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MEDICALLY IMPORTANT

PARASITES

UNICELLULAR

PARASITOLOGY REVIEW

PHYLUM
SUBPHYLUM
CHARACTERISTICS
Sarcomastigophora Sarcodina
Single celled
(amoeba)
Move by pseudopodia
T and C stages
Asexual reproduction
Mastigophora Same as Sarcodina
(flagellates)
But move by flagella
Some have T stage only
Blood and tissue
flagellates (APET)

MEDICALLY IMPORTANT
PARASITES

AMOEBA

UNICELLULAR
PHYLUM
Ciliaphora (ciliates)

CHARACTERISTICS
Single celled
Target tissue and blood cells
T and C stages
Asexual reproduction

Apicomplexa (sporozoa)

Same as the ciliates


Other stages include:
Oocyst,sporocyst, sporozoite,
merozoite,trophoaoite,schizont,
and gametocyte (OSSMTSG)

Entameoba histolytica
CYST
1-4 nuclei
4 nuclei in mature cyst
Cental karyosome/bulls eye
karyosome
Cigar-shaped chromatoidal
bodies

TROPHOZOITE
ONE nucleus (all trophozoite
uncer Sarcodina)
Fine, peripheral chromatin
Cental karyosome/bulls eye
karyosome
Ingested RBC (none for E.
disparr)
Pseudopodia
Progressive, directional
motility

Pathogenesis of E. histolytica

Ingestion of mature cyst


Excystation in the small intestine
Undergo binary fission
Trophozoite invade the membrane of cecum
(flask-shaped ulcer) bloody-mucoid stool
From the cecum trophozoite migrates to other
organ e.g. liver or encystation occur
Liver and lung amoebiasis

NONPATHOGENIC AMOEBAE

Entamoeba hartmanni

Entameoba coli
CYST
CB have splintered ends/
whisk-broom appearance

TROPHOZOITE
Largest among the amoeba

Eccentric karyosome

Eccentric karyosome

8 nuclei in mature cyst

Plenty of vacuoles with


ingested bacteria
(Dirty-looking cytoplasm)

NONPATHOGENIC AMOEBAE
Endolimax nana

Nucleus with eccentric karyosome


No peripheral chromatin
comma-shaped CB, if present
Ground glass cytoplasm
Punched-out /crossed-eyed nuclei

Iodamoeba butschlii

NONPATHOGENIC AMOEBAE

Glycogen mass, to 2/3 of its size


Stained deep-mahogany brown by Lugols Iodine
With achromatin granules
Cyst and trophozoite only possess a single nucleus
Mistaken as Blastocystis hominis

Resembles E. histolytica but non-pathogenic


Central or eccentric karyosome
No ingested RBC
small race
With rice grain like CB

Entamoeba gingivalis
Oral cavity
With ingested WBC
No cyst form

Opportunistic Amoebae
Naegleria fowleri
Primary amoebic encephalitis (PAM)-purulent
spinal fluid without bacteria
Swimming
3 stages:

Opportunistic Amoebae
Acanthamoeba sp.
Granulomatous amoebic encephalopathy
Associated with amoebic keratitis
With spine-like projections
Acanthopodia (locomotion)

Amoebic trophozoite
Flagellate trophozoite
cyst

A. culbertsoni- slightly wrinkled ectocyst


A. polyphaga-delicate looking acanthopodia
A. castellani-wrinkled ectocyst
A. astronyxis-folded ectocyst

FLAGELLATES

Giardia lamblia
Travellers diarrhea/ Leningrads curse
Trophozoite

Pear-shaped
Falling-leaf motility
Old man with eyeglasses
4 pairs of flagella

Cyst
Retracted cytoplasm
Axonemes appear as four pairs of curved
bristles

Giardia lamblia
Giardiasis
Carpet the intestinal mucosa---duodenum and
jejunum
Steatorrhea and Jaundice

Laboratory Diagnosis:
String test
Duodenal aspirate-specimen of choice
DFS

Chiomastix mesnili
Trophozoite

Pear-shaped trophozoite
Assymetric
Spiral groove at the posterior end
Cytosome (hour-glass appearance)
Exihibits boring/cork-screw motility

Cyst
Lemon-shaped
Cytostome present
Single nucleus

Trichomonas spp.

