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STOOL
EXAMINATION
DEFINITION
Stool examination by Dr. Priyanka
Buragohain
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• Human feces is called as stool.
• faeces / feces is plural of latin term faex
meaning RESIDUE.
• It is the waste residue of indigestible materials
of an animal’s digestive tract expelled through
the anus during defecation.
• Meconium is newborn’s first feces.
• SCATOLGY or CAPROLOGY is the study of
feces.
COMPOSITION
Stool examination by Dr. Priyanka
Buragohain
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• ¾ water, ¼ solid
• Undigested and unabsorbed food
• Intestinal secretions, mucous
• Bile pigments and salts
• Decomposed products
• Bacteria and inorganic material
• Epithelial cells, leukocytes.
PRECAUTION BEFORE COLLECTION
Stool examination by Dr. Priyanka
Buragohain
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• Patient should avoid the following things for at
least 48 hours before collection of stool:
• Mineral oils, bismuth, non absorbable anti
diarrhoeal drugs, antimalarial drugs, antibiotics,
etc
• Pt. Should not have barium swallow examination
before stool R/E
• Avoid iron containing drugs, meat, fish etc for
atleast 48 hours before stool for occult blood.
• In constipated patients use only non residual
purgative
COLLECTION
Stool examination by Dr. Priyanka
Buragohain
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• Universal precautions
• Pt. is asked to pass stool in a clean container.
• Stool should be collected in a steralized, wide mouthed container.
• Loose/last/portion containing mucus, blood etc is to be collected
in a wide mouthed bottle.
• Should be uncontaminated with urine or any other body
secretions.
• >2gm is required.
• Properly named and always a fresh sample should be tested.
• Liquid stool to be examined within ½ hour
• Solid stool to be examined within 1 hour.
• If delayed store in a refrigerator.
COLLECTION CONTD...
Stool examination by Dr. Priyanka
Buragohain
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• 3 samples of stool within 10 days to exclude
false negatives.
• 2 samples to be examined on alternate days
after normal defaecation and 1 sample after a
purgative for certain worms.
• Formalin is the best preservative. It kills the
bacteria but ptreserves the protozoa and
helminthes.
• For culture no preservatives to be used
Stool examination by Dr. Priyanka
Buragohain
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TYPES OF EXAMINATION
Stool examination by Dr. Priyanka
Buragohain
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• PHYSICAL EXAMINATION: colour, volume,
consistency, odour, mucus, pus, concreations,
helminths.
• CHEMICAL EXAMINATION: reactions, occult blood,
fat, carbohydrate, protein, etc
• MICROSCOPIC EXAMINATION:
pus cells, macrophages, RBCs,
remnants of food,
crystals, bacteria,
yeasts, molds, protozoa, helminths.
• STOOL CULTURE:
Stool examination by Dr. Priyanka
Buragohain
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Physical examination
• AMOUNT
• CONSISTENCY
• COLOUR
• ODOUR
• REACTION
• MUCUS
• CONCRETION
• BLOOD
• PUS
• FOOD REMNANT
• UNDIGESTED TABLETS
MACROSCOPIC EXAMINATION
Stool examination by Dr. Priyanka
Buragohain
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AMOUNT
• Normal is 150 g to 200 g/day
• Increased in steatorrhoea, diarrhoea, indigestion
of carbohydrate.
CONSISTENCY OR FORM
Stool examination by Dr. Priyanka
Buragohain
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• Normal is soft but formed
• Excessively hard/scybala- habitual constipation
• Flattened or ribbon like-intake of excess of
mineral oil, carcinoma of rectum, stricture of
rectum
• Soft, mushy, liquid and voluminous- diarrhoea,
intake of purgatives
• Small numerous, largely mucus and blood with
small amount of stool- dysenteries
• Rice watery without fecal matter- Cholera
12
Stool examination by Dr. Priyanka Buragohain
Colour
Stool examination by Dr. Priyanka
Buragohain
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• Dark Grey- excessive cocoa or chocolate
ingestion
• Reddish or blackish brown- large amount of
fruits
• Green – ingestion of green leafy vegetables,
administration of calomel due to biliverdin
• Red – Beat ingestion fresh blood
• Yellow – rhubarb or senna ingestion, normal
stool
Stool examination by Dr. Priyanka
Buragohain
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• Clay – obstructive jaundice, barium meal x-ray
• Tarry black – haemorrhage in stomach/upper
intestine
• Dark brown to bright red – bleeding in rectum
or sigmoid colon
• Red streaks of blood on the surface of faeces-
haemorrhoids, fissures, carcinoma ,ulcerative
colitis
Odour
Stool examination by Dr. Priyanka
Buragohain
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• Normal odour is aromatic due to indole and
skatole
• Increased- excessive protein ingestion
• Sour rancid- fatty acid in milk indigestion (in
children and adults), normal in infants
• Putrid- severe diarrhoea of malignancy,
gangrenous dysentry
Reaction
Stool examination by Dr. Priyanka
Buragohain
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• Normal is neutral
• Ph varies from 6.9 to 7.2
• pH is dependent on bacterial fermentation
and putrefaction in the bowel.
• Alkaline – excess protein ingestion
• Acidic – excess carbohydrate ingestion
Mucus
Stool examination by Dr. Priyanka
Buragohain
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• Small quantity of mucin is normal
• Small quantity – faeces from small gut
• Excessive quantity – infection of intestine
• Entirely mucus with little or no faeces and
streaks of blood- dysentery, ileo colitis,
intussusception
Concretion
Stool examination by Dr. Priyanka
Buragohain
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• In infants whitish curds may be found
• Gall bladder stones may be rarely found
Blood
Stool examination by Dr. Priyanka
Buragohain
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• Absent in normal faeces
• Formed stool with streaks of blood – lesion in
sigmoid colon, rectum or anal canal
• Liquid stool with bright red blood, pus and
mucus- bacillary dysentery, ulcerative colitis
• Semi formed stool with deep tarry black
blood- melena
• Loose stool with deep cherry red blood-
melena
Pus
Stool examination by Dr. Priyanka
Buragohain
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• Normally absent
• Pus with blooded mucus- ulcerative colitis,
bacillary dysentery, ulcerative carcinoma
Stool examination by Dr. Priyanka
Buragohain
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• GROSS FOOD REMNANTS MAY IN NORMAL
STOOL
• UNDIGESTED TABLETS MAY BE FOUND
CHEMICAL EXAMINATION
Stool examination by Dr. Priyanka
Buragohain
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Chemical Examination Of Stool
Stool examination by Dr. Priyanka
Buragohain
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• Acidity/basicity
• Fats
• Nitrogen
• Stercobilinogen
• Coproporphyrin
• Occult blood
• Reducing substances
• N. B : most commonly used chemical
examination of stool is pH, occult blood and
reducing substances
Stercobilinogen
Stool examination by Dr. Priyanka
Buragohain
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• Normal is 40 to 280 mg/day
• Average is 150 mg/day
• Dependent on amount of bilirubin passing to
intestine(jaundice)
Occult blood
Stool examination by Dr. Priyanka
Buragohain
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• Detect blood which is present in amount or form
not visible macroscopically
• Normally nil
• Abnormal presence in condition of occult
haemorrhage in the GI tract
• BENZIDINE TEST
• GUAIAC TEST
• ORTHOTOLIDINE TEST
• Most commonly used test is benzidine test
Stool examination by Dr. Priyanka
Buragohain
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BENZIDINE TEST
• 4 gm benzidine in 100 ml of glacial acetic acid
• Emulsify pea sized bit of faeces in 5 ml of
water.
• Mix 1 ml emulsion and 1 ml of reagent in test
tube
• Add several drops of 35 H2O2
• Blue colour indicates positive reaction
• Trace- faint blue colour after 1
min
• 1+ - definite blue green slowly
• 2+ - green blue rapidly
• 3+ - blue almost immediately
• 4+ - dark blue immediately
Stool examination by Dr. Priyanka
Buragohain
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GUAIAC TEST
• Less sensitive
• With loss of 20 to 30 ml of
blood all test will be positive
• Guaiac reagent consist of 1 gm
Guaiac in 5 ml of 95% ethanol.
• Make a small smear of feces on
a filter paper
• Add 2 to 3 drops of gum guaiac
solution +
2 to 3 drops of glacial acetic acid
+ 2 to 3 drops of 3% H2O2
Stool examination by Dr. Priyanka
Buragohain
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• Trace- faint blue green in 1 min
• 1+ light blue slowly
• 2+ clear blue rapidly
• 3 + deep blue almost immediately
• 4+ deep blue immediately
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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ORTHOTOLIDINE TEST
• Intermediate sinsitivity
• Smear the stool on a filter paper with an
applicator
• Pipette a few drops of the reagent on to the
filter paper(orthotolidine barium peroxide 200
mg+ glacial acetic acid 5 ml)
• After 30 sec examine for a blue colour
• Blue green colour within 30 sec means
positive test
Stool examination by Dr. Priyanka
Buragohain
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Interpretation
• Gastric disease eg chronic ulcer and
malignancy
• Intestinal diseases eg dysentery, typhoid fever,
carcinoma
• Haemorrhoids
• During instrumentation
Faecal reducing substance test
Stool examination by Dr. Priyanka
Buragohain
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• To diagnose lactose intolerance
• Sample of 5 gm stool is needed
• Sample needs to be delivered to the
laboratory as soon as possible, preferably
within 1 hr , cause lactose in the stool will
normally be broken down by chemical
processes within 2-4 hrs after the specimen is
produced.
