Industrial training report

1. REPORT ON STUDENT INDUSTRIAL EXPERIENCE SCHEME(SIWES)
TRAINING PROGRAMME AT
PETROLEUM TRAINING INSTITUTE EFFURUN, DELTA STATE
BY
EHIMEN, EJOR PRINCESS
MATRIC NO: ECU/2019/SCN/017/806
SUBMITTED TO THE DEPARTMENT OF MICROBIOLOGY, FACULTY OF
BIOLOGICAL SCIENCES, EDWIN CLARK UNIVERSITY, KIAGBODO,
DELTA STATE.
IN PARTIAL FUFILLMENT OF THE REQUIREMENT FOR THE
AWARD BACHELOR OF SCIENCE(BS.c) DEGREE IN
MICROBIOLOGY SUPERVISOR
SUPERVISED BY:

2. DEDICATION
I sincerely dedicate this report to GOD Almighty for giving me the wisdom, knowledge,
understanding, strength and patience to start, continue and complete my industrial training
program. Also with utmost gratitude to my lovely parents MR and MRS OMOROGBE and my
siblings for their support during my industrial training

3. ACKNOWLEDGEMENT
I give thanks to GOD Almighty for his mercies upon my life, seeing me through my industrial
training program and the writing of this report. I also want to acknowledge the

4. ABSTRACT
The Student Industrial Work Experience Scheme established by the Federal Government of
Nigeria was aimed at exposing student of higher institutions to acquire industrial skills and
practical experience in their approved course of study and also to prepare the students for the
industrial work situation which they are likely to meet after graduation. This technical report is
based on the experiences gained during my six months of industrial training at Petroleum
Training Institute/Clinical/Medical laboratory Efferun Delta state. This report highlights how
patients are being managed and also the several test carried out for patients such as Pregnancy
test (PT), Packed cell volume test (PCV), Widal (Typhoid) Test, Culture methods, Serial dilution. I
was opportune to work in the department of clinical and biology lab.

5. TABLE OF CONTENT
TITLE PAGE
DEDICATION -------------------------------------------------------------------------------------
ACKNOWLEDGEMENT--------------------------------------------------------------------------
ABSTRACT----------------------------------------------------------------------------------------
TABLE OF CONTENTS--------------------------------------------------------------------------
CHAPTER ONE
1.0 INTRODUCTION
1.1 OVERVIEW OF SIWES-------------------------------------------
1.2 BRIEF HISTORY OF PETROLEUM TRAINING INSTITUTE (PTI)
1.3 DEPARTMENTS IN PETROLEUM TRAINING INSTITUTE
CHAPTER TWO
2.0 THE LABORATORY (MICROBIOLOGY LABORATORY & CLINICAL LABORATORY)
2.1 INTRODUCTION TO THE LABORATORY
2.2 SAFETY RULES IN THE LABORATORY
2.3 APPARATUS IN THE LABORATORYAND THEIR USES

6. CHAPTER THREE
3.0 THE MICROBIOLOGY LAB
3.1 DILUTION TECHNIQUES AND PIPETTING
3.2 MEDIA PREPARATION
3.3 CULTURE METHODS
3.4 MICROSCOPIC EXAMINATION OF THE MAIN GROUPS OF MICRORGANISM
3.5 GRAM STAINING
3.6 METHODS OF ISOLATION OF PURE CULTURE
CHAPTER FOUR
4.0 THE CLINICAL LABORATORY AND VARIOUS TEST PERFORMED
4.1 WIDAL TEST
4.2 PREGNANCY TEST
4.3 URINALYSIS TEST
4.4 PACKED CELL VOLUME TEST
4.5 MALARIA PARASITE TEST
CHAPTER FIVE
5.1 RELEVANCE OF SIWES

7. 5.2 RECOMMENDATIONS
5.3 CONCLUSION
REFERENCES

8. CHAPTER ONE
1.0 THE HISTORY AND SCOPE OF SIWES
The Student Industrial Work Experience Scheme (SIWES), also known as Industrial
Training is a compulsory skills training programme designed to expose and prepare
students of Nigerian Universities, Polytechnics, Colleges of Education, Colleges of
Technology and Colleges of Agriculture, for the industrial work situation they’re
likely to meet after graduation.
The scheme also affords students the opportunity of familiarizing and exposing
themselves to the needed experience in handling equipment and machinery that
are usually not available in their institution.
Before the establishment of the scheme, there was a growing concern among
industrialists, that graduates of institutions of higher learning lacked adequate
practical background studies preparatory for employment in industries.
Thus, employers were of the opinion that the theoretical education in higher
institutions wasn’t responsive to the needs of the employers of labour.
SIWES introduction, initiation and design was done by the Industrial Training Fund
(I.T.F) in 1993 to acquaint students with the skills of handling employer’s equipment
and machinery.
The Industrial Training Fund (I.T.F) solely funded the scheme during its formative
years. However, due to financial constraints, the fund withdrew from the scheme
in 1978.
The Federal Government, noting the significance of the skills training handed the
management of the scheme to both the National Universities Commission (N.U.C)
and the National Board for Technical Education (N.B.T.E) in 1979.
The management and implementation of the scheme was however reverted to the
I.T.F by the Federal Government in November, 1984 and the administration was

9. effectively taken over by the Industrial Training Fund in July 1985, with the funding
solely borne by the Federal Government.
1.1. OBJECTIVES OF SIWES IN NIGERIA
Provides the avenue for students in institutions of higher learning to acquire
industrial skills and experiences in their course of study. Prepare the students for
the industrial work situation they’re likely to meet after graduation.
Expose students to work method and techniques in handling equipment and
machinery that may not be available in their institutions.
Make the transition from school to the world of work easier and enhance students
contact for later job placement.
Provides students with an opportunity to apply their knowledge in real work
situations thereby bridging the gap between theory and practice
Enlist and strengthens employers involvement in the entire educational process
and prepare students for employment after graduation.
1.2. BRIEF HISTORY OF PETROLEUM TRAINING INSTITUTE (PTI)
Petroleum Training Institute (P.T.I.) in Effurun, Delta State was established in 1973
by the federal government of Nigeria as a prerequisite for the membership in the
Organization of Petroleum Exporting Countries (OPEC) to train indigenous middle-
level manpower to meet the labour force demands of the oil and gas industry in
Nigeria and the West African sub region. It awards General Welding Certificates,
ND (National Diploma) and HND (Higher National Diploma) certificates. It also has
an affiliation with the standard laboratory which is a subsidiary of the standard
hospital located in Warri. As an oil industrial bound institute, the need for other
departments has over time become of great importance as well as doing everything
to meet up with world standard and also venture into other aspects of sciences.

