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Stool Exam - Routine, Conc. Method - Students'

The document discusses methods for diagnosing parasitic infections through laboratory examination of stool samples, including microscopic examination of fresh or concentrated stool samples to identify parasites, eggs, larvae, cysts, or trophozoites, or detection of immune responses to parasites. Proper collection, handling, and processing of stool specimens is important for generating reliable results. A variety of concentration techniques can be used to recover more parasites from stool samples with light infections.

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Anastasia
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284 views

Stool Exam - Routine, Conc. Method - Students'

The document discusses methods for diagnosing parasitic infections through laboratory examination of stool samples, including microscopic examination of fresh or concentrated stool samples to identify parasites, eggs, larvae, cysts, or trophozoites, or detection of immune responses to parasites. Proper collection, handling, and processing of stool specimens is important for generating reliable results. A variety of concentration techniques can be used to recover more parasites from stool samples with light infections.

Uploaded by

Anastasia
Copyright
© © All Rights Reserved
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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STOOL EXAMINATION –

ROUTINE & CONCENTRATION


METHOD
By: BROMARIE M. OLARTE, RMT, MLS (ASCPi)
PARASITOLOGY

It is the area of biology concerned with the


phenomenon of dependence of one living organism on
another.

Medical Parasitology is concerned primarily with


animal parasites of humans and their medical
significance, as well as their importance in human
communities.
DIAGNOSTIC PARASITOLOGY

Laboratory Diagnosis
Accurate diagnosis of parasitic infections can help decrease the
prevalence of incidence of a parasitic condition. A parasitology
laboratory should be able to:

1. Confirm a clinical impression that the condition has a


parasitic nature.
2. Rule out a diagnosis
3. Aid a clinician in the choice of proper medication
4. Help in monitoring the effect of a treatment regimen
DIAGNOSTIC PARASITOLOGY

Laboratory Diagnosis
The ability of the parasitology lab to generate reliable results is
dependent on the following:

• Proper collection, handling and processing of specimen


prior to collection.
• The skill of the laboratory analyst (examiner)
• Quality of the equipment used in the examination
DIAGNOSTIC PARASITOLOGY

Diagnosis of parasitic infections is done by:

• Demonstration of parasites – provides a definitive diagnosis


(e.g., adults, eggs, larvae, cysts, oocysts and trophozoites)

• Detection of host immune response to the parasites – provides a


presumptive evidence of infection
(e.g., antibodies and antigen)
DIAGNOSTIC PARASITOLOGY
• Among the specimens available for parasitic examinations, stool is
most studied.
• Some intestinal parasites pass diagnostic stages in specimens other
than feces; it may then be necessary to examine the following
specimens to properly diagnose the infection:
Sputum  Sigmoidoscopic material
Duodenal or biliary aspirates  Urine
Vaginal and urethral exudes  Perianal material
Blood
• Specimens must be fresh and in sufficient amounts.
COLLECTION AND PROCESSING OF
SPECIMENS FOR PARASITIC
DETERMINATION
TYPES OF FECAL SPECIMENS
1. Normally Passed Feces
- Formed, mushy or both in general; can be expected to contain helminth eggs and larvae
and protozoan cysts, but usually NOT protozoan trophozoites.

2. Purged Specimens
- Some intestinal protozoa and some helminths are more likely to be found in mushy
rather than in formed stools. Purgation increases the possibility of finding these
organisms. The purpose of administering a cathartic is to
produce a mushy and not a watery stool.
Examples of Cathartics:
Saline cathartics, sodium sulfate, buffered phosphor – soda, Cascara sagrada (one of the
best); Castor oil, mineral oil, bismuth, magnesia compounds – unsatisfactory (obscure
the organism).
GENERAL RULES IN COLLECTING FECAL
SPECIMENS
1. Containers
– Disposable, clean, leak – proof, should have a capacity
of about half a pint (0.473 L), wide – mouthed
containers made of waxed cardboard or plastic with a
tight – fitting lid.

