UNIVERSITY OF DAR ES SALAAM

COLLEGE OF NATURAL AND APPLIED SCIENCES

DEPARTMENT OF MOLECULAR BIOLOGY AND
BIOTECHNOLOGY
PRACTICAL REPORT 3
GROUP 9
COURSE CODE: MC 132
COURSE DIRECTOR: DR. LUCY NAMKINGA
COURSE NAME: PRACTICALS IN EUKARYOTIC ORGANISMS
DATE OF PRACTICAL: 17 – 06 - 2021
DATE OF SUBMISSION: 05 – 07 – 2021
S/N NAME Registration No.
01 MTUI MONICA AUGUST 2020-04-08256
02 BASSO VANESSA ANTHONY 2020-04-00658
03 LEMA ASSAJIDAH ALLY 2020-04-13553
04 SIMION ELISHA WILLIAM 2020-04-11666
05 JONASI JOHANSENI N 2020-04-02826
06 MSOBA AMINA HUSSEIN 2020-04-13020
07 NYAMBACHA MADELINE PIUS 2020-04-09949
08 YOHANA ELICIAN 2020-04-12509
09 SAYALE HURUMA MOSSES 2020-04-11204
10 MCHARO JONATHAN ABINERI 2020-04-06606

PARTICIPANTS:
ABSTRACT

The aim of this practical was diagnosis of malaria which are transmitted by female
anopheles’ mosquito.

During diagnosis blood smear used to check for malaria disease. Sample of blood is placed in
a glass slide prepared, and observed under microscope. There are two types of smear which
are thick smear, for white blood cell and thin smear for red blood cells.

Blood smear test can help to diagnose malaria as indicate presence of parasite in the blood
and confirm that one has Malaria. Some blood test can take several days to complete results
while others produce results in less than 15 minutes.

Malaria is treated with prescription drugs to kill the parasite; the type of drugs and length of
treatment will vary depend on which type of malaria parasite a person has. Also depend on
symptoms, age or whether a person is pregnant.

Plasmodium falciparum is mainly found in Africa and it most’ dangerous compare to P.

vivax, P. malariae and P. Ovale
 INTRODUCTION

Plasmodium is the genus of parasitic protozoa having four species, Plasmodium falciparum,
Plasmodium malariae, Plasmodium vivax and Plasmodium ovale that cause malaria to
human. The disease at uncomplicated stage has symptoms like fever, vomiting myalgia,
headache, arthralgia and weakness. P. falciparum causes more fatal malaria characterized by
acute renal failure, acute respiratory distress syndrome and cerebral malaria and severe
anemia.

Plasmodium is carried from one person to another through b by female anopheles’ mosquito
(vector). The geographical distribution of malaria cases depends on the favorable
environment for vector multiplication, for anopheles’ mosquito the places with stagnant water
experience more malaria transmission. In sub Saharan Africa the common parasite is P.ovale
while P.falciparum is predominant in the world. However, the distribution can be affected by
climatic changes and people mobility.

In laboratory the presence of plasmodium parasites is detected rapidly by using malaria test
kit. Microscopic method is used to identify the number of parasites and their species. Two
types of smear, thick smear and thin smear which are stained using giemsa staining then
observed by microscope. In thick smear we detect the presence of parasites and number of
parasites is counted against white blood cells and in thin smear the number of parasites is
counted against red blood cells. Thin smear is fixed to ensure the red blood cells are not
destructed during preparation of smear.

Infection by P. falciparum and P. malariae does not cause enlargement and change in shape
of red blood cells. P. ovale and P. vivax cause changes in size and shape of red blood cells. In
red blood cells trophozoite, schizonts or merozoites of plasmodium can be seen in the slide.
The infection of multiple red blood cells is common for the case of P. falciparum than in
other species. The parasite president Maurer’s cleft and 2 chromatin dots. In trophozoite
stage, seldom and dark pigment can be seen in peripheral blood, the Maurer’s cleft less
visible. In schizonts stage 8 to 24 merozoites are seen clumped in one mass. In gametophyte
the red blood cells are less distorted and the parasite has crescent shape and chromatin in
single mass.
Malaria can be prevented by preventing multiplication of vectors by draining stagnant water,
cutting glass and using biological control. It can also be done by using chemicals such as
pesticide to kill mosquito in homes. Treatment also can be provided depending on severity of
the disease, species of the parasite and geographical location. This is due to the resistance
behavior to some malaria drugs.
 METHODOLOGY

