1 Notes in Clinical Parasitology Lecture (Module 1) Francis Cedrick J.

Victorino, RMT, MSMT(c)

PARASITOLOGICAL SPECIMEN J. MOUTH SCRAPPINGS

A. STOOL • Entamoeba gingivalis
• Ova of intestinal parasites: • Trichomonas tenax
1. ___________________________
2. ___________________________ K. NASAL DISCHARGE – Naegleria fowleri
3. ___________________________
4. ___________________________ STOOL EXAMINATION
5. ___________________________ COLLECTION:
6. ___________________________ • Acceptable amount: ________
7. ___________________________ • Container: Clean, watertight
8. ___________________________ o Patients name and ID number
• Rhabditiform larva of _____________________ o Physicians name
__________. o Date and time of collection
• Cyst and trophozoite of: • Number of Recommended Stool Sample:
1. ___________________________ o Recommended: ___________________
2. ___________________________ _______________.
3. ___________________________ o Amoebiasis: ______________________
• Oocysts of _____________________________. _______________.

B. URINE •ADULTERANTS:
• Ova of ________________________________. o Urine
• _________________ of Trichomonas vaginalis o Toilet water – with _________________
o Toilet paper
C. SPUTUM o Water – destroys __________________ and
• Ova of ________________________. __________________
• Trophozoite of Entamoeba histolytica, o Barium, bismuth, mineral oil – before
_________________________. administration or _____________ after
• Filariform larva of ________________________ administration
• Larva of _______________________________, o Antibiotics or antimalarial drugs – ________
__________________________. after administration
PRESERVATION
D. BLOOD • Examination:
• Ring form trophozoite and gametocytes: o Watery stool: ____________________
1. ___________________________ o Semi-formed Stool: _______________
2. ___________________________ o Formed stool: ____________________
• Microfilariae of Filarial worms • Storage:
• Blood flagellates o Room temp: _____________
o Ref temp (3-5C): __________
E. CEREBROSPINAL FLUID o DO NOT FREEZE or PLACED IN INCUBATOR
• Naegleria fowleri • FIXATIVES
• Acanthamoeba spp. o Preserves morphology of protozoa and
• Taenia solium _________________________ prevent further development of certain
helminth eggs and larva.
F. DUODENAL ASPIRATE o Recommended Ratio: ______________
• Methods of Collection: _________________.
1. Nasogastric intubation
2. Enteric capsule test - _______________ A. FORMALIN
• _______________________ trophozoite • All-purpose fixative
• Cryptosporidium spp. • __________________ – protozoan cysts
• Isospora belli • __________________ – helminth egg and larva
• Strongyloides stercoralis _______ and ________ • Advantages: easy to prepare, long shelf-life,
• Ova of ____________________ and __________ preserves specimen for several years
________________. • Disadvantages:
o does not preserve parasite morphology
G. PERIANAL SWAB adequately in permanent smears
• ____________________________ o ___________________ are usually not
• Taenia spp. ova recovered
o Morphologic details of ________ and
H. EYE SPECIMEN ________ fade overtime
• Acanthamoeba spp. - _____________________.
• Loa loa - aka _________________.

I. SKIN SNIPS - ___________________________.

Original Intellectual Resource Material of Francis Cedrick J. Victorino, RMT, MSMT(c)