Species:
Trichomonas vaginalis - GUT
Trichomonas tenax - oral cavity
Trichomona hominis intestines

Frothy, creamy, mucopurulent vagiunal


discharge
Dysuria
Most common STP
Urine must be collected properly to avoid
fecal contamination

Dientamoeba fragilis
Irregular shape with
leaf-like pseudopodia
1-2 nuclei
Karyosome is
tetracoccic/fragmented
Progressive motility
Co-infection with
Enterobius vermicularis

HEMOFLAGELLATES
Leishmania donovani
Trypanosoma gambiense
Trypanosoma rhodesiense
Trypanosoma cruzi

HUMAN
(Leishmania sp. and T. cruzi)

(Trypanosoma sp.)

TRYPOMASTIGOTE

AMASTIGOTE

INSECT

PROMASTIGOTE

Pyriform shape
Undulating membrane (1/2 of its length)
Jerky-tumbling motility
Found in urine

Trichomoniasis

Trophozoite stage only


T. vaginalis is the only pathogenic

Sandfly (vector for Leishmania)

Trichomonas vaginalis

Tse-tse fly and Reduviid bugs (vector for


Trypanosoma sp.)

EPIMASTIGOTE

Leishmania donovani

Trypanosoma gambiense
Trypanosoma rhodesiense

TRYPANOSOMES
Trypanosoma gambiense
Mid-and West African sleeping sickness

Trypanosoma rhodesiense
East african sleeping sickness

Trypanosoma cruzi
Chagas disease

Trypomastigote infective stage to man


Tse-tse fly (Glossina sp.)
S/S of acute infection:
Winterbottoms sign
Kerandel sign

Sleeping Sickness
Parasite invades the CNS

Dx:
Blood (fever), BMA, LN juice, CSF

Trypanosoma cruzi
Metacyclic trypanosomes-infective stage to
man
Reduviid bugs (Panstrongylus, Triatoma, Rhodnius)
Stages of development:
APET-man
AET-insect

Reduviid/kissing bugs
Chagas disease

Trypanosoma cruzi
Diagnosis
Romanas sign-edema of face
Demo of Amastigote in tissues
Demo of trypomastigote in blood-bulging
kinetoplast, spindle or C-shaped
Xenodiagnosis

Destruction of the RES


cardiomegaly

CILIATES
Balantidium coli
Largest protozoa pathogenic to man
cecum
Trophozoite :
Cilia and cytostome
Large kidney-shaped macronucleus
Small spherical micronucleus

Cyst
Thick cyst wall, hardly stained
2 nuclei similar to trophozoite

COCCIDIA
Undergo both sexual and asexual
reproduction
Cryptosporidium parvum
Isospora belli
Sarcocystis hominis
Plasmodium falciparum
Toxoplasma gondi
Plasmodium malariae
Plasmodium ovale
Plasmodium vivax

Life Cycle
SPOROGONY
SCHIZOGONY

Gametes
combine

Trophozoite

Zygote

Schizont

Merozoite

Gametocytes

Oocyst
(with sporoblas)
Sporocyst
(with sporozoites)
Sporocyst
(into mature oocyst)

Cryptosporidium parvum
Animal parasite
One of the major opportunistic parasite in AIDS patient
(Cyclospora cayetanensis-recent)
Infects the intestine (brush border)
Oocyst has 4 sporozoites BUT NO SPOROCYST
Schizont produces 8 falciform merozoites
Diagnosis

Sheaters Sugar Flotation method


DFS for ID of oocyst
Enterotest to recover oocyst
Modified AFS (red stained oocyst)

