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Protozoan parasites
At the end of this unit, the student
is able to:
 Classify the Protozoans
 Describe the morphology of each protozoa
 Explain the pathophysiology, life cycle, infective stages,
modes of transmission, epidemiology, prevention and
control
 Describe the diagnostic features of each parasite.
 Enumerate the different specimens used for identification
 Identify correctly the diagnostic features of each parasite.
 Perform the laboratory tests
 Discuss specimen collection,transport, processing,
preservation and disposal
General Characteristics
 They are single-celled organisms which are generally
classified according to their organelles of locomotion.
 Cilia, flagella & pseudopodia
 Considered as the simplest forms of animals
Organelles
 Nucleus
 Nucleulos
 Mitochondria
 Locomotory Organelles
Kingdom Protista
 Phylum Sarcomastigophora
 Subphylum Mastigophora
 (Superclass Rhizopoda)
Flagella (whip-like) = organ of locomotion
 Subphylum Sarcodina
 (Class Zoomastigophora)
Pseudopodia (hyaline foot like)= organ of locomotion
 Phylum Ciliophora
Cilia (hair like) = organ of locomotion
 Balantidiasis (Balantidium coli)
Kingdom Protista
 Phylum Apicomplexa
 w/ apical complex at the anterior end (apicoplast)
 Babesiosis (Babesia)
 Malaria (Plasmodium)
 Coccidian diseases including:
 Cryptosporidiosis (Cryptosporidium parvum)
 Cyclosporiasis (Cyclospora cayetanensis)
 Toxoplasmosis (Toxoplasma gondii)
 Phylum Microspora
 Spore-forming parasites
Protozoan parasites
Protozoan parasites
Protozoan parasites
Giardia lamblia Amoeba Amoeba feeding
Kingdom Protista
 Subkingdom Protozoa
 automonous unicellular eucaryotic organisms
possessing various degrees of cellular
specialization and some type of motility
 Organelles specialize in nutrient storage,
excretion, digestion, motility
 Phylum = based upon motility
Classification of Protozoa and Descriptions
 Size 2 - 100 micrometers
 Parasitic protozoa are most facultative anaerobes;
 Mostly heterotrophic, chemoorganotrophs
 Nutrients assimilated via phagocytosis, pinocytosis, diffusion
 Digestion may be both extracellular and intracellular
 Two life stages
 Trophozoiteactive pathogenic vegetative stage
 Cyst latent survival form
 Sites of pathogenesis
 Intestinal, Urogenital, Blood and Tissue
Pathogenic Protozoa - Characteristics
 Phylum Sarcodina
 Motility via Pseudopoda
 Extensions of the cell membrane into which the
cytoplasm flows
 Cyclosis
 No cell wall; thus, no uniform shape
 Shape changes with movement
 Reproduce asexually by mitosis; sexually by meiosis
 Trophozoite
 Motile vegetative stage
 Cyst
 Latent survival stage
Protozoa - Amoeba
 Phylum Ciliata
 Motility via cilia
 Cilia surround the cell; their co-ordinated movement
 Uses energy from the cell membrane
 Synchronized ciliary movement
 Two nuclei per cell
 Small micronucleus governs sexually reproduction
 Large macronucleus governs metabolism and
growth
 Both nuclei divide during asexual mitosis
 Intracellular organelles
 Function in digestion and excretion
 Only one ciliate pathogen affects humans
Protozoa - Ciliates
 Phylum Mastigophora
 Motility via flagella
 One or more whip-like filaments move in counter-
clockwise fashion creating rapid movement
 A flagellum is a complex organelle composed of
many smaller tubules
 Flagella attach to the cell membrane and derive
energy from movement from this membrane
 Most numerous of the protozoan types
 Several are pathogens of humans
 All reproduce asexually; some reproduce sexually
Protozoa - Flagellates
 Phylum Sporozoa
 No established means of movement
 Thus, not motile
 All are intracellular parasites in a variety of hosts
 Several parasitic species affect humans
 Most are in the Class Coccidia
 All lack intracellular organelles
 All have three things in common
 No motility
 Sexual and Asexual mode of Reproduction
 Intracellular parasites
Protozoa - Sporozoans
Classification of Protozoan Parasites:
Sarcomastigophora
Sarcodina Acanthamoeba castellani, Endolimax nana
Entamoeba coli, Entamoeba dispar
Entamoeba gingivalis, Entamoeba histolytica
Iodamoeba butschlii, Nagleria fowleri
Mastigophora Chilomastix mesnili, Dientamoeba fragilis
Giardia lamblia,Trichomonas (homini, tenax
,vaginalis)
Leishmania (braziliensis, donovani, tropica)
Trypanosoma brucei complex, Trypanosoma cruzi
Ciliophora Balantidium coli
Apicomplexa Bebesia spp., Cryptosporidum hominis, Cyclospora
cayetansis, Isospora belli, Plasmodium spp.,
Toxoplasma
Microspora Enterocytozoon bineusi, Encephalitozoon spp.
