PARASITOLOGY LESSON 1 AND 2 2B

 Accidental or incidental host-

Host other than the normal one
PARASITOLOGY
that is harboring a parasite
 Parasites
 Carrier- An asymptomatic host
 Frequency and distribution
that harbors a parasite and is
 Parasite-host relationship
capable of transmitting it to
 Life cycle (transmission,
others
infective stage, diagnostic stage
TERMINOLOGY
 Helminths- Worms that include
TERMINOLOGY
nematodes (roundworms),
PARASITE-HOST (BIOLOGICAL)
cestodes (tapeworms), and
RELATIONSHIPS
trematodes (flukes)
 Symbiosis- The living together
 Nematode- Roundworm
of unlike organisms.
 Trematode- Fluke; with hookers
 Commensalism- Beneficial to
or suckers
one member and
 Cestode- Tapeworm
harmless/neutral to another
 Trophozoite- Developmental
 Mutualism -Two organisms
stage of protozoa
mutually benefit from each other
 Cyst- Thick-walled stage of
 Parasite -An organism that
protozoa resistant to adverse
obtains its nutrients from
conditions
another organism (the host)
 Larva- Juvenile stage of a
while harming the host
parasite
 Parasitism -One organism, the
 Ciliate- Protozoa motile by
parasite, lives in or on another,
means of cilia
depending on the latter for its
 Oocyst- Encysted form of an
survival and usually at the
egg
expense of the host
 Hydatid cyst- Larval stage of
Pathogenic -Parasite that has
Echinococcus granulosus
demonstrated the ability to cause
 Filariae- Blood or tissue
disease
roundworms
 Flagellate- Protozoa motile by
TYPE OF HOST
 Host- Living organism that means of flagella
harbors another organism  Gravid- Containing ova
 Definitive host- Host in which  Schizont
the adult sexual phase of - Immature schizont: Early
parasite development occurs stage of asexual sporozoan
 Intermediate Host-Host in trophozoite
which the larval asexual phase - Mature schizont: Developed
of parasite development occurs stage of asexual sporozoan
 Reservoir Host- Harboring trophozoite
parasites that are parasitic for TYPE OF HOST
 Host- Living organism that
humans and from which
harbors another organism
humans may become infected
 Definitive host- Host in which
the adult sexual phase of
parasite development occurs
PARASITOLOGY LESSON 1 AND 2 2B

 Accidental or incidental host- 4. Entry via drilling through the

Host other than the normal one skin
that is harboring a parasite 5. Unprotected sexual relations
 Carrier- An asymptomatic host 6. Mouth-to-mouth contact
that harbors a parasite and is 7. Droplet contamination
capable of transmitting it to 8. Eye contact with infected
others swimming water
 Zoonosis- An animal infection MAJOR BODY AREAS
or disease that humans ASSOCIATED
accidentally acquire 1. Gastrointestinal (GI)
 Hermaphroditic- Organism 2. Urogenital (UG)
capable of self-fertilization 3. Tracts
POPULATIONS AT RISK FOR 4. Blood and tissue
CONTRACTING PARASITES 5. Liver
1. Individuals in undeveloped 6. Lungs
areas and countries 7. CSF
2. Refugees 8. Eyes
3. Immigrants 9. Skin
4. Visitors from foreign countries 10. Extremities
5. Individuals who are PARASITIC DISEASE
immunocompromised CHARACTERISTICS
6. Individuals living in close 1. Diarrhea is the most frequent
quarters symptom, along with abdominal
7. Children who attend day care cramping, seen in
centers gastrointestinal tract infections.
PARASITIC DISEASE RISK 2. Other symptoms depend on the
FACTORS parasite and the site of
1. Unsanitary food infection.
handling/preparation (i.e., a. Intestinal obstruction, weight
contaminated meats and loss, and bloating
vegetables) b. Organ involvement with
2. Contaminated water for drinking ulcers, lesions, abscesses,
or recreational use enlargement, and edema
3. Immunocompromised c. Blood and tissue parasites
conditions resulting from can cause anemia, fever, chills,
disease states or poor nutrition bleeding, encephalitis,
4. Blood transfusion and organ meningitis
transplantation d. Elephantiasis, blindness and
5. Foreign travel to endemic vitamin deficiency
regions of the world SPECIMEN COLLECTION AND
MODE OF PARASITIC PROCESSING
TRANSMISSION 1. Diagnosis of parasite infections
1. Ingestion of contaminated food often depends on observing
or drink (water) parasite forms that include
2. Hand-to-mouth transfer protozoa, ova, larva, or adult
3. Insect bite (vector) forms.
PARASITOLOGY LESSON 1 AND 2 2B

