Dr. Abdullah A.

Hama Clinical Parasitology Lab 3

Practical part of Clinical Parasitology

Sulaimani Polytechnic University

College of Health and Medical Technology
Medical Laboratory Technique

Concentration Technique (Floatation method)

Dr. Abdullah Ahmed Hama

abdullah.hama@spu.edu.iq

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Dr. Abdullah A. Hama Clinical Parasitology Lab 3

General stool examination

Macroscopic Microscopic Others

Permanent Temporary

Direct saline smear Iodine smear Concentration techniques

(wet smear)

Floatation Sedimentation

Sat saline Saline

Formol ether
Zinc sulfate

Sheather’s sugar
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

General advantages and disadvantages of concentration methods:

Advantages
• Maximizes the numbers of organisms detected which may be too scanty to be seen by
direct microscopy.
• Worm eggs, larvae, and protozoan cysts may be recovered.

Disadvantages
• Most concentration methods destroy trophozoites stages.
• The purpose of concentrating feces is to increase possibility to finding ova, cyst, or larvae
in samples that not be able to seen by direct microscopy.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

Advantages & disadvantages of Floatation

& unfertilized eggs of A. lumbricoides

Dr. Abdullah A. Hama Clinical Parasitology Lab 3

Flotation Technique for Fecal Parasite

• This technique is predominantly used in parasitology laboratories. By exploiting the
density of the parasites, particularly eggs, it allows the parasites to float to the top of a
dense solution (final specific gravity of about 1.20) and can then be skimmed from the
top of the tube.

• Operculated eggs as well as schistosoma and infertile Ascaris eggs are not easily
recovered by this method.

• Also trophozoites are killed due to the high specific gravity and certain other fragile eggs
such as Hymenolepis nana become distorted.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

The Specimen Collection:

• Fresh fecal sample (liquid stool) preferably within 30 minutes of passage by prior arrangement with
lab.
• If a fecal sample is not properly collected and contaminated before examination, they will be of little
or no value for accurate diagnosis. This is especially true if protozoa are present.
• Amoebic trophozoites begin to degenerate 1-2 hours after passage, as do flagellate trophozoites.
• Cysts will deteriorate if the fecal specimens are left standing for many hours or overnight, especially
at high temperatures.
• Helminth eggs and larvae are less affected by the age of the specimen than are protozoa. Nevertheless,
changes may occur that could affect their identification. eg, hookworm larvae may become embryonated and
larvae may hatch from the eggs, this lead to confusion with Strongyloides larvae. Larvae themselves may
disintegrate thus making their identification difficult.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

• To ensure that good specimens are provided for examination, it is important to note
the following points:
1. A clean dry container must be used for the collection of fecal samples. Urine and water
will destroy trophozoites, if present, and the presence of dirt also causes identification
problems.
2. Ideally the specimen should be brought to the lab as soon as it is passed, to avoid
deterioration of protozoa and alterations of the morphology of protozoa and
helminths.
3. The specimen container should be clearly labelled with the patients name, date, and time
of passage of the specimen.
4. An amount of stool adequate for parasite examination should be collected and a repeat
sample collection requested if too little is supplied.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

Reagents for floatation techniques:

• The most commonly used reagent is zinc Sulfate

• Normal saline (0.85%).

Equipment:

• Fume hood, safety centrifuge, microscope with ocular micrometer, applicator

stick, Funnel filter – disposable (PML), caped centrifuge tube, Pasteur pipette,
Glass microscope slides, Cover slips (22 x 40 mm)
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

Procedure:
1. Mix 10-20g of feces with 10-12ml of saline by applicator sticks then mix well. Filter the
mixture through a commercial funnel filter or two layers of dampened surgical gauze into a
15ml disposable plastic centrifuge tube.
2. Centrifuge the suspension at 1500 rpm for five minutes. Decant the supernatant into
disinfectant.
3. Resuspend the sediment and recentrifuge in saline if there is excessive debris in the sample.
4. Resuspend and thoroughly mix the sediment in 12ml of zinc sulphate solution (specific
gravity, 1.18 to 1.20, as verified with a hydrometer).
5. Centrifuge for one minute at 2500 rpm. Place tube in a rack in a vertical position and slowly
add enough zinc sulfate with a dropper pipette to fill the tube so that an inverted meniscus
forms.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3

6- Without shaking the tube, carefully place coverslip on top of the tube so that its
underside rests on the meniscus. The meniscus should not be so high that fluid runs
down the side of the tube carrying parasites away from the cover glass.

7-Allow the tube to stand vertically in a rack with the coverslip suspended on top for
ten minutes.

8- Carefully lift the coverslip with its hanging drop containing parasite eggs and cysts
on the underside and put on a clean slide, liquid side down.

9- A small drop of iodine stain may be placed on the slide prior to adding the coverslip.
10- The slide is gently rotated after adding the coverslip to ensure a uniform mixture.
11- The slide is then thoroughly examined microscopically.
Dr. Abdullah A. Hama Clinical Parasitology Lab 3