Note: This report is the combination of 4 praticals that we have done with Dr Rafidah, that

means it have 4 covers pages and references.

FACULTY OF MEDICINE, BIOSCIENCE & NURSING

SCHOOL OF BIOSCIENCE

PRACTICAL REPORT (1)

MEDICAL PARASITOLOGY & ENTOMOLOGY (BMC 213)

Name: AISSATA DIAWARA

Student ID: BBSH 22106335

Course: Bachelor In Biomedical Science

Practical Title: Collection & Transportation of Fresh Faecal Specimens

Module Leader: Dr. Rafidah

Declaration: I ID No: bbsh22106335 confirm that I have read and understood the University
regulations concerning
plagiarism and that the work contained within this Practical Report (1) is my work inside the
meaning of the regulations.

Sign: Diawara.
1. INTRODUCTION
The faeces should be obtained as soon as possible, As this when the micro-organisms are most
active,It is good before the start of antibiotic treatement.The sample can be colleected early
like morning so that it can arrive at laboratory before middle day afternoon and be analyzed
this day
(same day).Stool that has solidified should rejected.Fresh fecal sample is good to a rectal
sample ,
Even thoughthe rectal swab is appropriate if the sample can not be taken directely or if the
transport of the stool to the lab is extenced.And we need to know panthogens can live up to a
week at room temperature,But more recommendation is refrigeration.
2. OBJECTIVE
The primary objective for analyzing faeces is to see if a kind of disease or parasite is affecting
the intestines. Many microscopic organisms that live in the intestinal region are required for
proper metabolism. But, if the intestines get contaminated with pathogenic bacteria or parasites,
it can
produce complications such as bloody diarrhoea, and testing stool can help determine the
reason.
3. MATERIALS
Specimen collector; red lids indicates
that it is used to collect only stool
Figure 1 Figure 2

Specimen collector; red lids indicates

that it is used to collect only stool
4. METHOD
1. Request the patient to pass the faeces sample into a waxed cardboard or plastic cup with a
tight-fitting lid. Sample collection in a matchbox or on plant leaves is not an acceptable option.
2. For a normal test, 20-40 grams of well-formed faeces or 5-6 table spoonful of watery stool
would be adequate.
3. Take a small sample of stool and add a drop of sodium chloride (NaCl).
4. Mix the solution properly and make sure there are no lumps and then cover it with a
coverslip.
5. Ingestion of medications before collecting a stool sample may affect with parasite
identification. Tetracycline, sulfonamides, antiprotozoal medicines, laxatives, antacids, castor
oil, magnesium hydroxide, barium sulphate, bismuth kaolin compounds, and hypertonic salts
are examples of such medications. These should not be taken 1-2 weeks before the stool sample
testing.
6. All collections must be appropriately labelled with the patient's name, age, gender, and
collection date and time.
7. Observe the stool mixed with NaCl solution under the microscope of either 4x or 10x.

5. RESULTS
From the findings that wasobserved during the experiment were mostly contaminations,
bacteriaand other debris. Below are some observations in the form of images:
Fig3

Debris Bacterie

Figure 3
Picture on top shows a type of bacteria found in the sample stool
 Specimen lumps

