Collection of Fecal Sample to Egg Hatching Assay Flashcards
TRUE OR FALSE:
In the collection of fecal sample, the container should be sterile, disposable, wide mouth with tight-fitting lid, transparent
(for physical examination of the specimen)
TRUE
This is very critical when collecting sample
Container
Enumerate the collection of fecal samples
- Container
- AVOID CONTAMINATION WITH URINE, WATER AND SOIL.
- LABEL
- HANDLE CAREFULLY BECAUSE IT IS A POTENTIAL SOURCE
OF INFECTION
Enumerate the samples which are unsuitable samples are from patients receiving:
Barium
Oil
Bismuth
Kaolin
Antibiotics
Enumerate the types of stool specimen
Dry and hard/ Scybalous
Ribbonlike
Mushy
Diarrheic
What type of stool specimen is spastic colitis?
Ribbonlike
What type of stool specimen does not retain the bowel cast?
Mushy
What type of stool specimen is
due to trapped gases?
Mushy
What type of stool specimen will readily flows upon tilting of the container because it is watery.
Diarrheic
In the preservation of stool samples, we should never ____
Incubate
In the preservation of stool samples, should be maintained at room temperature for how many minutes?
30 minutes
In the preservation of stool samples, Stool is allowed at room temperature for only how many minutes?
30 minutes
In the preservation of stool samples, Refrigeration should be at what temperature and maintained for how many days?
4-8 degree Celsius for 2-3 days
Chemical means of preservation:
It is a chemical used for the preparation of stained smears
Polyvinyl Alcohol
Chemical means of preservation:
It is prepared in the lab or purchased commercially
Polyvinyl Alcohol
Chemical means of preservation:
What are the composition of PVA?
Schaudinn’s fluid 93.5 mL
Glycerol 1.5 mL
Glacial acetic acid 5.0 mL
Polyvinyl alcohol, powdered 5.0 g
Chemical means of preservation:
5-10% of formalin is formalinized specimens be prepared thru?
- Wet mounts
- with or without iodine (direct fecal smear same
with wet mounts) - Concentration techniques
What is the Quality control for formalin?
Temperature favors development of helminth eggs
Chemical means of preservation:
Transcribe MIF
MERTHIOLATE-IODINE-FORMALIN
Chemical means of preservation:
MIF is also known as
Thimerosal Iodine Formalin (TIF)
Chemical means of preservation:
This will stain (Iodine) for direct wet mounts of fresh specimens
Thimerosal Iodine Formalin (TIF) or MIF
Enumerate the composition of MIF
Tincture of Merthiolate 200 mL
Formaldehyde 25 mL
Glycerol 5 mL
Distilled water 250 mL
Enumerate the composition of Lugol’s Iodine
Iodine 5 g
Potassium Iodide 10 g
Distilled water 100 mL
TRUE OR FALSE:
For the chemical MIF, in the solutions are mixed in the following proportion per gram
of feces which will serve as the dye
TRUE
How many mL of MF will be multiplied with Lugols Iodine?
9.4 mL MF X 0.6 mL Lugol’s
Chemical means of preservation:
It is for fresh materials obtained from intestinal mucosal linings
SCHAUDINN’S
What is the composition of Schaudinn’s?
HgCl2 solution Two parts
95 % ethyl alcohol One part
TRUE OR FALSE:
For the composition of Schaudinns
HgCl2 solution One part
95 % ethyl alcohol Two parts
FALSE
What is the advantage of using Sodium acetate- acetic acid formalin (SAF)?
Does not contain HgCl2
What is the disadvantage of using Sodium acetate- acetic acid formalin (SAF)?
Images of organisms are not sharp after staining
This chemical does not contain HgCl2?
Sodium acetate- acetic acid formalin (SAF)
Transcribe SAF
Sodium acetate- acetic acid formalin
In the Examination of fecal sample / Importance of Fecalysis:
It is used to detect what?
the presence of parasites
In the Examination of fecal sample / Importance of Fecalysis:
It is used to detect the evidence of ?
dysfunction of some parts of the GIT, liver and pancreas
In the Examination of fecal sample / Importance of Fecalysis:
(TRUE OR FALSE)It is used to detect the evidence of GIT bleeding?
TRUE
In the Examination of fecal sample / Importance of Fecalysis:
It is used to detect the evidence for the detection of excessive fats in the stool. The excessive fats in stool is known as?
Steatorrhea
In the Examination of fecal sample / Importance of Fecalysis:
It is used as a clue in the diagnosis of?
Medical and surgical diagnosis
What is the form and consistency of this stool, soft and formed?
