ML S MTA P 3

L E S SO N 2 : PRO TOZ OA

• Phylum Sarcomastigophora
o Amoeba & Flagellates
• Phylum Ciliaphora
o Ciliate
• Phylum Apicomplexa
o Sporozoa & Coccidia

INTESTINAL AND ATRIAL AMOEBA

• Entamoeba histolytica
• Entamoeba coli
• Endolimax nana
• Iodamoeba butschlii
• Entamoeba gingivalis

DEVELOPMENTAL STAGES NOTES 😊

• Non-motile • Laboratory Examination
• Usually infective o Stool (Fecal Smear) – E. histolytica, E. coli, E. nana,
• Found in FORMED feces I. butschlii
Cyst
• Resistant to damage o Oral Scrapings – E. gingivalis
• Usually smaller than trophozoite • Achromatic – non-stainable
• Best visualized using IODINE • Chromatoidal Bodies – stored food that can be stained
• Motile with iron hematoxylin → black
• Usually not infective • E. histolytica – an invasive parasite which can produce
• Found in LIQUID feces lesions and ulcerations, producing bleeding → blood
Trophozoite • Susceptible to damage; easily → RBC (HEMATOPHAGUS)
disintegrates • Trophozoite
• Usually larger than cyst o 1 nucleus
• Best seen using permanent stains o Pseudopodia – locomotion
(IRON HEMATOXYLIN → black)
▪ E. histolytica – long finger-like

DEVELOPMENTAL
AMOEBA HABITAT EFFECT TO HOST INFECTIVE STAGE MOT
STAGE
E. histolytica Pathogenic
E. coli Cyst & Ingestion / Fecal-
Colon Cyst
E. nana Non-pathogenic / Trophozoite oral route
I. Butschlii Commensal
E. gingivalis Buccal cavity Trophozoite Trophozoite Oral contact

MORPHOLOGICAL DIFFERENCES
CYST TROPHOZOITE
AMOEBA # OF CHROMATOIDAL CYTOPLASMIC
SIZE SIZE KARYOSOME MOTILITY
NUCLEI BODIES INCLUSIONS
Indistinct
Cigar or barr central Progressive
Entamoeba histolytica 10-20 um 1-4 12-60 um RBC
shaped “Bull’s aye directional
karyosome”
Splintered ends; Sluggish
Eccentric or
Entamoeba coli 10-35 um 1-8 with broomstick 15-50 um non-
off center
appearance directional
Blot like or
Endolimax nana 5-10 um 4 Small spherical 6-12 um Bacteria,
irregular
food
Surrounded
NONE vacuoles &
with
instead has a debris
achromatic
Iodamoeba butschlii 5-20 um 1 large glycogen 8-20 um
granules
mass → stain
(non-
with IODINE
stainable)
Food
Entamoeba gingivalis Unable to produce cyst 10-20 um vacuoles and
WBC

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CRITERIA FOR IDENTIFICATION OF

TROPHOZOITE 😊
1. Motility
2. Shape of pseudopodia
3. Cytoplasmic inclusions
4. Shape and location of
karyosome

DEVELOPMENTAL STAGES 😊

Metacyclic
Pre-cyst Cyst Trophozoite
trophozoite

Metacyclic
EXCYSTATION Cyst Trophozoite
trophozoite

ENCYSTATION Trophozoite Pre-cyst Cyst

ENTAMOEBA HISTOLYTICA

INTESTINAL AMEBIASIS
(stool exam)
1. ACUTE / SYMPTOMATIC
o Colon
o Bloody diarrhea, abdominal pain
o Patient passes out TROPHOZOITE in the feces
2. CHRONIC / ASYMPTOMATIC
o Stool consistency is NORMAL (FORMED)
o Patient are carriers
o Patient passes out CYST in feces

• Goes beyond the colon

• Trophozoite is TISSUE INVADING
EXTRAINTESTINAL AMEBIASIS
• (1) Hepatic amebiasis – Amebic Liver Abscess (ALA) → most common form
(serological tests)
• (2) Pulmonary amebiasis – lungs
• (3) Cerebral amebiasis – brain

