Medical Parasitology Lab

Medical Parasitology Lab.
Concentration techniques
Flotation Method
Concentration Methods
1. Sedimentation method
 Modified Formal- Ether sedimentation technique
 Acid- Ether sedimentation technique
2. Flotation method
 Saturated Salt Solution technique
 Sheather’s Sugar Centrifugal Flotation technique
 Zinc Sulphate Centrifugal Flotation technique
Raed Z. Ahmed, Medical Parasitology Lab.,2012

Flotation technique
 These method use the high specific gravity of a solution to
float the lighter ova and cyst. They can be improved by
centrifugation.
• Advantage:
• Easy to perform .
• Disadvantage:
• Delay in examination can result in distortion.
• Larvae and some fluke eggs do not concentrate.
• Frequent checking of specific gravity.

Flotation Methods
SATURATED SALT SOLUTION

Materials and Method
1. Boil granular sodium chloride in excess in water to produce a
saturated solution which when cooled has a specific gravity of
1.18 - 1.2.
2. Half fill a wide- mounted flat bottomed container with the
saturated salt solution.
3. Emulsify 1gm of feces in the solution and strain it to remove
the debris from the surface.
4. Pour the filtrate into meniscus and fill it to the top with
saturated salt solution.
5. Lay a glass slide or large coverslip over the top, avoiding any
bubbles being trapped.
6. Leave for 20 min before quickly inverting the slide.
7. Scan for ova using the 10x objectives.
Saturated Salt solution technique
 Advantages:
– It is cheap preparation using simple apparatus.
– It concentrates nematode ova well.
 Disadvantages:
– It doesn’t concentrate cysts.

Flotation Methods
SHEATHER’S SUGAR CENTRIFUGAL

FLOTATION TECHNIQUE
Materials and Method
 Sheather’s sugar solution:

– Table sugar -------------------------------------- 500gm
– Distilled water ---------------------------------- 320ml
– Phenol crystal ( melt in hot water bath) ----- 6.5gm

Procedures
1. Soften 1gm of feces with water to a soft.
2. Strain the aqueous suspension through a wire sieve.
3. Mix 1 part aqueous suspension with 2 part of Sheather's
sugar solution.
4. Pour into a centrifuge tube, centrifugation 1500 rpm for
10 minutes.
5. Pour the supernatant into a meniscus and add a sufficient
solution to bring the meniscus to the top.
6. Place a coverslip and wait for 10 minutes.
7. Examine under microscope.

Sheather’s Sugar solution technique
 Advantages:
– Reveals most nematode eggs and protozoan cyst.
 Disadvantages:
– Flukes eggs and tape worm eggs are not demonstrate well.
– Also most nematode larvae are not demonstrate well.

Intestinal Protozoa
Balantidium coli
Balantidium coli
 Is a parasitic species of ciliate protozoa that cause
balantidiasis or Balantidium dysentery.
 Balantidium coli has two developmental stages, a
trophozoite stage and a cyst stage.
 The cyst is the infective stage measures about 50 to 70 µm
in diameter, characterized by the presence of a large
kidney-shaped macronucleus.
 The trophozoite is oval with 2 nuclei and measures
approximately 50 to 100 µm long and 40 to 70 µm wide.

B.coli Cyst

B. coli Trophozoite

Intestinal Protozoa
Cryptosporidium parvum
Cryptosporidium parvum
 Infect human and most mammals.
 The infective stage is oocyst containing sporozoites
measuring 5u in diameter.
 The diagnostics stage is sporocyst containing 4 sporozoites.
 Diagnosis:
– Detecting oocyst in stool.
– Acid-fast stain.

Cryptosporidium parvum oocyst

Tissue Protozoa
Toxoplasma gondii
Toxoplasma gondii
• Habitat: reticulo-endothelial system, monocyte, and muscle
fiber and cause toxoplasmosis.
• Humans can acquire Toxoplasma gondii infection by ingestion
cyst or sporylated oocyst
– Cyst: precipitated in flesh of cow.
– Sporulated oocyst: cat feces.
o Intermediate host: Human.
o Definitive host: Cat.
• The tachyzoites are crescent-shaped and measure about 5 µm in
length.
• Diagnostic stages:
– Diagnosis also can be done by detection antibody IgG, IgM, IgE and
IgA.
Tachyzoites

Urogenital Protozoa
Trichomonas vaginalis
Trichomonas vaginalis
 Is the most common cause of vaginitis, inhabit the
urogenital system
 There is no cyst stage but only have trophozoite stage.
 The trophozoite measures about 15 x 10 µm. The
trophozoite has a single nucleus and four flagella and
undulating membrane.
 Diagnosis:
– Male: finding trophozoite in urethral prostatic discharge
– Female: finding trophozoite in vaginal discharge.

T. vaginalis Trophozoite

Intestinal Cestodes
Hymenolepis nana
Hymenolepis nana
 This is one of the most common cestodes of humans, especially
children. And it’s the smallest tape worm in man, also called
dwarf tape worm.
 Eggs measures 30-37u, and have double membrane and contain
six hooked oncosphere.
 The scolex of H. nana has four suckers and a short rostellum
with hooks.
 Infective stage: Eggs.
 Diagnosis:
– Stool examination to detect the eggs.

H. nana egg

H. nana adult worm

H. nana scolex

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Medical Parasitology Lab.

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

SATURATED SALT SOLUTION

Raed Z. Ahmed, Medical Parasitology Lab.,2012

SHEATHER’S SUGAR CENTRIFUGAL

 Sheather’s sugar solution:

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

Raed Z. Ahmed, Medical Parasitology Lab.,2012

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