UNIVERSITI TEKNOLOGI MARA

(BERTAM, PULAU PINANG)

Faculty of Health Sciences
Diploma in Medical Laboratory
Technology
______________________________________

CSI 153
CLINICAL PARASITOLOGY
PRACTICAL 3: MACROSCOPIC AND
MICROSCOPIC EXAMINATION
(DIRECT WET MOUNT) OF
STOOL SPECIMEN

NAME : NURUL FATINAJIHAH BINTI ZULIKIPLI

STUDENT ID : 2016625096

IC NUMBER : 981012115026

LECTURER’S NAME : MADAM NURHIDAYAH BINTI

 AB. RAHIM

INTRODUCTION

The collection of the stool specimen is essential for reliable stool

examination.Intestinal parasites passed the diagnostic stage in both feces and others
materials.Usually, diagnostic stage is at the egg stage and adult worm stage. But, it
is easier to found the eggs because it can be found on the feces. For the adult worm,
it can be found in small intestine.Examination of the stool specimen includes
macroscopic examination and microscopic examination (direct wet mount).

Macroscopic examination is the process where the stool was collected in tightly
capped container.Urine or other things must be avoided from contaminated the stool
specimens because it can affect the examination.Naked eyes was used to observe
the stool specimens in this experiment. The presence of barium, oil, blood, mucus,
and form was observed and recorded.

Microscopic examination or also known as direct wet mount is an easiest technique

which is mix the reagent with the stool before microscopic.The basic type of wet
mount that was used in this experiments is saline and iodine.Two drops of saline or
iodine was dropped on a glass slide with a pasteur pipette.Then, the stool specimens
was taken by using the end of wooden applicator stick and after that mix the
specimens with the saline or iodine on the glass slide before.Cover the slide by using
a short cover slip before examining under microscopic.Then,examination started
under 40X objective. When observe under the microscope, the
category,structure,morphology and motility of the stool was recorded.
A) MACROSCOPIC EXAMINATION

OBJECTIVE:
1. To collect stool specimen for practical.
2. To perform macroscopic examination on collected stool specimen.

METHODOLOGY:
1. A wide mouth container was collected from the lab assistant.
2. Stool specimen from human or animal (pets or farm animals) was collected by
using the container provided.
3. The containers was labelled.
4. The presence of barium and oil was examined and the result was recorded.
5. The presence of blood and mucus was examined and the result was recorded.
6. The form of stool was examined and the result was recorded.
7. Both specimens was brought to the practical held on the next day.

RESULTS:

Categorize : Animal
Age of subject: 3 years
Sex : Female
History : No

Barium Absent
Oil Absent
Blood Present
Mucus Present
Type 3 ( A sausage
Form shape with cracks in the
surface)
B) DIRECT WET MOUNT

OBJECTIVE:
To examine stool specimen for motile protozoa trophozite,protozoa cyst,helminth
eggs and larvae.

METHODOLOGY:
1. One drop of each of the following reagents was placed on a glass slide (one
reagent on one glass slide).
- Saline
- Iodine
2. A small amount of stool specimen was taken by using a wood applicator and
was mixed with the reagent on the glass slide.The mixture should be just thick
enough to read a newspaper print through the slide.
3. The specimen on the glass slide was covered with a cover slip.
4. Motile trophozoite, cyst, egg and larvae was examined under microscope and
the result was recorded.
5. The entire area under the cover slip was scanned by using objective 10X with
back and forth motion.
6. The suspicious parasite was confirmed by using objective 40X.

RESULTS:

Plant Fiber
 Category Artifacts (plant fiber)
Resemble hookworm
Structure
larvae
Morphology Square and yellowish
Motility Non-motile
Reagent Iodine

Crystals

Category Artifacts (crystals)

Structure Charcot-Leyden
Cylindrical shape and
Morphology
yellowish
Motility Non-motile
Reagent Iodine

Air bubble
 Category Artifacts (Air bubble)
Resemble protozoan
Structure
cyst
Oval shape and
Morphology
colourless
Motility Non-motile
Reagent Iodine

DISCUSSION

The stool was collected from a cow. Based on the macroscopic and microscopic
examination, there were some errors that may occur during this experiments and
affected the results.

In macroscopic examination, there was no presence of barium and oil in the stool
specimen but there was the presence of blood in the stool specimen. So, the stool
was not a normal stool and the presence of blood maybe because of the breakdown
of the blood products by intestinal acid.The form of this stool is Type 3 (a sausage
shape with cracks in the surface) according to the Bristol Stool Chart.This shows that
the stool was normal.

In microscopic examination, the results was not showing any morphology of the
parasite.We only saw the artifacts that contain similar figure to the parasite.From the
results,artifacts that showed is crystals,plant fiber and air bubbles.The presence of
the plant fiber maybe confused with Strongyloides or hookworm larvae and it also
presence because of the plant cannot digest well in the digestive system.

There were a few errors that occur during this experiments.Firstly is the presence of
the air bubble in microscopic examination.This happen because of the wrong
technique of placing the coverslip on the mixture of stool and reagent on glass
slide.When we place the coverslip, we need to do it from the end of slide with 45
degree angle.Let down the cover slip slowly so the air will removed from the
specimen.Other than that is the saline or iodine and the stool was dropped too much
on the glass slide so it can cause overlap when observe under microscope.
There were also some advantages and disadvantages of the direct wet mount or
microscopic examination.One of the advantages of direct wet mount is his process is
a quick preparation which is it can save our time and also energy.So that we can do
many direct wet mount in a given time.Another advantage from this experiments is it
has given natural colors to the specimen.We can see the colors when observe the
specimen under the microscop.The disadvantage from this experiments is we may
get inaccurate results because of too much reagent was mixed with the stool so, it
cause overlap.Other than that, the disadvantage is we can’t diagnose the
trophozoites because the stool specimen was kept for one day before the
experiments was started.

Based on this experiments,there are some steps that we need to focus on.Gloves
must be wear all the time when handling the stool to avoid contact and infection of
any bacteria from the stool.When we examine the specimen under microscope,move
the glass slide with zig-zag pattern to make sure the whole slide are observed.The
procedure of examine the stool must be done carefully and properly to get the good
results.

CONCLUSION

The macroscopic examination was performed by using the collected stool specimen
from a cow while the microscopic examination was performed by mixing the stool
with saline or iodine on a glass slide.The stool specimen was examined for motile
protozoa trophozite,protozoa cysts,helminth and larvae and only artifacts were found.
The objective of this experiment had been achieved succesfully.

REFERENCES:
 Laboratory Manual CSI 153 Clinical Parasitology, Pn. Nurhidayah binti Ab Rahim
 Lecture’s Note-Introduction to Parasitology
 http://www.med-chem.com/para-site.php?
url=procedures&subsection=microscopic&tertiary_section=direct_wet_smear
 http://www.microbehunter.com/making-a-wet-mount-microscope-slide/
 MONASTYRSKY, K. (n.d.). Gut Sense. Retrieved from Bristol Stool Form Scale:
http://www.gutsense.org/constipation/normal_stools.html
 Ghosh, S. (2013).Textbook of Medical Parasitology. Nepal: Jaypee Brothers
Medical Publishers(P) Ltd.