EXTRAINTESTINAL AND ZOONOTIC AMOEBA

• Entamoeba dispar
• Entamoeba hartmanni
• Entamoeba polecki

CYST
AMOEBA TROPHOZOITE NOTES
SIZE # OF NUCLEI
Currently, the closest morphologically
Entamoeba dispar 10-20 um
wise to E. histolytica
4 Bacteria, no RBCs Back then is called small-race
Entamoeba hartmanni 5-10 um histolytica → differentiate through
immunologic or molecular asatys
Entamoeba polecki May appear like E. histolytica but is zoonotic infecting pigs & monkeys

NOTES 😊
• These are species appearing like E. histolytica
• If you are not sure during fecal smear microscopy
report as → (+) E. histolytica/E. dispar

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FACULTATIVE AMOEBA

• Naegleria fowleri
• Acanthamoeba spp.
• Balamuthia mandrillaris

CYST
AMOEBA TROPHOZOITE NOTES
SIZE # OF NUCLEI
• Ameboflagellate → can
produce cyst and trophozoite
which has two forms: (1)
AMEBOID, (2) FLAGELLATED
• Thermophilic → can survive
• Measures 10-35 um in warm bodies of water
• With amoeboid and • Infective Stage: Amoeboid
flagellated forms trophozoite
• Organs for locomotion • Dx Stage: Trophozoite in CSF
o FLAGELLA and brain tissues; flagelalted
o LOBOPODIA forms occasionally in CSF
• Biphasic w/ ameboid • MOT: Diving or swimming in
Naegleria fowleri 7-15 um 1
(lobate pseudopodia) stagnant water
& flagellated (flagella) • MOE: Intranasal
forms • Cyst is NOT SEEN in brain
• Ameboid lobate • Cyst has a SMOOTH double
pseudopodia wall
• Rapidly motile • Flagellated forms with 2
flagella and is RAPIDLY
MOTILE
• Laboratory Diagnosis
o Primary Amebic
Menigoencephalitis
(PAM)
• Ubiquitous → can be found
everywhere
• Infective Stage: Cyst &
Trophozoite
• Dx Stage: Cyst and
Trophozoite in tissues and
CSF
• Cyst has a OUTER WRINKLED
• Measures 15-45 um double wall
• With spine/ filiform • Trophozoite has spine like
like projections process called
Acanthamoeba spp. 10-25 um 1
• Organ for locomotion ACANTHOPODIA which is
→ ACANTHOPHODIA SLOWLY MOTILE
• Slowly motile • Laboratory Diagnosis
o Granulomatous amebic
encephalitis (GAE)
o Amebic keratitis – soft
contact lens wearers
o Skin lesions
o Culture with NON
NUTRIENT AGAR (NNN)
seeded w/ E. coli
• Isolated from soil and dust from autopsy specimens of infected humans &animals
• Infective Stage: Cyst and Trophozoite
• Can enter through
Balamuthia mandrillaris o (1) Nasal passages
o (2) Ulcerated or broken skin
• Cyst and trophozoite in tissues
• Cyst is 10-25 um with DOUBLE CYST WALL

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FLAGELLATES

• Gardia lamblia
• Chilomastix mesnili
• Enteromonas hominis
• Retortamonas intestinalis
• Dientamoeba fragilis
• Pentatrichomonas hominis
• Trichomonas vaginalis
• Trichomonas tenax
• Leishmania
• Trypanosoma

DEVELOPMENTAL
FLAGELLATE HABITAT EFFECT TO HOST INFECTIVE STAGE MOT
STAGE
INTESTINAL
G. lamblia Pathogenic
Small intestine
C. mesnili
Non-pathogenic
R. intestinalis Cyst & Trophozoite
Ingestion
Non-pathogenic
E. hominis
Large intestine Pathogenic if ↑
D. fragilis
Non-pathogenic Trophozoite
P. hominis
ATRIAL
T. vaginalis GUT Pathogenic Sexual contact
Trophozoite
T. tenax Buccal cavity Non-pathogenic Oral contact
ATRIAL AND LUMINAL FLAGELLATES
CYST (DIAGNOSTIC STAGE)
FLAGELLATE
SHAPE AND SIZE # OF NUCLEI NOTES
Ovoid, ellipsoidal
Gardia lamblia 4 With pair of parabasal body
8-15 um
Lemon or Nipple Curved fibril alongside of cytostome,
Chilomastix mesnili 1
7-10 um usually referred to as a lemon/nipple cyst
Ellipsoidal Resembles cyst of E. nana – a binucleate
Enteromonas hominis 4
6-8 um condition
Ovoid or Pyriform Fibrils extend above nucleus giving it a
Retortamonas intestinalis 1
4-7 um bird’s beak fibrillar appearance