10. This necessitated the introduction of the science and laboratory technology (SLT)
department.
To a great extent, the likes of the biochemist, biologist and basically other life
sciences can now venture there in the opportunity presented in the oil company.
Furthermore, the need for a healthy atmosphere cannot be overemphasized as
staff and students need to know their health status and hence the introduction of
the PTI clinic.
1.3. DEPARTMENT IN PETROLEUM TRAINING INSTITUTE
The departments in the Petroleum Training Institute are as follows:
• Science Laboratory Technology Department (SLT)
• Industrial Safety And Environmental Technology Department (ISET)
• Petroleum And Natural Gas Processing Department (PNGPD)
• Electrical Engineering (EED)
• Petroleum Marketing And Business Studies (PMBS)
• Mechanical Engineering (MED)
• Petrochemical Engineering (PEG)
• Computer Science And Information Technology (CSIT)
• Petroleum Engineering And Geosciences (PEG)
• Department Of Welding And Offshore Technology (DWOT)

11. CHAPTER 2
2.0. THE LABORATORY (BIOLOGY LABORATORY & CLINICAL LABORATORY)
2.1. INTRODUCTION TO THE LABORATORY
The laboratory is quite equipped with adequate tools and apparatuses.
The clinical laboratory which is specific for various testing and result deduction is
provided with the appropriate equipment for the working condition to be effective.
I was also introduced to the medical laboratory scientists in carrying out certain
test. I was immediately briefed on the safety rules in the laboratory.
The biology lab is also well equipped with adequate tools apparatuses. Biological
science Laboratory Apparatus enrich the quality of life in numerous ways by
providing new solutions in health and materials and energy usage.
2.2. SAFETY RULES IN THE LABORATORY
Some of the safety rules include:
1. Do not touch or handle any equipment without the permission or supervision of
the technician.
2. Avoid placing your hands freely on equipment without the use of surgical gloves.
3. You must be properly suited at all times with laboratory coat.
4. Operation of equipment should be done with discretion and must be carried out
by skilled personnel.

12. 5. Report any incident or unusual occurrence to the appropriate authority.
6. DO NOT eat in the laboratory.
7. NO CHARGING of phones near working equipments.
2.3. APPARATUS IN THE BOTH LABORATORIES AND THEIR USES
Some apparatus in the laboratory and their uses are as follows:
Microscope: It is used to magnify objects and it is mainly used for viewing cells and
tissues to make them look larger. Thus it is possible to detect the presence of
bacteria, virus and other infections in the blood sample. They are also used to study
the sediments of urine to detect kidney problems.
Haematology Analyzer: this type of medical lab equipment is used for conducting
blood tests using the blood samples. It is possible to come up with blood count and
perform coagulation tests.

13. Urinalysis Analyzer: This medical lab equipment is mainly used for tracking illness
due to the infections in the urinary tract.
Immunoassay Analyzer: This type of medical lab equipment is used for the
diagnosis of various infectious diseases. Testing of cancer markers and cardiac
analysis are also done with this device.
Medical Autoclave: This type of medical lab equipment is mainly used for sterilizing
surgical and pharmaceutical items.

14. Micropipette: It is special laboratory equipment which is mainly used for measuring
liquids of smaller volume so that it can be transferred easily. The measurements
are specified in the pipette. By this, it is possible to transfer a smaller quantity of
liquid from one container to another.
Tourniquet: This is a constricting or compressing apparatus that is used to control
venous blood flow for a short period of time. It is used to make the vein more
prominent for venipuncture.

15. Syringe: A syringe is a medical device with a nozzle and a piston or bulb for
injecting fluids into or withdrawing fluids from the body.
Centrifuges: This type of lab equipment comes in various shapes and sizes. The
equipment is capable of separating the non-soluble material from the available
sample. Thus it is used for separating cells from a medium.

16. Magnetic Stirrers: This laboratory equipment serves as an automatic stirrer. A
magnetic bead stirs the solution
Water Bath: This laboratory equipment is used for transferring heat to a certain
volume of liquid. It is used for maintaining the required temperature of the cell
culture medium bottles.
Glasswares: This is the most essential lab equipment. Some of the types of
glassware used in laboratories are beakers, flasks etc. They are used for
transferring, storing and measuring liquids. Florence flask is a special type of
glassware used for heating liquids.

17. Test Tubes: These are common lab equipment used for collecting various samples.
It is also used for mixing and heating of certain chemical solution.
Incubator: This is a laboratory equipment that provides a condusive environment
for microbial growth.
Hot air oven: It is a laboratory equipment that uses heat to sterilize glasswares,
powders, materials containing oil, and metal equipment (scissors and blades).

18. Autoclave: This laboratory equipment provides a physical method of sterilization by killing
bacteria, viruses, and even spores present in the material put inside of the vessel
using steam under pressure.
Spectrophotometer: This laboratory equipment is used to measure the amount of
light absorbed by a sample.
.

19. CHAPTER THREE
3.0. THE MICROBIOLOGY LAB
A microbiology laboratory is a laboratory devoted to the culturing, examinations
and identifications of mircoorganisms including bacteria, fungi, algae, e.t.c. The
microbiology lab has a crucial role in effective infection prevention and control
(IPC). The microbiology lab is used to determine the most frequent microbes
causing healthcare-associated infections, and perform some basic typing of
microorganisms for epidermological examinations. Below are some basic activities
that are being carried out in the microbiology lab.
1) Media Preparation
2) Dilution Techniques and Pipetting
3) Bacterial Colony Morphology
4) Antimicrobial Chemicals
5) Bacterial Numbers
6) Use of the Microscope
7) Simple Staining
8) Gram Staining
3.1 DILUTION TECHNIQUES
Bacteria are often present in such huge numbers that they can be difficult to count.
Dilution requires the thorough mixing of a small, accurately measured sample with
a large volume of sterile water, saline or other appropriate liquid called the diluent
or a dilution blank.
MATERIALS NEEDED FOR DILUTION TECHNIQUES:
 Flask of blue water
 Nonsterile pipettes - 1, 5, and 10ml
 6 nonsterile test tubes
 Blue and green pi-pumps

20. THE PROCEDURE FOR DILUTION TECHNIQUE
Fig. 4
1. You will need a bottle of blue dye.
2. Prepare the following dilution blanks with tap water using a 10ml pipette and
the green pi-pump---4.5ml, 9ml, 9.5ml, 4ml, 3ml, 12ml .
3. Make 3 sets of dilution tubes as seen in the Fig. 4. Mix the tube contents.
 Dilution set 1: Transfer 0.5ml of blue water into the 4.5ml of water,
then 1ml of tube 1 into the next tube of 9ml water.
 Dilution set 2: Transfer 1ml of blue water into the 4ml of water, then
0.5ml of tube 1 into the next tube of 9.5ml.
 Dilution set 3: Transfer 1ml of blue water into the 3ml of water, then
0.5ml of tube 1 into the next tube of 12ml.
PIPETTING: A pipette (sometimes spelled pipet) is a laboratory tool commonly used
in chemistry, biology and medicine to transport a measured volume of liquid, often
as a media dispenser. Pipettes come in several designs for various purposes with
differing levels of accuracy and precision, from single piece glass pipettes to more
complex adjustable or electronic pipettes. Many pipette types work by creating a
partial vacuum above the liquid-holding chamber and selectively releasing this
vacuum to draw up and dispense liquid. Measurement accuracy varies greatly
depending on the instrument.
4.3 CULTURE METHODS
Microbial cultures are used to determine the type of organism, its abundance in
the sample being tested, or both. It is one of the primary diagnostic methods of
microbiology and used as a tool to determine the cause of infectious disease by

