Stool analysis

Lab-1 parasitology Part I

Dr-Salah alsaadon
 FECAL SPECIMENS
ANALYSIS

- Fecal specimens collection & transport

. Preservation
- Macroscopic examination
- Microscopic examination
. Direct wet mount

- Artifacts & Pseudoparasites

 STEPS OF STOOL ANALYSIS
 Stool specimen collection
 Stool specimen preservation
 Stool specimen examination
 Macroscopic examination
 Microscopic examination
 Direct smear (wet mount)
 Concentration techniques
 Concentration floatation
 Concentration sedimentation
Collect in a clean, dry, water-proof,
wide-mouth container

FECAL SPECIMENS COLLECTION

 FECAL SPECIMENS COLLECTION &
TRANSPORT

•NO contamination with H2O: could bring

free-living protozoa
•NO contamination with urine: destroys
trophozoites
•Collect before barium enema: obscures
organisms (7+ days)
•Collect before antibiotics: may decrease
number of organisms
 FECAL SPECIMENS COLLECTION &
TRANSPORT

Series of 3 specimens
within 7-10 days
on alternate days

are needed to exclude parasitic infection

 PRESERVATION

Specimen preservation is required if

not delivered to lab immediately:
► 3 parts fixative to 1 part stool
Record time placed in preservative ►

*Advantages:
- Preserve protozoan morphology
- Prevent development of worm eggs/larvae
 5 - 10% formalin:
wet mount/concentrate; cysts, eggs, larvae
are preserved for long time
 Polyvinyl mercuric chloride (PVA):
mercury chloride: fixative •
polyvinyl alcohol: resin aids adherence •

 Merthiolate-iodine-formalin (MIF):
wet- mount & concentration (Not permanent
stain); preserves protozoa/worms
 5% OR 10%
FORMALIN PVA

PERMENANT
STAINED SMEAR

CONCENTRATION

GENERAL PRECISE
MORPHOLOGY
MORPHOLOGY
 I

Items that should be observed by

macroscopic examination are:
 Consistency
 Color
 Blood
 Mucous parts
 Parasites or fragments
 :

 Liquid ……….. soft ………….. Formed

 Liquid and soft specimens:
 Examined within 1 hour of collection.
 Formed specimens:
 Examined within the same day of collection.
 Otherwise, refrigerated up to 24 hrs
 Not to be incubated (change diagnostic stage,
avoid bacterial growth(
 Trophozoites (protozoa motile form) ….. Liquid specimens

Trophozoites and cysts ……… might be found in soft

specimens.


Cysts ……… generaly found in formed speciemens.


Helminthes Eggs ……. May found in any type

 In liquid speciemns , dilution  chances of Egg recovery

 2- COLOR:

 Brown: normal
 Dark: possible bleeding -upper GI tract-
 Fresh blood: possible bleeding -lower GI
tract-. Select areas of stool with blood for
analysis.
 3- BLOOD:

4- Mucus:
Select areas of stool with blood
and/or mucous parts to examine
 Cestodes segments may
beneath stool.
or on foundbe

 Adult Entrobius & Ascaris may occasionally

be found on the surface or in the stool.
 in addition to normal specimen debris specimen may reveal
the following:
 Parasitic materials:
 Helminthes eggs/larvae
 Trophozoites and cysts of intestinal protozoa

 Coccidia, microsporidia Oocysts, spores.

 Non-parasitic materials:
 Fecal depris
 RBCs …….. Bleeding
 WBCs ……. Infection
 Fungi and other yeasts
 Plant cells,, Pollen grains, fungal spores ~ eggs, cysts
 Plant fibers, root hairs or animal hairs ~ helminthes larvae
MICROSCOPIC EXAMINATION

 Microscopic examination
 Direct smear (wet mount)

 Concentration techniques
 Concentration floatation

 Concentration sedimentation
 Advantages
 It is a fast, simple, procedure & provides a
quick answer when positive
 It provides an estimate of the parasitic burden
 It can be used as a screening test to check
trophozoite motility

Disadvantages:
Negative results should
be confirmed by concentration methods or
permanent stained smears
 MATERIAL:

 Microscope slides
 Cover slips
 Wooden stick
 Gloves
 0.9% Sodium chloride solution
 d’Antoni iodine
 Fresh stool
 PREPARATION:
 Use clean microscope slides
 Place a drop of saline

 Take a small amount of stool (a size of head of

match stick) with a wooden stick

 Mix stool with saline or iodine

 Put a 22 x 22 mm cover-slip & examine
 (CONT.):PREPARATION

Direct smear should be done with

saline and iodine:
saline:  wormdetect & eggs/larvae
motile protozoans liquid/softin
stools
 iodine: shows nuclear detail and
colorless protozoan cysts
 MICROSCOPY:

 correct illumination
 scan edges of cover-slip
 observe entire slide

 use references: pictures, size charts,

stained positive slides.
 MICROSCOPY:

 For helminth ova :

Use low power objective(10x)

 For protozoa:
Use high power objective (40x)
 COMMON MISTAKES:

1. Too much stool!

2. Air bubbles! on placing a cover-slip
After wet mount or concentration method you may find the egg (or
the following parasites by low powerlarva) of
10X
Flukes eggs
1 Nematodes 6
eggs & larva

3 2 7 8

4 9

1 1
5 1
3 1
0
1
5 1
4 1
Cestodes egg 2
VERMICULARIS

A. lumbricoides .2
S. stercolaris .3
larva
Hook worm .4
T. trichiura .5
D. latum .6
H. heterophyes .7
.Fasciola sp .8
S. mansoni .9
Paragonimus .10
.sp
S. japonicum .11
S. intercalatum .12
.Taenia sp .13
H. nana .14
H. diminuta .15
To demonstrate the nuclei of protozoan trophozoites and cysts you will
need to used Lugol’,s iodine and to examine the slide by high power
)x40(

tive 40
3
1
1
3 2 5
1 1
4 Iodine 5 4
6
1 Saline
8 1
6
7
1 9
7 8
1
9 1
1 2
1
40X
I. belli oocyst .1
A. .2
lumbricoides
3 E.Leucocytes
histolytica.3.4
13 1 cyst
E. histolytica .5
trophozoite
2 Red cells .6
15 5 S. stercolaris larva .7
14 E. coli cyst .8
Iodine 4 G. lamblia cyst .9
6 C. mesnili cyst .10
18 Saline Hook worm egg .11
16 G lamblia.12
trophozoite
E. coli cyst .13
7 I. buetschilii cyst .14
17 9 E. histolytica .15
8
cyst
19 E. nana cyst .16
T. trichiura egg .17
12 B. hominis .18
10
G. lamblia cyst .19
11
REQUIRED THAT FAECESIN FOUNDSTRUCTURES
FROM PARASITES.DIFFERENTIATION
STRUCTURES FOUND IN FECES THAT REQUIRED
DIFFERENTIATION FROM PARASITES.
ARTIFACTS