Stool Examination
By the end of the lecture, the
students should be able to:
1- Recognize the organisms detected by
microscopic examination of the faecal
Learning material .
Outcomes: 2- Understand faecal specimen
collection.
3- Outline Macroscopic examination of
stool samples.
4- Outline Microscopic examination of
stool samples.
Specimen collection precautions
• Clean wide mouth containers with tight fitting lids to
• Prevent accidental spillage
• Maintain moisture within the specimen
• No contamination with water or urine because
• Free living organisms in water may give false results
• Urine can destroy motile parasites
• Avoid collect specimens after barium or antibiotics
• Interfere with intestinal flora and protozoa identification
• Place the containers in plastic bags for transfer to the laboratory
• Must be carefully handled
• Potential source of infectious organisms (bacteria, fungi, viruses, and
parasites)
• Labeling with (patient’s name, identification number, physician’s name,
date and time of specimen collection).
Stool Examination:
Macroscopic Examination Microscopic Examination Additional techniques
Stool consistency Direct examination
Stool culture
techniques
Stool color Concentration
techniques
Blood Kato thick-smear
Permanent stained
technique.
Tapeworm proglottids smears
Adult worms Hatching of
schistosome Eggs.
India Ink Injection
Procedure
Macroscopic Examination
Stool consistency
Stool color
Blood
Tapeworm proglottids
Adult worms
A- Macroscopic Examination:
➢ Stool consistency:
- Liquid specimen must be examined within 30 minutes of passage, while soft ones
should be examined within one hour.
- Helminth eggs may be found with any consistency.
- Soft or liquid stool is highly suggestive of an amoebic infection.
- Trophozoites of the intestinal protozoa are usually found in liquid specimens,
while cysts are usually found in formed samples.
-The more liquid the stool, the more oocysts that are found in the specimen, e.g. C.
parvum infection.
➢ Stool color:
- Iron black
- Barium light tan to white
➢ Blood:
- Dark stool bleeding high in the small intestine.
- Fresh blood/ bright red bleeding at lower level
- Bloody specimens may indicate some parasites, e.g. S. mansoni.
- In certain parasitic infections blood and mucus may be present. e.g. amoebic
infection .
➢Tapeworm segments (proglottids)
- Tape worm segments may be found on the
sample or on the bottom of the collection
container.
➢ Adult worms:
- adult nematode worms .e.g. Ascaris
lumbricoides , E. vermicularis are occasionally
found on the surface or in the stool.
Microscopic Examination
Direct examination
Concentration techniques
Permanent stained smears
The microscopic examination of faecal material may detect:
1. Trophozoites and cysts of intestinal protozoa .
2. Oocysts of coccidia.
3. Helminth eggs and larvae.
4. RBCs ulceration or other hemorrhage.
5. WBCs inflammation.
6. Charcot-Leyden crystals disintegrating eosinophils
9. Fungi.
10. Plant cells, pollen grain may mimic some helminth eggs, protozoan cysts.
11. Plant fibers or root or animal hairs may mimic helminth larvae
Microscopic Examination:
Direct Concentration Permanent-
examination techniques stained
smears
• Sedimentation
• Direct wet smear
methods • Iron-hematoxylin
• Iodine smear
(Formalin-ether
• Trichrome
sedimentation)
• Special staining
• Flotation methods
(Zinc sulfate)
Technique of wet smear
a. Place one drop of saline on the centre of a clean slide.
b. Take about 2 mg. of stool, with an applicator stick, and
thoroughly emulsify it in saline.
c. Cover with cover slip and examine microscopically.
d. A drop of iodine can be put to stain protozoan cysts.
Direct Wet Smear:
• Can detect protozoan trophozoites, cysts, oocysts and
helminth eggs and larvae.
• Used mainly to detect motile protozoan trophozoites.
• When iodine is added it will kill trophozoites no
motility will be seen.
Direct wet smear:
Concentration Techniques
Sedimentation Methods Flotation Methods
Simple Saline Sedimentation
Formalin-Ether
Zinc Sulfate flotation
Sedimentation
Acid-Ether Sedimentation
Sedimentation Methods:
➢Simple Saline Sedimentation :
• Stool sample is mixed with saline in a tube.
• The tube is left undisturbed.
• The parasites will settle in the bottom of the tube.
➢Formalin-Ether Sedimentation Technique
• Formalin: fixation and preservation of the parasitic stages
• Ether: as an extractor of debris, fat and oils from the stool.
• fecal debris absorbs ether and becomes lighter than the
sediment.
• This technique leads to the recovery of all protozoan cysts, and
helminthic eggs and larvae.
Formalin-Ether Sedimentation
➢Acid-Ether Sedimentation Technique:
• The advantage of acids : dissolve mucus and other substances
resulting in a cleaner sediment.
Flotation Methods :
• It concentrates the eggs and cysts at the top because their
specific gravity is less than that of the suspending medium.
• technique detects protozoan cysts and some helminth
eggs and larvae.
Zinc Sulfate flotation
Permanent Stained Smears :
➢ For correct detection of most intestinal protozoa.
➢It allows permanent record of the protozoa identified.
➢May be used for consultation of specialists.
➢Various staining techniques are available
• Iron-hematoxylin.
• Trichrome.
➢Special staining techniques
• Modified Ziehl-Neelsen acid-fast stain.
Stool culture techniques:
➢For detection of Larvae of nematodes.
➢Examples: hookworms, S. stercoralis
➢Useful to :
a. Reveal their presence when they are scanty.
b. Identification of the adult parasite on the basis of larval
morphology.
c. Allow hatching of hookworm eggs, releasing larvae.
d. Allow development of the larvae for further differentiation.
Stool culture techniques
Egg studies: Kato-Katz thick film
Useful for:
• to detect egg burden
• determination of the intensity of infection.
• deciding on possible chemotherapy.
• evaluating the efficacy of the drugs administered.
Kato-Katz thick film
Hatching of Schistosoma Eggs
Tapeworm proglottids (India Ink Injection
Procedure)
1. This technique is …………………………..
2. Stool culture technique is used mainly to detect …………………………
Answer:
1. Hatching of Schistosoma Eggs
2. Larvae of nematodes
References
Diagnostic Medical Parasitology, Garcia, 2016