Normal
Enumerate the other form and consistency of this stool
o very soft and watery
o excessively hard and scybalous
o Rice water stool
o Pea-soup stool
o Flattened or ribbonlike
o Butter like
o Gaseous and fermentative
Characteristic of the consistency of Stool:
Cannot be punctured with an applicator stick
Hard
Characteristic of the consistency of Stool:
Maintains shape, can be punctured
Formed
Characteristic of the consistency of Stool:
Bottom side flattens in the container
Semi-formed
Characteristic of the consistency of Stool:
Can be cut with an applicator stick
Soft
Characteristic of the consistency of Stool:
Can be reshaped with applicator stick
Mushy
Characteristic of the consistency of Stool:
Stool shapes to container
Loose
Characteristic of the consistency of Stool:
Stool will flow slowly out of the container
Diarrheic
Characteristic of the consistency of Stool:
Fluid-like stool pours out of the container
Watery
What type of bristol stool chart:
Separate hard lumps, like
nuts (hard to pass)
Type 1
What type of bristol stool chart:
Sausage-shaped but lumpy
Type 2
What type of bristol stool chart:
Like a sausage but with
cracks on its surface
Type 3
What type of bristol stool chart:
Like sausage or snake,
smooth and soft
Type 4
What type of bristol stool chart:
Soft blobs with clear-cut
edges (passed easily)
Type 5
What type of bristol stool chart:
Fluffy pieces with ragged
edges, a mushy stool
Type 6
What type of bristol stool chart:
Watery, no solid pieces.
ENTIRELY LIQUID
Type 7
Color of Stool:
Light brown to dark brown due to stercobilin
Normal
Normal color of stool is due to
Stercobilin
Color of Stool:
Yellow is due to administration of _______ and ______ seen in antibiotic therapy
Santonin and Senna
Color of Stool:
What color is increased milk diet in infants?
Yellow
Color of Stool:
What color is presence of unchanged bilirubin?
Yellow
Color of Stool:
What color is the obstruction to bile flow?
Yellow
Color of Stool:
What color is ingestion of the large amounts of cornmeal?
Yellow
Color of Stool:
What color of the stool after ingestion of Barium meals?
Light Clay or putty color
Color of Stool:
What color of the stool that is due to the absence of bile pigments?
Light Clay or putty color
Color of Stool:
What color is the bleeding in the lower GIT (fresh blood)?
Reddish or Bloody
Color of Stool:
What color is the undigested beets and tomatoes?
Reddish or Bloody
Color of Stool:
What color is the bleeding in the upper GIT?
Dark red/ chocolate brown
Color of Stool:
What color is the increased intake of coffee, chocolate
and cherries, black berries
Dark Red / Chocolate brown
Color of Stool:
If the color of dark red stool when a person has a dengue, it is advised
not to eat ______ like chocolate
because this can cause a false _____ result because of the stool color.
dark foods
False positive
Color of Stool:
What is the color of the stool if it is associated with digestion of blood due to bleeding in the upper GIT?
Black/tarry
Color of Stool:
What is the color of the stool if it is increase intake of iron, Bismuth and charcoal
Black/Tarry
Color of Stool:
The color of the stool if black/ tarry, if you regularly take in medication like
_________, there is a tendency that
stool color
ferrous sulfate
Color of Stool:
The color of the stool Greenish may be due to what?
- Amoebiasis
- ingestion of vegetables like spinach
- presence of unchanged biliverdin
Color of Stool:
Due to cocoa and chocolate produce dark gray
stools
Gray
Color of Stool:
Gray color stool that is due to blueberry juices, carbon, iron and
bismuth that causes the color of ________ to _____ feces
gray to pitch black feces
Color of Stool:
Gray color stool with the presence of undigested fats, following
inadequate bile secretion
Light Gray
Color of Stool:
Due to Malabsorption syndrome
Gray
Color of Stool: Miscellaneous
1. Presence of drugs
2. Whitish discoloration or _____
3. Aluminum hydroxide prep`n and _______
4. Blue - _____ and ____
5. Orange-red- ______
- Bizarre colors
- speckling
- barium
- Methylene blue and dithiazanine
- Pyridium
Odor of Stool:
Tryptophan is due to ____ and ____
Indole and Skatole
Odor of Stool is dependent on
Amount of meat in the diet Putrefactive bacteria
Odor of Stool:
foul to offensive due to indole, skatole and butyric
acid
Normal
Odor of Stool:
Sour Odor
1. Normal for infants: ________ in
ingested milk
2. Adults - ______
- Fatty Acids
- Steatorrhea
Odor of Stool: severe diarrhea in
children
Sour odor to a putrid odor
Abnormal odor found in ulcerative and malignant
tumor of the lower bowel
Putrid Odor
Abnormal odor that indicates gas formation, fermentation of carbohydrate, unabsorbed fatty
acids
Sour/rancid odor
Abnormal odor that is usually in seen in alkaline stools,
putrefaction of undigested protein.
