATUS: Microscope, slide, geimsa stain Procedure: Blood of the patient is collected either through vein or thumb. A thick blood film is made on the slide with spreader and kept to dry after which it is stained with

geimsa stain and allow to air dry. Examine under microscope using oil immersion x 100 magnifications. If there is presence of malaria parasite it will look as shown below:

Blood smear showing ring state of malaria parasite 2.9 WIDAL (TYPHOD) TEST When a person is infected with pathogen organism the body immune system produces antibodies against the antigen present on the microbial cell with flagella (if present), these antibodies are usually detected 2 3 weeks after infection and reach their maximum concentration at 3 5 weeks. In many cases where the involving micro-organism induces a tebril reaction in the patient, the specificity on the antibody can be demonstrated by testing the patient serum against bacteria of known types and stain. Apparatus: Patients blood, tile, widal kit (antigen) hand pipette. Methods: 2 ml of blood is collected from the patient through the vein and dispensed into the test-tube and allowed to settle. The blood is then centrifuged to separate the serum from the plasma. The serum is drawn from the test-tube using hand pipette and then drop into eight places on the white clean tole. Shake the antigen suspension and put a drop or two of s. typhyi D, S paratyphic A, S paratyhi B and S paratyphi C all

have O and H antigen to the eight drop of the serum respectively and then mix over an area of 2cm, rock gently for 3-5 minutes and examine for agglutination. Agglutination shows positive while no reaction shows negative. It is shown in the stage below.

A A1

B B1

C C1

D D1

Tile

The eight drops of serum with paratyphi

Agglutination

No reaction

TE: D D1 A A1 B B1 = = = = = = S. Typhi 0c S. Typhi Hag S. Paratyphi 0a S. Paratyphi Hag S. Paratyphi Bc S. Paratyphi Hag

HEPATITIS TEST (HBSAG, HCV, AND HBC) Hepatitis B (HBSAG) is a viral disease and dangerous to the body.

Apparatus Contents of blood, test-tube and HBSAG strip Procedure After the blood has been collected from the patient (2 ml) it was suspended and allowed to settle. The settled blood was centrifuged and separated from the plasma. The strip is then dipped inside the serum test tube without touching the plasma. The serum will then rise in the tube through capillary action. Double line (i.e. test and control) shows positive while single line (control) shows negative, it is as shown below.

Test Control

Double line (positive)

Control Single line (negative)

HBsag strip showing positive

HBsag strip showing positive

HCV and HBc are carried out with the same respective strips using the same procedure. Double and single lines are also the same as in HBsag.

CHAPTER THREE 3.0 MICROBIOLOGY Microbiology laboratory is divided into bacteriology/parasitology. The activities carried out here are mainly concerned with investigation of reported cases of diseases caused by bacterial and parasite. 3.1 CONCEPT OF CULTURE
1. Chocolate Agar: It is an enriched medium for the inoculation of pus,

TYPES AND DEFINITION OF AGAR (MEDIUM) swab, sputum etc. It is prepared from nutrient agar by adding blood and heat for a while.
2. Sabouraud 4% Detrose Agar: It is a culture medium for pathogenic and

non-pathogenic fungi especially dermatophytes. It is used for HVS inoculation. It has a pH of 7.0 +0.2.
3. Cytene Lactose Electrotytes Deficiency (C. L. E. D): It is a differential

medium for the enumeration of urinary track pathogens. It is used for inoculating urine. It is a medium that has a deficiency of electrolyte.
4. Nutrient Agar: It is used for sensitivity and has a pH of 7.3 + 0.2. 5. Salmonella Shigela Agar:

It is highly differential medium for the

isolation of salmonella and shigella from clinical specimen. It is used for inoculation of stool. 3.4 PREPARATION OF AGAR (MEDIUM) Preparation of agar depends on what types of agar is to be prepared. But all have things in common that is the measurement and application of solvent which is usually, distilled water. Chocolate agar is prepared by adding fresh blood specimen to already sterilized broth, heat on a spirit lamp till it gives a chocolate colour and is poured in a Petri dish to solidify. The result media after pouring the specimen is referred to as blood agar.

Salmonella shigella agar is prepared by weighing 3g of S.S.A dissolved in 50 ml of distilled water and then sterilized by boiling and allowed for few minutes before pouring it in a Petri dish (plate). Nutrient agar is prepared by dissolving 3g of nutrient agar on 100 ml of distilled water mix well and then sterilized in an autoclave at 121oC for 15 minutes. Sabour and Dextrose agar is prepared by weighing 3g of S. D. A dissolved in 50 ml of distilled water and then sterilized in an autoclave at 121oC for 15 minutes. All agars are allowed to cool to 40oC before pouring into a sterilized Petridish and are allowed to gel or solidify. After gelling, they are allowed to dry in an incubator for 24 hours 3.3 CULTURE AND SENSITIVITY TEST To carry out sensitivity test, the bacteria insolated from the culture from the culture media (a sub cultural into the nutrient agar), various antibiotic known as tipped multiple antimicrobial disc which is kept at 2 8oC are placed at different location within the sub-cultural plate. Failure of the isolates to grow close to a specific antibiotic indicates that it is susceptible or sensitive to that antibiotic. Growth close to or burst around the antibiotic indicated to suppress bacteria diseases are administered by doctors and scientists. These antibiotics can be under gram positive and gram negative. The gram negative is mostly used for urinary track isolate. Examples of some antibiotics are Augmentrin, tetracycline, amoxylin, erythromycin, seprin etc. 3.4 STERILIZATION Sterilization can be defined as the process of dis-infecting materials/objects in the laboratory. METHOD OF STERILIZATION Boiling Method: The use of water bath/boiling water

