MLS 108 (LAB) – Parasitology (PRELIM)
L3: Microscopic Examinations of Stool
Microscopic Examinations
➔ Can reveal many elements present in a stool
specimen aside from parasites and other normal
fecal constituents.
➔ It is imperative to identify parasites apart from
artifacts.
1. Direct wet mounts of fresh or preserved material
2. Wet mounts of concentrates
3. Permanent stains
DIRECT WET MOUNTS (Direct Wet Preparation)
➢ a slide made by mixing a small portion of unfixed
stool (stool with no added preservatives) with saline
or iodine and subsequent examination of the
resultant mixture under the microscope
a. Direct Iodine Wet preparation - made by placing a
drop of iodine (Lugol’s or D’Antoni’s formula) on a
glass slide (a 3- × 2-inch size is suggested) and
mixing with a small portion of unfixed stool using a
wooden applicator stick or another mixing tool
b. enhances the detail of protozoan cysts
c. kills trophozoites
Types of Direct Wet Mount
0.85% NSS dense enough that newspaper print
can be just read through it; motility of
slow-moving amoeba – 15 seconds
Lugol’s – enhances visibility of nuclear
iodine (1:5) structures in protozoal cysts & for
glycogen inclusions; loss of
trophozoite motility & cyst refractility &
in recognizing chromatoidal bodies
Artifacts and Confusers
Microscopic Elements
1. White Blood Cells
a. Polymorphonuclear cells – indicates
inflammation; average size: 15 µm, are often
mistaken for amebic cysts, especially those of
Entamoeba histolytica (12 to 18 µm)
b. Eosinophils – immune response to a parasitic
infection • Red Blood Cells – indicates ulceration
or bleeding
2. Macrophages
- present on both bacterial and parasitic
infection
- Can be mistaken for protozoan trophozoites (E.
histolytica), only the macrophages ingest
polymorphonuclear WBCs
- significantly smaller (5 to 10 µm)
3. Charcot Leyden Crystals
- are released after disintegration of eosinophils.
- It may indicate hypersensitivity or parasitic
infection especially amebiasis
- typically found in stool or sputum specimens
4. Epithelial cells
- from the intestinal mucosa
5. Eggs of Arthropods, plant nematodes, and other
spurious parasites
- may be mistaken for human parasites
6. Fungal Spores Candida spp., yeast, and
yeast-like fungi
- mistaken for parasites
Microscopic Elements
Elements of Plant in Origin which resemble some
parasites:
❖ Plant cell fibers
❖ Pollen Grains
❖ Vegetables Spirals
❖ Starch Granules
❖ Vegetable cells
Plant and Animal Hair –may look like helminth larvae
➔ To work safely in this environment, laboratory
personnel must learn what hazards exist, the basic
safety precautions associated with them, and finally
apply the basic rules of common sense required for
everyday safety.
MLS 108 (LAB) – Parasitology (PRELIM)

Artifact Parasite Distinguishing Characteristics of Artifact

Epithelial cells amebic trophozoites lack the typical amebic trophozoite interior structures. For example, the
cytoplasm of epithelial cells is usually smooth and contains no inclusions

Plant cells Helminth & cestode Breaks in cell wall or wall too thick; lack of internal egg structure, defined
eggs; Protozoan embryo, or oncosphere; lack of characteristic operculum, spines, or knobs;
cysts size

Vegetable cells helminth eggs typically large and roundish oval to irregularly round in shape and may
measure up to 150 µm in size; thick cell walls are usually present; interior
portion of vegetable cells is unorganized and often appears to consist primarily
of large vacuole

Vegetable helminth larvae do not have a head or tail region; distinguished from parasitic forms by their
spirals ladder-like appearance (consists of a series of rungs that are spaced closely
together)

Pollen grains Taenia species or Size; lack of 6-hooked oncosphere; lack of embryo wall; shape
Trichuris trichiuria
eggs or Ascaris

Mushroom Taenia species or Lack of striations; lack of embryo; size

spores Enterobius species
eggs

Beaver bodies T. trichiura eggs/ Lack of bipolar plugs; incorrect internal structure; size
Diatoms helminth eggs
(henry)
MLS 108 (LAB) – Parasitology (PRELIM)
MLS 108 (LAB) – Parasitology (PRELIM)

Structures That Mimic Protozoan Parasites

Artifact Parasite Distinguishing Characteristics of Artifact

Polymorphonuclear Entamoeba histolytica cyst Granular or frothy cytoplasm; nuclear/cytoplasmic ratio 1:1;
leukocyte (PMN) presence of chromatin bands

Macrophage E. histolytica or Entamoeba coli Ingested PMNs, round red bodies; nuclear/cytoplasmic ratio
trophozoites 1:4 to 1:6; debris-filled cytoplasm

Squamous & Amebic trophozoites other than E. Large single nucleus; distinct cell border; smooth cytoplasm
columnar epithelial histolytica that stains poorly
cells

Yeasts Protozoan cysts including Size (4 to 8 µm ); thick refractile cell wall; cytoplasm uniform
Cyclospora, Cryptosporidium (4 to with little internal structure; no nucleus; presence of budding
6 µm) & Microsporidium species; forms
Entamoeba hartmanni (5 to 12
µm), Entamoeba nana (4 to 12
µm), and Entamoeba hominis (3
to 10 µm

Starch E. hartmanni and E. nana (both Round to angular form; refractility; no internal structure or
granules/cells measure 5 to 12 µm nucleus; nondescript mass located inside the cell is often
present and may resemble a nucleus staining
characteristics with iodine
MLS 108 (LAB) – Parasitology (PRELIM)

Other Artifacts

Type of Artifact Resemblance

● Yeasts Protozoan cysts (especially Endolimax nana)

● Fungal Conidia Helminths eggs
● Ingested mite eggs Helminths eggs
● Ingested plant nematode eggs Helminths eggs

● Ingested plant nematode larvae Nematode larvae

● Plant hairs Nematode larvae
● Fat globules, air bubbles, mucus Protozoan cysts

● Clumped or fused platelets Malarial parasites, especially the young trophozoite form
● Giemsa-stain precipitate Malarial parasites
● Red blood cell abnormalities, (HowellJolly Malarial parasites
bodies or Cabot’s rings