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(LECTURE)

2A PARASITOLOGY
P-04 Dr. Julius Capili | March 03, 2019

BABESIA
 Maltese-cross formation Sheaters Flotation Gold standard for
 often mistaken as Plasmodium falciparum technique identifying isospora
but differs in: Mod.Acid Fast Stain used to stain the
 lacks pigments in the cytoplasm isopora
 lacks of growing trophozoites
(sporozoites or merozoites) 2. Cryptosporidium parvum
 the vector are Ticks--- Ixodes  parasite that causes intestinal infection
scapularis among AIDS patients
 Animals (ex: deer) is the usual definitive  associated with watery, frothy, diarrhea
host with oocyst shed in feces
 Man (incidental Host) is infected by the bite  Detection: Sheath’s Sugar flotation,
of the intermediate host (Tick----IXODES) Modified Acid Fast Stain
and blood transfusion.
 Causes: NOTE: ISOSPORA AND CRYPTOSPORIDIUM both
o Headache and fever causes Gastroenteritis, intestinal malabsorption
o Hemolytic anemia with among AIDS patient but MOST COMMON is
hemoglobinuria in immunocompetent Crystosporidium.
host
o Red water fever on cattles (can also 3. Toxoplasma gondii
become a definitive host)  Associated with CNS disorder
 Definitive Host: CAT
COCCIDIA
 Intermediate host: MAN (source of
-literally refer to sporozoans which are infecting
trophozoites that has crescent
HIV/ AIDS patients
appearance in tissues)
o Schizogony (asexual) in variety of
nucleated cells of the definitive host.
 Lifecycle:
o Sporogony (sexual) in intestinal
1. Toxoplasma divide in tissues of man as
mucosa of the definitive host----
TACHYZOITES (actively dividing
infective oocyst excreted in feces.
trophozoites),
*INFECTIVE STAGE: Oocyst
2. Pseudocysts (group of BRADYZOITES) are
also formed
Babesia vs Malaria vs Coccidia 3. Females who acquire infection during
Babesia: Man is Incidental host pregnancy may transit infection to embryo
Malaria: Man is Intermidiate host resulting in fetal death, mental retardation in
Coccidia: Man maybe Definitive or newborn or blindness in later life.
Intermediate (depends on the lifecycle)
 Laboratory Diagnosis
1. Isospora belli a) Sabin fieldman dye test: Methylene blue
 Definitive host: MAN staining of Tachyzoites inhibited by prior
 Intermediate host: PIGS/CATTLES addition of patient serum containing
 Transmission: ingestion of sporulated oocyst antibodies to toxoplasma
in contaminated food and water.  Positive result: Colorless organisms
 Lab dx: Stool exam and Modified Acid Fast over a blue background
Stain - 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 2. PVA (Polyvinyl Alcohol-fixative)
1. Vaginal and urethral discharges, prostatice -preserves tropozoites and cysts for
sections and urine subsequent permanent staining
2. skin biopsy (Onchorera, Mansonella) , muscle 3. MIF (Methiolate-iodine-formaldehyde)
biopsy (Trichinella) -preserves all stages for wet mount
examinations
COLLECTION OF SPECIMENS 4. PAF (Phenol-alcohol-formaldehyde)
-preserves all stages for wet mount
 Containers examinations
 Contaminants and Drugs  Permanent stains cannot be made from feces
 Size of Specimen preserved in MIF and PAF solutions;
 Age of Specimen therefore, a portion of the specimen should
1. Size also be preserved in PVA-fixative
-entire fecal passage, if possible, or at least 20  Fecal specimens: normally passes fecal
to 30 grams or about 1 to 2 tablespoonful specimens, 2-3 days intervals
2. Contaminants or Interfering substances
a. Urine- destroys protozoan trophozoites  Container: plastic cups; defecate directly on
b. Water-destroys protozoan trophozoites the cup or in a cardboard (karton) to avoid
c. Dirt-interferes with examinations; may contamination
introduce free-living organisms that  Conduct stool examination before prescribing
might be confused with parasites drugs especially antibiotics
 Label of the specimen (eg name of patient)
Oily Interfere with examination; may should be placed on the side of the cup, not on
laxatives cause inaacurate or ineffective the top (takip)
performance of techniques
Bismuth Crystals in feces which interfere NEMATODES
with examinations - referred as ASCALMINTHES
Barium Abrasive action, which may destroy - they are round and embossed
organisms; interferes with
- they are huge parasites and multicellular
examinations
Kaolin Abrasive action may affect
compounds appearance of organisms. Wait for Characteristics:
7-10 days after these compounds 1. System of the parasite
have been given before collecting
specimens for parasitologic A. Integumentary system
examinations - known as SYNCYTIUM / CUTICLE
Antibiotics Cause decrease in numbers of Composed of 3 muscle bands/layers:
protozoa in intestinal tract; wait 1 to 1. INNERMOST- longitudinal
2 weeks after termination of therapy 2. MIDDLE LAYER- oblique
before collecting specimens 3. OUTER LAYER- circular
3. Age Movement:
- unpreserved specimens should reach the Adult parasite - Figure of 8
laboratory within 1-2 hours after passage Larva- Purposely motion (they can go anywhere)
-time and date of passage should be recorded

 PRESERVATIVES B. Nervous System


1. Formalin (5-10%) - has a nerve center referred to as the ganglia
-preserves eggs, larvae and cysts for wet seen in the esophageal area or the CIRCUM
mount examinations ESOHAGEAL RING

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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
- nerve center transmit nerve impulses or a. Bipartile: 2 lobes (ex. Necator
senses to the different parts of the parasites americanus)
through TRANSVERSE COMMISURE b. Tripartile: 3 lobes (ex. Ancylostoma
- Parasite would access into one organ referred spp.)
to as CHEMORECEPTOR 3. Cleft or the cleavage
Caudal Chemoreceptors: Phasmids 4. Spicule- most important because it is the
Other Chemoreceptors: Aphasmids penis of the parasite
- It is the copulatory organ
C. Circulatory System - For nematodes it contains one spicule and
- It is considered to be absent, the fluid inside other parasite contain multiple spicules
the parasite works the function of the blood - Some of the spicules are barbed or fused (ex.
N. americanus), Ancylostoma is plenty or
D. Digestive System unfused
- known as a Complete system:
Mouth or Buccal capsule Female Reproductive Organ
Esophagus Organ: Genital primordium/ Gynecophoral canal
Alimentary canal - Found/ along one side of the parasite,
Anus (female parasite)/Cloaca (male probably ¼ to ½ anterior to the body
parasite)
- Referring to the mouth or the buccal capsule Sexual position: inverted Y position
of the parasite it is accessorized.
 Dorsal Cutting Plates- ex. Necator The central dogma of the life cycle of
americanus nematodes:
 Teeth- ex. Ancylostoma spp.
1. Ova
 3 lips (trilobe)- ex. Ascaris
2. Larva
 Papilated lips- ex. Trichuris
3. Adult
E. Reproductive System
- Males separate from Female  Ova Stage
Ex. Ascaris lumbricoides male and Ascaris -Ova of the parasites would have 3 types
lumbricoides female 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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