Extremely foul odor
Abnormal odor seen in the ulcerations of
intestines and rectum, malignancy, syphilis,
dysentery and necrotic lesions of the intestines
Extremely foul odor
blood which is not visible to the naked eye and can only be
detected by chemical means only
Occult blood
Occult blood may be seen in
- Benign peptic ulcer
- Esophageal, stomach, duodenal bleeding
- Hemorrhoids
- Liver cirrhosis
- GIT carcinoma/ Rectal carcinoma
Enumerate the laboratory examination for occult blood test
- Benzidine test
- Guaiac’s test
- Hematest
A kit that we used in the blood
Hematest
Identify if False + or False -:
Peroxidase activity of substance or fecal material
o Bacteria
o Fresh fruits
o Vegetables
o Cauliflowers
o Chlorophyll of plants
False +
Identify if False + or False -:
Iron in the diet
False +
Identify if False + or False -:
Myoglobin in ingested meat
False +
Enumerate the Peroxidase activity of substance or fecal material seen in false + result in occult blood test
o Bacteria
o Fresh fruits
o Vegetables
o Cauliflowers
o Chlorophyll of plants
Identify if False + or False -:
Large amount of vitamin C. Interferes in the oxidation of _____
False -
Chromogen
Identify if False + or False -:
Breakdown of blood and its constituents
False -
Identify if False + or False -:
Hemorrhage in the upper GIT
False -
Identify if False + or False -:
Technical errors
False -
PATIENT PREPARATION PRIOR TO OCCULT BLOOD TEST: Meat free diet for _______ prior to the test
3-5 days
Microscopic Examination:
due to hemorrhagic disorder, ulcers and contamination
RBC
Microscopic Examination:
indicative of inflammation
WBC
Simplest and most frequently used
Direct Fecal Smear
DFS uses ______ and ______
0.85% sodium chloride/NSS and D’antonis solution/Lugol’s Iodine
Composition of DFS that is for the observation of the motility of trophozoites
0.85% sodium chloride/NSS
Composition of DFS that doesn’t demonstrate glycogen vacuole and nuclei
0.85% NaCl or NSS
Composition of DFS where cysts appear glistening and refractile
0.85% NaCl or NSS
Composition of DFS for protozoan cysts and helminth ova
D’antonis solution/Lugol’s Iodine
Composition of DFS that kills the trophozoites and helminth larva
D’antonis solution
Composition of DFS where chromatodail bars are fully seen
and for glycogen vacuole
D’antonis solution
USES OF COVERSLIP:
1. to avoid ______ of the preparation
2. to avoid _______ and contamination of objectives
3. to avoid ______ of the stain
- drying
- fogging
- rapid loss
Reagents used for Kato Thick Smear :)
o Distilled water 100 ml
o Glycerin 100 ml
o 3% malachite green 1 ml
o Use cellophane as cover slip
Coverslip used in KTS
Cellophane
Clearing time when using KTS
1. ______ at room temperature
2. ___________ in a dry incubation.
- 1 HOUR
- 20-30 minutes
ADVANTAGES OVER NSS AND D` ANTONIS
1. For _______ examination
2. Satisfactory for all kinds of ______________
- large scale
- helminth eggs
DISADVANTAGES OVER NSS AND D’ANTONIS
1. Unsuitable for ____________ stools
2. Unsuitable for _______________ and _____________
- diarrheic stools
- protozoan cysts and trophozoites
Detection of small number of parasites not detected using
DFS
Concentration techniques
A method used for Protozoan cysts, helminth eggs and larvae
Sedimentation Method
It is less efficient than flotation because protozoan cysts and
many eggs may not be seek
Sedimentation Method
Concentrate eggs, larvae and cysts of parasites, operculated
and schistosomal eggs
FORMALIN-ETHER TECHNIQUE
FECT uses _________
Ether (ethyl acetate)
Material can be examined hours or even days
after preparation
FECT
Not applicable for protozoan cysts
ACID-ETHER SEDIMENTATION
we may observe helminth eggs and larvae
acid-ether
Not recommended because it is time consuming
SIMPLE SEDIMENTATION
For Microfilariae
KNOTT CONCENTRATION TECHNIQUE
What specimen is used for Knott Concentration technique
Blood
Helminths eggs and protozoan cysts may either float or sink
depending on their specific gravity
Flotation Method
Flotation Method:
1. Helminth eggs, protozoan cysts and larvae - _________
2. Chemical solutions: (________) specific gravity that we
use
3. Eggs and cysts: ________