Stool examination by Dr. Priyanka
Buragohain
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Interpretation
• Negative/ trace- < 0.25 g/dl
• Suspicious(grade 1) – 0.25-0.5 g/dl
• Abnormal(grade 2-4)->0.5 g/dl
• Found in Carbohydrate malabsorption
• Tropical sprue
MICROSCOPIC
EXAMINATION
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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NEED FOR MICROSCOPIC EXAMINATION
• For the diagnosis of microscopic elements.
• Trophozoites and its movements are better seen
in unstained preparation of a fresh material.
• Cystic forms &Nuclear character are better seen
in stained preparation(iodine)
• Gycogen mass- stained with iodine
• Chromatoid bars- unstained preparation
• N.B – Both stained and unstained materials are
to be prepared
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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Need of concentration technique
• To see whether treatment of parasite is
successful
• To find ova of S. Mansoni or Taenia if few or
other ova and cyst are not seen in routine
examination
• To examine stool specimens from patients
who do not come from an area where a
particular parasite is found
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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FLOATATION TECHNIQUE
• Use solutions which have highier specific
gravity(zinc sulphate or Sheather’s sugar) than
the organisms to be floated so that the organisms
rise to the top and the debries sink to the
bottom.
• Advantage – produce a cleaner material than the
sedimentation technique
• Disadvantage – walls of eggs and cyst will often
collapse, hindering identification.
• Some parasite eggs do not float.
SEDIMETATION TECHNIQUE
• Use solutions of lower specific
gravity than the parasitic
organisms(formalin ethyl
acetate technique)
• Recommended for general
diagnostic laboratories due to
easy to perform and less prone
to technical error.
Stool examination by Dr. Priyanka
Buragohain
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Sedimentation techniques
Stool examination by Dr. Priyanka
Buragohain
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• Mix a small piece of stool with 10 ml of water or saline in a tube/ bottle
• Sieve the suspension into a beaker through a strainer with small holes.
• Pour the contents into a centrifuge tube
• Centrifuge at 2000-3000/rpm for 1 min
• Pour off the supernatant part
• Resuspend the deposit in clean water and add enough water to fill the
tube.
• Mix well and recentrifuge
• Pour off the supernatant part
• Resuspend in zinc sulphate solution, fill the tube with the solution
• Centrifuge at high speed for 1 min
• Transfer the contents from the surface of the tube to a slide, using a
bacteriological wire loop
• Add small drops of saline and mix
• Cover with a cover slip
• Examine under 10x and 40x objectives
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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Stainning methods
Wet mount
normal saline
Iodine solution
Buffered methylene blue solution
Eosin solution
Stainning for permanent preparation
Schaudinn’s fluid
Heidenhain’s Haematoxilin method
Trichome stain
Microscopic examination of wet mount
Stool examination by Dr. Priyanka
Buragohain
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Saline wet mount
Stool examination by Dr. Priyanka
Buragohain
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Iodine wet mount
Stool examination by Dr. Priyanka
Buragohain
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• Iodine kills the organisms, therefore
motility is lost.
• Used mainly to stain nuclei and
glycogen mass if present.
• Flagella becomes recognisable.
• Cyst can usually be specifically
identified in this method.
• Lugol’s iodine solution is used.
Stool examination by Dr. Priyanka
Buragohain
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Lugol’s iodine
• Its very strong
• Must be diluted about 5 times with distilled water
• Stain deteriorates quickly hence to be prepared every 2
weeks
• Contains:
Iodine crystals(powdered): 5 g
Potassium iodide
Distilled water
:10 g
: 10
• Potassium iodide is dissolved in distilled water and
iodine crystals are slowly added. Solution is filtered and
kept in a stoppered bottle of amber colour
Buffered methylene blue wet mount
• Stains only trophozoites of
amoeba
• It does not stain amoebic
cyst or trophozoites and cyst
of flagellates.
• Nucleus and the inclusions
such as RBC or yeast cells
stain dark blue
• Cytoplasm stains light blue
Stool examination by Dr. Priyanka
Buragohain
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Eosin wet mount
• Detection of trophozoites
and cyst
• They can be much more
easily detected against
the pink- red background
of eosin preparation
Stool examination by Dr. Priyanka
Buragohain
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Buragohain
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Buragohain
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Buragohain
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Cover with a cover slip
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Buragohain
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Buragohain
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Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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microscopic examination findings
• REMNANTS OF FOOD
• vegetable cells
• Muscle fibres
• Starch granules
• Fat globules
• Connective tissue/ elastic
fibres
• Mineral oil or castor oil
globules
• CELLS
• Epithelial cells
• Pus cells
• Macrophages
• Ghost cells
• Pyknotic bodies
• Eosinophills
• RBC
• Crystals
• Yeasts and molds
• Protozoa
• Helminthic parasites
Vegetable cell
• Sometimes causes
confusion with ova,
eggs, cyst or cell bodies
• IRREGULAR OUTER
MARGIN
Excess quantity is seen in
excess intake of
vegetables or
indigestion
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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Muscle fibres
• May confuse with Tinea segments
• Excess protein intake or indigestion
• Its excess excretion is called
Creatorrhoea(flesh-flow)
Starch granules
• Variable in size, round to
polygonal in shape,
colourless, circular or Y
shaped dot in the centre
• Confused with ova of
helminths
• Found in carbohydrate
dyspepsia
• Better seen in iodine
preparation
Stool examination by Dr. Priyanka
Buragohain
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Fat globules
• Appear similar to parasitic
cyst or cell bodies
• Emulsifying agents are
used to eliminate
confusion
• Confused with ova of
helminths
• Found in fat dyspepsia
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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Connective tissue/elastic fibres
• Confused with tinea
segments
• Signify indigestion
Stool examination by Dr. Priyanka
Buragohain
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Mineral oil/ castor oil globules
• When taken as purgative
• May be confused with ova of helminthes
Epithelial cells
• Excess presence due to
inflammatory
conditions of colon,
rectum, anal canal
Stool examination by Dr. Priyanka
Buragohain
63
Pus cells
• Commonly found in
normal stool, help to ease
the passage of stool
• Normally not visible to
human eye.
• If visible indicates disease
• Bacillary dysentery, UC,
acute Amoebic dysentery,
malignancy of rectum,
drug induced enterocilitis
Stool examination by Dr. Priyanka
Buragohain
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Macrophages
• Large mononuclear cells
with vesicular nucleus
and ingested materials
including RBC
• Confused with E.
Histolytica cyst or E. Coli
cyst
• Excess in Amoebic or
bacillary dysentery
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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Ghost cells
• Degerative form of
macrophages, epithelial
cells
• Its an enlarged/swollen
eosinophilic epithelial
cell with only
eosinophilic
cytoplasmic outline but
without a nucleus
• Characteristic of
bacillary dysentery
Stool examination by Dr. Priyanka
Buragohain
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Pyknotic bodies
• Nuclear remains of
tissue cells and
leucocytes
• Characteristic of acute
amoebic dysentery
Stool examination by Dr. Priyanka
Buragohain
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Eosinophils
• In intestinal allergy
• Diluting fluid used- Randolph’s diluting
fluid,Pilot’s stain
• Carbol chromotrope technique
• A measured quantity of the deposit is taken and
diluted with the diluting fluid 1:10 or 1: 20
according to the concentration of the residue and
counted in haemocytometer.
• Increased in allergic conditions, parasitic
infestation and drug allergy, ulcerative colitis
Stool examination by Dr. Priyanka
Buragohain
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RBCs
• seen in cases of ulcrative lesions of gut
• in bacillary dysentery – yellowish discrete
• Amoebic dysentery – greenish and in clumps
Crystals
• Fatty acid crystals
• Calcium oxalate crystals
• Triple phosphate crystals
• Charcot Leyden crystals
• Haemotoidin crystals
• Crystals of drugs
Stool examination by Dr. Priyanka
Buragohain
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• Charcot Leyden crystal:
• Slender and pointed at both ends,
Hexagonal bipyramidal structures
localised in the primary granues of
cytoplasm of eosinophils and
basophils
• Evidence of parasitic infiltrate eg
amoeba, ascaris, hookworm, fasciola
• diamond shaped or whetstone
shaped crystals
• Normally colourless, stained purplish-
red by trichome
• Vary in size and may be as large as 50
µm in length
• Found in UC, dysentery, malignant
ulcers, schistosomiasis etc
Stool examination by Dr. Priyanka
Buragohain
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• Haematoidin crystals:
• Ironless pigment derived from
haemoglobin and formed within
tissues(reticuloendothelial cells)
but found extracellularly after 5-7
days in foci of previous
haemorrhage.