Toxoplasma gondii
Toxoplasmosis
In adult and newborn
Resembles infectious mononucleosis, lymphadenopathies,
malaise, muscle pain, low-grade fever
Hypersensitivity
Mother to fetus transmission (tachyzoites) (transplacental
transfer and blood transfusion)

Morphology
Trophozoite pyriform of crescent shaped troph.
Tachyzoites 9acute) and bradyzoites (chronic)

Cat is the definitive host


Man is intermediate host

Isospora belli
Human coccidiasis
Asymptomatic and self-limiting
Mature oocyst-infective stage to man
Diagnosis:
DFS
Immature oocyst in stool
Leave at room temperature

Zinc Sulfate Method

Toxoplasma gondi
Diagnosis:
Complement fixation test
Sandwich ELISA
Sabin-Feldman dye test
Frenkel skin test
PCR

Malarial Parasites
Plasmodium falciparum
Most common in the Philippines

Plasmodium vivax
Second most common the Philippines

Plasmodium malariae
Occasionally seen in the Philippines

Plasmodium ovale
Endemic in Africa, West coast, Ethiopia

COMPARISON OF MALARIAL SPECIES


Plasmodium
Species

G-6-PD deficiency
Sickle cell anemia trait
PABA deficiency

Susceptibility to malarial infection


Duffy factor

Signs and symptoms are due to parasites coming


from the liver
P.vivax and P.ovale

Recrudescence
Signa and symptoms are due to surviving parasites
in RBC
P.falciparum and P.malariae

Number of
Merozoites
in Schizont

Other
Characteristic

Normal size
Multiple
infection
Maurers dot

Small, delicate
Double chromatin
dots
Applique forms
Dark pigments
Multiple parasites

12-36

Cresent-shaped
gametocyte
Ring and
gametocyte stages
only peripheral
blood

P.malariae
72 hours
Quartan LC

Normal size
Ziemanns dots
(rare)

Compact,
May assume band
form, or rosette

6-12

Wide range of
stages in peripheral
blood

P. Vivax
48 hours
Tertian life
cycle

Enlarged
Schuffners
dots

Amoeboid
Large vacuoles

12-24

Wide range of
stages in peripheral
blood, Infects
reticulocytes

P. Ovale
48 hours
Tertian life
cycle

Enlarged
Schuffners
dots

Compact
Golden-brown
(P.vivax), double
chromatin dots

6-16

Wide range of
stages in peripheral
blood

PATHOGENESIS
The infection mainly causes tissue hypoxia,
which is due to several pathway secondary to
the presence of the parasites, mainly inside
RBC.
The ERYTHROCYTIC MEROZOITES are the
pathogenic stage of the parasite.

Relapse-renewed parasitic and clinical


activity
Recurrence/ true relapse

Trophozoite

P. falciparum
36-48 hours
Tertian life
cycle

Malarial Parasites
Resistance to malarial infection:

Red Blood
Cell
Morphology

Manner of Transmission

Mosquito bite (Anopheles sp.)


Blood transfusion (no sporozoites)
Contaminated syringes and needles
Across the placenting during delivery

Pernicious anemia caused by severe


malarial infection
Algid malaria-low blood pressure, shock, renal
failure
Blackwater type-massive intravascular
hemolysis and hemoglobinuria
GI type-diarrhea and dysentery
Hemorrhagic type-oozing of blood in tiny
vessels of skin and mucous membranes

DIAGNOSIS of MALARIAL Infection


History of travel
Hepatosplenomegaly and or anemia
Malarial smear
Thin smear-examined within 15 min.
Thick smear-examined within 5 min (identification)
Trophozoite, schizonts, and gametocytes are seen in
PBS

Babesia microti

Infects RBC
Transmitted by ticks (Ixodes spp.)
Simultaneous transmission with B. burgdorferi
Mistaken for Plasmodia

Anemia
Malaise, fever, chills

QBC coated with acridine orange


WBC, platelets, Plasmodia, Trypanosoma,
Microfilariae
Blood collection for Malaria detection should be
made DURING FEBRILE STATES, antimalarial drugs
taken 4 days prior to blood collection will reduce
chances of recovering malarial parasites.