4. Fecal materials: __________
5. Optimal time for examination: ________
- (1.05 to 1.15)
- 1.12 to 1.23
- Float
- Sinks
- 5-20 minutes
ZINC SULFATE CENTRIFUGAL FLOTATION TECHNIQUE
1. Specific gravity: ______
2. 80% effective in detecting eggs and cysts in ________
3. Hookworm: __________
4. Ascaris (fertile): ________
5. Trichuris: ________
6. Giardia: ________
7. E. histolytica: ________
- 1.18
- light infections
- 1.005 does it floats
- 1.110 it floats
- 1.150 it floats
- 1.060 it floats
- 1.070
ZINC SULFATE CENTRIFUGAL FLOTATION TECHNIQUE:
________ ova don’t float because it cannot be detected, diagnostic stage may still be
viable but it destroys trophozoite
Operculated ova
Technique used for Cryptosporidium
SUGAR FLOTATION TECHNIQUE/SHEATHER’S SUGAR
FLOTATION
We can isolate here the rounded oocysts and crescent
sporozoites
SUGAR FLOTATION TECHNIQUE/SHEATHER’S SUGAR
FLOTATION
SUGAR FLOTATION TECHNIQUE/SHEATHER’S SUGAR
FLOTATION COMPOSITION:
o Sucrose - ______ g
o Tap water - ____ mL
o Phenol - _____ g
Sucrose: 500 g
Tap water: 320 mL
Phenol: 6.5g
uses sat. NaCl solution- 40g NaCl + 100 mL dist. H2O
WILLIE’S BRINE
Method used for the detection of Enterobius vermicularis and Taenia
CELLULOSE TAPE PREPARATION/GRAHAM SCOTCH TAPE METHOD
CELLULOSE TAPE PREPARATION/ GRAHAM SCOTCH TAPE METHOD :
________ negative results
Collection: _______ before the patient washes or defecates
_________________ are available
4 consecutive
Collection: morning
Commercial collection kits
getting a sample directly into the rectum, it is called as the
________ because the sticky part of the scotch tape
will be attached to the rectum and will serve as the coverslip
Graham Scotch Method
A technique that estimates worm burden
Degree of infection
Hookworms, Ascaris, Trichuris
EGG COUNT TECHNIQUE
A technique for the degree of infection
Egg count technique
Enumerate the parasites that can be seen when using the egg count technique
Hookworms, Ascaris, Trichuris
DIRECT SMEAR EGG COUNT/ BEAVER DIRECT SMEAR
METHOD:
1. 1.5 mg feces (______) = epg
2. 2 mg feces (______) = epg
- 667
- 500
STOLL’S EGG COUNTING TECHNIQUE DILUTION EGG
COUNT- _______ NaOH
0.1 N
THICK SMEAR EGG COUNT- KTS for ________
SCHISTOSOMES
DILUTION-FILTRATION EGG COUNT- for _______
SCHISTOSOMES
EGG HATCHING:
The counts can be reported as eggs per direct smear
DIRECT SMEAR EGG COUNT
EGG HATCHING:
It uses the dilution egg count
DIRECT SMEAR EGG COUNT
EGG HATCHING:
Direct Smear egg count was proposed by ______ and ________
Stoll and Hausheer
THICK SMEAR EGG COUNT:
The Kato cellophane covered thick smear as modified by
______ and _______
Martin and Beaver
THICK SMEAR EGG COUNT:
For simultaneous qualitative and quantitative diagnosis of the
common intestinal nematode infections and routine
quantitative diagnosis of __________________ and
____________ infections
Schistosoma mansoni and Schistosoma japanicum infections
EGG HATCHING:
For simultaneous qualitative and quantitative diagnosis of the
common intestinal nematode infections
THICK SMEAR EGG COUNT
EGG HATCHING:
Dilution-Filtration Egg count was proposed by _______
Bell
EGG HATCHING:
Schistosome eggs can be concentrated filtering a suspension of feces through filter paper on which the eggs are stained and counted
Dilution-Filtration Egg count
EGG HATCHING:
The procedures are complicated and time
consuming and special equipment is required.
Dilution-Filtration Egg count
EGG HATCHING:
the tool being used is like the Neubauer counting chamber
Dilution-Filtration Egg count
__________ is a laboratory tool used to determine
a given parasite’s resistance to extant drug therapy.
Egg hatch assay (EHA)
Fresh eggs are incubated from the parasite of interest and
serial dilutions of the drug of interest are applied. The
percentage of eggs that hatch or die is determined at each
concentration and a drug response curve may be plotted.
EGG HATCHING TECHNIQUE/ASSAY
EGG HATCHING TECHNIQUE/ASSAY:
The data can then be transformed and analyzed to give further statistics such as an _______
ED50
The technique is labor intensive, expensive and can take
some tine however an egg hatch assay will give and accurate
and reliable result
EGG HATCHING TECHNIQUE/ASSAY