(LECTURE)
2A PARASITOLOGY
P-04 Dr. Julius Capili | March 03, 2019
BABESIA
 Maltese-cross formation
 often mistaken as Plasmodium falciparum
but differs in:
 lacks pigments in the cytoplasm
 lacks of growing trophozoites
(sporozoites or merozoites)
 the vector are Ticks--- Ixodes
scapularis
 Animals (ex: deer) is the usual definitive
host
 Man (incidental Host) is infected by the bite
of the intermediate host (Tick----IXODES)
and blood transfusion.
 Causes:
o Headache and fever
o Hemolytic anemia with
hemoglobinuria in immunocompetent
host
o Red water fever on cattles (can also
become a definitive host)
COCCIDIA
-literally refer to sporozoans which are infecting
HIV/ AIDS patients
o Schizogony (asexual) in variety of
nucleated cells of the definitive host.
o Sporogony (sexual) in intestinal
mucosa of the definitive host----
infective oocyst excreted in feces.
*INFECTIVE STAGE: Oocyst
Babesia vs Malaria vs Coccidia
Babesia: Man is Incidental host
Malaria: Man is Intermidiate host
Coccidia: Man maybe Definitive or
Intermediate (depends on the lifecycle)
1. Isospora belli
 Definitive host: MAN
 Intermediate host: PIGS/CATTLES
 Transmission: ingestion of sporulated oocyst
in contaminated food and water.
 Lab dx: Stool exam and Modified Acid Fast
Stain
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Sheaters Flotation Gold standard for
technique identifying isospora
Mod.Acid Fast Stain used to stain the
isopora
2. Cryptosporidium parvum
 parasite that causes intestinal infection
among AIDS patients
 associated with watery, frothy, diarrhea
with oocyst shed in feces
 Detection: Sheath’s Sugar flotation,
Modified Acid Fast Stain
NOTE: ISOSPORA AND CRYPTOSPORIDIUM both
causes Gastroenteritis, intestinal malabsorption
among AIDS patient but MOST COMMON is
Crystosporidium.
3. Toxoplasma gondii
 Associated with CNS disorder
 Definitive Host: CAT
 Intermediate host: MAN (source of
trophozoites that has crescent
appearance in tissues)
 Lifecycle:
1. Toxoplasma divide in tissues of man as
TACHYZOITES (actively dividing
trophozoites),
2. Pseudocysts (group of BRADYZOITES) are
also formed
3. Females who acquire infection during
pregnancy may transit infection to embryo
resulting in fetal death, mental retardation in
newborn or blindness in later life.
 Laboratory Diagnosis
a) Sabin fieldman dye test: Methylene blue
staining of Tachyzoites inhibited by prior
addition of patient serum containing
antibodies to toxoplasma
 Positive result: Colorless organisms
over a blue background
- Tachyzoites resist the dye.
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**Principle: INHIBITION MECHANISM (trophozoites a. Wheatley’s trichrome stain
don’t absorb dye) b. Iron Hematoxylin stain
c. Modified Acid fast stains
(Cryptosporidium, cyclospora, Isospora)
b) IFA
Positive result: Fluorescence (emission of C. Concentration methods for protozoa cysts and
light) helminthes eggs and larva
Ex of Dye used: 1. Sedimentation techniques
 Phycocyanin RED a. Acid ether concentration
 Texas Red fluorescence b. formalin ether concentration
 FITC Green Gold -dissolves debris, lipid and CHOs
fluorescence 2. Flotation technique
a. Brine flotation (NaCl)
c) TORCH Test b. Zinc sulfate centrifugal flotation techniques
 Toxoplasma, Rubella, CMV, Herpes (specific gravity of 1.18)
c. Sheathers sugar flotation
LABORATORY METHOS -best for Cryptosporidium
A. Examination of blood
1. Detects agents of malaria, babesiosis, D. Culture methods for protozoa
trypanosomiasis, leishmanasis and 1. Culture media for intestinal amoeba
filariasis a. Boeck Dorbohlav’s Diphasic medium
2. Include thick and thin blood films (modified by Dobell and Laidlaw)
3. Concentration techniques (Quantitative b.Las’s Casein Hydrolysate serum medium
Buffy Coat/QBC) c. Balamuth’s Aqueous Egg Yolk infusion
a. Preparation of buffy coat d. Cleveland Colliers medium
smears (L. donovani, e. Feinberg medium
trypanosomes, microfilaria) f. Whittington medium
b. Knott’s concentration g. Modified Diamond’s medium
membrane filtration h. PYGC medium with antibiotics
(detects microfilaria) i. Nove McNeal Nicolle (NNN) medium for
c. Fluorochrome acridine Teismania and Trypanosoma
orange in microhematocrit j. Hockmeyer;s medium- visceral leishmania
centrifuge format (QBC k.Weinman’s medium-Trypanosoma
blood parasite detection gambiense
method) l.Toboes Diphasic medium- hemoflagellates