21. letting the agent multiply in a predetermined medium. For example, a throat
culture is taken by scraping the lining of tissue in the back of the throat and blotting
the sample into a medium to be able to screen for harmful microorganisms, such
as Streptococcus pyogenes, the causative agent of strep throat. Furthermore, the
term culture is more generally used informally to refer to "selectively growing" a
specific kind of microorganism in the lab.
There are several types of bacterial culture methods that are selected based on the
agent being cultured and the downstream use.
Broth cultures:
One method of bacterial culture is liquid culture, in which the desired bacteria are
suspended in a liquid nutrient medium, such as Luria Broth, in an upright flask. This
allows a scientist to grow up large amounts of bacteria for a variety of downstream
applications.
Liquid cultures:
These are ideal for preparation of an antimicrobial assay in which the experimenter
inoculates liquid broth with bacteria and lets it grow overnight (they may use a
shaker for uniform growth). Then they would take aliquots of the sample to test for
the antimicrobial activity of a specific drug or protein (antimicrobial peptides).
Liquid cultures of the cyanobacterium Synechococcus PCC 7002. As an alternative,
the microbiologist may decide to use static liquid cultures. These cultures are not
shaken and they provide the microbes with an oxygen gradient.
Agar plates:
Microbiological cultures can be grown in petri dishes of differing sizes that have a
thin layer of agar-based growth medium. Once the growth medium in the petri dish
is inoculated with the desired bacteria, the plates are incubated at the optimal
temperature for the growing of the selected bacteria (for example, usually at 37
degrees Celsius, or the human body temperature, for cultures from humans or
animals, or lower for environmental cultures). After the desired level of growth is
achieved, agar plates can be stored upside down in a refrigerator for an extended
period of time to keep bacteria for future experiments.

22. There are a variety of additives that can be added to agar before it is poured into a
plate and allowed to solidify. Some types of bacteria can only grow in the presence
of certain additives. This can also be used when creating engineered strains of
bacteria that contain an antibiotic-resistance gene. When the selected antibiotic is
added to the agar, only bacterial cells containing the gene insert conferring
resistance will be able to grow. This allows the researcher to select only the colonies
that were successfully transformed.
Agar based dipsticks:
Miniaturised version of agar plates implemented to dipstick formats, eg. Dip Slide,
Digital Dipstick show potential to be used at the point-of-care for diagnosis
purposes. They have advantages over agar plates since they are cost effective and
their operation does not require expertise or laboratory environment, which
enable them to be used at the point-of-care.
Stab cultures:
Motile and non-motile bacteria can be differentiated along the stab lines. Motile
bacteria will grow out from the stab line while non-motile bacteria are present only
along the stab line.
Stab cultures are similar to agar plates, but are formed by solid agar in a test tube.
Bacteria is introduced via an inoculation needle or a pipette tip being stabbed into
the center of the agar. Bacteria grow in the punctured area. Stab cultures are most
commonly used for short-term storage or shipment of cultures.
Culture collections:
Microbial culture collections focus on the acquisition, authentication, production,
preservation, catalogueing and distribution of viable cultures of standard reference
microorganisms, cell lines and other materials for research in microbial
systematics. Culture collection are also repositories of type strains.
3.2 MEDIA PREPARATION
Bacteria and fungi are grown on or in microbiological media of various types. The
medium that is used to culture the microorganism depends on the microorganism
that one is trying to isolate or identify. Different nutrients may be added to the
medium, making it higher in protein or in sugar. Various pH indicators are often
added for differentiation of microbes based on their biochemical reactions: the

23. indicators may turn one color when slightly acidic, another color when slightly
basic. Other added ingredients may be growth factors, NaCl , and pH buffers which
keep the medium from straying too far from neutral as the microbes metabolize.
In this exercise, you will make all-purpose media called trypticase soy broth and
trypticase soy agar. These 2 media----one a liquid and the other a solid---are the
exact same formula save for the addition of agar agar (really- agar agar), an extract
from the cell walls of red algae.
The old way to make media was by the cookbook method--- adding every
ingredient bit by bit. The only time that is done today is when making a special
medium to grow a certain finicky organism, where particular growth factors,
nutrients, vitamins, and so on, have to be added in certain amounts. This medium
is called a chemically defined medium (synthetic). Fortunately, the most common
bacteria that we want to grow will do nicely with media that we commonly use in
lab. Some of our media is bought, but most is produced in the prep area behind the
lab. Since this type of medium has some unknown ingredients, or sometimes
unknown quantities it is called complex media.
It is really very simple to make complex media these days:
rehydrate the powder form of the medium
stir and boil the agar medium to get the agar powder dissolved (if making an agar
medium rather than a broth medium)
distribute the medium into tubes
autoclave to sterilize the tube media
autoclave the agar medium for plate production and then pour into sterile petri
dishes
3.3. STERILIZATION AND THE AUTOCLAVE
When microbiological media has been made, it still has to be sterilized because of
microbial contamination from air, glassware, hands, etc. Within a few hours there
will be thousands of bacteria reproducing in the media so it has to be sterilized
quickly before the microbes start using the nutrients up. The sterilization process is
a 100% kill, and guarantees that the medium will stay sterile UNLESS exposed to
contaminants by less than adequate aseptic technique to exposure to air.

24. Media sterilization is carried out with the autoclave, basically a huge steam cooker.
Steam enters into a jacket surrounding the chamber. When the pressure from the
steam is at a certain point in the jacket, a valve allows the steam to enter the
chamber. The pressure will go up over 15 pounds per square inch (psi): at this point
the timer begins to count down--- usually for 15 minutes, depending on the type of
media. The high pressure in a closed container allows the temperature to go above

25. the highest temperature one could get by just boiling, around 121⁰C. Therefore,
the parameters for sterilization with an autoclave are 121⁰C at >15 psi for 15
minutes. Fifteen minutes is the thermal death time for most organisms (except
some really hardy sporeformers).
The prepared media is distributed in different ways, depending on the form one is
making. Broths and agar deeps are dispensed into tubes and then sterilized. Agar
slant tubes are sterilized and then the rack is tilted to allow the agar to solidify in a
slanted fashion. Agar medium to be be poured into plates is sterilized in a flask, and
then poured afterward. Not all media or solutions can be sterilized via an autoclave.
Certain high-protein solutions such as urea, vaccines, and serum will denature in
the extreme heat, and so they may have to be filter-sterilized without heat. You will
be making slant and broth media, but not plate media in this lab.
MATERIALS NEEDED (Per table)
2 plastic weigh boats
1 test tube rack
1-1 liter Erlenmeyer flask
1 pipet pump
1 graduated cylinder several nonsterile glass10 ml pipets
1 spatula
28 medium, nonsterile test tubes
1 jar agar powder 15 green caps
1 jar nutrient trypticase soy broth powder
15 yellow caps
1 magnetic stir bar
1 pipet disposal jar
THE PROCEDURE
Refer to the diagram below for the entire production:

26. Figure 1: General procedure to make media
Begin making the TSB (broth) by pouring 250ml of distilled water into a 500ml or
1L flask. Put in the stir bar and turn on the stir plate so that the surface is just
disturbed. Add 3.25 grams of the TSB powder to this flask and allow it to dissolve
(will happen quickly). No heat need be applied at this stage.
Once the powder is dissolved, pipet out 5ml green cap.
Green caps are always used for TSB.
With the remaining solution (about 100ml) still stirring, add 2 grams of agar
powder.
The next step will require you to apply heat to the mixture. Before you do this,
however, you should be aware that agar has a strong tendency to boil over when it
reaches 100⁰C. Someone in your group should be watching the flask at all times
once you see steam coming off of it. At the first sign that the mix is near boiling,
REMOVE it from the hot plate (paper towels around the flask neck). DO NOT simply
turn off the heat, letting the flask sit there. The metal plate retains a significant
amount of heat, and turning off the heat will not prevent the flask from boiling
over. Folded paper towels allow you to grasp the flask neck tightly, yet not burn
your hand.
Have you read step 4? OK, then you can turn on the heat to setting 9 (not High).
Make sure that the magnetic bar is stirring the solution.
Upon boiling, the agar dissolves, it will turn clear, deeper tan. Remove it from the
heat and pipet out 5ml aliquots into 15 tubes for slants (will not be BE slants until
removed from autoclave and tilted to the side to solidify). Cover the slant tubes
with yellow caps. THE REST OF THE AGAR MEDIUM IN THE FLASK WILL BE POURED
INTO 1 LARGE FLASK FOR THE CLASS.

27. From this point on, yellow caps will be used for nutrient agar slants.
3.4 MICROSCOPIC EXAMINATION OF THE MAIN GROUP OF MICRORGANISM:
The major groups of microorganisms—namely bacteria, archaea, fungi (yeasts and
molds), algae, protozoa, and viruses. Briefly explained below.
BACTERIA
They have a variety of shapes, including spheres, rods, and spirals. Individual cells
generally range in width from 0.5 to 5 micrometres (μm; millionths of a metre).
Although unicellular, bacteria often appear in pairs, chains, tetrads (groups of four),
or clusters. Some have flagella, external whiplike structures that propel the
organism through liquid media; some have capsule, an external coating of the cell;
some produce spores—reproductive bodies that function much as seeds do among
plants. One of the major characteristics of bacteria is their reaction to the Gram
stain. Depending upon the chemical and structural composition of the cell wall,
some bacteria are gram-positive, taking on the stain’s purple colour, whereas
others are gram-negative.
Microscopic view of a bacteria cell(salmonella)
Through a microscope the archaea look much like bacteria, but there are important
differences in their chemical composition, biochemical activities, and
environments. The cell walls of all true bacteria contain the chemical substance
peptidoglycan, whereas the cell walls of archaeans lack this substance. Many
archaeans are noted for their ability to survive unusually harsh surroundings, such
as high levels of salt or acid or high temperatures. These microbes, called

28. extremophiles, live in such places as salt flats, thermal pools, and deep-sea vents.
Some are capable of a unique chemical activity—the production of methane gas
from carbon dioxide and hydrogen. Methane-producing archaea live only in
environments with no oxygen, such as swamp mud or the intestines of ruminants
such as cattle and sheep. Collectively, this group of microorganisms exhibits
tremendous diversity in the chemical changes that it brings to its environments.
ALGAE
The cells of eukaryotic microbes are similar to plant and animal cells in that their
DNA is enclosed within a nuclear membrane, forming the nucleus. Eukaryotic
microorganisms include algae, protozoa, and fungi. Collectively algae, protozoa,
and some lower fungi are frequently referred to as protists (kingdom Protista, also
called Protoctista); some are unicellular and others are multicellular.
Unlike bacteria, algae are eukaryotes and, like plants, contain the green pigment
chlorophyll, carry out photosynthesis, and have rigid cell walls. They normally occur
in moist soil and aquatic environments. These eukaryotes may be unicellular and
microscopic in size or multicellular and up to 120 metres (nearly 400 feet) in length.
Algae as a group also exhibit a variety of shapes. Single-celled species may be
spherical, rod-shaped, club-shaped, or spindle-shaped. Some are motile. Algae that
are multicellular appear in a variety of forms and degrees of complexity. Some are
organized as filaments of cells attached end to end; in some species these filaments
intertwine into macroscopic, plantlike bodies. Algae also occur in colonies, some of
which are simple aggregations of single cells, while others contain different cell
types with special functions.
Microscopic view of an algae.
FUNGI

29. Fungi are eukaryotic organisms that, like algae, have rigid cell walls and may be
either unicellular or multicellular. Some may be microscopic in size, while others
form much larger structures, such as mushrooms and bracket fungi that grow in
soil or on damp logs. Unlike algae, fungi do not contain chlorophyll and thus cannot
carry out photosynthesis. Fungi do not ingest food but must absorb dissolved
nutrients from the environment. Of the fungi classified as microorganisms, those
that are multicellular and produce filamentous, microscopic structures are
frequently called molds, whereas yeasts are unicellular fungi.
Microscopic view of fungi (aspergillus).
PROTOZOA
Protozoa, or protozoans, are single-celled, eukaryotic microorganisms. Some
protozoa are oval or spherical, others elongated. Still others have different shapes
at different stages of the life cycle. Cells can be as small as 1 μm in diameter and as
large as 2,000 μm, or 2 mm (visible without magnification). Like animal cells,
protozoa lack cell walls, are able to move at some stage of their life cycle, and ingest
particles of food; however, some phytoflagellate protozoa are plantlike, obtaining
their energy via photosynthesis. Protozoan cells contain the typical internal
structures of an animal cell. Some can swim through water by the beating action of
short, hairlike appendages (cilia) or flagella. Their rapid, darting movement in a
drop of pond water is evident when viewed through a microscope.

30. Microscopic view of protozoa.
3.5 GRAM STAINING
Gram staining is a common technique used to differentiate two large groups of
bacteria based on their different cell wall constituents. The Gram stain procedure
distinguishes between Gram positive and Gram negative groups by coloring these
cells red or violet. Gram positive bacteria stain violet due to the presence of a thick
layer of peptidoglycan in their cell walls, which retains the crystal violet these cells
are stained with. Alternatively, Gram negative bacteria stain red, which is
attributed to a thinner peptidoglycan wall, which does not retain the crystal violet
during the decoloring process.
How Gram Staining Work
Gram staining involves three processes: staining with a water-soluble dye called
crystal violet, decolorization, and counterstaining, usually with safanin. Due to
differences in the thickness of a peptidoglycan layer in the cell membrane between
Gram positive and Gram negative bacteria, Gram positive bacteria (with a thicker
peptidoglycan layer) retain crystal violet stain during the decolorization process,
while Gram negative bacteria lose the crystal violet stain and are instead stained
by the safranin in the final staining process. The process involves three steps:
Cells are stained with crystal violet dye. Next, a Gram's iodine solution (iodine and
potassium iodide) is added to form a complex between the crystal violet and iodine.
This complex is a larger molecule than the original crystal violet stain and iodine
and is insoluble in water.
A decolorizer such as ethyl alcohol or acetone is added to the sample, which
dehydrates the peptidoglycan layer, shrinking and tightening it. The large crystal
violet-iodine complex is not able to penetrate this tightened peptidoglycan layer,
and is thus trapped in the cell in Gram positive bacteria. Conversely, the the outer
membrane of Gram negative bacteria is degraded and the thinner peptidoglycan