2. Specimen types include stools for mercury. Compared to PVA,

(most common), tissue, urine, these preservatives do not
sputum, and blood. provide the quality of
a. Stool samples should be free preservation of intestinal
of antimicrobial agents or other protozoa
substances that inhibit parasite c. Optimal detection of
growth. Barium (from enemas) parasites often requires
can obscure parasites during concentration of specimen.
microscopic examination. - Gross examination of stool
 At least 3 grams of fecal sample may detect adult forms,
on three consecutive days are particularly helminths
required for most parasite (worms).
analyses. - Concentration procedures
 Because urea and acidic pH for feces remove debris that
inhibit some parasites and could obscure parasites.
distort their morphology, stool Barium is not removed
should be free of urine. during concentration
 Liquid stools are best to detect procedures.
trophozoites, whereas formed - Fecal concentration
stools are best to detect ova methods
and cysts.  Fecal concentration methods:
3. Specimen types include stools 1. Formalin-ethyl acetate
(most common), tissue, urine, sedimentation:
sputum, and blood. - 3-4 grams of stool are
b. Stool preservatives suspended in 5 or 10% formalin
 Stool specimens should not be - suspension is filtered using
frozen, and unpreserved gauze into a 15 mL centrifuge
specimens should not be stored tube
at room temperature longer - either 0.85% NaCl or 5 or 10%
than a couple of hours.
formalin is added to fill the tube
 Formalin (5 or 10%) is an all-
- centrifuged at 500 X g for 10
purpose preservative to
minutes, discard supernatant
preserve stool specimens for
- wash repeatedly until the
concentration procedures.
supernatant is clear
 Polyvinyl alcohol (PVA) is a
- after the last wash, resuspend
mercury-containing preservative
for preparing permanent stained sediment in about 9 mL of
smears. formalin
 Sodium acetate formalin - 4 - 5mL aliquot of ethyl acetate
(SAF) is a mercury-free is added; the tube is shaken
preservative that can be used to vigorously for at least 30
preserve stool samples for both seconds
concentration and permanent - loosen cap slightly to release
stained smears. pressure in the tube, centrifuge
 Less toxic preservatives again, and four layers should be
generally substitute zinc sulfate visible (from top to bottom):
ethyl acetate, plug of fecal
PARASITOLOGY LESSON 1 AND 2 2B

debris, formalin, and fecal - procedure is similar to the zinc

sediment sulfate procedure, except
- loosen the plug from the side of sucrose is used in place of zinc
the tube by applicator stick, the - sucrose solution has a specific
top three layers are poured off, gravity of 1.25-1.27
sediment is resuspended in - Sheather sugar flotation
formalin and used for wet procedure is generally
mounts. recommended for
2. Zinc sulfate flotation: Cryptosporidium and some ova.
- 3-4 grams of stool are 4. Blood concentration methods
suspended in 5 or 10%
formalin a) The Knott method uses low-
- suspension is filtered using speed centrifugation to
gauze in a 15 mL centrifuge concentrate blood samples
tube suspected of containing minimal
- centrifuged at 500 X g for 10 numbers of parasites.
minutes, after centrifugation, b) Buffy coat slides are used for
the supernatant is discarded Leishmania or Trypanosoma
- wash repeatedly until the detection.
supernatant is clear, after Various stains are used for
the last wash, the sediment microscopic detection of stool, tissue,
is resuspended in about 3 and blood parasites.
mL of 33% aqueous solution a) Saline wet mounts are quick
of zinc sulfate and easy to perform and will
- the specific gravity of the allow trophozoite motility and
zinc sulfate should be helminth ova and larvae to be
adjusted to 1.20 in formalin seen.
fixed stools or to 1.18 in b) Iodine wet mounts are useful for
fresh (nonformalinized) the detection of larvae, ova, and
stools protozoan cysts in stool
- after resuspending the fecal samples.
material, the tube is filled to c) Permanent stained smears are
within 3-4 mm of the top, used to enhance parasite
tube is centrifuged for 2 morphology and to allow for
minutes at 500 X g future study. Stained fecal
- two layers will result: a small smears are important in the
amount of sediment and a identification of Entamoeba
layer of zinc sulfate. 1 or 2 histolytica.
drops of the surface film Permanent stained smears are used to
should be removed with a enhance parasite morphology and to
loop before removing the allow for future study. Stained fecal
tube from the centrifuge smears are important in the
- the liquid is examined for identification of Entamoeba histolytica.
- Iron hematoxylin stain - used
parasites.
3. Sheather sugar flotation: when enhanced detail is
needed; but it is difficult to
 PARASITOLOGY LESSON 1 AND 2 2B