buble
Bacteria

Debris

Figure 4

6. DISCUSSION
1. If looking for trophozoites, stool specimen must be transported very rapidly to the laboratory
to avoid disintegration of trophozoites. Stool samples should be examined
within 30 minutes of collection of the specimen and not receipt of the specimen in the
laboratory. Stool specimens should never be frozen and thawed or placed in an incubator
because parasitic forms deteriorate very rapidly.2.For permanent fixation of the stool specimen,
10% formol-saline (prepared by adding 1 00
ml formaldehyde to 900 ml of 0.85% sodium chloride) is a well-known preservative. Polyvinyl
alcohol (PVA) is a widely used preservative because the performance of concentration
procedures and preparation of permanent stained smears are both possible with this.
Based on results:
After a long observation under the microscope with x10 by observing the used stool mixed
with sodium chloride.I had to observe bacteria on the above images.The bacteria appeared as
a long structure with more intensity of color than the rest of the stool sample.On the above
picture, you can see the rest of the sample being clearer which is the stool deposits ,bulb and
some debris .You can also see that there is already a bacterial infection based on the irregularly
shaped structures that I saw on the lab images. I did not see any presence of egg morphology
and usually this also depends on the type of infection.on the other hand the appearance of
crystalline shape can mean that there is a bacterial infection.
In the process of collecting the stool sample, you are not allowed to ingest anything, either in
solid or liquid form, because this is usually how bacteria enter the subject's body and cause an
infection. Immediately after completing the experiment, you should be sure to remove your
gloves and any other lab clothing you were wearing and wash your hands with detergents
before leaving the lab.
And After observing the stool l had stained with sodium chloride,l can conclude that the
individual had a bacterial infection.From the picture l tool. l have labeled the bacteria which is
appearing as an a long structure like most of the sample is stool from the individual that l
collected.I was not able to observe the presence of eggs and puss from the sample.
The most important thing to note when you are carrying out this experiment is that you should
not eat or drink during stool collection as thus many causeinfection.

7. CONCLUSION
Additionally, the experiment will benefit from the integrity of your faeces samples being
ensured
by adhering to these collection, transit, and storage procedures, which will ultimately result in
trustworthy results. It will be simpler to make or prescribe treatments for the patient to stop any
sickness or ailment brought on by these microorganisms by using the approach of this
investigation.

8. QUESTION:
1. What is the importance of specific faecal specimen collector for parasitological analysis?
Answer: To observe the morphology and to find the pathogenesis of the parasite that is located
at the intestinal tract.
2. Why is it important to observe the faecal specimen as soon as the sample is reached to the
lab?
Answer: To avoid any false results during the experiment as the parasites in it does not live

REFERENCES
• Aryal, S. (2020). Collection and transport of stool specimens. [online] Microbe Notes.
Available at: https://microbenotes.com/collection-and-transport-of-stool-specimens/.
• Cary, S. G., & Blair, E. B. (1964). New transport medium for shipment of clinical
specimens I: fecal specimens. Journal of bacteriology, 88(1), 96-98.
 FACULTY OF MEDICINE, BIOSCIENCE & NURSING
SCHOOL OF BIOSCIENCE

PRACTICAL REPORT (2)

MEDICAL PARASITOLOGY & ENTOMOLOGY (BMC 213)

Name: AISSATA DIAWARA

Student ID: BBSH 22106335
Course: Bachelor In
Biomedical Science

Module Leader: Dr. Rafidah

Practical Title: Microscopic examination of fecal specimens by using ‘saline wet mount
method’
Aim: To prepare saline wet mount for observation of fecal specimens
1.Introduction:
Wet Mount Microscopy is a very simple technique that is applied widely in clinical
laboratories. e.g. Saline wet mount and iodine wet mount for stool examination, potassium
hydroxide (KOH) mount of skin, nails, hairs for fungal elements, vaginal wet mount for vaginal
yeast infection ( candidiasis), trichomoniasis, and bacterial vaginosis. Saline wet mount
microscopy of the stool or stool wet mount microscopy is the simplest and basic method for
the study of feces and is applicable in every medical laboratory even in a small setup. Saline
wet mount microscopy uses for the following purposes.
2.Objective:

1. To observe live trophozoites ( e.g. Entamoeba histolytica/dispar, Girdia lablia,

Trichomonas, etc.) and larvae of parasites ( e.g. Strongyloides stercoralis) are motile
except inactive forms.
2. To find out eggs, cysts, oocysts of different parasites ( Helminths, protozoa, and
coccidian parasites).
3. To determine the presence of leukocytes and erythrocytes in a fecal smear.
4. It also gives clues towards the motility of bacteria (Shigella non-motile causing
bacillary dysentery whereas Vibrio cholerae shows darting motility which is causative
agent cholera).
5. It also remarks the presence of fungal elements ( yeast cells, hyphae, or fungal spores).
6. It also visualizes the presence of non-parasitic structures like Charcot Leyden crystal,
muscle fibers, fat globules, starch cells, vegetable fibers, hair, etc.