ATRIAL AND LUMINAL FLAGELLATES

TROPHOZOITE
FLAGELLATE
# OF NUCLEI # OF FLAGELLA MOTILITY NOTES
Trophozoite is bilaterally
Falling, leaf-like, kite-
Gardia lamblia 2 8 (4 pairs) symmetrical; old man with
like
eyeglasses
3 – external Stiff, rotary, corkscrew,
Chilomastix mesnili 1 Shepherd’s crook appearance
1 – internal spiral jerky
3 – anterior
Enteromonas hominis 1 Jerky slow directional Rare
1 – posterior
1 – anterior Rapid directional
Retortamonas intestinalis 1 Rare
1 - posterior corkscrew

Shepherd’s crook appearance (NEMATODE) Shepherd’s crook appearance (BACTERIOLOGY)

• Mansonella streptocerca • Mycobacteria kansasi (photochromogen)

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Gardia lamblia
• Note axostyle
that does not
go beyond the
body

FLAGELLATES WITHOUT CYSTIC FORMS

Dientamoeba fragilis
(luminal)
Pentatrichomonas hominis
(luminal)
Trichomonas texax
Trichomonas vaginalis
• Previously an amoeba but now a flagellate
• Trophozoite is 5-15 um
• With multiple hyaline, leaf-like pseudopodia
• Karyosome is composed of a cluster of 4-8 granules; often referred to as
TETRAKARYOSOME
• 80% has 2 nucleus; 20% has 1 nucleus
• P. hominis (commensal)
o 5-15 um
o Large intestine
o Ingestion
• T. tenax (commensal)
o 5-12 um (smallest)
o Buccal cavity
o Oral contact
• T. vaginalis (pathogenic)
o 7-23 um (largest)
o Vagina
o Sexual contact
• With RAPID JERKY motility
• Equipped with:
o Undulating membrane → enables to move in a circular manner; responsible
for rotatory motion
o 5 flagella
• Axostyle EXTENDS BEYOND the body

NOTES 😊 NOTES 😊
• Gardia lamblia → causes giardiasis, lambiasis or • Giardiasis
traveler’s diarrhea, can cause GAY BOWEL SYNDROME o Stool exam → fats are seen
among homosexual men o Compared to E. vermicularis it is not tissue
• Enterobius vermicularis → may carry D. fragilis invading and will NEVER cause bloody diarrhea
trophozoites in its eggs
• T. vaginilis → can cause trichomoniasis [intense
itchiness, burning sensation, foul smelling vaginal
discharge], vaginitis, urethritis, prostatitis

HAEMOFLAGELLATES – BLOOD AND TISSUE FLAGELLATES

AMASTIGOTE / LEISHMANIA
PROMASTIGOTE / LEPTOMONAS
EPIMASTIGOTE / CRITHIDIA
TRYPOMASTIGOTE / TRYPANOSMOA
DEVELOPMENTAL STAGES
(APET)
Oval shaped with nucleus & kinetoplast but NO undulating membrane and flagellum
Elongated with nucleus, kinetoplast, flagellum but NO undulating membrane
With nucleus kinetoplast, flagellum & undulating membrane
Kinetoplast is ABOVE the nucleus
With nucleus kinetoplast, flagellum & undulating membrane
Kinetoplast is BELOW the nucleus