2. Labels
- The stool specimen submitted should include the
following information:  Requested procedure
 Patient’s name  Presumptive diagnosis
 Age  Prior infections
 Sex  Travel history
 Date of collection & hour of passage
 Requesting physician
GENERAL RULES IN COLLECTING FECAL
SPECIMENS
3. Intake of drugs/ medicinal substances

Anti – diarrheals
Antacids
Bismuth found to leave
Barium crystalline residues
Laxatives
Antibiotics
Gallbladder dyes

- Stool samples should be collected a week after the


last intake of any of these drugs.
GENERAL RULES IN COLLECTING FECAL
SPECIMENS
4. Amount of stool to be collected is dictated by
the techniques which will be used. A routine
stool examination usually requires a
thumb – sized specimen of formed stool or
about 5 – 6 tablespoons of watery stool.

5. Contamination with toilet water, urine, and


soil must be prevented since these can destroy
protozoan trophozoites.
GENERAL RULES IN COLLECTING FECAL
SPECIMENS

6. Age of the stool sample is very important for


diarrheic specimens since the trophozoites it
may contain are likely to die within 30 minutes
to 1 hour after passage.

7. Delay in examination of specimens may


require preservation to ensure that parasites are
present in the identifiable stage.
GENERAL RULES IN COLLECTING FECAL
SPECIMENS

8. Temporary storage of fecal samples in a refrigerator


(3 to 5 deg C) is acceptable, but prolonged
refrigeration can bring about desiccation.
Trophozoites are killed by refrigeration, although
helminth eggs and protozoan cysts are not damaged
generally.

NEVER FREEZE STOOL SAMPLES.


NEVER KEEP THEM IN INCUBATORS.
GENERAL RULES IN COLLECTING FECAL
SPECIMENS

9. Due to intermittent passage of certain parasites and


the limitations of diagnostic procedures,
only one – third to one – half of the species present
are detected in a single fecal specimen, thus
collecting a multiple fecal specimen is
recommended.
• Three normally passed stool specimens, spaced at
2 – 3 days intervals should be examined before other
procedures such as catharsis and sigmoidoscopy are
considered.
METHODS OF EXAMINATION

MACROSCOPIC OR PHYSICAL EXAMINATION


• Stool samples are submitted to the laboratory in the fresh state or as preserved
samples. If stools are fresh, the laboratory can classify the consistency of the
samples as to:
 Formed  Loose
 Semi – formed  Watery
 Soft
• Protozoan trophozoites are generally observed in soft or liquid stool.
• Protozoan cysts are often found in formed or semi – formed samples.
• Helminth eggs and larvae can be found in any type of consistency.
STOOL CONSISTENCY

• HARD – resists puncture

• FORMED – can be punctured

• SOFT – can be cut

• MUSHY

• LOOSE – can be reshaped

• WATERY - “pours”

• DIARRHEIC “flows”
METHODS OF EXAMINATION

MACROSCOPIC OR PHYSICAL EXAMINATION


• The color of the stool can be indicative of the presence of the parasite.
• Presence of blood should always be reported.
• Dark – colored blood suggests bleeding high up in the GI tract, while bright –
red blood means bleeding in at the lower level.
• Blood and mucus in soft watery stools may possibly yield the presence of
trophozoites.
• Ingestion of some compounds may impart a characteristic color to the stool (e.g.,
black color with iron intake)
METHODS OF EXAMINATION
MICROSCOPIC EXAMINATION

1. White blood cells – inflammation, immune response to a parasitic infection


2. Red blood cells – ulcerations or bleeding
3. Macrophages – bacterial and parasitic infections; can be mistaken as trophozoites
4. Charcot – Leyden crystals – hypersensitivity, parasitic infections (amebiasis)
5. Epithelial cells
6. Eggs of arthropods, plant nematodes and other spurious parasites
7. Fungal spores from Candida spp,. yeast and yeast – like fungi
8. Elements of plant origin – plants cells/ fibers, pollen grains, vegetable spirals, starch
granules
9. Plant and animal hairs – may resemble helminth larvae
DIRECT FECAL SMEAR
What you need:
– about 2 mg of stool sample
– 0.85% sodium chloride solution (NSS)
– Coverslip