B. Examination of Fecal Specimen E. Additional techniques for examination of

1. Specimens maybe submitted to the ecterice parasites
laboratory either fresh or in appropriate a. Cellulose tape technique for pinworm
preservative (Scoth tape method)
2. Stool fixatives: b. Egg studies
a. 10% formalin 1. direct smear method of Beaver
b. Scaudinn’s solution 2. Stoll dilution egg count
c. Polyvinyl alcohol 3. Kato Thick smear
d. Merthiolate-Iodine formalin (MIF) c. Nematode culture and recovery techniques
e. Sodium acetate formlain (SAF) 1. Harda-Mori filter paper culture
3. Direct Wet mount 2. Baermann funnel technique
a. Saline mount (strongloides)
b. Iodine mount
4. Permanent stains

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F. Examination of other specimens
1. Vaginal and urethral discharges, prostatice
sections and urine
2. skin biopsy (Onchorera, Mansonella) , muscle
biopsy (Trichinella)
COLLECTION OF SPECIMENS
 Containers
 Contaminants and Drugs
 Size of Specimen
 Age of Specimen
1. Size
-entire fecal passage, if possible, or at least 20
to 30 grams or about 1 to 2 tablespoonful
2. Contaminants or Interfering substances
a. Urine- destroys protozoan trophozoites
b. Water-destroys protozoan trophozoites
c. Dirt-interferes with examinations; may
introduce free-living organisms that
might be confused with parasites
Oily Interfere with examination; may
laxatives cause inaacurate or ineffective
performance of techniques
Bismuth Crystals in feces which interfere
with examinations
Barium Abrasive action, which may destroy
organisms; interferes with
examinations
Kaolin Abrasive action may affect
compounds appearance of organisms. Wait for
7-10 days after these compounds
have been given before collecting
specimens for parasitologic
examinations
Antibiotics Cause decrease in numbers of
protozoa in intestinal tract; wait 1 to
2 weeks after termination of therapy
before collecting specimens
3. Age
- unpreserved specimens should reach the
laboratory within 1-2 hours after passage
-time and date of passage should be recorded
 PRESERVATIVES
1. Formalin (5-10%)
-preserves eggs, larvae and cysts for wet
mount examinations
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2. PVA (Polyvinyl Alcohol-fixative)
-preserves tropozoites and cysts for
subsequent permanent staining
3. MIF (Methiolate-iodine-formaldehyde)
-preserves all stages for wet mount
examinations
4. PAF (Phenol-alcohol-formaldehyde)
-preserves all stages for wet mount
examinations
 Permanent stains cannot be made from feces
preserved in MIF and PAF solutions;
therefore, a portion of the specimen should
also be preserved in PVA-fixative
 Fecal specimens: normally passes fecal
specimens, 2-3 days intervals
 Container: plastic cups; defecate directly on
the cup or in a cardboard (karton) to avoid
contamination
 Conduct stool examination before prescribing
drugs especially antibiotics
 Label of the specimen (eg name of patient)
should be placed on the side of the cup, not on
the top (takip)
NEMATODES
- referred as ASCALMINTHES
- they are round and embossed
- they are huge parasites and multicellular
Characteristics:
1. System of the parasite
A. Integumentary system
- known as SYNCYTIUM / CUTICLE
Composed of 3 muscle bands/layers:
1. INNERMOST- longitudinal
2. MIDDLE LAYER- oblique
3. OUTER LAYER- circular
Movement:
Adult parasite - Figure of 8
Larva- Purposely motion (they can go anywhere)
B. Nervous System
- has a nerve center referred to as the ganglia
seen in the esophageal area or the CIRCUM
ESOHAGEAL RING
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- nerve center transmit nerve impulses or
senses to the different parts of the parasites
through TRANSVERSE COMMISURE
- Parasite would access into one organ referred
to as CHEMORECEPTOR
Caudal Chemoreceptors: Phasmids