31. layer of Gram negative cells is unable to retain the crystal violet-iodine complex and
the color is lost.
A counterstain, such as the weakly water soluble safranin, is added to the sample,
staining it red. Since the safranin is lighter than crystal violet, it does not disrupt the
purple coloration in Gram positive cells. However, the decolorized Gram negative
cells are stained red.
How To- Staining Protocol and Concerns:
Reagents:
Crystal violet (primary stain)
Iodine solution/Gram's Iodine (mordant that fixes crystal violet to cell wall)
Decolorizer (e.g. ethanol)
Safranin (secondary stain)
Water (preferably in a squirt bottle)
1) Make a slide of cell sample to be stained. Heat fix the sample to the slide by
carefully passing the slide with a drop or small piece of sample on it through a
Bunsen burner three times.
2) Add the primary stain (crystal violet) to the sample/slide and incubate for 1
minute. Rinse slide with a gentle stream of water for a maximum of 5 seconds to
remove unbound crystal violet.
3) Add Gram's iodine for 1 minute- this is a mordant, or an agent that fixes the
crystal violet to the bacterial cell wall.
4) Rinse sample/slide with acetone or alcohol for ~3 seconds and rinse with a gentle
stream of water. The alcohol will decolorize the sample if it is Gram negative,
removing the crystal violet. However, if the alcohol remains on the sample for too
long, it may also decolorize Gram positive cells.
5) Add the secondary stain, safranin, to the slide and incubate for 1 minute. Wash
with a gentle stream of water for a maximum of 5 seconds. If the bacteria is Gram
positive, it will retain the primary stain (crystal violet) and not take the secondary
stain (safranin), causing it to look violet/purple under a microscope. If the bacteria
is Gram negative, it will lose the primary stain and take the secondary stain, causing
it to appear red when viewed under a microscope.

32. 3.6. METHOD OF ISOLATION OF PURE CULTURE
The following points highlight the most common methods used for obtaining pure
culture of microorganisms. The methods are: 1. Streak Plate Method 2. Pour Plate
Method 3. Spread Plate Method.
Streak Plate Method
This method is used most commonly to isolate pure cultures of bacteria. A small
amount of mixed culture is placed on the tip of an inoculation loop/needle and is
streaked across the surface of the agar medium (Fig. 16.13). The successive streaks
“thin out” the inoculum sufficiently and the micro-organisms are separated from
each other. It is usually advisable to streak out a second plate by the same
loop/needle without reinoculation. These plates are incubated to allow the growth
of colonies. The key principle of this method is that, by streaking, a dilution gradient
is established across the face of the Petri plate as bacterial cells are deposited on
the agar surface. Because of this dilution gradient, confluent growth does not take
place on that part of the medium where few bacterial cells are deposited.
Presumably, each colony is the progeny of a single microbial cell thus representing
a clone of pure culture. Such isolated colonies are picked up separately using sterile
inoculating loop/needle and re-streaked onto fresh media to ensure purity.
VARIOUS METHODS OF STREAKING
Pour Plate Method:
This method involves plating of diluted samples mixed with melted agar medium
(Fig. 16.14). The main principle is to dilute the inoculum in successive tubes
containing liquefied agar medium so as to permit a thorough distribution of
bacterial cells within the medium. Here, the mixed culture of bacteria is diluted
directly in tubes containing melted agar medium maintained in the liquid state at a
temperature of 42-45°C (agar solidifies below 42°C). The bacteria and the melted

33. medium are mixed well. The contents of each tube are poured into separate Petri
plates, allowed to solidify, and then incubated. When bacterial colonies develop,
one finds that isolated colonies develop both within the agar medium (subsurface
colonies) and on the medium (surface colonies). These isolated colonies are then
picked up by inoculation loop and streaked onto another Petri plate to insure
purity.
Pour plate method has certain disadvantages as follows:
(i) The picking up of subsurface colonies needs digging them out of the agar
medium thus interfering with other colonies, and
(ii) The microbes being isolated must be able to withstand temporary exposure to
the 42-45° temperature of the liquid agar medium; therefore this technique proves
unsuitable for the isolation of psychrophilic microorganisms. However, the pour
plate method, in addition to its use in isolating pure cultures, is also used for
determining the number of viable bacterial cells present in a culture.

34. Plate Method:
In this method (Fig. 16.15), the mixed culture or microorganisms is not diluted in
the melted agar medium (unlike the pour plate method); it is rather diluted in a
series of tubes containing sterile liquid, usually, water or physiological saline.
A drop of diluted liquid from each tube is placed on the center of an agar plate and
spread evenly over the surface by means of a sterilized bent-glass-rod. The medium
is now incubated.
When the colonies develop on the agar medium plates, it is found that there are
some plates in which well-isolated colonies grow. This happens as a result of
separation of individual microorganisms by spreading over the drop of diluted
liquid on the medium of the plate.
The isolated colonies are picked up and transferred onto fresh medium to ensure
purity. In contrast to pour plate method, only surface colonies develop in this
method and the microorganisms are not required to withstand the temperature of
the melted agar medium.

35. CHAPTER FOUR
4.0. THE CLINICAL LABORATORY AND VARIOUS TESTS PERFORMED
In the clinical/medical laboratory, various tests were performed but the major test
were that of widal(typhoid test) and bilirubin test.
4.1. WIDAL TEST
Introduction, Principle, Procedure, Result interpretation, Applications and
limitations
INTRODUCTION
Widal test is a serological test which is used for the diagnosis of enteric fever or
typhoid fever. The test was developed by Greembaum and Widal in 1896. Typhoid
or enteric fever is caused by a gram negative bacteria Salmonella enterica
(Salmonella Typhi or Salmonella Paratyphi), found in the intestine of man.
Salmonella paratyphi also causes Typhoid but of a milder form.
Salmonella possess O antigen on their cell wall and h antigen on their flagella. On
infection, these antigens stimulate the body to produce specific antibodies which
are released in the blood. The Widal test is used to detect these specific antibodies
in the serum sample of patients suffering from typhoid using antigen-antibody
interactions. These specific antibodies can be detected in the patient’s serum after
6 days of infection (fever).
Salmonella Typhi possesses O antigen on the cell wall and H antigen on flagella.
Salmonella Paratyphi A and S. Paratyphi B also possess O antigen on their cell wall
and but have AH and BH antigen on their flagella respectively.
PRINCIPLE OF WIDAL TEST:
Widal test is an agglutination test in which specific typhoid fever antibodies are
detected by mixing the patient’s serum with killed bacterial suspension of
Salmonella carrying specific O, H, AH and BH antigens and observed for clumping

36. ie. Antigen-antibody reaction. The main principle of Widal test is that if homologous
antibody is present in patient’s serum, it will react with respective antigen in the
suspension and gives visible clumping on the test slide or card.
FOR WIDAL TEST:
i) Fresh serum, stored at 2-8° Serum should not be heated or inactivated.
ii) The complete kit containing five vials containing stained Salmonella antigen
S. Typhi———-O antigen
S. Tyhhi———- H antigen
S. Paratyphi —–AH antigen
S. Paratyphi —–BH antigen
iii) Widal positive
control
iv) Widal test card or
slide
v) Applicator stick
PROCEDURE OF WIDAL TEST
Widal test can be done in two ways-one is rapid test on slide and another is tube
test in which result may be obtained after one night of incubation.
I. RAPID SLIDE TEST:
• Clean the glass slide or test card supplied in the kit well and makes it dry.