• Occurs as refractile, yellow- brown
and orange-red granules
• Characteristically as rhomboid
plates arranged in a radial pattern,
so called hematoidin burrs.
Stool examination by Dr. Priyanka
Buragohain
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Yeast and molds
• Yeast are normally
present
• Excess in cases of AIDS
• Molds are rare but may
be seen in
immunodeficiency
conditions
Stool examination by Dr. Priyanka
Buragohain
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Parasites
Stoolexaminationby Dr. Priyanka Buragohain 80
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
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Stool
83
protozoa
examination by Dr. Priyanka
Buragohain
Entamoeba histolytica
TROPHOZOITE STAGE
• Identified by motility and
presence of ingested RBC
• Shape : constantly changing
position
• Size :ranges from 18 to 40 µm
,average being 20 to 20 µm
• Cytoplasm :divisible in two
portion
• Nucleus :spherical in shape
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain 85
RBC appear yellowish green inside the
endoplasm
Nucleus is not visible but a faint outline may be
detected
Endoplasm shows bluish or ground glass
appearance
Eccentric nucleus with karyosome (a small dot
at the centre surrounded by a clear halo),
nuclear membrane,
Linen network having a spoke like radial
arrangement
PRE CYSTIC STAGE
• Size :small in size,10 to 20
µm
• Shape :round or slightly
ovoid with blunt
pseudopodium
• Free from ingested RBC and
other materials
• Nucleus : large nucleus
• Retains the characteristics
of trophozoite.
Stool examination by Dr. Priyanka
Buragohain
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CYSTIC STAGE
• Size : 6 to 9 µm / 12 to 15
µm
• Shape : round, surrounded
by a highly refractile
membrane called cyst wall
• Nucleus :quadrinucleate
• Clear and hyaline cytoplasm
• Nuclear structure retainning
the character of trophozoite
Stool examination by Dr. Priyanka
Buragohain
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Stool examination
Burago
by Dr. Priyanka
hain
88
Saline mounting
•Chromatid bodies are seen as round
refracile bars
•Cyst wall smooth and thin
•Glycogen bar not visible
•Outlines of nuclei may be visible
•Iodine mounting
•Body of the parasite stains yellow to light
brown
•Nucleus is clearly seen with a karyosome
•Cytoplasm is smooth and hyaline
appearance
•Glycogen mass stains brown
Iron haematoxillin stain
•Chromatid body and nucleus stain jet
black
•Cytoplasm stains bluish or greyish
•Glycogen mass gets dissolved in the
process of stainning and remains as a
vacuole
Entamoeba coli
Stool examination by Dr. Priyanka
Buragohain
83
TROPHOZOITE STAGE
• Largest amoeba 20-40 µm in diameter
• Sluggishly motile
• Cytoplasm not clearly defined
• Opaque endoplasm packed with food
vacuoles with bacteria and others but
no RBC
• Nucleus visible in unstainned
preparation
• In stainned prep nucleus shows large
eccentric karyosome surrounded by
broader halo and coarse chromatin
ranules linning nuclear membrane
• CYST
• 15- 20 µm in diameter
• Rounded body
• Octanucleate
• Largen glycogen mass in
binucleate stage
• Chromatoid body if present
are in slender filaments or
pointed threads
• Glycogen mass and
chromatoid bodies are absent
in mature cyst
Stool examination by Dr. Priyanka
Buragohain
84
Endolimax nana
• Commensal in large intestine of
man
• Trophozoites are smaller in
size(8-9 µm in diameter)
• Sluggish in motility
• Cytoplasmic inclusions contain
bacteria and food particles but
no RBC
• Nucleus has irregular karyosome,
eccentric and in contact with
nuclear membrane
Stool examination by Dr. Priyanka
Buragohain
85
CYST
• Cyst are oval
• Same size as the
trophozoites
• Number nuclei are 1-4
• Mature cyst are
quadrinucleate
• Chromatoid bodies and
glycpgen mass are not seen
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Buragohain
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Difference between different trophozoites
Stool examination by Dr. Priyanka
Buragohain
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Difference between different cyst
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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OTHER NON PATHOGENIC AMOEBIDA SPECIES
Trophozoite of E. hartmani
Trophozoite of Iodamoeba butschlii
Cyst of blastocystis hominis
Stool examination by Dr. Priyan ka 96
Balantidium coli
•Largest protozoal parasite
•Pig is the common reservoir
•Two stages
•Trophozoite and encysted stage
TROPHOZOITE
•Oval body
•60-70 µm in length and 40-50 µm breadth
•Body is covered with a delicate pellicle showing
longitudinal striations
•Cilia are short and delicate, of uniform length, on mouth
are longer called adoral cilia
•Thin layer of ectoplasm and granular endoplasm
•Groove at the anterior end(peristome) leading to a
mouth(cytostome) terminating in a short funnel shaped
gullet(cytopharynx) extending upto 1/3rd of the body
•At the posterior part permanent anus called cytophage is
situated
•2 nuclei: kidney shaped macronucleus, round
micronucleus in the concavity of micronucleus
•2 contractile vacuole, many food vacuole
Buragohain
CYST
• Smaller than trophic, 50-60
µm in diameter
• Cytoplasm is granular
contains the macronucleus,
micronucleus, refractile
body
• Contractile vacuole
• Thick transparent double
layered wall
Stool examination by Dr. Priyanka
Buragohain
91
Giardia lambia
• TROPHOZOITE
• Flat view: a tennis or badminton racket
• Side view: longitudinally split pear
• Dorsal surface is convex and the ventral
surface in concave with a sucking disc
• Size is 14 × 7 µm
• Anterior end is broad and rounded,
posterior end tapers to a sharp point
• Bilaterally symmetrical and all organs are
paired.
• 2 axostyles, 2 nuclei, 4 pairs of flagella
Exist in 2 phase: trophozoite and cyst
Stool examination by Dr. Priyanka
Buragohain
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CYST
• Oval in shape
• Size is 12 × 7 µm
• Axostyle lie diagonally like dividing wall
within the cyst wall
• 4 nuclei lie clustered at one end, lie in
pairs at opposite poles
• Remains of flagella and sucking disc
may be seen in cytoplasm
• Acid causes the parasite to encyst
Stool examination by Dr. Priyanka
Buragohain
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Stool examination by Dr. Priyanka
Buragohain
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What we can see in microscopic
examinationof stool ?
ADULT WORM
LARVA
OVA
CYST
Stool examination by Dr. Priyanka
Buragohain
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HELMINTHES
PLATYHELMINTHES NEMATHELMINTHES
CESTODE
TREMATODE
NEMATODE
TAPEWORM
Taenia solium
Taenia saginata
ECHINOCOCCUS
Stool examination by Dr. Priyanka
Buragohain
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G. Hominis
F. Buski
F. Hepatica
C. Sinensis
P
. westermani • A. Lumbricoids
• T
. Trichiura
• A. Duodenale
• E. Vermicularis
•S. Stercoralis
Stool examination by Dr. Priyanka
Buragohain
97
ADULT WORM
• TAENIA SAGINATA/SOLIUM
• DIPHYLLOBOTHRIUM
• HYMENOLEPIS
• DYPILIDIUM
LARVAL STAGE
• ECHINOCOCCUS: Hydatid
cyst
• SPIROMETRA
• HYMENOLEPIS
• TAENIA SOLIUM
• MULTICEPS:
Taenia solium
• Segments of Tape worm or single
segment may be found
• White in colour, semi transparent.
• May be 3 – 10 m long/ 1-3 cm
segments maybe upto 24 m
• Variable length(1000-2000
proglottids)
• When stool is allowed to dry up
the pieces of segments will roll
upand appear as round worm,
moistening the segments will
restore the shape
• Head is quadrate in outline, has 4
circular suckers
• Head is absent of
rostellum/hooklets
Stool examination by Dr. Priyanka
Buragohain
98
Eggs of T. saginata
• Spherical and brown in colour
• 31-43µm in diameter
• Thin outer transparent
shell(remnants of yolk mass),
causes egg to clump together
• Inner embryophore is brown,
thick walled, radially striated
• Contains an oncosphere(14-
20µm), with 3 pairs of hooklets
• Doesnot float in saturated
solution of common salt
Stool examination by Dr. Priyanka
Buragohain
99
Taenia solium
• 2-3 metres long(800-900
proglottides)
• Scolex is 1mm in diameter,
globular in outline, 4 circular
suckers,
• Head with rostellum armed
with a double row of alternating
large and small hooklets,
shaped like daggers or Arabian
poniards
• Segments are shed in chains of
5-6 at a time, not single.
