Lab Discussion-1
Lab Discussion-1
Lab Discussion-1
Parasitology
MedBio4: Medical Parasitology
1st Semester, A.Y. 24-25
Laboratory Diagnosis
Example:
Dysentery is a disease that is
characterized by diarrhea and contains
both blood and mucus (macroscopic
examination).
Example:
Dysentery is a disease that is
characterized by diarrhea and Bacillary dysentery Amoebic dysentery
contains both blood and mucus
(macroscopic examination). Feces: Feces:
no forms of trophozoites
bacteria are present
Diagnosis through clinical signs
identified as
and stated symptoms, but
trophozoites
confirmed by microscopic
examination.
Laboratory Diagnosis
Procedures:
EPA Method 1623
Modified Method 1623
Procedure:
Collect using auger/spade -
20 cm deep
Put in sterile plastic bags
Sent to lab for immediate
processing
Amphizoic - organism that can exist either
as a parasite or as free-living organism
Collection and Processing:
Human Biological Samples
a. Urine
Trichomonas vaginalis (trophozoites)
and Schistosoma haematobium (eggs)
Schistosoma haematobium
b. Stool
Consistency:
Enterobius vermicularis
scotch tape
tongue depressor
glass slide
PPE (gloves and mask) - Highly infective
Enterobius vermicularis
scotch tape
tongue depressor
glass slide
PPE (gloves and mask) - Highly infective
Collect early morning and put deep sputum Procedure for deep sputum: rinse mouth with
water and expel the sputum by breathing and
in sterile containers coughing at 2-minutes interval
Why deep sputum? High sens/concentrated
d. Sputum
Acanthamoeba spp.
Laboratory Diagnossis
of Parasitic Diseases
MedBio4: Medical Parasitology
1st Semester, A.Y. 24-25
Outline
1. Parasitic (morphological) diagnosis
Macroscopic
Microscopic
2. Immunological diagnosis
Antibody detection
Antigen detection
3. Molecular diagnosis
4. Culture
5. Animal inoculation (Xenodiagnosis)
Morphological Diagnosis
Macroscopic and Microscopic
Morphological Diagnosis
A. Macroscopic examination
Ex. stool specimen examined
with the naked eye for:
occult blood
parasite component
color
consistency
B. Microscopic examination
Ex. blood, stool, urine (stained or
unstained preparation)
Morphological Diagnosis
B. Microscopic examination
Elements that may be found in stool specimens
White blood cells
Polymorphonuclears (PMNs)
Eosinophils
Macrophages
Red blood cells
Charcot-Leyden crystals
Epithelial cells
Eggs of arthropods
Fungal spores
Elements of plant origin
Animal hairs
Morphological Diagnosis
Special stains and corresponding parasites
Temporary stain Permanent stain in histopathologic slides
Giemsa/field
Eosin
stain
Iron-Hematxylin
Lugol’s iodine solution
stain
Modified
Sargeaunt’s stain
Ziehl-Neelson stain
Phenol-Auromine
Burrow’s stain
method
Acridine orange
Permanent Stain
PVA-preserved specimen
Iron-hematoxylin Stain
stains intestinal protozoa Images captured from a trichrome stained smear were submitted
helminth egg and larvae may be obscured to DPDx for diagnostic assistance. The smears were made from a
good stain for fresh, PVA, or SAF-preserved polyvinyl-alcohol (PVA) preserved fecal specimen, but no other
fecal smears patient or specimen information was given. What is your
diagnosis? Based on what criteria? What valuable information
Wheatley's Trichrome Stain would have been useful if provided?
one of the most commonly used stain for
intestinal protozoa
stain for fresh and PVA-preserved but not
w/ SAF-preserved smears
Acid-Fast Stain
Cryptosporidium,
Isospora,
Cyclospora
Immunodiagnosis
A. Antibody detection
serum
Ab is produced in response to a
particular parasitic infection
Ab may persist for a long period of
time in the serum after an infection
has ended
unable to distinguish between past
or present infection
Immunodiagnosis
A. Antibody detection
However, parasites can be low in numbers during pre-patent and chronic periods
of infection, hence, microscopic examination may yield false negative results.
Xenodiagnosis
A. Macroscopic examination
Ex. stool specimen examined with
the naked eye for:
occult blood
parasite component
color
consistency
B. Microscopic examination
Ex. blood, stool, urine, mucus
(stained or unstained preparation)
Examination of Stool
B. Microscopic examination
Elements that may be found in stool specimens
White blood cells
Polymorphonuclears (PMNs)
Eosinophils
Macrophages
Red blood cells
Charcot-Leyden crystals
Epithelial cells
Eggs of arthropods
Fungal spores
Elements of plant origin
Animal hairs
Biosafety
Potential risks with stool specimens includes:
ingestion of eggs or cysts,
skin penetration by infective larvae, and
infection by non-parasitic agents found in stool and biologic fluids.
clean, wide-mouthed
containers with tight
fitting lid
retain moisture, prevent
spillage, and
contamination
3 specimens within 10
days (Amebiasis - 6 in
14 days)
Collection of Stool or Fecal Sample
protozoan: 3 - 4 weeks
helminth: 1 - 2 weeks
Taenia sp.: 5 - 6 weeks
How many stool samples should be collected when following the
typical O&P collection protocol?
A. 1
B. 2
C. 3
D. 4
Important factors in stool analysis
A. Intake of drugs/medicinal
All of these drugs have been
substances
found to leave crystalline
1. antacids residues.