3.Materials:
A. Reagents.
1. 0.85% NaCl
B. Supplies

1. Microscope slides 2.Coverslips

3.Pasteur pipettes
C. Equipment
1. Binocular microscope Oculars should be 10X. Some may prefer 5X.

2. Magnetic stirrer mixed stir bar

 Specimen: Stool sample

4.Methods.
When doing this treatment, put on gloves.
1. Put a drop of NaCl 0.85% on the glass slide.
2. Collect a very small amount of faeces (about the amount that would fit on the tip of an
applicator).
fully emulsify the stool in the saline (stool should not stick when placed into the specimen).
3. Cover the saline suspension with a coverslip.
4. Use a 10X objective to systematically scan the suspensions. The entire region of the coverslip
should be inspected.
5. Use the 40X objective for a more in-depth examination if you notice anything strange. Even
if nothing alarming is visible, at least one-third of the coverslip should be looked at with the
40X objective.
Saline wet mount: It is used to find cysts, protozoan trophozoites, and worm eggs or larvae.
Then again.

5.Results/Observation:

cyst

Cyst
6.REPORTING RESULTS/DISCUSSION
After a long observation under the microscope with x10 by observing the used feacal specimens
mixed with sodium chloride.I had to observe bacteria on the above images.The bacteria
appeared as a long structure with more intensity of color than the rest of the stool sample.
On the above picture, you can see the rest of the sample being clearer which is the feacal
specimens ,bulb and some debris .You can also see that there is already a bacterial infection
based on the irregularly shaped structures that I saw on the lab images. I did not see any
presence of egg morphology and usually this also depends on the type of infection.on the other
hand the appearance of crystalline shape can mean that there is a bacterial infection.

7.Conclusion:
Finally, Saline wet mount preparation for stool uses analyzing a stool specimen in coprology
(study of feces). It utilizes a physiological saline solution (0.85% NaCl ) as an isotonic media
to maintain the cellular structure of the various organisms as well as our cells that are found in
stool.

8. Questions:
1. Mention about the saline wet mount?
Ans: saline wet is made by 2mg of feces in a drop of saline placed on a clean glass slide.the
smear is then examined under a microscope.Saline wet mount is used for the detection of the
trophozoites and cyst of protozoa and eggs and larvae of helminths

2. Give some examples of helminth eggs seen in wet mount Method?

Answer: regarding the detection of specific intestinal helminths direct wet mount microscopy
reported 12.9% (48/372)hookworm 5.37%(20/372) Ascaris lumbricoides and 0.54%(2/372)
hymenolepis nana.but Taenia species and Strogyloides stercorallis were not detected by this
method
3. What is the advantages of Saline wet mount?
Answer: saline wet mount is used for the detection of trophozoites and cyst of protozoa and
eggs and larvae of helminths.it is particularly useful for the detection of live motile trophozoites
of E.histolytica ,Giardia lamblia,and Balantidium coli.

References Books:
1. Medical Parasitology by Abhay R. Satoskar, Gary L. Simon, Peter J. Hotez and
Moriya Tsuji
2. Atlas of Medical Helminthology and protozoology -4th edn -P.L. Chiodini,
A.H. Moody, D.W. Manser
3. Markell and Voge’s medical parasitology-9th edition.
 FACULTY OF MEDICINE, BIOSCIENCE & NURSING
SCHOOL OF BIOSCIENCE

PRACTICAL REPORT (3)

MEDICAL PARASITOLOGY & ENTOMOLOGY (BMC 213)