Materials used

 Microscope
 Grass slides
 Giemsa stain
 Distilled water
 Methanol
 Blood sample

Preparation of thick blood smear

 Three drops of blood were placed on in the centre of the slide and spread around
evenly with the corner of another slide about a circle of 1cm was made
 smear was allowed to dry free from flies and dust, the slide was not heated to prevent
the damage of parasite
 The Giemsa buffer was added to stain the smear, smear was stained for about 30
minutes
 Slide was careful rinsed with distilled water
 Slides were allowed to completely to dry at the average time of 15 minutes
 Lastly, the slide was observed on the microscope

Preparation of thin blood smear

 Three drops of blood were placed in the centre of the pre cleaned blood, labelled
slide, near its frosted end
 Another slide was placed at an angle 30-45 on top of the drop allowing blood to
spread along the contact line of the two slides
 upper slide was quickly pushed toward the unfrosted end of the lower slide
 Slide was fixed with methanol and allowed to dry for about one minute
 smear was stained with Giemsa for about 30 minutes
 The slide was allowed too completely dry
 Followed by observation under the microscope.
 RESULTS

ACTIVITY ONE: PREPARATION OF THIN AND THICK SMEAR

Thick and thin smear was prepared to determine if parasite was present and confirm the
Plasmodium species present respectively.

TYPE OF SMEAR OBSERVATION

THICK SMEAR

THIN SMEAR
ACTIVITY TWO: MICROSCOPIC OBSERVATION OF PLASMODIUM

TYPE OF SMEAR DIAGRAM OBSERVATION

Few blue stained cells
THICK SMEAR were observed and other
cells retained their colour
after addition of giemsa
stain.

Ring-form morphology
THIN SMEAR was observed in few cells
and possessing chromatin
dots.
 DISCUSSION

The red blood cells affected by malaria parasite are stained blue by giemsa stain while the
unaffected cells remain unstained. Microscopy test for malaria provides three valuable
informations concerning the blood sample collected. First, determination of the presence of
malaria parasites in the thick blood smear (diagnosis). Then, the thin smear can be used to
determine the malaria specie and also the percentage of red blood cells affected by the
parasite.

The red blood cells affected by malaria parasite are stained blue purpish and red orange by
giemsa stain while the unaffected cells did not form the blue color due to absence of the
parasite nucleus. This is due to fact that Gimsa is the differential stain that contain the azure
and methlene blue a basic dye that binds to the acidic nucleus producing the blue color, Also
contain the eosin that is the acidic buffer dye that is attracted to the cytoplasmic and
cyotoplasmic granules which are alkaline producing red orange coloration .

Microscopy test for malaria provides three valuable informations concerning the blood
sample collected.First, determination of the presence of malaria parasites in the thick blood
smear (diagnosis). Then, the thin smear can be used to determine the malaria specie and also
the percentage of red blood cells affected by the parasite.

The species is determined through observation of characteristics in the

trophozoite,schozoint,and gametocytes stage forexample p.falciparum form rings in the
peripheral of red blood cell(accole,oppalique) thus used as the distinguishing feature of
P.falciparum.also Gmsa is used in the diagnosis of malaria because is easily available ,easy
to prepare and to use .

RECOMMENDATION
The practical on plasmodium was well conducted on the preparation of smears.

During observation there was shortage of microscope and the quality of the image was poor.
The number of people per group was large and so there was poor participation.

We suggest that the number of microscopes should be increased as well as the lens should be
changed so as to increase active participation and the number of people per group should be
reduced.

CONCLUSION.
Generally, this practical is most applicable clinically in diagnosis of malaria. Since
microscopic examination of thick and thin blood smears is the easiest and most reliable test
for malaria. Thick blood smears are most useful for detecting the presence of parasites
because they examine larger sample of blood. Thin smears help doctor to discover what
species of malaria is causing the infection.

In addition to that, its better to use universal precautions while preparing the smear for
malarial parasites; use gloves, use only disposable needles, wash hand, handle and dispose
the sharp instruments and other materials contaminated with blood careful to avoid injury.
 REFERENCES.

 Moody AH, Chiodin PL, Methods for detection of blood parasites. 2000
 Lee SH, Kara UA, Koay E, Lee MAA, Lam S, Teo D. New strategies for diagnosis and
screening malaria. 2002
 Adeoye GO, Nga IC. Comparison of Quantitative Buffy Coat technique QBC with
Giemsa-stained thick film for diagnosis of malaria. Parasitology International 2007.
 Cabezos J, Bada JL. The diagnosis of malaria by thick film and the QBC. 1993
 White NJ, Malaria Manson’s Tropical Disease, Philadelphia PA, WB Saunder 2003