Stool examination by Dr. Priyanka
Buragohain
100
• EGGS
• Same as T. Solium
• 30x40 µm sized egg
• Pale yellow
• Thick radially striated
embryophore with 6
hooklets inside
Stool examination by Dr. Priyanka
Buragohain
101
Echinococcus granulosus
• Commonly called dog
tapeworm/ hydatid worm
• Man harbours the larval form,
not the adult
• Larva found wiyhin the hydatid
cyst, scolex of the future adult
worm remains invaginated
within a vesicular body.
Stool examination by Dr. Priyanka
Buragohain
102
Stool
Hymenolepis diminuta
examination by Dr. Priyanka
Buragohain
103
Gastrodiscoides hominis
• Prevalent in Assam and Bengal
• Pyriform in shape
• Measures 5-10 mm × 4-6 mm
• Body has 2 parts: anterior conical and
posterior hemispherical portion which
is hollowed out ventrally to form a
concave disc
• Acetabulum is postero terminal,
situated ventrally
• Notch at posterior end
• Eggs are ovoid, operculated, 130×60
µm, immature when oviposited
Stool examination by Dr. Priyanka
Buragohain
104
Fasciola hepatica
• Eggs
• Large, operculated, ovoid , brownish
yellow(bile stained)
• Size is 140× 80 µm
• Contains a large unsegmentad ovum
in a mass of yolk cells
• Excreted with bile into duodenum
and then passed out along with the
faeces
• Does not float in saturated common
salt sol.
• Can develop only in water
Stool examination by Dr. Priyanka
Buragohain
105
Fasciola buski
• Reported in Assam, bengal,
china, thailand and other
oriental regions
• Largest trematode(2-7.5 length,
8-20 mm bredth, 0.5-3 mm
thickness)
• Elongated and oval in shape
• Resembles F. Hepatica but does
not possess any cephalic cone
• Each worm lay 25,000 eggs per
day
• Eggs are indistinguishable from
F. hepatica
Stool examination by Dr. Priyanka
Buragohain
106
Clonorchis sinensis
EGGS
• Yellowish brown
• Flask shaped
• Operculated
• Possess a terminal hook like
spine(resembling an electric bulb)
• Small in size(35×20µm)
• Ciliated embryo (oviposited stage)
• Do not float in saturated solution
of common salt
Stool examination by Dr. Priyanka
Buragohain
107
Paragonimus westermani
• Golden brown in colour
• Oval in shape with flattened
opercula
• 80- 55 µm
• Each egg contains an
unsegmented ovum
surrounded by yolk cells
• Prevalent in Assam
• Found in sputum and faeces
Stool examination by Dr. Priyanka
Buragohain
108
Ascaris lumbricoids
• Large round worms may
be males and females
or both
• Pinkish in colour
• 0.3-0.4 cm in thickness
• 15-25 cm long
• Males are shorter than
females
• Have curved tapering
tail
Stool examination by Dr. Priyanka
Buragohain
109
• UNFERTILISED EGGS
• Do not float on floatation method
• Size- 40×70 µm
• Yellow in colour
• Elongated
• Mammilated thin shell, ovum
containing refractile yolk globules
occupying the whole inside space
• May be confused with veg cell
• FERTILISED EGGS
• Float on floatation method
• Size- 40×70 µm
• Yellow in colour
• Oval or round
• Thick mammillated coat and
single celled ovum inside
Stool examination by Dr. Priyanka
Buragohain
110
Enterobius vermicularis
• Small round worm or thread
like worm or pin
worm(spindle shaped)
• White coloured
• 0.5-1 cm long
• Tail pointed
• Males smaller than females
and posterior body is curved
and sharply truncated(found
only after purgation)
Stool examination by Dr. Priyanka
Buragohain
111
• EGGS
• Colourless(not bile
stained)
• 20×50 µm
• Assymetrical, Oval
planoconvex,
• thin transparent
shelled,
• contain coiled tadpole
likelarva inside
• Floats in saturated salt
solution
Stool examination by Dr. Priyanka
Buragohain
112
Ancylostoma duodenale
• Hook worm
• Small greyish white or pink
coloured cylindrical
• 1-1.5 cm long
• One end is curved like a hook
• 6 teeth, 4 hook like on ventral
surface and 2 knob like on the
dorsal surface
Stool examination by Dr. Priyanka
Buragohain
113
EGGS
• Oval or elliptical in shape
• 40x60 µm sized egg
• Colourless(not bile
stained)
• Surrounded by a
transparent hyaline
shelled membrane
• Contains 4 segmented
ovum inside.
• Floats on saturated
solution of salt
Stool examination by Dr. Priyanka
Buragohain
114
Trichuris trichiura
• Whip worm
• Looks like a tiny whip
with a handle and a lash
• 3-5 cm in length
• White coloured
Stool examination by Dr. Priyanka
Buragohain
115
• 25x50 µm sized egg
• Brown coloured
• Double shelled, outer
one is bile stained
• Thick shelled, barrel
shaped with mucus plug
at both pole
• Single ovum
• floats in saturated
solution of common salt
Stool examination by Dr. Priyanka
Buragohain
116
Strongyloids stercoralis
• Adult worm:
• Females are readily
discovered than males
• 2.5mm ×40-50 µm
(females)
• Posterior extrimity is
pointed
• Males are shorter and
broader than females
Stool examination by Dr. Priyanka
Buragohain
117
Stool examination by Dr. Priyanka
Buragohain
118
• Eggs
• Eggs are conspicuous within the body in a
single line
• 55-30µm
• Thin shelled
• Transparent, oval
• Contain larva ready to hatch. It is the larva not
the eggs are found in stool.
• Rhabditiform larvae
• Develop directly from
gravid females
• Short mouth, double-
bulb oesophagus
• Filariform larvae
• Longer and slender
• Short mouth and
cylindrical oesophagus
Stool examination by Dr. Priyanka
Buragohain
119
Stool examination by Dr. Priyanka
Buragohain
120
Helminthes that float/do not float on
saturated solution of saline
Stool examination by Dr. Priyanka
Buragohain
121
• A. Duodenale
• N. Americanas
• E. Vermicularis
• H. Nana
• A. Lumbricoids
• T
. Trichuria
• H. Diminuta
Eggs that float Eggs that do not float
• A. Lumbricoids
• T
. solium
• T
. Saginata
• Trematodes
• F. Buski
• F. Hepatica
• C. Sinensis
Stool examination by Dr. Priyanka
Buragohain
122
Stool examination by Dr. Priyanka
Buragohain
123
Stool examination by Dr. Priyanka
Buragohain
124
Plant hairs can be confused for larvae
oh hookworm or Strongyloides stercoralis
Plant hair resembling S. strongiloids
Stool examination by Dr. Priyanka
Buragohain
125
Parasite of earth worm
Pollen grain resemblening to
fertile egg of A. lumbricoids
Stool examination by Dr. Priyanka
Buragohain
126
Bee pollen resembling t. Trichuria egg
Stool examination by Dr. Priyanka
Buragohain
127
STOOL CULTURE
• Used to detect the presence of disease
causing(pathogenic )bacteria
• Help to diagnose an infection of digestive
system
• Used in conjunction with stool test
• Reference range for stool culture is negative
Stool examination by Dr. Priyanka
Buragohain
128
Most common bacteria for culture of stool
• Campylobacter species
• Salmonella species
• Shigella species
• Yersinia species
• Vibrio species(travel history)
• Some bacteria cause illness by producing
toxins(PCR, Antigen test are to be done with stool
test)
• Escherichia coli
• Clostridium difficile
Stool examination by Dr. Priyanka
Buragohain
129
Collection
• Specimen collected via rectal swab(in infants)
• Sterile collection container not required
• No detergent or preservative should be present in
the container
• Specimen should be immediately transported to
the laboratory
• If transport is delayed by longer than 2 hours,
transport media(eg Cary- Blair) is recommended
• Samples must be sent in a sealed, leak-proof
container marked with a biohazard sticker
Stool examination by Dr. Priyanka
Buragohain
130
Medias used commonly
• MacConkey agar- Salmonella species, shigella
species
• Eosin methylene blue agar
• Triple sugar iron(TSI)- differentiate salmonella
and shigella
• Sabouraud agar
• Hekteon enteric agar
• Selenite broth
Stool examination by Dr. Priyanka
Buragohain
131
Summary
Stool examination by Dr. Priyanka
Buragohain
132
• Definition of stool
• Composition of stool
• Precaution and collection of
sample
• Physical examination
• Chemical examination
• Microscopic examination
• Artifacts
• Stool culture
THANK YOU !!!
Stool examina Dr. iyan
uragoh

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routineexaminationofstool-170326170848.pptx

  • 2. DEFINITION Stool examination by Dr. Priyanka Buragohain 2 • Human feces is called as stool. • faeces / feces is plural of latin term faex meaning RESIDUE. • It is the waste residue of indigestible materials of an animal’s digestive tract expelled through the anus during defecation. • Meconium is newborn’s first feces. • SCATOLGY or CAPROLOGY is the study of feces.