Name: AISSATA DIAWARA

Student ID: BBSH 22106335
Course: Bachelor In
Biomedical Science

Module Leader: Dr. Rafidah

Practical Title: Thin Blood Film preparation and Wright’s Stain for Parasitological
Examination
Aim: To prepare thin blood film for Wright’s stain
1.Introduction:
The most accurate and consistent method for determining the presence of almost all blood
parasites is now stained blood films. They offer a permanent record and can be delivered to a
reference lab for advice or to confirm a diagnosis. Typically, a lab technician will check for
blood parasites, two different types of blood films, a thin film and a thick film—are prepared.
The thick film can be formed on different microscope slides, or both can be made on the same
slide with the thin film on one end and the thick film on the other. Preparing a thin film on one
slide, a thick film on another, and a combination of thin and thick films on a third slide is
advised when malaria is suspected. Within minutes, the thin film can become discoloured. If
the patient has a high level of parasitaemia, the thin film can be stained in a matter of minutes
and will allow for a quick diagnosis of malaria; the thick film can be stained in a matter of
hours and will allow for a quicker diagnosis of a lighter infection; and the combination film is
stained several hours later when the blood has had more time to dry out, allowing for a better
differential stain. The fast diagnosis is then confirmed using the combo film, which is also
preserved as a permanent record. Any blood parasite can be found using this three-slide
method.
Thin blood films can be stained with Wright's stain to look for blood parasites, however it is
not as effective as
Giemsa is used to stain thick films. Using buffered water with a pH of 6.8, the staining
reaction—which is relatively comparable to that of Giemsa—is accomplished. The fixative and
stain are combined in the stain.

2. Objective:
Commonly, Thin Blood Film preparation and Wright’s Stain are prepared for the diagnosis as
well as prognosis of any abnormalities (if present). The Thin blood smear is prepared for
studying the morphology of the blood cells and for the identification of microbial agents. The
Thick blood smears are prepared for detecting the Blood parasites such as Plasmodium spp.
(Malaria parasite), Wuchereria bancrofti & Brugia malayi (Lymphatic filariasis), Leishmania
donovani or other species of Leishmania (Leishmaniasis), Babesia spp. (Babesiosis)

3. Principle
The combination thick-thin blood film provides both options on one glass slide, and the slide
can be
stained as either a thick or thin blood film. If fixed prior to staining, then the smear will be read
as a
thin blood film; if RBCs are lysed during staining, the preparation will be read as a thick blood
film
(parasites, platelets, WBCs). This combination blood film dries more rapidly than the
traditional
thick blood film, thus allowing staining and examination to proceed with very little waiting
time for
the slide(s) to dry (practical 1, 2).

4. Materials:
Specimen:
Fresh blood
Reagents and supplies
Glass slides (1 by 3 in., or larger if you prefer) alcohol washed, Glass marker, Blood collection
supplies (if applicable), Paper with newsprint-size print, Applicator sticks, Coplin jars, Staining
rack, disposable pipettes, Microscope, Timer (Stop watch), 1 h. Wright’s stain, Phosphate
buffer.
Systematically, prepare both a thick and a thin blood film (this can be done on the same slide).
Note: if both are made on the same slide, start by preparing the thin film.
For a thick blood film
Touch the drop on a glass slide (see top picture)

Then spread the blood evently with the corner of another slide (see centre picture) to make a
square or a circular patch of moderate thickness (it should be possible to read through it). Dry
the slide, while protecting it from dust, flies and ants.
For a thin blood film
The drop of blood should be smaller than for the thick film. Apply the edge of another glass
slide to the top of the drop of blood at an angle of 45°, allowing the blood to spread along its
edge, then push the spreader slide forward keeping it at the same angle (see bottom picture).

A properly made thin film should consist of an unbroken layer of red blood cells with the
'tongue' of the film not touching the edge of the slide. The thin film must be dried immediately
by waving the slide from side to side or by holding it for a few seconds in front of a fan: this
ensures a good preservation of the shape of the cells.
5. Staining:
1. Place air-dried films on a level staining rack.
2. Use a pipette to cover the surface of the slide with stain, adding stain drop by drop. Count
the number of drops needed to cover the surface. Let stand 1 to 3 min (optimal staining time
to obtain correct color range and intensity] will vary with each batch of stain).
3. Add to the slide the same number of drops of buffered water as was used of stain for step
V.B.2 and mix the stain and water by blowing on the surface of the fluid.
4. After 4 to 8 min, flood the stain from the slide with buffered water. Do not pour the stain off
before flooding, or a precipitate will be deposited on the slide.
5. Wipe the underside of the slide to remove excess stain.
6. Let air dry in a vertical position.
6. Results:
A blood cell is a cell produced through hematopoiesis and found mainly in the blood. Major
types of blood cells include red blood cells (erythrocytes), white cells (leukocytes), and
platelets (thrombocytes).