MEDICALLY IMPORTANT
PARASITES

MEDICALLY IMPORTANT
PARASITES

MULTICELLULAR
PHYLUM
Aschelminthes

SUBPHYLUM
Nematoda
(roundworms)

Platyhelminthes

Trematoda
(flukes)

CHARACTERISTICS
Adult of both sexes
Eggs, larva, adult
With of w/o intermediate
host

PHYLUM

SUBPHYLUM
Cestoda
(tapeworms)

Whip worm
Cecum
Ingestion of embryonated eggs
Adult male
Tail end is coiled 360 degrees

Female adult
Tail end is more or less straight

Egg
Foot-ball or barrel shaped with
protruding bipolar mucus plugs

CHARACTERISTICS
Ribbon-like body
Hermaphroditic
Egg,larva,adult
Mammals and insects are
intermediate hosts

Bilaterally symetric
Most are hermaphroditic
Egg,miracidium,cercaria,a
dult (EMCA)
Intermediate host (fish,
snails,crabs)

Trichuris trichiura

MULTICELLULAR

Trichinella spiralis

Trichina worm
pigs and rats are source of infection
Encysted larva-infective stage
Male and Female adult are thin and
threadlike
Male
Pair of conspicuous conical papillae

Female
Larger than male adult
Vulva about from anterior end

Encysted larva
Tightly coiled larvae between muscle
fibers enclosed by a thick capsule

Dx: muscle biopsy, Bach-mann


intradermal test, Bentonite
flocculation test

C.
philippinen
sis
T. spiralis

Eggs are peanut shaped, with flattened


bipolar plugs similar to those of Trichuris
With typical and atypical eggs

S.
stercoralis

* Eggs similar to those of hookworms but not


commonly seen in feces, eggs are ovoid
containing developed larva, commonly
known as Chinese Lantern Ova

Hookworm
Ova

* Eggs of all hookworm species are ALIKE;


ovoid with thin shell containing 2-8 germ
cells

Larva often encysted in striated muscles

Rhabditiform Larva

Filariform Larva

First stage larva


Feeding stage
With open mouth
Shorter and robust
Not infective

Second stage Larva


Non-feeding stage
With Closed mouth
Longer and slender
infective

Worm

Rhabditiform larva

Hookworm

With long buccal


cavity
With small genital
primordium

S. stercoralis

With short buccal


cavity
With large genital
primordium

Filariform larva
Sheathed
With pointed tail
With short
esophagus

Unsheathed
With notched/bifid
tail end
With longer
esophagus

N. americanus

No teeth, but with semi-lunar


cutting plates

A. duodenale

With two pairs of teeth

A. caninum

With three pairs of teeth

A. braziliense

With one pair of teeth

Male N. americanus

Male A. duodenale

With fused and barbed


copulatory burse
With bipartite/bidigitate
dorsal rays

With Tripartite and


Tridigitate dorsal rays
Copulatory spicules are not
fused

Important Properties of Microfilaria


1. It exhibits periodicity
2. Microfilaria may be sheathed or unsheathed

PARASITE

UNSHEATHED
MICROFILARIA

O.
volvulus

Nuclei do not
extend to the tip
of tail

M. ozzardi

Nuclei extend to
the tip of tail

M.
perstans

Nuclei do not
extend to the tip
of tail

MICROFILARIAE
PARASITE

SHEATHED
MICROFILARIA

W.
bancrofti

Nuclei do not extend


to the tip of the tail

B. malayi

With two distinct


terminal nuclei

L. loa

Nuclei extend to the


tip of the tail

Morphology of Ova
1.

The protozoan parasite with motility comparable to E. histolytica


a) E. coli b) E. nana c) G. lamblia d) B. coli d) D. fragilis

2.

The hookworm specie with the most number of buccal teeth


a) N. americanus b) A. caninum c) A. duodenale d) none of these

3.

The protozoan parasite with macronucleus and micronucleus


a) B. coli b) E. nana c) I. butschlii d) D. fragilis e) none of these

4.