obtain consistent staining f) Tissue: Naegleria,

results. Acanthamoeba, and
- Trichome stain (Wheatley or Leishmania
Gomori) is the most commonly g) Urine: Schistosoma and
used stain for fecal parasite Trichomonas
study. h) Sputum: Ascaris and
- Modified acid-fast stain is for Strongyloides
Cryptosporidium and Isospora. DIAGNOSTIC TESTS
- Modified trichrome stains for 1. Direct fluorescent antibody:
microsporidia: microsporidia Used to identify Giardia lamblia,
are not easily stained; stain Cryptosporidium, Trichomonas
concentration and the staining vaginalis
time is increased. Alternatively, 2. Latex Agglutination/Direct
a hot stain can be used. In the agglutination test: Used to
Weber green stain, diagnose leishmaniasis and
microsporidia stain pink (oval, Chagas disease
1-3 um) and the background is 3. Indirect fluorescent antibody
green. With the Ryan blue stain, (IFA)
the microsporidia also stain 4. ELISA: Used to identify Giardia
pink, but the background is blue lamblia, Cryptosporidium,
COLLECTION METHODS: Toxoplasma gondii
a) The cellophane (Scotch) tape 5. Rapid immunochromatography
method is used to collect technique
Enterobius vermicularis 6. DNA probes and polymerase
(pinworm) eggs from the chain reactions (PCR) are used
perirectal area. to diagnose parasite infections
b) The EnteroTest® (string test) is TREATMENT OPTIONS
used to obtain duodenal 1. Antiparasitic medications
contents for parasitic 2. Change diet
examination. 3. Vitamins supplements
c) Sigmoidoscopy is used to 4. Fluid replacement
collect colon material. 5. Blood transfusion
5. Sample types and associated 6. Bed rest
parasites PREVENTION AND CONTROL
a) Feces: Giardia, 1. Information, education and
Cryptosporidium, Entamoeba, campaign
Ascaris, Enterobius, etc. 2. Use of pesticides and other
b) Blood: Plasmodium, chemicals
Leishmania, Trypanosoma, and 3. Use of protective clothing
microfilariae 4. Use of protective netting
c) Skin: Onchocerca 5. Proper water treatment
d) Vaginal or urethral: 6. Good personal hygiene
Trichomonas 7. Proper sanitation practices
e) Eye scrapings: Acanthamoeba 8. Proper handling, cooking and
food preparation
 PARASITOLOGY LESSON 1 AND 2 2B