Vittaforma cornea,Trachipleistophora hominis
Pleistophora spp., Brachiola vesicularum,
Microspordium spp.
Intestinal
 Amebiasis .......................... Entamoeba histolytica
 Giardiasis .......................... Giardia lamblia
 Balantidiasis .......................... Balantidium coli
 Crytosporidosis ...................... Cryptosporidium parvum
 Urogenital
 Trichomoniasis ....................... Trichomonas vaginalis
 Blood and Tissue
 Malaria ................................... Plasmodium spp
 Meningoencephalitis ............... Naegleria fowleri
 Toxoplasmosis ........................ Toxoplasma gondii
 Trypanosomiasis
 African Sleeping Sickness ... Trypanosoma brucei
 Chagas Disease .................. Trypanosoma cruzi
 Leishmaniasis
 visceral leishmaniasis( Kala-azar) ... Leishmania donovani
 cutaneous leishmaniasis .......... Leishmania topica/braziliensis
Protozoan Diseases
AMOEBAS
Intestinal and Luminal Protozoa
 Constantly changing shapes, move by
ending out fingerlike extensions of their cell
membranes called pseudopods or “false-
feet” projections
 the name comes from the Greek word for
“change” is an example of a sarcodine,
found in fresh water or salt water
 six species have been established as parasites of man:
Entamoeba histolytica
Entamoeba coli
Entamoeba gingivalis
Dientamoeba fragilis
Endolimax nana
Iodamoeba butschlii
Intestinal and Luminal Protozoa
 Stages of Development
Cyst – infective stage
Trophozoite – vegetative stage
General Statements:
 All amoebas are capable of producing cyst and
trophozoite except for Entamoeba gingivalis
and Dientamoeba fragilis.
 Found in the colon except Entamoeba
gingivalis which is found in the oral cavity
 All are commensals except Entamoeba
histolytica.
Entamoeba histolytica
A) Trophozoite Stage (I & H preparation)
 shape: irregular
 shape: 10– 60 micra
 structures:
- cytoplasm is grayish violet with a clear
demarcation between the ectoplasm
and endoplasm
Protozoan parasites
Protozoan parasites
Protozoan parasites
Protozoan parasites
Entamoeba histolytica
B. Cystic Stage
 shape: spherical
 size: 5 – 20 micra
 structures:
- smooth, definite, relatively thin cystic
wall
Entamoeba histolytica
B. Cystic Stage
the number of nuclei range from 1-4,
rarely more, with a centrally located
karyosome
in the younger cyst, the chromatoidal
bars with rounded ends and glycogen
mass may be present
Protozoan parasites
Protozoan parasites
Protozoan parasites
Entamoeba histolytica
Diseases:
Amoebiasis
Amebic dysentery
Amebic hepatitis
Pulmonary amoebiasis
Entamoeba histolytica
Entamoeba histolytica
Diagnosis:
 Microscopy
 Antigen detection
 Serology
 PCR
 Imaging studies
Entamoeba histolytica
Treatment:
 metronidazole
 tetracycline with diiodohydroxyquin
Entamoeba coli
A. Trophozoite Stage (I & H
preparation)
 shape: condensed, thickened,
rounded mass
 size: 15 – 50 micra
Entamoeba coli
A. Trophozoite Stage (I & H preparation)
 structures:
 cytoplasm is grayish violet; there is no clear
demarcation between the ectoplasm and
endoplasm
 the endoplasm is highly vacuolated containing
bacteria
 the nucleus is spherical with thick nuclear
membrane lined with coarse, irregular chromatin
granules or plaques; the karyosome is moderately
large and eccentrically located
Protozoan parasites
Protozoan parasites
Entamoeba coli
B. Cystic Stage
 shape: spherical
 size: 10 – 33 micra
Entamoeba coli
B. Cystic Stage
 structures:
 smooth, definite, thick cystic wall
 number of nuclei range from 1 – 8 rarely
more with an eccentric karyosome
 in younger cysts, chromatoidal bars with
splintered ends and glycogen mass
Protozoan parasites
Endolimax nana
A.Trophozoite Stage (I & H preparation)
 shape: irregular
 size: small, 6 – 15 micra
Endolimax nana
A.Trophozoite Stage (I & H preparation)
 structures:
 cytoplasm is grayish-violet, vacuolated
which may contain bacteria
 there is small, spherical, single nucleus
with a large irregular, central or eccentric
karyosome
Protozoan parasites
Endolimax nana
B. Cystic Stage
 shape: oval
size: 6 – 10 micra
Endolimax nana
B. Cystic Stage
 structures:
 smooth, definite cystic wall
 number of nuclei ranges from 1-4
 I & H preparation
 cytoplasm is grayish, granular
 nuclei are visible as darkly stained dots
 chromatoidal bodies occasionally seen as
deeply stained, small, slightly curved rods
Protozoan parasites
Protozoan parasites
Entamoeba gingivalis
A.Trophozoite Stage (I & H preparation)
 shape: irregular
 shape: 5 – 35 micra
 structures:
 clear demarcation between the clear
ectoplasm and granular endoplasm
Entamoeba gingivalis
 endoplasm contains food vacuoles with
bacteria and rarely rbc
 possesses a single nucleus which is
spherical in shape, karyosome is near the
center
 have multiple pseudopodia with
progressive directional movement similar
to E. histolytica
Protozoan parasites
Dientamoeba fragilis
A.Trophozoite Stage (I & H preparation)
 shape: irregular
 size 3 – 22 micra
A.Trophozoite Stage (I & H preparation)
 structures:
cytoplasm is granular with vacuoles
containing bacteria
nuclei, commonly 2 in number, with
karyosome consisting of 4-8 chromatin
granules
Dientamoeba fragilis
Protozoan parasites
Iodamoeba buetschlii
A.Trophozoite Stage (I & H preparation)
 shape: irregular
 shape: 6 – 25 micra
Iodamoeba buetschlii
A.Trophozoite Stage (I & H preparation)
 structures:
 granular, vacuolated cytoplasm and
vacuoles may contain bacteria and yeast
cells
 single, spherical nucleus with a large
central or eccentric karyosome
surrounded by an unstained halo
Protozoan parasites
Iodamoeba buetschlii
B. Cystic Stage
 shape: irregular
 size: 6 – 15 micra
Iodamoeba buetschlii
B. Cystic Stage
 structures:
smooth, definite cystic wall
have a single nucleus, rarely with 2
nuclei
large glycogen vacuole
Iodamoeba buetschlii
B. Cystic Stage
I & H preparation
violet-gray granular cytoplasm
nucleus is visible
large glycogen vacuole is unstained
chromatoidal bodies are usually
absent; if present, they are seen as
deeply stained small rods
Protozoan parasites
Protozoan parasites
Soil andWater Amoebas
A) Trophozoite Stage (I & H preparation)
endoplasm is briefly granular with
ingested rbc which are bluish black in
color
single, spherical nucleus; nucleus
membrane is lined by fine, regularly
arranged chromatin granules;
karyosome is relatively small and
centrally located
Symptoms:
intermittent diarrhea
abodominal cramps
vomiting
general body malaise
 Amebiasis( Amebic dysentery)
- colitis with diarrhea(somtimes bloody) accompanied by abdominal pain
and cramping: numerous bloody stools per day
 extraintestinal amebiasis a complication of intestinal amebiasis in
which the trophozoites enter the blood and are carried
to other organs; most often the liver
 these patients have fever, leukocytosis, rigors
 Parasite : Entamoeba histolytica
 Trophozoite : sarcodine protozoa which slowly move among the crypts
of the large intestine and are shed into the feces
 Cyst : latent survival stage which is significant in transmission;
spherical containing four characteristic nuclei; infective stage
 Pathogenesis the trophozoites bind to colonic epithelial cells and secrete a
cytotoxin which alters cell membrane permeability causing the cells to take in
calcium and lyse; necrosis and inflammation also accompanies the lysis of
neutrophils, lymphocytes and monocytes
Pathogenic Protozoa - Entamoeba histolytica
 Diagnosis microscopic observation of cyst in the feces
 the trophozoite is seen occasionally
 multiple concentrated stools specimens may be required since cysts
are distributed randomly