  • 3. COMPOSITION Stool examination by Dr. Priyanka Buragohain 3 • ¾ water, ¼ solid • Undigested and unabsorbed food • Intestinal secretions, mucous • Bile pigments and salts • Decomposed products • Bacteria and inorganic material • Epithelial cells, leukocytes.
  • 4. PRECAUTION BEFORE COLLECTION Stool examination by Dr. Priyanka Buragohain 4 • Patient should avoid the following things for at least 48 hours before collection of stool: • Mineral oils, bismuth, non absorbable anti diarrhoeal drugs, antimalarial drugs, antibiotics, etc • Pt. Should not have barium swallow examination before stool R/E • Avoid iron containing drugs, meat, fish etc for atleast 48 hours before stool for occult blood. • In constipated patients use only non residual purgative
  • 5. COLLECTION Stool examination by Dr. Priyanka Buragohain 5 • Universal precautions • Pt. is asked to pass stool in a clean container. • Stool should be collected in a steralized, wide mouthed container. • Loose/last/portion containing mucus, blood etc is to be collected in a wide mouthed bottle. • Should be uncontaminated with urine or any other body secretions. • >2gm is required. • Properly named and always a fresh sample should be tested. • Liquid stool to be examined within ½ hour • Solid stool to be examined within 1 hour. • If delayed store in a refrigerator.
  • 6. COLLECTION CONTD... Stool examination by Dr. Priyanka Buragohain 6 • 3 samples of stool within 10 days to exclude false negatives. • 2 samples to be examined on alternate days after normal defaecation and 1 sample after a purgative for certain worms. • Formalin is the best preservative. It kills the bacteria but ptreserves the protozoa and helminthes. • For culture no preservatives to be used
  • 7. Stool examination by Dr. Priyanka Buragohain 7
  • 8. TYPES OF EXAMINATION Stool examination by Dr. Priyanka Buragohain 8 • PHYSICAL EXAMINATION: colour, volume, consistency, odour, mucus, pus, concreations, helminths. • CHEMICAL EXAMINATION: reactions, occult blood, fat, carbohydrate, protein, etc • MICROSCOPIC EXAMINATION: pus cells, macrophages, RBCs, remnants of food, crystals, bacteria, yeasts, molds, protozoa, helminths. • STOOL CULTURE:
  • 9. Stool examination by Dr. Priyanka Buragohain 9 Physical examination • AMOUNT • CONSISTENCY • COLOUR • ODOUR • REACTION • MUCUS • CONCRETION • BLOOD • PUS • FOOD REMNANT • UNDIGESTED TABLETS
  • 10. MACROSCOPIC EXAMINATION Stool examination by Dr. Priyanka Buragohain 10 AMOUNT • Normal is 150 g to 200 g/day • Increased in steatorrhoea, diarrhoea, indigestion of carbohydrate.
  • 11. CONSISTENCY OR FORM Stool examination by Dr. Priyanka Buragohain 11 • Normal is soft but formed • Excessively hard/scybala- habitual constipation • Flattened or ribbon like-intake of excess of mineral oil, carcinoma of rectum, stricture of rectum • Soft, mushy, liquid and voluminous- diarrhoea, intake of purgatives • Small numerous, largely mucus and blood with small amount of stool- dysenteries • Rice watery without fecal matter- Cholera
  • 12. 12 Stool examination by Dr. Priyanka Buragohain
  • 13. Colour Stool examination by Dr. Priyanka Buragohain 13 • Dark Grey- excessive cocoa or chocolate ingestion • Reddish or blackish brown- large amount of fruits • Green – ingestion of green leafy vegetables, administration of calomel due to biliverdin • Red – Beat ingestion fresh blood • Yellow – rhubarb or senna ingestion, normal stool
  • 14. Stool examination by Dr. Priyanka Buragohain 14 • Clay – obstructive jaundice, barium meal x-ray • Tarry black – haemorrhage in stomach/upper intestine • Dark brown to bright red – bleeding in rectum or sigmoid colon • Red streaks of blood on the surface of faeces- haemorrhoids, fissures, carcinoma ,ulcerative colitis
  • 15. Odour Stool examination by Dr. Priyanka Buragohain 15 • Normal odour is aromatic due to indole and skatole • Increased- excessive protein ingestion • Sour rancid- fatty acid in milk indigestion (in children and adults), normal in infants • Putrid- severe diarrhoea of malignancy, gangrenous dysentry
  • 16. Reaction Stool examination by Dr. Priyanka Buragohain 16 • Normal is neutral • Ph varies from 6.9 to 7.2 • pH is dependent on bacterial fermentation and putrefaction in the bowel. • Alkaline – excess protein ingestion • Acidic – excess carbohydrate ingestion
  • 17. Mucus Stool examination by Dr. Priyanka Buragohain 17 • Small quantity of mucin is normal • Small quantity – faeces from small gut • Excessive quantity – infection of intestine • Entirely mucus with little or no faeces and streaks of blood- dysentery, ileo colitis, intussusception
  • 18. Concretion Stool examination by Dr. Priyanka Buragohain 18 • In infants whitish curds may be found • Gall bladder stones may be rarely found
  • 19. Blood Stool examination by Dr. Priyanka Buragohain 19 • Absent in normal faeces • Formed stool with streaks of blood – lesion in sigmoid colon, rectum or anal canal • Liquid stool with bright red blood, pus and mucus- bacillary dysentery, ulcerative colitis • Semi formed stool with deep tarry black blood- melena • Loose stool with deep cherry red blood- melena
  • 20. Pus Stool examination by Dr. Priyanka Buragohain 20 • Normally absent • Pus with blooded mucus- ulcerative colitis, bacillary dysentery, ulcerative carcinoma
  • 21. Stool examination by Dr. Priyanka Buragohain 21 • GROSS FOOD REMNANTS MAY IN NORMAL STOOL • UNDIGESTED TABLETS MAY BE FOUND
  • 22. CHEMICAL EXAMINATION Stool examination by Dr. Priyanka Buragohain 22
  • 23. Chemical Examination Of Stool Stool examination by Dr. Priyanka Buragohain 23 • Acidity/basicity • Fats • Nitrogen • Stercobilinogen • Coproporphyrin • Occult blood • Reducing substances • N. B : most commonly used chemical examination of stool is pH, occult blood and reducing substances
  • 24. Stercobilinogen Stool examination by Dr. Priyanka Buragohain 24 • Normal is 40 to 280 mg/day • Average is 150 mg/day • Dependent on amount of bilirubin passing to intestine(jaundice)
  • 25. Occult blood Stool examination by Dr. Priyanka Buragohain 25 • Detect blood which is present in amount or form not visible macroscopically • Normally nil • Abnormal presence in condition of occult haemorrhage in the GI tract • BENZIDINE TEST • GUAIAC TEST • ORTHOTOLIDINE TEST • Most commonly used test is benzidine test
  • 26. Stool examination by Dr. Priyanka Buragohain 26 BENZIDINE TEST • 4 gm benzidine in 100 ml of glacial acetic acid • Emulsify pea sized bit of faeces in 5 ml of water. • Mix 1 ml emulsion and 1 ml of reagent in test tube • Add several drops of 35 H2O2 • Blue colour indicates positive reaction
  • 27. • Trace- faint blue colour after 1 min • 1+ - definite blue green slowly • 2+ - green blue rapidly • 3+ - blue almost immediately • 4+ - dark blue immediately Stool examination by Dr. Priyanka Buragohain 27
  • 28. GUAIAC TEST • Less sensitive • With loss of 20 to 30 ml of blood all test will be positive • Guaiac reagent consist of 1 gm Guaiac in 5 ml of 95% ethanol. • Make a small smear of feces on a filter paper • Add 2 to 3 drops of gum guaiac solution + 2 to 3 drops of glacial acetic acid + 2 to 3 drops of 3% H2O2 Stool examination by Dr. Priyanka Buragohain 28
  • 29. • Trace- faint blue green in 1 min • 1+ light blue slowly • 2+ clear blue rapidly • 3 + deep blue almost immediately • 4+ deep blue immediately Stool examination by Dr. Priyanka Buragohain 29
  • 30. Stool examination by Dr. Priyanka Buragohain 30 ORTHOTOLIDINE TEST • Intermediate sinsitivity • Smear the stool on a filter paper with an applicator • Pipette a few drops of the reagent on to the filter paper(orthotolidine barium peroxide 200 mg+ glacial acetic acid 5 ml) • After 30 sec examine for a blue colour • Blue green colour within 30 sec means positive test
  • 31. Stool examination by Dr. Priyanka Buragohain 31 Interpretation • Gastric disease eg chronic ulcer and malignancy • Intestinal diseases eg dysentery, typhoid fever, carcinoma • Haemorrhoids • During instrumentation
  • 32. Faecal reducing substance test Stool examination by Dr. Priyanka Buragohain 32 • To diagnose lactose intolerance • Sample of 5 gm stool is needed • Sample needs to be delivered to the laboratory as soon as possible, preferably within 1 hr , cause lactose in the stool will normally be broken down by chemical processes within 2-4 hrs after the specimen is produced.