9. Avoidance of unprotected lumbricoides. It is OK to use the

sexual relations genus by itself if you mean to
RULES IN WRITING SCIENTIFIC refer to multiple species within the
NAMES genus. Example: "There are many
In scientific writing (and in science in species of Ascaris that are
general), common names are rarely affected by exposure to soil."
used. The Latin binomial (i.e. "scientific 6. If you must use a common name,
name") is used instead. There are first define it in terms of the
several important rules regarding the scientific name. For example,
use of scientific names. “Ascaris lumbricoides (Giant
1. Scientific names are always intestinal roundworm)".
italicized. When hand-writing 7. Incorrect formatting of scientific
text, you can underline them names may result in a point
instead, but there is no excuse deduction
not to italicize when using a SPECIMEN COLLECTION
word processor. Example: 1. Diagnosis of parasite infections
Ascaris lumbricoides or Ascaris often depends on observing
lumbricoides parasite forms that include
2. The genus is always protozoa (cyst and trophozoites)
capitalized. and helminths (ova, larva,
3. The species is never proglottids or adult forms)
capitalized, even when it refers 2. Specimen types include stools
to the name of a place or (most common), tissue, urine,
person. Be careful because sputum, and blood.
word processors will often a) Stool samples should be free of
"auto-correct" a species name antimicrobial agents or other
and incorrectly capitalize it or substances that inhibit parasite
change the spelling. Example: growth. Barium (from enemas)
Ascaris lumbricoides can obscure parasites during
4. In its first use within a particular microscopic examination.
document, the genus is always - At least 2 - 5 grams of fecal
written in full. In subsequent uses, sample on three consecutive
the genus can be abbreviated days in 10 days required for
using the first initial and a period. most parasite analyses or up to
In the case where two species 6 specimens in 14 days is
from different genera have the acceptable.
same abbreviation for their genus, - Because urea and acidic pH
prevent confusion by writing out inhibit some parasites and
the full genus. Example: on first distort their morphology, stool
use, write Ascaris lumbricoides should be free of urine.
and reserve A. lumbricoides for - Liquid stools are best to detect
subsequent references. trophozoites, whereas formed
5. A species name is never used stools are best to detect ova
without a genus or genus and cysts
abbreviation. Example: write
Ascaris lumbricoides or A.
lumbricoides but never just
 PARASITOLOGY LESSON 1 AND 2 2B

Specimen types include stools (most removed during concentration

common), tissue, urine, sputum, and procedures.
blood.  Fecal concentration methods
b. Timing MACROSCOPIC EXAMINATION
 process immediately 1. COLOR
 Watery to loose – with Brown Normal color is due to urobilin
trophozoites (30 minutes) (stercobilin)
 Semi-formed – 1 hour Black or Melena – Upper gastrointestinal
 Formed - within 2 hours Tarry bleeding, iron therapy, charcoal,
c. Stool preservatives bismuth, digested blood
 Stool specimens should not be Red Lower gastrointestinal bleeding,
pyridium compounds, beets and
frozen, and unpreserved
food coloring, rifampicin,
specimens should not be stored
bleeding low down in the
at room temperature longer intestinal tract
than a couple of hours. Pale Bile duct obstruction, barium
 Formalin (5 or 10%) is an all- Yellow,
purpose preservative to White, Gray
preserve stool specimens for Yellow Milk, diet, corn meal, rhubardb,
concentration procedures. senna, santonin, fats
 Polyvinyl alcohol (PVA) is a Green Biliverdin, green vegetables,
mercury-containing preservative antibiotics, spinach, calomel,
for preparing permanent stained cooked green, unchanged
smears. biliverdin, meconium, porphyrins
 Sodium acetate formalin Pale Steatorrhea
(SAF) is a mercury-free
preservative that can be used to 2. CONSISTENCY
preserve stool samples for both Formed Normal
concentration and permanent Bulky or frothy Poor fat digestion
stained smears. Hard Constipation
 Less toxic preservatives Flattened and Obstruction in lower
ribbon-like bowel
generally substitute zinc
Semisolid Digestive upset, mild
sulfate for mercury. Compared
diarrhea or after taking
to PVA, these preservatives do laxative
not provide the quality of Watery Bacterial infection or
preservation of intestinal after taking purgative,
protozoa. Viral
d. Optimal detection of Bulky Steatorrhea
parasites often requires pale/Frothy/Foam
concentration of specimen. y
 Gross examination of stool may Rice watery stools
Cholera
detect adult forms, particularly Mucoid Common causes can be
helminths (worms). constipation,
 Concentration procedures for dehydration, Crohn's
feces remove debris that could disease, and irritable
bowel syndrome
obscure parasites. Barium is not
Rounded Prolonged constipation;
scybalous masses goat dropping stool
 PARASITOLOGY LESSON 1 AND 2 2B

