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Hymenolopsis nana

Hymenolepsis is now Vampirolepsis. It’s another name dwarf  tapeworm  because of being the smallest (3-4 cm) tapeworm to infect humans. Above picture is showing an egg of Vampirolepsis nana in saline wet mount of stool.

Identification features

1. Spherical or oval in shape
2. Diameter 35-45µm
3. colorless
4. It floats in saturated common salt solution.
5. Oncosphere is also colorless  and shows 3 pairs of hooklets.
6. Space between outer membrane and embryophore  has filled with yolk granules and polar filaments.

Key notes

1. Differentiating point from Hymenolepsis diminuta: H . diminuta egg is larger, outer shell is yellowish and devoid of polar filaments.
2. There is a little difference between normal and physiological saline. Physiological saline is  0.85% NaCl where as normal saline 0.9% NaCl.
3. Gram’s iodine  is not applicable for staining parasitic organisms and for this D’Antoni’s iodine uses.
4. Oil immersion examination is also preferred in parasitology for the permanent stained smear of parasites.
5. Intestinal protozoa can not conform on the basis of a wet mount alone and thus permanent stained smears requires to confirm the specific identification of suspected organisms.

Principle of saline wet mount of stool

Saline wet mount preparation for stool is the simplest and basic method of the analyzing a stool specimen in  coprology (study of feces). It utilizes a physiological  saline solution (0.85% NaCl ) as an isotonic media to maintain the cellular structure of the various organisms that are found in stool and that we like to examine.

Requirements for saline wet mount

• Physiological saline (  0.85% NaC)
• Specimen: stool
• Sterile bamboo sticks or low cone on the end of a wooden applicator stick
• Clean and grease free slides and
• Cove slips(22- by 22-mm)
• Microscope
• Gloves

Saline wet mount of stool Preparation

1. Take a clean and grease free slide.
2. Add one drop of physiological saline and then add  a stool equivalent to a match stick head (2 mg)  with the help of  stick.
3. Mix it properly and apply a cover slip over a uniform suspension  without creating bubbles.
4. Note: If a fresh stool specimen is received and if blood and mucus are present, the specimen should be examined as a direct mount making sure to sample the bloody areas.
5. Examine the entire 22- by 22-mm coverslip systematically with the low power objective (10X ) and low light intensity.
6.  If any suspicious objects encounter, examine with the high dry objective (40X).

//Eggs of Hymenolypsis  nana in saline wet mount of stool under the microscope as shown in video-

Result Interpretation

Presence of active trophozoite/s: Motile retractile bodies

Cyst , oocyst, egg, inactive trophozite/s, larvae: Retractile bodies and finally focus at high dry power field as shown above video

Limitations of saline wet mount for stool examination

1. Due to lack of stain, it is difficult to get morphological details.
2. Inappropriate preparation of the smear may hide parasites
3. Improper adjustment of the microscope in relation to the objective may create problems.

References

1. Medical Parasitology by Abhay R. Satoskar, Gary L. Simon, Peter J. Hotez and Moriya Tsuji
2. Atlas of Medical Helminthology and ptotozoology -4th edn  -P.L.  Chiodini, A.H. Moody, D.W. Manser
3. Merkell and voge’s medical parasitology
   9th edition.
4. Parasitology: 12th edition
   By K. D. Chatterjee
5. District laboratory practice in Tropical countries –Part-I .
   By Monica cheesbrough.
6. Isenberg clinical microbiology procedures Handbook
   2nd edition. Vol. 2
7. http://www.med-chem.com/para-site.php
8. https://www.cdc.gov/dpdx/hymenolepiasis/index.html