Title: EXAMINATION OF PARASITES IN BLOOD

OBJECTIVES
1.To examine blood parasites under a light microscope.
2.To determine the presence of parasites in blood.
INTRODUCTION
The examination of parasites in blood,"blood parasitology" is a critical aspect of diagnosing
and managing parasitic infections in the field of medical microbiology.Blood parasites are
organisms that infect human blood and that of other animals leading to various significant
diseases like Trypanosomiasis and filariasis. Some of the most common blood parasites
include Trypanosoma species, Microfilariae,Chagas disease and plasmodium species causing
malaria. These parasites are transmitted to humans mainly through bites of infected vectors
like female anopheles mosquitoes,ticks and tsetseflies.They can also lead to severe morbidity
and mortality if not promptly identified and treated. Other blood parasites like babesiosis
pose significant health risks globally,particularly in tropical and subtropical regions.

MATERIALS
1.Human blood sample,
2.Thin and thick blood smear
3.microscope and slides
4. Blood lancet
5.Absolute alcohol,
6.Buffer P.H of 7.4
7.Gloves
8.Wright's, Leishman's or Giemsa's stain.

METHODOLOGY
i)Draw blood from a finger of your choice.
ii)Place a drop of thin blood near the sides of a smooth slide(Slide A) and smear thick blood
on the central position of the other slide(Slide B).
iii)Incline another slide(SlideC) at an angle of 30° adjascent to slide A to produce a thin blood
smear usually a smooth tongue.
iv)Air dry the smear.
v)Fix the thin smear in Absolute ethanol by flooding.Do not fix the thick smear.
vi)Stain using Giemsa's stain for 10 minutes.
vii)Rinse off the Giemsa's stain using Buffer solution
viii)Place the slide with a thin blood smear on the stage of a microscope for observation.

RESULTS

fig 1(a) fig 1(b);neutrophils

fig 1(c);monocytes fig1(d);neutrophils
DISCUSSION
°The examination of the thin blood smear,revealed the presence of various leukocytes
including neutrophils,monocytes,eosinophils and lymphocytes. However, no blood parasites
were detected in the specimen sample. At 4 times(x4) objective lens magnification,we
observed a broad view of the blood smear. This helped in ensuring the smear was evenly
spread and stained. At (x40) magnification which is high magnification power,the
morphology of individual blood cells like erythrocytes and leucocytes was observed.Since the
blood being examined had no noticeable parasites,at 100 times objective lens magnification
neutrophils,monocytes, lymphocytes and eosinophils were observed. Neutrophils were
characterized with 3 lobes, a multi-lobed nucleus and granular cytoplasm. They were the
most abundant type of white blood cells in the blood sample and they play a crucial role in
the body's defense against bacterial infections. The presence of neutrophils indicated a
normal immune response as they are essential for phagocytosis and destruction of
pathogens. Monocytes on the other hand, were the largest leukocytes with a kidney-shaped
nucleus and also contained granules. Monocytes usually differentiate into macrophages and
dendritic cells in tissues,where they perform phagocytosis and antigen presentation. Their
presence suggested ongoing immune surveillance and tissue repair processes in the human
body.

Eosinophils had a large, bi-lobbed nucleus and red orange granules present in the cytoplasm.
You could easily identify them because they had 2 observable lobes. Their presence in the
blood smear despite the absence of parasites, indicated an allergic response or a parasitic
infection that had been resolved. Also, lymphocytes were observed as small cells that had a
large round nucleus and scant cytoplasm. Generally, lymphocytes were critical for adaptive
immunity. They included the B-cells, T-cells and natural killer cells responsible for producing
antibodies, regulating immune responses and directly killing infected cells. The absence of
blood parasites in the smear significantly indicated that the patient or individual did not have
active parasitic infections at the time of examination. This meant that the patient was not
exposed to those infections or had successfully cleared any previous infections. The presence
of the observed leukocytes indicated a well functioning immune sytem .They also provided
insights into the patient's immune status and the potential underlying conditions. While no
parasites were detected, the presence of eosinophils might have warranted further
investigations to rule out other causes of eosinophilia such as allergies or other non-parasitic
infections.
Using a thin blood smear allows for detailed observation of the parasites within the blood
cells,which is essential for species identification and understanding the infection's impact on
the cells. However,thick smears are essential for initial detection especially when parasite
levels are low(low Parasitemia). A thin blood smear can be fixed with methanol to preserve
the cell structure while a thick blood smear cannot be fixed with methanol,allowing
erythrocytes to lyse making it easier to see parasites. A thick blood smear was more
sensitive for detecting low levels of parasitemia while a thin blood smear was less sensitive
for detecting low levels of parasitemia. Giemsa's stain a differential stain contained a mixture
of azure,methylene blue and eosin dyes. Azure and methylene blue are basic dyes that stain
acidic components like the nucleus giving them a blue-purple color while Eosin is an acidic
dye that stains basic components like cytoplasm in red. The buffer solution was used to
maintain the PH around 6.8 to 7.4,which was necessary for proper binding of the dyes to the
cellular components. This ensured that staining was effective and the cellular structures were
clearly visible hence a more effective examination of parasites in blood.

CONCLUSION
In conclusion,the examination of blood parasites brings new and formidable tasks in
diagnosing parasitic infections and coming up with possible treatments and control
measures for various diseases.The combination of Giemsa’s stain and thin blood smears
provides a reliable method for the detailed examination and identification of blood parasites.
For comprehensive diagnostic purposes, it is recommended to use both thin and thick blood
smears to balance the need for detailed morphological study and sensitive detection of
parasitemia. This dual approach enhances the accuracy and reliability of parasitic blood
examinations, contributing to better diagnosis and treatment of parasitic infections.

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