Red blood cell

platelets

8. Discussion / Interpreting Results

The blood cells are red cells, white cells, and platelets. In this figure we can identify red blood
cells whose carry oxygen to all parts of the body and platelets who are tiny cells that have a big
job in stopping bleeding.
Smears will be evaluated on the following criteria:
1. Alternative thick and thin smear through out the length
2. Thickness
3. Proper labelling
4. Staining quality
5. Morphology of parasites if any

Erythrocyte – The color turns yellowish to red.

Neutrophils – The result varies depending on the specific location, such as the following:

• Granules – reddish
• Nucleus – dark purple
• Cytoplasm – pale pink

Eosinophils – For eosinophils, the following changes in colors are observed:

• Nucleus – blue
• Cytoplasm – blue
• Granules – red to orange

Basophils – For basophils, the following changes in colors can be observed:

• Nucleus – Purple to dark blue

• Granules – Dark purple

Monocytes – These changes in color are observed:

• Nucleus – dark purple

• Cytoplasm – Mosaic pink and blue

Lymphocytes – These changes in color are evident:

• Nucleus – dark purple

• Cytoplasm – Sky blue

Platelets – The granules appear violet to purple

9. Questions:
1. Describe the optimal area for identifying the parasites on a well-stained smear.
Answer: thick smears allow a more efficient detection of parasites (increased sensitivity)
however they do not permit an optimal review of parasite morphology
2. Describe the characteristics of the intracellular and extracellular parasites on a well-stained
smear.
Answer:

3. What are the basic components in Romanosky stain

Answer: Romanowsky stains are neural stains composed of a mixture of oxidized methylene
blue (azure)dyes and eosin Y.The azure are basic dyes that bind acid nuclei and result in a blue
to purple color

References:
1. Dunning, K., & Safo, A. O. (2011). The ultimate Wright-Giemsa stain: 60 years in the
making. Biotechnic & Histochemistry, 86(2), 69-75.
2. Houwen, B. (2002). Blood film preparation and staining procedures. Clinics in laboratory
medicine, 22(1), 1-14.
3. OKTIYANI, Neni, MUHLISIN, Ahmad, ROEBIAKTO, Erfan, et al. Utilization of
Alternative Buffer Solutions for Staining Thin Blood smears by the Giemsa, Wright stain and
Romanowsky method. Tropical Health and Medical Research, 2023.
 FACULTY OF MEDICINE, BIOSCIENCE & NURSING
SCHOOL OF BIOSCIENCE

PRACTICAL REPORT (4)

MEDICAL PARASITOLOGY & ENTOMOLOGY (BMC 213)

Name: AISSATA DIAWARA

Student ID: BBSH 22106335
Course: Bachelor In
Biomedical Science

Module Leader: DR Rafidah

Title: Slide examination (Ascaris lumbricoids fertilized and unfertilized eggs Enterobius

vermicularis Ancylostoma duodenale.,)

Aim: To examine slides of Ascaris lumbricoidis fertilized and unfertilized eggs, Enterobius

vermicularis Ancylostoma duodenale.

1.Introduction:
parasitic roundworm is its usual name. It can grow up to 35 cm long, which is what causes
ascariasis. is common in tropical locations and unhygienic places.
Infection is brought on by eating food that has been contaminated with faeces that contain
Ascaris eggs.
Infections typically have no symptoms, especially if there are few worms present. However,
they could also be accompanied by fever, diarrhoea, and inflammation, and if the worms spread
to other regions of the body, they might cause serious issues. Entobius vermicularis More
prevalent in wealthy countries in the chilly and temperate climates. sickness that affects kids
more often than adults. It keeps its mouth parts firmly affixed to the caecal mucosa. Adults
have some innate resistance, therefore it is not noticeable in them. Transmission is mainly
through ingestion, inhalation and most commonly auto infection. to Have a typical bend of the
anterior part of the body thus-hook worms. Hook worms are mainly classified as old world and
new world. Old world – Ancylostoma duodenale. New world – Necator americanus.
Adult worm inhabits in the small intestine of man (jejunum). Man is the only definitive host.
They are grey in color but appear brownish-red due ingested blood

2.Material:
Microscopes, mounted positive slides and Oils for oil immersion objective

3.Procedure:
(Microscopy)
1. Initially the examined slide placed on the stages of microscope and systematically scan with
the 10X objective.
2. The entire coverslip area should be examined. If you see something suspicious, use the 40X
objective for more-detailed study.
3. At least one-third of the coverslip should be examined with the 40X objective, even if
nothing suspicious has been seen.
4.Observation:

1.

