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Concentration techniques
2. Concentration techniques :
Advantages
Maximizes the numbers of organisms detected which may
be too scanty to be seen by direct microscopy alone.
Worm eggs, larvae, and protozoan cysts may be recovered.
Disadvantages
Destroys trophozoite stages. Most concentration methods
destroy trophozoites stages.
Concentration Methods
1. Sedimentation method
Modified Formal- Ether sedimentation technique
Acid- Ether sedimentation technique
2. Flotation method
Saturated Salt Solution technique
Sheathers Sugar Centrifugal Flotation technique
Zinc Sulphate Centrifugal Flotation technique
Sedimentation Methods
Procedures
1. Emulsify 1 gm. of feces in 7 ml of 10% formalin in a
centrifuge tube.
2. Strain the suspension through a brass wire sieve, and
collect in beaker.
3. Pour the filtrate into a 15 ml boiling tube and add 3 ml
of ether, then mix well 15 sec on vortex or whirlimixer
or 1 min by hand.
4. Transfer the ether- formalin suspension back into the
washed centrifuge tube, and centrifuge at 3,000 rpm for
1 min.
Raed Z. Ahmed, Medical Parasitology Lab.,2012
Procedures
(cont.)
5. Loosen the fatty layer and debris at the top of the tube
with an applicator stick and invert the tube quickly to
discard the supernatant.
6. On righting the tube, a few drops only should remain
with the sediment, mix the sediment well and transfer
one drops onto a glass slide and cover it with coverslip.
7. Scan the whole coverslip using 10x objective, turning
into 40x for confirmation of identification of parasites.
Sedimentation Methods
Procedures
1. Mix thoroughly 1 gm. feces with 3 ml of 15% of
hydrochloric acid and then mix well.
2. Add and additional 5-6 ml of 15% HCl and mix.
3. Strain the suspension through a wire sieve into beaker.
4. Place suspension in a glass centrifuge tube and make up
to the 10 ml with distilled water.
5. Add 4 ml of ether, stopper the tube and shake
vigorously 20 -30 sec using vortex.
Procedures
(cont.)
Intestinal Protozoa
Giardia lamblia
Raed Z. Ahmed, Medical Parasitology Lab.,2012
Giardia lamblia
It is the most common flagellate of the intestinal tract that
cause giardiasis, Traveler's diarrhea.
There is two diagnostic stages for Giardia lamblia :
1. Cyst (infective stage).
2. Trophozoite (motile form, motility by flagella).
Diagnosis:
. Stool examination to see cyst stage, or trophozoite
stage if the sample is fresh.
Life cycle