  • 33. Stool examination by Dr. Priyanka Buragohain 33 Interpretation • Negative/ trace- < 0.25 g/dl • Suspicious(grade 1) – 0.25-0.5 g/dl • Abnormal(grade 2-4)->0.5 g/dl • Found in Carbohydrate malabsorption • Tropical sprue
  • 34. MICROSCOPIC EXAMINATION Stool examination by Dr. Priyanka Buragohain 34
  • 35. Stool examination by Dr. Priyanka Buragohain 35 NEED FOR MICROSCOPIC EXAMINATION • For the diagnosis of microscopic elements. • Trophozoites and its movements are better seen in unstained preparation of a fresh material. • Cystic forms &Nuclear character are better seen in stained preparation(iodine) • Gycogen mass- stained with iodine • Chromatoid bars- unstained preparation • N.B – Both stained and unstained materials are to be prepared
  • 36. Stool examination by Dr. Priyanka Buragohain 36
  • 37. Stool examination by Dr. Priyanka Buragohain 37 Need of concentration technique • To see whether treatment of parasite is successful • To find ova of S. Mansoni or Taenia if few or other ova and cyst are not seen in routine examination • To examine stool specimens from patients who do not come from an area where a particular parasite is found
  • 38. Stool examination by Dr. Priyanka Buragohain 38
  • 39. Stool examination by Dr. Priyanka Buragohain 39 FLOATATION TECHNIQUE • Use solutions which have highier specific gravity(zinc sulphate or Sheather’s sugar) than the organisms to be floated so that the organisms rise to the top and the debries sink to the bottom. • Advantage – produce a cleaner material than the sedimentation technique • Disadvantage – walls of eggs and cyst will often collapse, hindering identification. • Some parasite eggs do not float.
  • 40. SEDIMETATION TECHNIQUE • Use solutions of lower specific gravity than the parasitic organisms(formalin ethyl acetate technique) • Recommended for general diagnostic laboratories due to easy to perform and less prone to technical error. Stool examination by Dr. Priyanka Buragohain 40
  • 41. Sedimentation techniques Stool examination by Dr. Priyanka Buragohain 41 • Mix a small piece of stool with 10 ml of water or saline in a tube/ bottle • Sieve the suspension into a beaker through a strainer with small holes. • Pour the contents into a centrifuge tube • Centrifuge at 2000-3000/rpm for 1 min • Pour off the supernatant part • Resuspend the deposit in clean water and add enough water to fill the tube. • Mix well and recentrifuge • Pour off the supernatant part • Resuspend in zinc sulphate solution, fill the tube with the solution • Centrifuge at high speed for 1 min • Transfer the contents from the surface of the tube to a slide, using a bacteriological wire loop • Add small drops of saline and mix • Cover with a cover slip • Examine under 10x and 40x objectives
  • 42. Stool examination by Dr. Priyanka Buragohain 42
  • 43. Stool examination by Dr. Priyanka Buragohain 43 Stainning methods Wet mount normal saline Iodine solution Buffered methylene blue solution Eosin solution Stainning for permanent preparation Schaudinn’s fluid Heidenhain’s Haematoxilin method Trichome stain
  • 44. Microscopic examination of wet mount Stool examination by Dr. Priyanka Buragohain 44
  • 45. Saline wet mount Stool examination by Dr. Priyanka Buragohain 45
  • 46. Iodine wet mount Stool examination by Dr. Priyanka Buragohain 46 • Iodine kills the organisms, therefore motility is lost. • Used mainly to stain nuclei and glycogen mass if present. • Flagella becomes recognisable. • Cyst can usually be specifically identified in this method. • Lugol’s iodine solution is used.
  • 47. Stool examination by Dr. Priyanka Buragohain 47 Lugol’s iodine • Its very strong • Must be diluted about 5 times with distilled water • Stain deteriorates quickly hence to be prepared every 2 weeks • Contains: Iodine crystals(powdered): 5 g Potassium iodide Distilled water :10 g : 10 • Potassium iodide is dissolved in distilled water and iodine crystals are slowly added. Solution is filtered and kept in a stoppered bottle of amber colour
  • 48. Buffered methylene blue wet mount • Stains only trophozoites of amoeba • It does not stain amoebic cyst or trophozoites and cyst of flagellates. • Nucleus and the inclusions such as RBC or yeast cells stain dark blue • Cytoplasm stains light blue Stool examination by Dr. Priyanka Buragohain 48
  • 49. Eosin wet mount • Detection of trophozoites and cyst • They can be much more easily detected against the pink- red background of eosin preparation Stool examination by Dr. Priyanka Buragohain 49
  • 50. Stool examination by Dr. Priyanka Buragohain 50
  • 51. Stool examination by Dr. Priyanka Buragohain 51
  • 52. Stool examination by Dr. Priyanka Buragohain 52
  • 53. Cover with a cover slip Stool examination by Dr. Priyanka Buragohain 53
  • 54. Stool examination by Dr. Priyanka Buragohain 54
  • 55. Stool examination by Dr. Priyanka Buragohain 55
  • 56. Stool examination by Dr. Priyanka Buragohain 56 microscopic examination findings • REMNANTS OF FOOD • vegetable cells • Muscle fibres • Starch granules • Fat globules • Connective tissue/ elastic fibres • Mineral oil or castor oil globules • CELLS • Epithelial cells • Pus cells • Macrophages • Ghost cells • Pyknotic bodies • Eosinophills • RBC • Crystals • Yeasts and molds • Protozoa • Helminthic parasites
  • 57. Vegetable cell • Sometimes causes confusion with ova, eggs, cyst or cell bodies • IRREGULAR OUTER MARGIN Excess quantity is seen in excess intake of vegetables or indigestion Stool examination by Dr. Priyanka Buragohain 57
  • 58. Stool examination by Dr. Priyanka Buragohain 58 Muscle fibres • May confuse with Tinea segments • Excess protein intake or indigestion • Its excess excretion is called Creatorrhoea(flesh-flow)
  • 59. Starch granules • Variable in size, round to polygonal in shape, colourless, circular or Y shaped dot in the centre • Confused with ova of helminths • Found in carbohydrate dyspepsia • Better seen in iodine preparation Stool examination by Dr. Priyanka Buragohain 59
  • 60. Fat globules • Appear similar to parasitic cyst or cell bodies • Emulsifying agents are used to eliminate confusion • Confused with ova of helminths • Found in fat dyspepsia Stool examination by Dr. Priyanka Buragohain 60
  • 61. Stool examination by Dr. Priyanka Buragohain 61 Connective tissue/elastic fibres • Confused with tinea segments • Signify indigestion
  • 62. Stool examination by Dr. Priyanka Buragohain 62 Mineral oil/ castor oil globules • When taken as purgative • May be confused with ova of helminthes
  • 63. Epithelial cells • Excess presence due to inflammatory conditions of colon, rectum, anal canal Stool examination by Dr. Priyanka Buragohain 63
  • 64. Pus cells • Commonly found in normal stool, help to ease the passage of stool • Normally not visible to human eye. • If visible indicates disease • Bacillary dysentery, UC, acute Amoebic dysentery, malignancy of rectum, drug induced enterocilitis Stool examination by Dr. Priyanka Buragohain 64
  • 65. Macrophages • Large mononuclear cells with vesicular nucleus and ingested materials including RBC • Confused with E. Histolytica cyst or E. Coli cyst • Excess in Amoebic or bacillary dysentery Stool examination by Dr. Priyanka Buragohain 65
  • 66. Stool examination by Dr. Priyanka Buragohain 66 Ghost cells • Degerative form of macrophages, epithelial cells • Its an enlarged/swollen eosinophilic epithelial cell with only eosinophilic cytoplasmic outline but without a nucleus • Characteristic of bacillary dysentery
  • 67. Stool examination by Dr. Priyanka Buragohain 67 Pyknotic bodies • Nuclear remains of tissue cells and leucocytes • Characteristic of acute amoebic dysentery
  • 68. Stool examination by Dr. Priyanka Buragohain 68 Eosinophils • In intestinal allergy • Diluting fluid used- Randolph’s diluting fluid,Pilot’s stain • Carbol chromotrope technique • A measured quantity of the deposit is taken and diluted with the diluting fluid 1:10 or 1: 20 according to the concentration of the residue and counted in haemocytometer. • Increased in allergic conditions, parasitic infestation and drug allergy, ulcerative colitis
  • 69. Stool examination by Dr. Priyanka Buragohain 69 RBCs • seen in cases of ulcrative lesions of gut • in bacillary dysentery – yellowish discrete • Amoebic dysentery – greenish and in clumps