HAEMOFLAGELLATES 😊 HAEMOFLAGELLATES 😊
• (1) Morphologically indistinguishable • (4) Intracellular parasites
• (2) Needs vector: arthropod transmitted o L. donovani → visceral (spleen, liver, BM, spleen)
o Sandflies: Genus Phlebotomus o L. tropica → skin
• (3) MOT: Vector bite, skin inoculation o L. braziliensis → mucous membranes of nose,
• (4) Intracellular parasites mouth pharynx, larnyx

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INFECTIVE
DEVELOPMENTAL
HAEMOFLAGELLATE VECTOR STAGE DISEASE CAUSED NOTES
STAGES
• Most severe
with the
Kala-azar Fever / involvement of
Visceral internal
leishmaniasis organs
Dumdum Fever/
• Fever
Leishmania donovani Death fever
• Anemia → BM
Tropical
involvement
splenomegaly /
Black disease • Hepatospleno
megaly
• Blackening of
skin
• Develop of
highly
disfiguring
leprosy like
lesions
Phlebotomus Amastigote particularly on
Promastigote Muco-cutaneous
spp. Promastigote the oronasal
Leishmaniasis
and
Leishmania braziliansis Espundia, Uta, pharyngeal
Chiclero Ulcer mucosa →
usually sa face
which leads to
deformity
• Deforming of
nose “TAPIR
NOSE’
• Widespread
Cutaneous lesions (not
Leishmaniasis/ painful) that
Delhi Ulcer takes about 1
Leishmania tropica
Baghdad Boil, year to heal
Aleppo • Lesions leave
ugly scar on
healing

NOTES 😊
• DIAGNOSIS
o DX Stage: Amastigote / Leishmania
o L. donovani → examination of BM, liver, spleen and lymph node aspirate
o L. tropica → skin biopsy
o L. braziliensis → tissue biopsy
• LIFE CYCLE
o MAN—Amastigote; VECTOR—Promastigote

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DEVELOPMENTAL
HAEMOFLAGELLATE VECTOR INFECTIVE STAGE DISEASES CAUSED
STAGE
Bruzi Complex
Trypanosoma brucei West African sleeping
gambiense Glossina spp. Metacyclic sickness
Epimastigote
trypanosome
Trypanosoma brucei (Tse tse fly) Trypomastigote East African sleeping
(Transitory stage)
rhodesiense sickeness

Chaga’s disease
(1) ACUTE Phase –
Kissing / Assassin Bug Chagoma &
Reduviid Metacyclic Romana’s sign
Trypanosoma cruzi All 4 stages
Triatoma trypanosome (2) CHRONIC Phase –
Panstrongylus spp. cardiomegaly,
vital organs are
affected
NEW WORLD TRYPANOSOMIASIS
OLD WORLD TRYPANOSOMIASIS
SOUTH AMERICAN TRYPANOSOMIASIS
AFRICAN SLEEPING SICKNESS
CHAGA’S DISEASE

• Lesion at bite site: Trypanosoma chancre

• Blood invasion → fever
o Spx: Blood • Lesion at bite site: Chagoma
• Lymph node → Winterbottom sign (enlargement of lymph • Acuter Phase
node) o Unilateral conjunctivitis or orbital edema called
o Spx: Lymph node aspirate Romana’s sign
• CNS → Kerandel’s sign o Examine blood
o Spx: CSF o Dx Stage: Amastigote and Typomastigote
• RAPIDLY FATAL → East African Sleeping Sickness
• LONGER COURSE → West African Sleeping Sickness

CHARACTERISTICS OF BRUCEI COMPLEX 😊 DEVELOPMENTAL STAGES 😊

• (1) Morphologically indistinguishable
• (2) Arthropod borne (Tse tse flies; Glossina spp.)
• (3) MOT: Skin inoculation
• (4) Epimastigote (2-3 days) → Metacyclic trypanosome
(infective stage)
• (5) Extracellular parasites (blood → lymph node → CNS)
• (1) Epimastigote – develops in the VECTOR but after 2-3
days becomes METACYCLIC TRYPANOSOME (infective
stage) → only a transitory stage; not permanent
• (2) Trypomastigote – develops in MAN (Dx stage)