Enterobius vermicularis (male

2.

Enterobius vermicularis(female)
3.

Ancylostoma caninum(female)

4.

Ascaria ambricoides, fertilized egg wm

5. Discussion
If we analyse the figures above , then we can see Unfertilized eggs are elongated and larger
than fertile eggs (up to 90 μm in length). Their shell is thinner and their mammillated layer is
more variable, either with large protuberances or practically none. Unfertile eggs contain
mainly a mass of refractile granules.

• Infertile
Measure: 88-99 um by 39-44 um
Longer and narrower w/ thin shell andirregular mammilated coating filled withrefractile
granules
Produced by immature worms
Color: yellowish brown

• Fertile
Measure: 45-70 um by 35-50 um
Thick, transparent, hyaline shell with athicker outer layer as a supportingstructure with a
vitelline, lipodial, innermembrane w/c is highly impermiable
Ovoid with a mass of protoplasm atoviposition w/c develop into a larvae inabout 14 days

• Enterobius vermicularis
In the above fig we can see for Enterobius an adult female is longer than adult male .
An adult male measures 2-4mm in length and 0.1-0.2mm breadth when an adult female is 8-
12mm in length and 0.3-0.5 mm in breadth.The posterior third of the body of male is curved,
sharply shortened and possess a exposed terminal copulatory spicule when the The posterior
end of male is extremely straight and drawn out into a long, tapering and finely pointed tail,
which is 1/3rd the length of the worm.

• Ancylostoma duodenale
After observation we can see the female Ancylostoma measure about 10 to 13 mm x 0.6 mm
while the males measure about 8 to 10 mm x 0.5 mm and the female gonopore is separate and
is located at the junction of the posterior and in the middle third. The male has a cloaca where
the ejaculatory duct opens.

6. Conclusion

The fertilized egg of Ascaris lumbricoides is the egg cell that has undergone fertilization. It
has a more rounded shape, and its shell is significantly thick. The unicellular embryo inside
the fertilized egg develops into an infective larva (L2). In comparison, the unfertilized egg of
Ascaris lumbricoides is the egg cell that has not undergone fertilization. It has an elongated
shape, and its shell is thin. Moreover, it does not contain a developing embryo. Both fertilized
and unfertilized eggs of Ascaris lumbricoides can pass through stool. However, the main
difference between fertilized and unfertilized egg of Ascaris lumbricoides is the structure and
the presence of a developing embryo.
is concluded that E. vermicularis occurs more frequently in uninflamed appendices. It may be
a cause of symptoms resembling acute appendicitis although the mechanism for this does not
involve mucosal invasion by the parasite.
Question:

1. Mension about the identified parasite?

Answer
A parasite is a living thing that inhabits its host and feeds off or at the expense of it. Protozoa,
helminths, and ectoparasites are the three main groups of parasites that can harm humans.
Diagnosis is possible for parasitic disorders such filariasis, malaria, and babesiosis. A drop of
blood is tested by placing it on a microscope slide. The slide is then dyed, and a microscope is
used to view it.

7. References:
• O'lorcain, P., & Holland, C. V. (2000). The public health importance of Ascaris
lumbricoides. Parasitology, 121(S1), S51-S71.
• WORLD HEALTH ORGANIZATION. Bench aids for the diagnosis of intestinal
parasites. World Health Organization, 2019.
• Kanaan, A. L., Dayoub, A., & Kabakli, R. Prevalence of intestinal helminths infection
in patients reporting to the microbiological laboratory at Tishreen University Hospital,
Lattakia, Syria, during the years 2016-2017.