The nematode egg flattened on one side containing motile larva is


produced by:
a) T. trichiura b) C. philippinensis c) E. vermicularis d) none

5.

Sheathed microfilaria with two terminal nuclei is seen in


a) W. bancrofti b) L. loa c) M. ozzardi d) B. malayi

P. westermani

Broadly ovoidal ; with flattened but prominent operculum


and with abopercular portion opposite the operculum and
contains undeveloped miracidium

C. sinensis

Broadly ovoid with wide operculum and abopercular knob,


contains developed miracidium
Eggs are said to resemble AN OLD FASHIONED ELECTRIC
BULB

O. felineus
H. heterophyes

Same with C. sinensis but elongately ovoidal

M. yokogawai

Same with H. heterophyes but thin shelled

F. hepatica, F.
buski and E.
ilocanum

Operculated and contains undeveloped miracidium

With narrow operculum and no abopercular knob ; thick


shelled

Adult Schistosoma

P. westermani
F. hepatica
F. buski
E. ilocanum

S. Japonicum

With smooth skin

S. Mansoni

With coarse tuberculations

S. haematobium

With fine tuberculations

Assumes a COFFEE BEAN


APPEARANCE
With CEPHALIC CONE & HIGHLY
BRANCHED intestinal ceca
Same with F. hepatica but NO
CEPHALIC CONE
Oral sucker with spines
CIRCUMORAL DISK WITH SPINES

C. sinensis

With DENDRITIC TESTES and


GRANULAR VITELLARIA

O. felineus

With LOBED TESTES and TRANSVERSE


VITELLARIA

H. heterophyes

With central 3rd sucker GENITAL


SUCKER

M. yokogawai

Without GENITAL SUCKER

OPERCULATED EGGS

Dipyllidium caninum

1. SCOLEX/ HEAD
1.1 spatulate /almond shaped/spoon shaped and with 2
false suckers BOTHRIA
* Diphyllobothrium latum
1.2 Globular with 4 true suckers sometimes with
rostellum and hooklets
WITH HOOKS:
* T. solium
* D. caninum
* H. nana
* E. granulosus
WITHOUT HOOKS
* T. saginata
* H. diminuta

2. SEGMENTS/PROGLOTTIDS
2.1 immature
2.2 mature
* T. solium with accessory ovarian lobe
* D. caninum with bilateral genital pores
2.3 gravid/ripe
* T. solium
elongated uterus with lateral
branches ( 8-15 )
* T. saginata 15-30 branches
* D. latum
rosette shaped uterus
* E. granulosus
loosely twisted uterus
* D. caninum uterus disintegrated into Packets
* H. nana
saccular uterus
* H. diminuta Saccular uterus

1.

The trematode specie that produces non-operculated egg with


unnoticeable lateral spine
a) S. haematobium b) S. japonicum c) P. westermani

2.

The adult fluke whose oral sucker is equipped with spines


a) E. granulosus b) E. ilocanum c) P. westermani d) none

3.

Egg packets are characteristic of


a) T. solium b) T. saginata c). D. caninum d) A. caninum

4.

Spatulate of spoon shaped scolex is observed in


a) F. hepatica b) D. latum c) D. caninum d) F. buski

5.

Eggs are said to resemble an old fashioned electric bulb


a) C. sinensis b) O. felineus c) M. yokogawai d) H. heterophyes

1.

Swimmers itch is due to cercarial penetration of what group


of trematodes?
a) Schistosoma b) Echinostoma c) Taenia d) none of these

2.

Microcytic hypochromic type of anemia may be induced by


a) D. latum b) Hookworms c) Pinworm d) Pudoc worm

3.

The so-called mystery disease is due to


a) E. vermicularis b) C. philippinensis c) T. trichiura

4.

Ingestion of T. solium eggs can lead to an infection known as


a) sparganosis b) Taeniasis c) Cysticercosis d) none of these

5.