NOTES 😊
• Trypanosoma cruzi
o Infective stage is transmitted when bug bites and defecates on the wound made by its bite
o Passed out in bug’s feces → metacyclic trypanosome
o Trypomastigates in “C” – “U” shape
o Laboratory detection of Chaga’s disease: Xenodiagnoses

ADDITIONAL INFORMATION
E. histolytica (Trophozoite) Cytoplasm with ground glass appearance
E. nana (Trophozoite) May mimic E. hartmanni and D. fragilis
I. butschlii Intestinal amoeba with BASKET NUCLEUS
G. lamblia Entero test (+) / String test (+)
E. hominis Resembles E. nana cyst
R. intestinalis Resembles C. mesnili cyst
E. dispar May appear like E. histolytica but will produce no symptoms and NOT invasive
Flies and Cockroaches Mechanical vector of E. histolytica
E. histolytica (Trophozoite) Produces FLASK SHAPED ulcers

Intestinal pathogen, non-invasive but can cause malabsorption syndrome in the host
G. lamblia
May induce excretion of FATS in feces – STEATORRHEA

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CILIATE

• Common parasite of pigs

Balantidium coli
• Habitat: Large intestine
• Regarded as the largest intestinal protozoa → even under LPO cyst and
trophozoite can be seen
• Cyst has MACRONUCLEUS (kidney shaped) for vegetation and MICRONUCLEUS
(spherical) for reproduction
• Trophozoites show directional, tumbling motility
• Organ for locomotion: CILIATE
• CYTOSTOME – cell mouth → food ingestion
• CYTOPYGE – cell anus → waste excretion
• Causes balantidiasis or balantidial dysentery, a tissue invading protozoa →
produce lesions & lesions with wide neck and round bases
• May also cause BLOODY STOOL
• Stool exam
• Dx Stage: Cyst, Trophozoite

COCCIDIAN / SPOROZOAN

• Plasmodium falciparum
• Plasmodium malariae
• Plasmodium ovale
• Plasmodium vivax
• Plasmodium knowelsi

MALARIAL PARASITES
GENERAL CHARACTERISTICS
• Intraerythrocytic
o Infect RBCs
o P. malariae → Mature or older RBCs
o P. vivax & P. ovale → Young or juvenile or immature RBCs (e.g., reticulocytes)
o P. falciparum P. knowelsi → RBCs of all ages
• Arthropod-borne female Anopheles mosquitoes
o Male Anopheles can NEVER be a vector
• MOT
o Primary mode: Bite of arthropod (skin inoculation)
o Other modes of transmission: Transplacental (mother to fetus)/vertically, blood transfusion
• Infective Stage
o Man: SPOROZOITES (introduced through bites)
o Vector: GAMETOCYTES (male & female; ingested during blood sucking; immature sex cells)
• Life cycle has 2 phases
o (1) SEXUAL or SPOROGONY cycle – happens in the vector
o (2) ASEXUAL or SCHIZOGONY cycle – happens in man
NOTE: Kapag nainfect ka, it will not go directly to the RBCs but the liver cell first
▪ (2.1) EXO-ERYTHROCYTIC cycle – within hepatic/liver cells
▪ (2.2) ERYTHROCYTIC cycle – within RBCs
▪ NOTE: No exoerythrocytic stage during vertical and blood transfusion
• Requires 2 hosts
o Definitive Host: FEMALE ANOPHELES MOSQUITO – host for the sexual/sporogony cycle
o Intermediate Host: MAN – host for the asexual cycle/schizogony
• Diagnosis: Examination of Thick and Thin Blood Smear (Gold Standard)
o THICK SMEAR is for RAPID DIAGNOSIS to determine if blood is (+) for malaria and must be DEHOMOGLOBINIZED prior to
staining
o THIN SMEAR is for SPECIES IDENTIFICATION and must be FIXED WITH ALCOHOL prior to staining (GIEMSA – stain for blood
parastes)
o Best time to collect blood for Malaria is at the height of fever

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MALARIAL PARASITES
GENERAL CHARACTERISTICS
• Paroxysm
o Classical symptom of malaria
o Characterized by fever, chills, and sweating