37. • Label the circles (1, 2, 3, 4, 5 and 6) in the test card as O, H, AH, BH, Negative
control and Positive control
• Place a drop of undiluted test serum in each of the four labelled circle (1, 2,
3 and 4) i.e. O, H, AH and BH and place a drop of Negative control serum in
circle 5 and Positive control in circle 6.
• Place a drop of antigen O, H, AH and BH in circle 1, 2, 3, and 4 respectively
and no antigen in circle 5 and O/H antigen in circle 6.
• Mix the content of each circle with a separate wooden applicator stick and
spread to fill the whole area of the individual circle.
• Rock the test card for a minute and observe for agglutination.
NOTE: if agglutination is visible within 1 minute, proceed for quantitative slide test
or tube test for the quantitative estimation of the titre of the antibody.
II. QUANTITATIVE SLIDE TEST:
• Clean the test card and make it dry
• Put 0.005 ml, 0.001 ml, 0.02ml, 0.04ml and 0.08ml of undiluted serum in 1st,
2nd, 3rd, 4th and 5th circles respectively in the test card.
• Add a drop of appropriate antigen suspension which showed agglutination
in rapid slide test, to each of the above circles.
• Mix the contents of each circle with a separate wooden applicator stick.
• Rock the slide slowly for 1 minute and observe for agglutination.
• The titre of the antibody is the highest dilution of serum up to which there is
clear agglutination.
• Repeats step 1 to 6 with all the antigens, which showed agglutination in rapid
slide test.

38. The serum volume in the quantitative slide test corresponds approximately to the
tube test is given below:
Fig.1: table showing the quantitative slide test
Circle Serum volume Antigen drop Approx.
titre
test tube
1st 0.08 ml 1 drop 1 : 20
2nd 0.04 ml 1 drop 1 : 40
3rd 0.02 ml 1 drop 1 : 80
4th 0.01 ml 1 drop 1 : 160
5th 0.005 ml 1drop 1 : 320
III. QUANTITATIVE TUBE TEST:
• Take a set of 8 clean dry test tubes (Kahn tubes) and label as 1, 2, 3, 4, 5, 6,
7 and 8 for O antibody detection.
Similarly, take 3 sets of 8 test tubes and label then as 1, 2…8.
• Dilute the serum samples as follows:
• Pipette into the tube No.1 of all sets 1.9 ml of isotonic saline.
• To each of the remaining tubes (2 to 8) add 1.0 ml of isotonic saline.
• To the tube No.1 tube in each row add 0.1 ml of the serum sample to be
tested and mix well.
• Transfer 1.0 ml of the diluted serum from tube no.1 to tube no.2 and mix
well.

39. • Transfer 1.0 ml of the diluted sample from tube no.2 to tube no.3 and mix
well. Continue this serial dilution till tube no.7 in each set.
• Discard 1.0 ml of the diluted serum from tube No.7 of each set.
• Tube No.8 in all the sets serves as a saline control. Now the dilution of the
serum sample achieved in each set is as follows: Tube No. : 1 2 3 4 5 6 7 8
(control) Dilutions 1:20 1:40 1:80 1:160 1:320 1:640 1:1280.
• 4. Add a drop of appropriate widal test antigen to all the test tubes
• 5. Mix well and incubate at 37°C for 16-20 hours and examine for
agglutination.
• 6. Antibody titre is the highest dilution of serum showing clear agglutination.
Fig.2: table showing the dilution result for various tubes
Test
tube
1 2 3 4 5 6 7 8
Dilution 1 : 20 1 : 40 1 : 80 1 :
160
1 :
320
1 :
640
1 :
1280
Control
(saline)
Result interpretation of Widal test:
Antibody titre greater than 1 : 80 is considered significant and usually suggests
positive test for Salmonella infection. Low titres are often in normal individuals
A single positive is less significant than the rising antibody titre, since rising titre is
considered to be a definite evidence of infection.
Applications of Widal test:
Rapid test for screening typhoid fever in endemic areas
When culture facilities is not available, Widal test is very reliable

40. Use for both Salmonella Typhi and Salmonella Paratyphi
LIMITATIONS OF WIDAL TEST:
Widal test is time consuming to find antibody titre and often times when diagnosis
is reached it is too late to start an antibiotic regimen.
Widal test may be falsely positive in patients who have had previous vaccination or
infection with S.Typhi.
Widal test cannot distinguish between a current infection and a previous infection
or vaccination against typhoid.
Widal test shows cross-reactivity with other Salmonella species.
False positive Widal test results are also known to occur in typhus, acute falciparum
malaria (Particularly in children), chronic liver disease associated with raised
globulin levels and disorders such as rheumatoid arthritis, myelomatosis and
nephrotic syndrome.
Widal test should be interpreted in the light of baseline titers in a healthy local
population. The antibody levels found in a healthy population however, may vary
from time to time and in different areas, making it difficult to establish a cut off
level of baseline antibody in a defined area and community.
Severe hypoproteinaemia may also prevent a rise in 0 and H antibody titres. False
negative Widal tests may be due to antibody responses being blocked by early
antimicrobial treatment or following a typhoid relapse.
In low typhoid endemic areas, weak and delayed O and H antibody responses limit
the usefulness of the Widal test. Variations also exist between laboratories in the
performance and reading of Widal tests which compromise further the reliability
of the test.

41. The World Health Organization (WHO) has said that due to the various factors that
can influence the results of a Widal test, it is best not to rely too much on this test.
RESULT INTERPRETATION OF WIDAL TEST
Result Interpretation of Widal Test
Positive: Agglutination within a minute
Negative: No agglutination indicative of absence of clinically significant levels of the
corresponding antibody in the patient serum.
The sample which shows the titre of 1:100 or more for O agglutinations and 1:200
or more for H agglutination should be considered as clinically significant (active
infection).
Demonstration of 4-fold rise between the two is diagnostic.
H agglutination is more reliable than O agglutinin.
Agglutinin starts appearing in serum by the end of 1st week with sharp rise in 2nd
and 3rd week and the titre remains steady till 4th week after which it declines.
Rising antibody titre is more convincing evidence of infection than positive tests
alone. It is preferable to test two specimens of sera at an interval of 7 to 10 days to
demonstrate a rising antibody titre.
4.1. PREGNANCY TEST
A blood pregnancy test is usually a quantitative blood test that shows how much
human chorionic gonadotropin(HCG) is in your blood. This is the type of blood
pregnancy test that most people are referring to when they talk about blood tests
for pregnancy. To test your blood for HCG, a simple blood sample is taken from one
of your veins, usually in the arm, through a procedure called a venipuncture.
HCG is a hormone secreted in pregnancy and detectable beginning a week to two
weeks (or more) after conception, depending on the test. Whether using a home
pregnancy test that uses urine or a blood pregnancy test from your doctor, the