2 Notes in Clinical Parasitology Lecture (Module 1) Francis Cedrick J. Victorino, RMT, MSMT(c)

B. POLYVINYL ALCOHOL (PVA)

• with plastic powder - ____________
• Combined with Schaudinn solution
o ___________________
o ___________________
o ___________________
• Advantages:
o Used for permanent stained smear
preparation
o Trophozoite, cysts, most helminth eggs, may
be detected C. MODIFIED ACID-FAST STAIN
o Suited for concentration techniques • Important for permanent stain procedure for
• Disadvantage: ___________________ detection of oocysts of:
1. _______________________
C. SODIUM ACETATE FORMALIN (SAF) 2. _______________________
• alternative for PVA with Schaudinn Solution 3. _______________________
• used for concentration techniques and and permanent
stained smears
• Advantages:
• Easy to prepare, long shelf-life
• Used for preparing smears for smears to be stained
with modified acid-fast stain to detect coccidian
oocysts
• Disadvantage:
• Trophozoites is not clear in permanent stained smears
D. MODIFIED IRON HEMATOXYLIN
D. MODIFIED POLYVINYL ALCOHOL (Modified PVA)
• With incorporation of _____________________.
• Replaced mercury-based preservatives with
________________ or _________________.
PROCESSING OF STOOL SPECIMEN
• Advantage:
MACROSCOPIC EXAMINATION:
o Used for concentration techniques and
• COLOR
permanent stained smears
o Normal color: ___________.
• Disadvantage:
o Black: ________, _________, ________,
o Preservation of protozoan morphology is not
__________.
as good as ___________________.
o Red: __________, __________
o Green: __________________, ________
PERMANENT STAIN
____________.
A. TRICHROME STAIN (Wheatley Modification)
o Clay/Pale - ___________; ___________
• most widely used
_________
• stains distinct features (cytoplasm, nucleus) of
• CONSISTENCY
parasite
o Soft or Liquid: _____________________
• stains background
o Formed: _________________________
o Liquid or Formed: _________________,
___________
• Blood or Mucus in Loose or Liquid stool:
______________________________.
• Bright red blood on formed stool: ___________
and _____________.
• Presence of adult worm and/or proglottids
• Presence of mucus
MICROSCOPIC EXAMINATION:
• Scan all microscopic fields
• Low Power Objective (LPO)
B. IRON HEMATOXYLIN o ______________________
• Reveals excellent morphology of ____________ o ______________________
_______________. • High Power Objective (HPO)
• ______________________ of parasites are also o ______________________
stained clearer and sharper than trichrome. o ______________________
o ______________________
o ______________________
• Normal Microscopic Structures: vegetable hair,
cells, spirals, starch granules

Original Intellectual Resource Material of Francis Cedrick J. Victorino, RMT, MSMT(c)

3 Notes in Clinical Parasitology Lecture (Module 1) Francis Cedrick J. Victorino, RMT, MSMT(c)

DIRECT SALINE WET MOUNT B. EGG COUNTING TECHNIQUE

• Useful in detection of _____________________ • Quantitative procedure
• ______________________________________ - • Carried out to:
stains trophozoites o Determine _______________________
• Preparation: o To assess effectiveness of anti-helminthic
o Small portion of stool + drop of _______ drugs
_______________________ • Two Methods:
o Emulsify on glass slide evenly then place I. Stoll Dilution Technique
coverglass • most widely used dilution egg-counting technique
o Examine under LPO and HPO • Special Equipment: Stoll flask (with _____ mL and
DIRECT IODINE WET PREPARATION _____ mL marking)
• Iodine: • Reagent: _____________________
o _________________
o _________________
• Enhances detail of protozoan ___________.
• Causes disappearance of protozoan
______________.
• Preparation:
o Small amount of stool + drop of Iodine
o Emulsify evenly on glass slide then place
coverglass
o Examine under LPO and HPO

DIAGNOSTIC PROCEDURE IN STOOL SPECIMEN

A. KATO THICK SMEAR
• Uses larger amount of stool and _____________
____________ in replace of coverglass
• Reagents:
o Glycerine - __________________
o Malachite Green - _________________
__________.
• Advantages:
o Economical
o Employs simple procedure II. Kato-Katz Technique
o Applicable for thick shelled eggs - • Aka ___________________________________
___________, ____________. • A measured amount of stool is sieved through a wire
• Disadvantages: mesh and pressed under cellophane paper soaked in
o Not suited for __________________ glycerine-malachite green solution.
o Not for _______ or ______________ • Useful for assessing the intensity of infection with
o Not for thin shelled eggs - ___________, _____________________ and common
_______________. __________________________
• Disadvantage: cannot be performed using
________ and ____________ stool sample.
• MATERIALS:
o Wire mesh
o Template
o Spatula
o Slides
o Cellophane
o Glycerine-malachite green solution

Original Intellectual Resource Material of Francis Cedrick J. Victorino, RMT, MSMT(c)

4 Notes in Clinical Parasitology Lecture (Module 1) Francis Cedrick J. Victorino, RMT, MSMT(c)

C. CULTURE
• Not commonly performed to detect parasites
• Parasites that can be isolated:
o Entamoeba histolytica
o Trichomonas vaginalis
o Leishmania spp.
o Trypanosoma cruzi
o Toxoplasma gondii
o Hookworms & Strongyloides stercoralis -
__________________, Copro culture

• HARADA-MORI TECHNIQUE
• Aka _____________________________
• Useful for recovery of larva of ______________, and
__________________________.

• PROCEDURE:
1. Smear stool in the center of a strip of filter paper
2. Add about 7 mL of distilled water in a test
tube/conical tube
3. Insert the filter paper strip into the tube
4. Stand the tube at 25-28 C for 10 days

Original Intellectual Resource Material of Francis Cedrick J. Victorino, RMT, MSMT(c)