The trematode specie that can cause TB like symptoms in a


host
a) D. latum b) E. granulosus c) F. hepatica d) none of these

REFRIGERATION
5% Formalin
10% Formalin
Polyvinyl Alcohol

Merthiolate Iodine
Formaldehyde
Sodium Acetate Formaldehyde

1.
2.
3.
4.
5.
6.
7.

Stool is examined to detect helminth egg, larva, adult and Protozoan


cyst and trophozoites
Acidic property of Urine kills motile protozoan trophozoites
Water promotes growth of other organisms and allows helminth
eggs to hatch
Formed feces usually contains protozoan cyst and helminth eggs
while liquid feces contains protozoan trophozoites
Protozoan stages appear in feces at intervals so if stool exam yielded
a negative result, stool must be again tested
Stool exam should be carried out ASAP, since protozoan trophozoites
may disintegrate
Motile Larva in fresh stool is diagnostic of S. stercoralis, however if
stool has been kept at room temperature for several hours/days,
hookworm eggs may hatch thus releasing also motile larva

SEDIMENTATIO
N

FLOTATION

Acid Ether Concentration


technique

Zinc Sulfate Centrifugal


Flotation

Formalin Ether Concentration


technique

Brines Flotation

Merthiolate Iodine Formalin


Concentration

Sheathers Flotation

3-5 degC for 4 hrs.


Protozoan cyst
Helminth eggs and Larva
Protozoan Cyst & Trophozoites,
used for Trichrome Stain

Liquid fixative with long shelf


life

1.

In the macroscopic exam, color and stool consistency is


noted

1.

In the preparation of DIRECT FECAL SMEAR


NSS, iodine may be used

3.

Permanent stains are used to study NUCLEAR STRUCTURES.


Examples are:
3.1 Iron Hematoxylin stain
3.2 Gomoris Trichrome
3.3 Modified AF stain

4. In the KATO THICK SMEAR Preparation, cellophanes are used


to cover the smear

1. MODIFIED KATO TECHNIQUE


2. STOLL DILUTION TECHNIQUE
3. FAUST MALONEY EGG HATCHING

6.

7.

8.

Stains for Blood Parasites include:


6.1 Giemsa stain
6.2 Wrights
6.3 Delafields Hematoxylin
Unstained blood preparation may also reveal Microfilaria ,
Trypanosoma and Malarial Parasites
KNOTTS concentration technique is used for Microfilaria

1.

Blood is examined for Plasmodium, Trypanosoma, Leishmania and


Microfilaria of Filarial worms

2.

Blood collection for Malaria detection should be made DURING FEBRILE


STATES, antimalarial drugs taken 4 days prior to blood collection will reduce
chances of recovering malarial parasites.

3.

T. gambiense and T. rhodesiense may be recovered in the blood only during


FEBRILE states, while during the SLEEPING stage CSF is examined instead.

4.

Blood collection for Microfilaria detection should take into consideration the
PERIODICITY of the suspected filarial specie.

5.

THICK SMEAR for malaria detection is used for RAPID DIAGNOSIS and needs
to be DEHEMOGLOBINIZE, while THIN SMEAR is used for SPECIE
IDENTIFICATION and should be FIXED with alcohol

Sabin Feldman Dye test

Toxoplasma gondii

Casonis test

Hydatid disease
/Echinococcus granulosus

Bachman Intradermal test Trichinella spiralis


Moan Hemagglutination
Technique

Entamoeba histolytica

Circumoval precipitin Test Schistosoma

1.

Blood collection for Malaria detection should be made


a) at the height of fever b) before fever
c) after fever d) at any time of the day

2.

Definite diagnosis of pinworm infection is through


a) urine exam b) perianal swab exam c) stool examn

3.

Sabin feldman dye test is used for


a) T. spiralis b) Schistosoma c) T. gondii

4.

Thick smear for Malaria is for


a) rapid diagnosis b) specie identification

5.

The following stains allow nuclear study of Protozoan


parasites except:
a) iodine b) Iron hematoxylin c) Trichrome d) none of
these