2. anti-diarrheals
3. barium - samples should be collected a
4. bismuth week after
5. laxatives
A barium enema is a type of x-ray that allows your doctor to see your colon
and rectum. It is also called a colon x-ray or lower GI exam. Barium enemas
can help diagnose changes to your large intestine, such as your colon and
rectum.
Important factors in stool analysis
B. Intake of antibiotics decreases the number of
protozoans for several weeks
D. Contamination w/ toilet
destroy trophozoites and may
water, urine, or soil
contain free-living organisms
Important factors in stool analysis
E. Age of stool sample Liquid specimen should be
examined within 30mins and
formed within 1 hour
Normal:
Adult: brown
New born infants: Black (meconium)
Breast feed infants: scrambled egg
Infant feed on animal milk: “curd like”
CONTENT WARNING:
POOP PICS AHEAD!!!
Gross/Macroscopic - Color
Meconium: Baby’s first stool
First stool a baby will pass thick
Green, tar-like substance
Lines the intestines of the fetus
First bowel movement within a few hours
after birth.
Breastfed Stool
Yellow
Runny
Small seed like objects in the stool
Often called baby poop mustard
Gross/Macroscopic - Color
Changes in stool color that persist can be a serious matter and should always be
investigated by a physician.
Gross/Macroscopic - Color
Abnormal:
Clay or white
absence of bile pigment (bile obstruction) or
diagnostic study using barium
´Black or tarry
drug (e.g., iron),
bleeding from upper gastrointestinal tract (e.g.,stomach, small intestine),
diet high in red meat and
dark green vegetables (e.g., spinach)
Gross/Macroscopic - Color
Abnormal:
Red
bleeding from lower gastrointestinal tract (e.g., rectum),
hemorrhoids
some foods
red gelatin,
tomato juice or soup
large amounts beets
Pale
malabsorption of fats,
diet high in milk and milk products and low in meat
Gross/Macroscopic - Consistency
1 - resists puncture
2 - can be punctured
3 - can be cut with applicator
4 - can be reshaped
5 - shaped into container
6 - can flow
7 - can pour
Gross/Macroscopic - Consistency
Normal: Formed, soft, semisolid or mushy
Abnormal:
Hard, dry, constipated stool
Dehydration, decreased intestinal motility resulting from lack of fiber in
diet, lack of exercise, emotional upset, laxative abuse
Diarrhea
Increased intestinal motility (e.g., irritation of the colon by bacteria)
Cleary watery, loose mixed with mucus and blood
Gross/Macroscopic - Consistency
Classification of the form, (appearance in a toilet) of feces into seven groups.
The form of the stool depends on the time it spends in the colon.
Chart breakdown
Types 1 and 2 indicate constipation;
Types 3 and 4 are usually the most comfortable to pass,
Types 5-6 tend to be associated with urgency, while
Type 7 is diarrhea.
Gross/Macroscopic - Shape
Normal:
cylindrical (contour of rectum) about 2.5 cm (1 inch) in diameter in adults
Abnormal:
narrow, pencil-shaped, or stringlike stool
obstructive conditional of the rectum
Gross/Macroscopic
Amount
Normal:
varies with diet
about 100 to 400 g per day
Size
Normal:
healthy piece of feces is about one foot long.
Gross/Macroscopic
Frequency
Normal:
three times a day to once every three days.
average person poops about once a day.
Odor
Normal:
aromatic, affected by ingested food and person’s own bacterial flora
Abnormal
Pungent (infection, blood, sloughed tissue)
Gross/Macroscopic - Odor
What makes feces smell so bad?
Specifically, the bacteria produce various compounds and gases that lead to the
infamous smell of feces.
Gut flora produce compounds such as indole, skatole, and thiols (sulfur
containing compounds), as well as the inorganic gas hydrogen sulfide
The bad smell of feces will usually be reduced by eating more natural foods that
do not contain any artificial flavors or chemicals
Gross/Macroscopic - Odor
What makes feces smell so bad?
Specifically, the bacteria produce various compounds and gases that lead to the
infamous smell of feces.
Gut flora produce compounds such as indole, skatole, and thiols (sulfur
containing compounds), as well as the inorganic gas hydrogen sulfide
The bad smell of feces will usually be reduced by eating more natural foods that
do not contain any artificial flavors or chemicals
Gross/Macroscopic - Constituents
Normal:
water (about 75%).
dead bacteria that helped us digest our food, living bacteria,
undigested food residue (known as fiber),
cellular linings, sloughed epithelial cells
substances released from the intestines (such as mucus) and the liver
fat, protein, dried constituents of digestive juices (e.g., bile pigments),
inorganic matter (e.g., calcium, phosphates)
Abnormal:
pus: bacterial infection
mucus: inflammatory condition
blood: gastrointestinal bleeding
large quantities of fat: malabsorption
foreign objects: accidental ingestion
Techniques
STH
Mass epidemiologic
studies
Techniques
C. Concentration Techniques
Sedimentation
Flotation
Preservation
Formalin
5% - protozoan cyst
10% - helminth eggs and larvae
FECT (formalin-ether/ethyl acetate
concentration technique)
Preservation
Schaudinn’s solution
fresh stool for staining the stool
smear
mercuric choride (toxic)
Preservation
Merthiolate-iodine-formalin (MIF)
intestinal protozoans, helminth eggs, and larvae
merthiolate and iodine (stain)
formalin (preservative)
wet mount using preserved stools