• Routine method of stool examination primarily useful in the detection of motile


protozoan trophozoites , in this preparation, trophozoites may appear very pale
and transparent.
• Protozoan cysts can also be seen in DFS with the addition of a weak iodine
solution to temporarily stain and demonstrate the nuclei.
• Helminth eggs and larvae can also be detected .
CONCENTRATION TECHNIQUE

• Recommended in cases of light infections, or if there is a need to recover more


parasites.

Sedimentation – parasites has a higher SG than the reagent, therefore the parasite
will sink at the bottom.

Flotation – parasite has a lower SG and will therefore float to the surface of the
preparation.

• Mounts prepared from flotation techniques are cleaner than those from
sedimentation.
FORMALIN ETHER CONCENTRATION TECHNIQUE

1.In a suitable container, thoroughly mix a portion of stool specimen about the size of a walnut into
10mL of saline solution. Mix thoroughly.

2.Filter the emulsion through fine mesh gauze into a conical centrifuge tube.

3.Centrifuge the suspension at relative centrifugal force (RCF) of 600 g (about 2000 rpm) for no less
than 10 minutes. The suspension should yield about 0.75mL of sediment for fresh specimens and
0.5 mL for formalinized feces.

4.Decant the supernatant and wash the sediment with 10 mL of saline solution. Centrifuge again and
repeat washing until supernatant is clear.

5.After the last wash, decant the supernatant and add 10 mL of 10% formalin to the sediment. Mix
and let stand for 5 minutes to effect fixation.
FORMALIN ETHER CONCENTRATION TECHNIQUE

6. Add 1 to 2 mL of ethyl acetate, Stopper the tube and shake vigorously.


7.Centrifuge at 450 g RCF (about 1500 rpm) for 10 minutes. Four layers should result
as follows – ETHER, PLUG OF DEBRIS, FORMALIN, SEDIMENT.
8.Free the plug of debris from the side of the tube by ringing with an applicator stick.
Carefully decant the top three layers.
9.With a pipette, mix the remaining sediment with the small amount or remaining fluid
and transfer one drop each to a drop of saline and iodine on a glass slide. Cover with
a coverslip and examine microscopically for the presence of parasitic forms.
FORMALIN ETHER CONCENTRATION TECHNIQUE
GROUP ACTIVITY (DUFFY)
Discuss and make a short presentation regarding the following method of
STOOL PRESERVATION. Include the advantages and disadvantages of
each. 30 mins preparation time. Reporting will start at 10:30 AM .

GROUP 1 – FORMALIN
GROUP 2 – MERTHIOLATE – IODINE FORMALDEHYDE (MIF)
GROUP 3 – SCHAUDINN’S SOLUTION
GROUP 4 – POLYVINYL ALCOHOL
GROUP 5 – SODIUM ACETATE – ACETIC ACID FORMALIN
GROUP ACTIVITY (KELL)
Discuss and make a short presentation regarding the following method of STOOL
EXAMINATION. Include the procedure, specimen required and the advantages of each
examination. 30 mins preparation time. Reporting will start at 10:30 AM .

GROUP 1 – KATO THICK SMEAR

GROUP 2 – ACID ETHER CONCENTRATION TECHNIQUE

GROUP 3 – FORMALIN ETHYL ACETATE CONCENTRATION TECHNIQUE

GROUP 4 – ZINC SULFATE FLOTATION TECHNIQUE VS SHEATHER’S


SUGAR FLOTATION

GROUP 5 – CELLULOSE TAPE/ SCOTH TAPE METHOD


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