Steaming Method: Pressurized steam eg autoclaves. Hot air Method: Eg. Hot air method for stile bottles. Flaming Method of sterilization: Heating the object to real hot or passing through flame, for metals. 3.5 HIGH VAGINAL SWAB (HVS) This test is done on the search for bacteria infection eg. Staphyloccus, aueus, Candida albica. The process involves microscopy, culture and sensitivity. PROCEDURE: HVS hand glove are worn; sterilized swab stick is inserted into the inner vagina and rotate randomly to collect enough fluid for microscopic, culture and sensitivity. Inoculation is done, smear for microscopy and sensitivity after growth. 3.6 PREGNANCY TEST This is a test used to determine whether a woman is having a baby in her womb or not. Procedure: Collect 2 ml of the patients urine and insert a pregnancy test strip into the urine. Allow it to stay for 3 seconds and remove the strip. The urine then move the strip through capillary action and left for 5 minutes. The result is either positive (double line) or negative (single line). 3.7 URINALYSIS It is a test used to detect abnormal substance in the urine such as ketone, bilirubin, ascorbic acid glucose etc. It is dine using urine and uristick. PROCEDURE

The urine sample is collected, the test strip is flooded with the urine and left for 2 seconds, the likely colour change in the embodied strip is marked and recorded. 3.8 URINE MICROSCOPY This is examination of urine microscopically. The process needs apparatus which are slide, test tube, urine sample, centrifuge machine and microscopes. PROCEDURE Some quantity of urine is been poured into a test tube and spun for 5 minutes in a centrifuge machine at high speed, then after 5 minutes, the machine is stopped and the supernatural been discarded so that the urine sediment will mix together thoroughly. The urine sediment is placed on the slide and examined under microscope with low and high power magnifications respectively. The content in the urine sediment is identified using guides. The following cells may be seen namely: epithelia cell, pus cells, yeast cell, R. B. C. east etc. 3.9 URINE CULTURE AND SENSITIVITY This is done to check an infection in urine by growth of organism(s) in inoculated urine. Apparatus: Patients urine, prepared chocolate agar, Bunsen burner, wire loop. Procedure The wire loop is flamed and used to collect a pinch of urine and smeared on a portion in the chocolate agar. It is then covered and dropped in an incubator for 24 hours. After that the growth of the organism is examine. If there is, it is isolated and sub-cultured I a nutrient agar plat for sensitivity. Sensitivity now is the application of anti-microbial disc to determine drug(s) that can cure the disease.

Now after sub-culture, anti microbial disc is then applied. Such ant- microbial are CPX (Ciproflaxin), Sptrin, AG (Augnentin). After 24 hours it is then examined, if there is a clear space around the anti microbial disc it then means that the microbial disc is sensitive to the disease but if there is no clear space it means that the drug is resistive to the disease and cannot be used to cure the sickness. 3.10 STOOL CULTURE AND SENSITIVITY This is done to check for infection in a stool by growth of organism (8) in an inoculated stool. Apparatus: Stool, prepared chocolate agar, Bunsenn burner, wire loop, incubator etc. Procedure The wire loop is flamed and use to collect a ball of stool and smeared in a portion in the chocolate agar plate. It is then covered and dropped in an incubator for 24 hours. After the hour given the growth of the organism is examined, if there is, it is isolated and sub cultured in a salmonella shigella agar for sensitivity. Anti microbial drugs (disc) are placed on the sub cultured sample and left for another 24 hours. Such anti microbial drugs are tetracycline etc. Finally, after the hour given the isolated sample (sub cultured sample) is then examined. If there is a clear space in the portion where the anti microbial discs are in contact with isolated sample it means that the drugs are sensitive to the disease but if there is no clear space, it is resistive. That is to say sensitive drugs cure whilre resistive drugs do not cure disease.

CHAPTER FOUR 4.0 CHEMICAL PATHOLOGY This is the scientific study of disease and their causative agent. 4.1 FASTING AND RANDOM BLOOD SUGAR The above test is done to check the amount of sugar in the blood, that is, to check whether a patient is having diabetes or not. The different between fasting blood sugar and random blood is that in fasting food sugar (FBS) blood is collected before the patient eats anything while random blood sugar (RBS) is collected two hours after eating. Apparatus: Glucose buffer reagent, sample (blood), standard, distilled water, specto-photometer. Procedure TEST Glucose buffer Sample Standard Distilled water .. 2ml . .. STANDARD 2ml 2ml . BLANK 2ml . 2ml

Mix the above well and incubate at 37oC for 10 minutes at room temperature, read absorbent and zero blank. RESULT: The colour is read with the aid of photospectometer or spectophotometer at a wavelength at a wavelength of 500 mm. The intensity of the colour formed is directly proportional to the glucose amount present in the sample. CALCULATION OD Optical density of test x concentration OS Optical density of standard solution

CONCLUSION The Students Industrial Work Experience Scheme is a very useful programme in students life. This is because it has been noticed that it exposes the student to several practical knowledge especially industrially. During the course of the training with Mount Arafat Hospital, the experience gained cannot be overemphasized. So many things were learnt as stated in the early page of the report in addition to the basic knowledge of the industrial practice of theoretical information received in the lecture rooms. The training gave an opportunity where students can acquire more practical skills and interact with people. In whole, the scheme organized is highly appreciated because it has really made students understand the happening in his/her field of study to practice it more. RECOMMENDATION It is hereby recommended that students unit Federal Polytechnic Idah, the ITF visiting officials should make a provision for transport allowances. Also, they should see that students are paid their allowances at the appropriate time and should also avoid the omission of students names in vouchers.