42. results will be based on measurements of this hormone in your urine or blood. Both
types of tests are very reliable, producing about 99% accuracy when used correctly.
Difference Between Blood Test And Urine Test: A pregnancy blood test is more
sensitive than a urine test and can offer more information. Most blood tests can
detect slightly lower amounts of HCG, which means they can tell if you're pregnant
a few days earlier. Urine tests tend to require slightly more HCG to read positive,
making false-negative results a bit more likely with urine tests, although the rates
are quite low overall.
Urine HCG Test
• Needs slightly more HCG for a positive reading
• Qualitative test
• Tells you if you are pregnant or not, not how much HCG is in your blood
• Accurate a few days to a week after a missed period
Blood HCG Test
• Slightly more sensitive than urine tests
• Can be qualitative or quantitative
• Can track hCG levels over time
• Can be accurate before a missed period
Qualitative Tests: Urine tests are qualitative, meaning they either detect HCG in
your blood, giving a positive reading, or they don't, which is a negative reading. You
may also have a qualitative blood HCG test to measure HCG in your blood. Again,
the results are very clear: yes, you are pregnant because we found HCG in your
blood or, no, you are not pregnant because we didn't find it.
Urine pregnancy tests can usually detect pregnancy by about 10 days after
conception. Blood pregnancy tests can pick up HCG in the blood six to eight days
past ovulation. It's recommended to wait a week (or at least a few days) after a
missed period before taking a urine pregnancy test for optimal efficacy.
Quantitative Test: Another option is getting a quantitative HCG blood test (also
called the beta HCG test). These tests can give your doctor or midwife more
information than just whether or not you're pregnant. Quantitative HCG tests can

43. pick up even tiny amounts of the hormone and measure exactly how much HCG is
in your blood. This information can be compared over time.
In general, your HCG levels will nearly double about every two days in early
pregnancy. By having multiple blood tests about 48 hours apart, your doctor can
track your HCG levels to get a better read on your pregnancy, if needed. These serial
blood tests can help your provider monitor your pregnancy for miscarriage or
ectopic pregnancy as well as the possibility that you’re carrying multiples.
Due to stress, expense, and other factors, these tests are not done routinely (or
needed) for every pregnant woman.Talk to your doctor or midwife if you think that
a blood test for pregnancy is right for you. If your specific circumstances don't
warrant a blood test, you should be able to rely on the results of your home
pregnancy tests (HPT) instead.
When a Blood Test Is Used: In routine pregnancies with healthy women, at-home
urine tests are more than sufficient and effective. Blood tests are primarily used in
doctors' offices, specifically, if there is a potential issue with the pregnancy or some
other complicating factor (such as multiples), in which the greater sensitivity of the
blood test is needed.
Blood tests may be ordered for higher risk pregnancies, during fertility treatments,
when urine tests read negative but a period hasn't come, to confirm or rule out a
possible miscarriage or multiples, and to diagnose a potential ectopic or tubal
pregnancy, as well as due to other pregnancy complications.
Efficacy of Urine vs. Blood Pregnancy Tests
Urine tests are low-cost, painless, easy-to-use, can be done in the privacy of one's
home, and are very accurate (in most scenarios), which is why they are the standard
test used for most pregnancies. A blood pregnancy test is warranted due to
complicating factors. These tests are also very accurate, providing quick results that
can give doctors more detailed information about your pregnancy.
Interpreting the Results: Normal results from a quantitative HCG blood pregnancy
test would show that HCG levels are rising rapidly during the first trimester of

44. pregnancy and then declining slightly after around 10 weeks. At around 16 weeks,
the HCG levels stabilize for the remainder of the pregnancy. Abnormal results, on
the other hand, can mean a variety of things.
Higher HCG Than Expected: A higher than normal level of HCG may indicate:
• Infection or malignant tumors of the uterus
• More than one fetus (for example, twins or triplets)
• Non-cancerous tumors of the uterus
• Ovarian cancer
• Sometimes, just an indication of a normal pregnancy, farther along in
gestation
• Testicular cancer (in men)
Lower HCG Than Expected: Lower than normal levels of HCG may mean:
• Ectopic pregnancy
• Incomplete or complete miscarriage
• Possible death of the fetus.
4.1. URINALYSIS TEST
A urinalysis is a group of physical, chemical, and microscopic tests. The tests detect
and/or measure several substances in the urine, such as byproducts of normal and
abnormal metabolism, cells, cellular fragments, and bacteria.
Urine is produced by the kidneys, two fist-sized organs located on either side of the
spine at the bottom of the ribcage. The kidneys filter wastes out of the blood, help
regulate the amount of water in the body, and conserve proteins, electrolytes, and
other compounds that the body can reuse. Anything that is not needed is
eliminated in the urine, traveling from the kidneys through ureters to the bladder
and then through the urethra and out of the body. Urine is generally yellow and
relatively clear, but each time a person urinates, the color, quantity, concentration,
and content of the urine will be slightly different because of varying constituents.

45. Many disorders may be detected in their early stages by identifying substances that
are not normally present in the urine and/or by measuring abnormal levels of
certain substances. Some examples include glucose, protein, bilirubin, red blood
cells, white blood cells, crystals, and bacteria. They may be present because:
1. There is an elevated level of the substance in the blood and the body responds
by trying to eliminate the excess in the urine.
2. Kidney disease is present.
3. There is a urinary tract infection present, as in the case of bacteria and white
blood cells.
A complete urinalysis consists of three distinct testing phases:
1. Visual examination, which evaluates the urine’s color and clarity
2. Chemical examination, which tests chemically for about 9 substances that
provide valuable information about health and disease and determines the
concentration of the urine
3. Microscopic examination, which identifies and counts the type of cells, casts,
crystals, and other components such as bacteria and mucus that can be present
in urine
See below for details on each of these examinations.
A microscopic examination is typically performed when there is an abnormal
finding on the visual or chemical examination, or if a healthcare practitioner
specifically orders it.
Abnormal findings on a urinalysis may prompt repeat testing to see if the results
are still abnormal and/or may be followed by additional urine and blood tests to
help establish a diagnosis.
How is the sample collected for testing?
One to two ounces of urine is collected in a clean container. A sufficient sample is
required for accurate results.

46. Urine for a urinalysis can be collected at any time. In some cases, a first morning
sample may be requested because it is more concentrated and more likely to detect
abnormalities.
Sometimes, you may be asked to collect a “clean-catch” urine sample. For this, it is
important to clean the genital area before collecting the urine. Bacteria and cells
from the surrounding skin can contaminate the sample and interfere with the
interpretation of test results. With women, menstrual blood and vaginal secretions
can also be a source of contamination. Women should spread the labia of the
vagina and clean from front to back; men should wipe the tip of the penis. Start to
urinate, let some urine fall into the toilet, then collect one to two ounces of urine
in the container provided, then void the rest into the toilet.
A urine sample will only be useful for a urinalysis if taken to the healthcare
provider’s office or laboratory for processing within a short period of time. If it will
be longer than an hour between collection and transport time, then the urine
should be refrigerated or a preservative may be added.
4.2. PACKED CELL VOLUME
A lower number of the PCV means that the RBC count loss is due to reasons such
as blood loss, cell destruction and less bone marrow production. Increased PCV can
generally mean that a person is dehydrated and there is a higher number of RBC
production.
By looking at the tube out of the centrifuge, you can get an idea of the WBC content
as well. This buffy coat normally lies between the plasma and red cell layer. (This
shouldn't be counted as a part of the PCV test).
The layer of plasma should also be examined for lipemia, hemolysis in addition to
icterus.
Preparation For The PCV Test