  • 70. Crystals • Fatty acid crystals • Calcium oxalate crystals • Triple phosphate crystals • Charcot Leyden crystals • Haemotoidin crystals • Crystals of drugs Stool examination by Dr. Priyanka Buragohain 70
  • 71. • Charcot Leyden crystal: • Slender and pointed at both ends, Hexagonal bipyramidal structures localised in the primary granues of cytoplasm of eosinophils and basophils • Evidence of parasitic infiltrate eg amoeba, ascaris, hookworm, fasciola • diamond shaped or whetstone shaped crystals • Normally colourless, stained purplish- red by trichome • Vary in size and may be as large as 50 µm in length • Found in UC, dysentery, malignant ulcers, schistosomiasis etc Stool examination by Dr. Priyanka Buragohain 71
  • 72. • Haematoidin crystals: • Ironless pigment derived from haemoglobin and formed within tissues(reticuloendothelial cells) but found extracellularly after 5-7 days in foci of previous haemorrhage. • Occurs as refractile, yellow- brown and orange-red granules • Characteristically as rhomboid plates arranged in a radial pattern, so called hematoidin burrs. Stool examination by Dr. Priyanka Buragohain 72
  • 73. Yeast and molds • Yeast are normally present • Excess in cases of AIDS • Molds are rare but may be seen in immunodeficiency conditions Stool examination by Dr. Priyanka Buragohain 73
  • 75. Stool examination by Dr. Priyanka Buragohain 75
  • 76. Stool examination by Dr. Priyanka Buragohain 76
  • 78. Entamoeba histolytica TROPHOZOITE STAGE • Identified by motility and presence of ingested RBC • Shape : constantly changing position • Size :ranges from 18 to 40 µm ,average being 20 to 20 µm • Cytoplasm :divisible in two portion • Nucleus :spherical in shape Stool examination by Dr. Priyanka Buragohain 84
  • 79. Stool examination by Dr. Priyanka Buragohain 85 RBC appear yellowish green inside the endoplasm Nucleus is not visible but a faint outline may be detected Endoplasm shows bluish or ground glass appearance Eccentric nucleus with karyosome (a small dot at the centre surrounded by a clear halo), nuclear membrane, Linen network having a spoke like radial arrangement
  • 80. PRE CYSTIC STAGE • Size :small in size,10 to 20 µm • Shape :round or slightly ovoid with blunt pseudopodium • Free from ingested RBC and other materials • Nucleus : large nucleus • Retains the characteristics of trophozoite. Stool examination by Dr. Priyanka Buragohain 80
  • 81. CYSTIC STAGE • Size : 6 to 9 µm / 12 to 15 µm • Shape : round, surrounded by a highly refractile membrane called cyst wall • Nucleus :quadrinucleate • Clear and hyaline cytoplasm • Nuclear structure retainning the character of trophozoite Stool examination by Dr. Priyanka Buragohain 81
  • 82. Stool examination Burago by Dr. Priyanka hain 88 Saline mounting •Chromatid bodies are seen as round refracile bars •Cyst wall smooth and thin •Glycogen bar not visible •Outlines of nuclei may be visible •Iodine mounting •Body of the parasite stains yellow to light brown •Nucleus is clearly seen with a karyosome •Cytoplasm is smooth and hyaline appearance •Glycogen mass stains brown Iron haematoxillin stain •Chromatid body and nucleus stain jet black •Cytoplasm stains bluish or greyish •Glycogen mass gets dissolved in the process of stainning and remains as a vacuole
  • 83. Entamoeba coli Stool examination by Dr. Priyanka Buragohain 83 TROPHOZOITE STAGE • Largest amoeba 20-40 µm in diameter • Sluggishly motile • Cytoplasm not clearly defined • Opaque endoplasm packed with food vacuoles with bacteria and others but no RBC • Nucleus visible in unstainned preparation • In stainned prep nucleus shows large eccentric karyosome surrounded by broader halo and coarse chromatin ranules linning nuclear membrane
  • 84. • CYST • 15- 20 µm in diameter • Rounded body • Octanucleate • Largen glycogen mass in binucleate stage • Chromatoid body if present are in slender filaments or pointed threads • Glycogen mass and chromatoid bodies are absent in mature cyst Stool examination by Dr. Priyanka Buragohain 84
  • 85. Endolimax nana • Commensal in large intestine of man • Trophozoites are smaller in size(8-9 µm in diameter) • Sluggish in motility • Cytoplasmic inclusions contain bacteria and food particles but no RBC • Nucleus has irregular karyosome, eccentric and in contact with nuclear membrane Stool examination by Dr. Priyanka Buragohain 85
  • 86. CYST • Cyst are oval • Same size as the trophozoites • Number nuclei are 1-4 • Mature cyst are quadrinucleate • Chromatoid bodies and glycpgen mass are not seen Stool examination by Dr. Priyanka Buragohain 86
  • 87. Difference between different trophozoites Stool examination by Dr. Priyanka Buragohain 87
  • 88. Difference between different cyst Stool examination by Dr. Priyanka Buragohain 88
  • 89. Stool examination by Dr. Priyanka Buragohain 95 OTHER NON PATHOGENIC AMOEBIDA SPECIES Trophozoite of E. hartmani Trophozoite of Iodamoeba butschlii Cyst of blastocystis hominis
  • 90. Stool examination by Dr. Priyan ka 96 Balantidium coli •Largest protozoal parasite •Pig is the common reservoir •Two stages •Trophozoite and encysted stage TROPHOZOITE •Oval body •60-70 µm in length and 40-50 µm breadth •Body is covered with a delicate pellicle showing longitudinal striations •Cilia are short and delicate, of uniform length, on mouth are longer called adoral cilia •Thin layer of ectoplasm and granular endoplasm •Groove at the anterior end(peristome) leading to a mouth(cytostome) terminating in a short funnel shaped gullet(cytopharynx) extending upto 1/3rd of the body •At the posterior part permanent anus called cytophage is situated •2 nuclei: kidney shaped macronucleus, round micronucleus in the concavity of micronucleus •2 contractile vacuole, many food vacuole Buragohain
  • 91. CYST • Smaller than trophic, 50-60 µm in diameter • Cytoplasm is granular contains the macronucleus, micronucleus, refractile body • Contractile vacuole • Thick transparent double layered wall Stool examination by Dr. Priyanka Buragohain 91
  • 92. Giardia lambia • TROPHOZOITE • Flat view: a tennis or badminton racket • Side view: longitudinally split pear • Dorsal surface is convex and the ventral surface in concave with a sucking disc • Size is 14 × 7 µm • Anterior end is broad and rounded, posterior end tapers to a sharp point • Bilaterally symmetrical and all organs are paired. • 2 axostyles, 2 nuclei, 4 pairs of flagella Exist in 2 phase: trophozoite and cyst Stool examination by Dr. Priyanka Buragohain 92
  • 93. CYST • Oval in shape • Size is 12 × 7 µm • Axostyle lie diagonally like dividing wall within the cyst wall • 4 nuclei lie clustered at one end, lie in pairs at opposite poles • Remains of flagella and sucking disc may be seen in cytoplasm • Acid causes the parasite to encyst Stool examination by Dr. Priyanka Buragohain 93
  • 94. Stool examination by Dr. Priyanka Buragohain 94
  • 95. What we can see in microscopic examinationof stool ? ADULT WORM LARVA OVA CYST Stool examination by Dr. Priyanka Buragohain 95
  • 96. HELMINTHES PLATYHELMINTHES NEMATHELMINTHES CESTODE TREMATODE NEMATODE TAPEWORM Taenia solium Taenia saginata ECHINOCOCCUS Stool examination by Dr. Priyanka Buragohain 96 G. Hominis F. Buski F. Hepatica C. Sinensis P . westermani • A. Lumbricoids • T . Trichiura • A. Duodenale • E. Vermicularis •S. Stercoralis
  • 97. Stool examination by Dr. Priyanka Buragohain 97 ADULT WORM • TAENIA SAGINATA/SOLIUM • DIPHYLLOBOTHRIUM • HYMENOLEPIS • DYPILIDIUM LARVAL STAGE • ECHINOCOCCUS: Hydatid cyst • SPIROMETRA • HYMENOLEPIS • TAENIA SOLIUM • MULTICEPS:
  • 98. Taenia solium • Segments of Tape worm or single segment may be found • White in colour, semi transparent. • May be 3 – 10 m long/ 1-3 cm segments maybe upto 24 m • Variable length(1000-2000 proglottids) • When stool is allowed to dry up the pieces of segments will roll upand appear as round worm, moistening the segments will restore the shape • Head is quadrate in outline, has 4 circular suckers • Head is absent of rostellum/hooklets Stool examination by Dr. Priyanka Buragohain 98
  • 99. Eggs of T. saginata • Spherical and brown in colour • 31-43µm in diameter • Thin outer transparent shell(remnants of yolk mass), causes egg to clump together • Inner embryophore is brown, thick walled, radially striated • Contains an oncosphere(14- 20µm), with 3 pairs of hooklets • Doesnot float in saturated solution of common salt Stool examination by Dr. Priyanka Buragohain 99