Other Chemoreceptors: Aphasmids
C. Circulatory System
- It is considered to be absent, the fluid inside
the parasite works the function of the blood
D. Digestive System
- known as a Complete system:
Mouth or Buccal capsule
Esophagus
Alimentary canal
Anus (female parasite)/Cloaca (male
parasite)
- Referring to the mouth or the buccal capsule
of the parasite it is accessorized.
 Dorsal Cutting Plates- ex. Necator
americanus
 Teeth- ex. Ancylostoma spp.
 3 lips (trilobe)- ex. Ascaris
 Papilated lips- ex. Trichuris
E. Reproductive System
- Males separate from Female
Ex. Ascaris lumbricoides male and Ascaris
lumbricoides female
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Difference between Male and Female
MALE FEMALE
Curved tails Pointed tails
Smaller Larger - for procreation
Male Reproductive Organ
Basic Unit structure: Gubernaculum
- seen at the posterior most portion
Parts of gubernaculum
1. Bursa – lobed part of gubernaculum
2. Bursal rays – inside the bursa
a. Bipartile: 2 lobes (ex. Necator
americanus)
b. Tripartile: 3 lobes (ex. Ancylostoma
spp.)
3. Cleft or the cleavage
4. Spicule- most important because it is the
penis of the parasite
- It is the copulatory organ
- For nematodes it contains one spicule and
other parasite contain multiple spicules
- Some of the spicules are barbed or fused (ex.
N. americanus), Ancylostoma is plenty or
unfused
Female Reproductive Organ
Organ: Genital primordium/ Gynecophoral canal
- Found/ along one side of the parasite,
probably ¼ to ½ anterior to the body
Sexual position: inverted Y position
The central dogma of the life cycle of
nematodes:
1. Ova
2. Larva
3. Adult
 Ova Stage
-Ova of the parasites would have 3 types
1. Unfertile ova- cannot proceed to the next
developmental life cycle stages.
2. Mature ova- potential ova, that needs to get to
the embryonated stage.
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3. Embryonated ova- INFECTIVE stage. Causes Embryonation requirements:
infection
1. Soil
 Larva stage: - Ascaris
1. Rhabditiform larva - Trichuris
- early stage. - Hookworm
- Feeding/ vegetative stage 2. Water
- Open mouth - Capillaria philipinensis
- Shorter but broader 3. None
2. Filariform Larva - Enterobius vermicularis
- Non-feeding/ non-vegetative stage
When the infective stage is the filariform larva, the
- Closed mouth
mode of transmission would be skin penetration.
- INFECTIVE stage(e.g. hookworms)
- Slender but longer If the infective stage is the embryonated ova, MOT
would be ingestion.
Larval stage of development will always be the same
among the nematodes 2 CLASSES OF NEMATODES

 Adult Stage: 1. Aphasmids/amphids/adenophorea

- Stage where parasites can be sexually - lack caudal chemoreceptors but have
differentiated. cephalic chemoreceptors.
- Male parasites are smaller and curved - Trichinella spiralis small
tails while females are bigger and have intestines
pointed tails. - Capillaria
- Diocophyma renale- kidneys
Classification Female adults according to ova laid: - Trichuris trichuria- large intestines
1. Oviparous- lay immature ova 2. Phasmids/Sercentea
2. Oviviparous- lay readily infective ova (e.g. - Hookworms
Enterobius vermicularis) - Ascaris small intestines
3. Viviparous/Larviparous- doesn’t lay ova, they - Stongyloides
lay larva. - Enterobius vermicularis- large
intestines
Classification Female adults depending on the - Filarial worms- tissues
uterine orientation:

1. Single- 1 ovary and oviduct(e.g. Trichuris

trichuria and Trichinella spiralis)
2. Bifurcated 2 pairs of ovary and oviduct (e.g.
Hookworms, Ascaris, Enterobius)

Molting
transition period of the larva going to adult stage.
 Ascaris lumbricoides- 3 moltings from
rhabditiform to filariform and 1 molting from
filariform to adult.

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