47. There isn't any special preparation required for the PCV test. If you are anxious
about the test, it is better to talk to the doctor and let him/her know. Also, any
medications that you've been taking have to be relayed to the doctor. If there are
any medical problems which are underlying too, you need to fast before taking a
test.
Uses Of Packed Cell Volume Test
A low PCV implies that the patient has a low number of red blood cells and is
suffering from anaemia. The doctor may ask the patient to undergo further tests to
determine the underlying causes of anaemia. Treatment will be given accordingly.
Measuring The PCV Test:
The PCV test is calculated with the help of an automated analyser which means that
it isn't directly measured. By multiplying the red cell count with the mean cell
volume, doctors get the final amount. PCV is slightly less accurate than the
hematocrit as they include small amounts of the plasma from the blood that is
generally trapped in between two red cells.
By tripling the haemoglobin concentration and dropping the units, an estimated
hematocrit can be determined in percentage.
The PCV can also be determined with the help of the of a capillary tube and the
centrifuging heparinised blood in it at around 10000 RPM for roughly five minutes.
This process helped in separating the blood into different layers, and the volume of
the total packed RBC divided by the blood sample's total volume gives the final
amount of the PCV. Since a tube is also used, it can be used to measure the lengths
lying between certain layers.
There is also another way to measure the levels of haematocrit, and this is through
optical methods such as spectrophotometry. With the help of differentials, the
differences between the optical densities of the sample flowing through the glass
tubes at isosbestic wavelengths and the product containing the luminal diameter
along with the hematocrit can create a linear relationship.
When A Low PCV Reading Occurs

48. There are certain conditions that contribute to the low reading in the PCV. These
include:
• Nutritional deficiencies of iron or vitamin (B12 or folate) and mineral deficiencies
• Bleeding
• Inflammatory conditions, for example,
• Kidney diseases
• Haemolysis, which is the situation where the RBCs are destroyed prematurely by
the immune system. This occurs due to certain organ damages and inherited
abnormalities of the RBCs
• Liver cirrhosis
• Medicines - including that of chemotherapy
• Abnormalities of RBCs or haemoglobin containing disorders such as
myelodysplastic syndrome, lymphoma, bone marrow disorders and myeloma
One of the most common causes of heightened PCV readings is that of dehydration.
With adequate fluid intake, the levels return to normal, but it can also create a
condition known as polycythaemia where there are more RBCs.
4.3. MALARIA PARASITE TEST
This is to determine the presence of malaria parasite in blood. Malaria is an
infectious disease caused by infection with single-celled parasites of the genus
Plasmodium. The parasite is transmitted to humans by the bite of the female
Anopheles mosquito. People with malaria often experience chills, fever, and flu-like
illness. Serious cases of malaria can result in death if left untreated. Four species of
Plasmodium parasites cause malaria in humans Plasmodium falciparum,
Plasmodium vivax, Plasmodium ovale, and Plasmodium malariae. Each causes a
different form of the disease. P. vivax and P. ovale cause the mildest forms; P.
falciparum, the severest and most deadly form. Other Plasmodium species infect
primates, rodents, birds, and lizards. Several of these species, particularly those

49. that infect rodents, have been used in experimental studies and for testing malaria
drugs and vaccines.
MATERIALS NEEDED: Cotton wool, 70% alcohol, sterile lancet, Glass slide, Giemsa
stain, microscope.
SAMPLE: Blood.
METHOD: Microscopic method using thin and thick smears.
PROCEDURES
Massage the patients thumb.
Disinfect with cotton wool dipped in 70% alcohol.
Use a sterile lancet to prick the thumb and wipe of the first blood that
comes out.
With the blood that comes out next, make a thick and thin smear of
blood on the glass slide and allow to dry.
Flood with Giemsa stain and allow to stain for 10 minutes.
Rinse the stain with distilled water and allow to dry.
Examine under the microscope using oil immersion objective lens
x100.
RESULT
 The thin smear is used to know the kind of Plasmodium species that is
present in the blood. A thick smear is used to check if there is any parasite in
the blood.

50. Fig1.2. Thick and thin smear of blood for malaria parasite test.
 A ring-form structure observed indicates the presence of malaria
parasite (either Plasmodium malariae or Plasmodium falciparum).
Fig1.3. Ring form of Plasmodium falciparum viewed under the microscope.

51. CHAPTER FIVE
5.1. RELEVANCE OF SIWES
The SIWES program provides students with the opportunity to work in areas
relating to their field of study which enables them to relate the theoretical
knowledge with practical work which will help them in their future jobs or
profession.
During my industrial training program, I have was exposed to various laboratory
tests which could not really be explained in lecture halls. I got firsthand experience
on how to run various tests, proper ways to collect samples, various culture
methods and analysis, even how to operate machines and laboratory equipment
not found in the schools laboratory. All this, theory would not have been able to
explain the way I understood with the practical.
5.2. RECOMMENDATIONS
Having successfully completed the SIWES training, I recommend that:
 Students on industrial training should take their training seriously because
the experience and the things they will be taught will help them a lot when
they find jobs after graduation.
 All university students should actively participate in the scheme to give them
an opportunity to familiarize and expose themselves to the needed
experience in handling working laboratory equipment that are not available
in our various institutions.
 Institutions should help students secure places for the training in time to
enable them complete their training before the duration of training expires.
 The University and departments should endeavor to send supervisors to the
various industries as this will go a long way to encourage students as well as
the industry based supervisors.
 Also, ITF should offer monthly financial assistance or other welfare packages
to students on attachment so as to help them carter for some of their
expenses such as transportation.

52. 5.3. CONCLUSION
The SIWES program has benefited me by providing a pre-professional working
experience and also serving as a bridge between theory and practical application.
It has also given me an insight into the adulthood life in the sense that I get to be
punctual, accountable and reliable.
The training has helped me in my career pursuit as a potential microbiologist,
improving my practical knowledge and marketability.
As such, SIWES should be taken seriously by everyone involved i.e. the students,
tertiary institutions, industries, Industrial Training Fund (ITF) and the Federal
Government.
It has also helped me solidify my theoretical base as well as my practical base in
microbiology.

53. REFERENCES
(December 16, 2015) Lerma E. Urinalysis. Medscape Reference. Available online at
http://emedicine.medscape.com/article/2074001-overview#a2. Accessed April
2016.
Arora, D. R. And Arora, B. B (2010). Medical Parasitology. 3rd
Edition, India. CBS
Publishers and Distributors PVT ltd.
Clarke, W. and Dufour, D. R., Editors (2011). Contemporary Practice in Clinical
Chemistry, AACC Press, Washington, DC, Pp 397-408.
Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed.
McPherson R, Pincus M, eds. Philadelphia, PA: Saunders Elsevier: 2011, Chapter 28.
Kasper DL, Braunwald E, Fauci AS, Hauser SL, Longo DL, Jameson JL eds. (2005)
Harrison’s Principles of Internal Medicine, 16th Edition, McGraw Hill. Pp 249-251,
1647-1649, 1718-1720.
Tietz Textbook of Clinical Chemistry and Molecular Diagnostics. Burtis CA, Ashwood
ER, Bruns DE, eds. 4th edition, St. Louis: Elsevier Saunders; 2006, 808-812.