  • 100. Taenia solium • 2-3 metres long(800-900 proglottides) • Scolex is 1mm in diameter, globular in outline, 4 circular suckers, • Head with rostellum armed with a double row of alternating large and small hooklets, shaped like daggers or Arabian poniards • Segments are shed in chains of 5-6 at a time, not single. Stool examination by Dr. Priyanka Buragohain 100
  • 101. • EGGS • Same as T. Solium • 30x40 µm sized egg • Pale yellow • Thick radially striated embryophore with 6 hooklets inside Stool examination by Dr. Priyanka Buragohain 101
  • 102. Echinococcus granulosus • Commonly called dog tapeworm/ hydatid worm • Man harbours the larval form, not the adult • Larva found wiyhin the hydatid cyst, scolex of the future adult worm remains invaginated within a vesicular body. Stool examination by Dr. Priyanka Buragohain 102
  • 103. Stool Hymenolepis diminuta examination by Dr. Priyanka Buragohain 103
  • 104. Gastrodiscoides hominis • Prevalent in Assam and Bengal • Pyriform in shape • Measures 5-10 mm × 4-6 mm • Body has 2 parts: anterior conical and posterior hemispherical portion which is hollowed out ventrally to form a concave disc • Acetabulum is postero terminal, situated ventrally • Notch at posterior end • Eggs are ovoid, operculated, 130×60 µm, immature when oviposited Stool examination by Dr. Priyanka Buragohain 104
  • 105. Fasciola hepatica • Eggs • Large, operculated, ovoid , brownish yellow(bile stained) • Size is 140× 80 µm • Contains a large unsegmentad ovum in a mass of yolk cells • Excreted with bile into duodenum and then passed out along with the faeces • Does not float in saturated common salt sol. • Can develop only in water Stool examination by Dr. Priyanka Buragohain 105
  • 106. Fasciola buski • Reported in Assam, bengal, china, thailand and other oriental regions • Largest trematode(2-7.5 length, 8-20 mm bredth, 0.5-3 mm thickness) • Elongated and oval in shape • Resembles F. Hepatica but does not possess any cephalic cone • Each worm lay 25,000 eggs per day • Eggs are indistinguishable from F. hepatica Stool examination by Dr. Priyanka Buragohain 106
  • 107. Clonorchis sinensis EGGS • Yellowish brown • Flask shaped • Operculated • Possess a terminal hook like spine(resembling an electric bulb) • Small in size(35×20µm) • Ciliated embryo (oviposited stage) • Do not float in saturated solution of common salt Stool examination by Dr. Priyanka Buragohain 107
  • 108. Paragonimus westermani • Golden brown in colour • Oval in shape with flattened opercula • 80- 55 µm • Each egg contains an unsegmented ovum surrounded by yolk cells • Prevalent in Assam • Found in sputum and faeces Stool examination by Dr. Priyanka Buragohain 108
  • 109. Ascaris lumbricoids • Large round worms may be males and females or both • Pinkish in colour • 0.3-0.4 cm in thickness • 15-25 cm long • Males are shorter than females • Have curved tapering tail Stool examination by Dr. Priyanka Buragohain 109
  • 110. • UNFERTILISED EGGS • Do not float on floatation method • Size- 40×70 µm • Yellow in colour • Elongated • Mammilated thin shell, ovum containing refractile yolk globules occupying the whole inside space • May be confused with veg cell • FERTILISED EGGS • Float on floatation method • Size- 40×70 µm • Yellow in colour • Oval or round • Thick mammillated coat and single celled ovum inside Stool examination by Dr. Priyanka Buragohain 110
  • 111. Enterobius vermicularis • Small round worm or thread like worm or pin worm(spindle shaped) • White coloured • 0.5-1 cm long • Tail pointed • Males smaller than females and posterior body is curved and sharply truncated(found only after purgation) Stool examination by Dr. Priyanka Buragohain 111
  • 112. • EGGS • Colourless(not bile stained) • 20×50 µm • Assymetrical, Oval planoconvex, • thin transparent shelled, • contain coiled tadpole likelarva inside • Floats in saturated salt solution Stool examination by Dr. Priyanka Buragohain 112
  • 113. Ancylostoma duodenale • Hook worm • Small greyish white or pink coloured cylindrical • 1-1.5 cm long • One end is curved like a hook • 6 teeth, 4 hook like on ventral surface and 2 knob like on the dorsal surface Stool examination by Dr. Priyanka Buragohain 113
  • 114. EGGS • Oval or elliptical in shape • 40x60 µm sized egg • Colourless(not bile stained) • Surrounded by a transparent hyaline shelled membrane • Contains 4 segmented ovum inside. • Floats on saturated solution of salt Stool examination by Dr. Priyanka Buragohain 114
  • 115. Trichuris trichiura • Whip worm • Looks like a tiny whip with a handle and a lash • 3-5 cm in length • White coloured Stool examination by Dr. Priyanka Buragohain 115
  • 116. • 25x50 µm sized egg • Brown coloured • Double shelled, outer one is bile stained • Thick shelled, barrel shaped with mucus plug at both pole • Single ovum • floats in saturated solution of common salt Stool examination by Dr. Priyanka Buragohain 116
  • 117. Strongyloids stercoralis • Adult worm: • Females are readily discovered than males • 2.5mm ×40-50 µm (females) • Posterior extrimity is pointed • Males are shorter and broader than females Stool examination by Dr. Priyanka Buragohain 117
  • 118. Stool examination by Dr. Priyanka Buragohain 118 • Eggs • Eggs are conspicuous within the body in a single line • 55-30µm • Thin shelled • Transparent, oval • Contain larva ready to hatch. It is the larva not the eggs are found in stool.
  • 119. • Rhabditiform larvae • Develop directly from gravid females • Short mouth, double- bulb oesophagus • Filariform larvae • Longer and slender • Short mouth and cylindrical oesophagus Stool examination by Dr. Priyanka Buragohain 119
  • 120. Stool examination by Dr. Priyanka Buragohain 120
  • 121. Helminthes that float/do not float on saturated solution of saline Stool examination by Dr. Priyanka Buragohain 121 • A. Duodenale • N. Americanas • E. Vermicularis • H. Nana • A. Lumbricoids • T . Trichuria • H. Diminuta Eggs that float Eggs that do not float • A. Lumbricoids • T . solium • T . Saginata • Trematodes • F. Buski • F. Hepatica • C. Sinensis
  • 122. Stool examination by Dr. Priyanka Buragohain 122
  • 123. Stool examination by Dr. Priyanka Buragohain 123
  • 124. Stool examination by Dr. Priyanka Buragohain 124
  • 125. Plant hairs can be confused for larvae oh hookworm or Strongyloides stercoralis Plant hair resembling S. strongiloids Stool examination by Dr. Priyanka Buragohain 125
  • 126. Parasite of earth worm Pollen grain resemblening to fertile egg of A. lumbricoids Stool examination by Dr. Priyanka Buragohain 126 Bee pollen resembling t. Trichuria egg
  • 127. Stool examination by Dr. Priyanka Buragohain 127
  • 128. STOOL CULTURE • Used to detect the presence of disease causing(pathogenic )bacteria • Help to diagnose an infection of digestive system • Used in conjunction with stool test • Reference range for stool culture is negative Stool examination by Dr. Priyanka Buragohain 128
  • 129. Most common bacteria for culture of stool • Campylobacter species • Salmonella species • Shigella species • Yersinia species • Vibrio species(travel history) • Some bacteria cause illness by producing toxins(PCR, Antigen test are to be done with stool test) • Escherichia coli • Clostridium difficile Stool examination by Dr. Priyanka Buragohain 129
  • 130. Collection • Specimen collected via rectal swab(in infants) • Sterile collection container not required • No detergent or preservative should be present in the container • Specimen should be immediately transported to the laboratory • If transport is delayed by longer than 2 hours, transport media(eg Cary- Blair) is recommended • Samples must be sent in a sealed, leak-proof container marked with a biohazard sticker Stool examination by Dr. Priyanka Buragohain 130
  • 131. Medias used commonly • MacConkey agar- Salmonella species, shigella species • Eosin methylene blue agar • Triple sugar iron(TSI)- differentiate salmonella and shigella • Sabouraud agar • Hekteon enteric agar • Selenite broth Stool examination by Dr. Priyanka Buragohain 131
  • 132. Summary Stool examination by Dr. Priyanka Buragohain 132 • Definition of stool • Composition of stool • Precaution and collection of sample • Physical examination • Chemical examination • Microscopic examination • Artifacts • Stool culture
  • 133. THANK YOU !!! Stool examina Dr. iyan uragoh