Small calibre Constipation; - loosen cap slightly to release

stools Hirschsprung’s disease pressure in the tube, centrifuge
again, and four layers should be
MICROSCOPIC EXAMINATION visible (from top to bottom):
 Microscope – the most ethyl acetate, plug of fecal
important tool debris, formalin, and fecal
 Microns – u or um defined as a sediment
unit measuring 0.001 millimeter - loosen the plug from the side of
DIRECT WET PREPARATION – the tube by applicator stick, the
Unfixed stool top three layers are poured off,
1. Saline wet mounts sediment is resuspended in
 are quick and easy to perform formalin and used for wet
and will allow trophozoite mounts
motility and helminth ova and 2. Zinc sulfate flotation – based
larvae to be seen on differences between specific
 0.85% saline gravity and the sample debris
2. Iodine wet mounts - 3-4 grams of stool are
 are useful for the detection of suspended in 5 or 10% formalin
larvae, ova, and protozoan - suspension is filtered using
cysts in stool samples
gauze in a 15 mL centrifuge
 Lugol’s or D’ Antoni’s formula
tube
CONCENTRATION METHODS- -
- centrifuged at 500 X g for 10
ability to detect small number of
minutes, after centrifugation, the
parasites (fixed or unfixed)
supernatant is discarded
1. Formalin-ethyl acetate
- wash repeatedly until the
sedimentation – principle:
supernatant is clear, after the
specific gravity
last wash, the sediment is
- 3-4 grams of stool are
resuspended in about 3 mL of
suspended in 5 or 10% formalin
33% aqueous solution of zinc
- suspension is filtered using
sulfate
gauze into a 15 mL centrifuge
- the specific gravity of the zinc
tube
sulfate should be adjusted to
- either 0.85% NaCl or 5 or 10%
1.20 in formalin fixed stools or
formalin is added to fill the tube
to 1.18 in fresh
- centrifuged at 500 X g for 10
(nonformalinized) stools
minutes, discard supernatant - after resuspending the fecal
- wash repeatedly until the
material, the tube is filled to
supernatant is clear within 3-4 mm of the top, tube is
- after the last wash, resuspend centrifuged for 2 minutes at 500
sediment in about 9 mL of Xg
formalin - two layers will result: a small
- 4 - 5mL aliquot of ethyl acetate amount of sediment and a layer
is added; the tube is shaken of zinc sulfate. 1 or 2 drops of
vigorously for at least 30 the surface film should be
seconds removed with a loop before
 PARASITOLOGY LESSON 1 AND 2 2B

removing the tube from the concentration and the staining

centrifuge time is increased. Alternatively,
- the liquid is examined for a hot stain can be used. In the
parasites Weber green stain,
3. Sheather sugar flotation: microsporidia stain pink (oval,
- procedure is similar to the zinc 1-3 um) and the background is
sulfate procedure, except green. With the Ryan blue
sucrose is used in place of zinc stain, the microsporidia also
- sucrose solution has a specific stain pink, but the background
gravity of 1.25-1.27 is blue.
- Sheather sugar flotation -
procedure is generally SPECIMEN COLLECTION
recommended for TECHNIQUES
Cryptosporidium and some ova. 1. The cellophane (Scotch) tape
4. Blood concentration methods method is used to collect
a) The Knott method uses low- Enterobius vermicularis
speed centrifugation to (pinworm) eggs from the
concentrate blood samples perirectal area.
suspected of containing minimal 2. The EnteroTest® (string test)
numbers of parasites. is used to obtain duodenal
b) Buffy coat slides are used for contents for parasitic
Leishmania or Trypanosoma examination
detection. 3. Sigmoidoscopy is used to
SPECIMEN COLLECTION AND collect colon material.
PROCESSING OTHER SPECIMEN COLLECTION
PERMANENT STAINED SMEARS Sample types and associated
 are used to enhance parasite parasites
morphology and to allow for a) Feces: Giardia,
future study Cryptosporidium, Entamoeba,
 stained fecal smears are Ascaris, Enterobius, etc.
important in the identification of b) Blood: Plasmodium,
Entamoeba histolytica. Leishmania, Trypanosoma, and
- Iron hematoxylin stain - used microfilariae
when enhanced detail is c) Skin: Onchocerca
needed; but it is difficult to d) Vaginal or urethral:
obtain consistent staining Trichomonas
results. e) Eye scrapings: Acanthamoeba
- Trichome stain (Wheatley or f) Tissue: Naegleria,
Acanthamoeba, and
Gomori) is the most commonly
Leishmania
used stain for fecal parasite
g) Urine: Schistosoma and
study.
Trichomonas
- Modified acid-fast stain is for
h) Sputum: Ascaris and
Cryptosporidium and Isospora.
Strongyloide
- Modified trichrome stains for
microsporidia: microsporidia
are not easily stained; stain
PARASITOLOGY LESSON 1 AND 2 2B