(｡•̀ᴗ-)✧
• Plasmodium vivax and Plasmodium ovale – Can infect young,
immature parasites
o Bata pa kaya VOVO pa
• All malarial parasites have ROUND/OVAL gametocytes except P.
falciparum → CRESECENT, SAUSAGE, BANANA shaped
gametocytes

DIFFERENTIATION OF MALARIAL PARASITES

Plasmodium falciparum Plasmodium vivax Plasmodium malariae Plasmodium ovale
Young, immature, Young, immature,
Intraerythrocytic RBCs of all ages juvenile RBCs juvenile RBCs
Mature RBCs
Can infect: (Fokfok parasite) i.e., Reticulocytes i.e., Reticulocytes
(VOVO parasite) (VOVO parasite)
Appearance of
No change in size Enlarged Reduced in size Enlarged
Parasitized RBCs
RBC Stippling
When inside the
RBC, the parasite Schuffner's dots /
Maurer's spots Schuffner's dots Ziemann's dots
consumes James's dots
(Ferdinand Marcos) (Vilma Santos) (Manila Zoo)
hemoglobin and (Orange Juice)
produces
stippling

Length of Asexual
Cycle
36-40 hours 48 hours 72 hours 48 hours
Rupturing of
RBCs every:
Stages Seen in Ring and gametocyte Ring, gametocytes, merozoite
the Blood (Dx stage) ALL STAGES MAY BE SEEN

Malignant tertian
(MOST SEVERE MALARIA
Diseases Caused → kase hindi choosy; it Benign tertian Quartan tertian Ovale malaria
can infect almost 7.5M
RBCs or half of it)

(｡•̀ᴗ-)✧ PLASMODIUM KNOWLESI (｡•̀ᴗ-)✧ (｡•̀ᴗ-)✧ ADDITIONAL INFORMATION (｡•̀ᴗ-)✧

• The 5th human malaria causes SIMIAN malaria
• 24 hrs cycle
• Infect any RBCs regardless of age; infected RBCs are
usually normal in size
• No Schuffner’s dots; with 16 merozoites and round
gametocytes
• Young/early stages mimic P. falciparum
• Mature/late stages mimic P. malariae
• Emerging spp.
• Paroxysm – classical sign of malaria characterized by
chills, fever and sweating
• DUFFY NEGATIVE individuals may show resistance to P.
vivax infection
• P. falciparum → Associated with G6PD deficiency
• P. vivax, P. ovale → May cause relapse due to persistence
of hypnozoites
o NORMAL: Cryptozoic schizont → merozoites (liver
cell) → rupture
o In VOVO parasites – it does not rupture and the last
stage is called HYPNOZOITES (dormant stage)
• Individual with SICKLE CELL ANEMIA trait may show
resistance to P. falciparum
• Blackwater fever → complication that may develop
following P. falciparum infection characterized by rapid
loss of hemoglobin through the urine

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(つ✧ω✧)つ OTHER PROTOZOA (つ✧ω✧)つ

• Cyclospora cayatenensis → a cyanobacteria like body that shows AUTO-FLUORESCENCE

• Babesia microti → intraerythrocytic parasite transmitted thru TICK BITES (Ixodes spp.) and can develop trophozoites with
MALTESE CROSS appearance; may resemble P. falciparum
• Cryptosporidium parvum → can cause mild diarrhea among healthy people but fatal diarrhea in those who are
immunocompromised
• Oocyst is both the infective and diagnostic stage of C. parvum, C. cayatenensis (DIAGNOSIS: Stool exam)
• Toxoplasma gondii → a protozoan parasite requiring feline/cats as the definitive host, can be detected thru SABIN FELDMAN
dye test (serologic test)
o MAN serves as its intermediate host
o Infect INTESTINAL EPITHELIAL CELLS
o MOT:
▪ (1) Transplacental
▪ (2) Ingestion of oocyst passed out in cat’s feces
▪ (3) Ingestion of infective cysts from the meat of contaminated animals
• TORCHES – can be transmitted transplacentally; cause congenital infection
o Toxoplasma
o Rubella (German measles)
o Cytomegalovirus
o Herpes
o Syphilis

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