 pathogenic amoeba must be distinguished from commensal amoeba
 new test are designed to detect fecal antigen and trophozoite DNA
 extraintestinal complications can be confirmed via biopsy or
serological tests for antibody
 Epidemiology
 reservior is infected humans, both asymptomatic and symptomatic
 from 10 - 50 % of world population is infected
 prevalence of infection in the U.S. is 1 - 2 %
 asymptomatic individuals are chronic carriers
 person -to - person, indirect, fecal-oral mode of transmission
 food, water, fomites
 fecal comtamination of fresh vegetables, water
 flies and cockroaches also transmit the cysts mostly to food and
fomites
Entamoeba histolytica
 Disease : Meningoencephalitis
 destruction of the brain tissue causeing symptoms of
frontal headache, sore throat, fever, stiff neck, blocked nose
with alter senses of taste and smell, and Kernig’s sign(a sign
of meningitis in which the patient cannot flex the leg)
 cerebrospinal fluid is purulent and may contain
erythrocytes
 leads to rapid death in 4 - 5 days
 Acanthomoeba can also enter the eye and cause keratitis
and corneal ulcers
 Parasite : Naegleria fowleri or Acanthamoeba spp.
 trophozoite is a free living amoeba found in soil and water
 cyst
Naegleria fowleri/ and Acanthamoeba
 Pathogenesis most infections occur when trophozoites in
water enter the nasal passages when people
swim; they then invade the nasal mucosa and extend into
the brain
 Diagnosis:
trophozoites ( in wet mounts or stained smears) in
nasal discharge, cerebrospinal fluid, or corneal scrapings
 Epidemiology - this organism is an environmental opportunist
acquired via contact with water, soil , or dust
 major cause of a disease called PAM
 has 2 long flagella at one end
 most cases are contracted in lakes and
swimming pools
 probably the flagellated trophozoites are
forced deep into the nasal passages
when the victim dives in water
 found in USA, Czechoslovakia, Mexico,
Africa, New Zealand, Australia
Protozoan parasites
PAM
 cause chronic infection of the skinor CNS in
immunocompromised persons
 has small spiky acanthopodia
 causes keratitis
 GranulomatousAmoebic Encephalitis
Protozoan parasites
Protozoan parasites
Naegleria Acanthamoeba
Trophozoite with
broad pseudopods
Actively motile
Forms the flagellate
stage
Single-walled cyst
Does not encyst in
tissues
Trophozoite with
filamentous pseudopods
Sluggishly motile
Does not form flagellate
stage
Double-walled cyst
May encyst in tissues
Protozoan parasites
Prevention and Control
 Avoid swimming or jumping into bodies of warm
fresh water
 Avoid swimming in thermally polluted water (water
around power plants)
 Do not swim in areas posted as "no swimming."
 Hold the nose shut or use nose clips when jumping
or diving into bodies of fresh water.
Balantidium coli
 Balantidiasis : a dysentery (watery stools with blood and pus)
accompanied by nausea, anorexia, tenesmus, and
abdominal pain and tenderness
 Parasite : Balantidium coli
 Trophozoite ,an actively motile ciliate; highly specialized for
reproduction and food intake (cytostome)
 Cyst, a latent survival stage is significant in transmission;
infective stage with one large nucleus
Pathogenic Protozoa - Balantidium coli
 Pathogenesis : ulceration of the intestinal
mucosa especially in the large intestine sometimes
compounded by bacterial infection
 no extraintestinal invasion as seen in amebiasis
 Epidemiology : reservoir is swine and humans
 fecal-oral transmission involving water and food
 Diagnosis : microscopic observation of cysts in the feces
 this trophozoite is very large and often seen also
Protozoan parasites
Protozoan parasites
Cysts
 Trophozoite
Giardia lamblia
Scientific classification
Domain:Eukaryota
Phylum: Metamonada
Order: Diplomonadida
Family: Hexamitidae
Genus: Giardia
Species: G. lamblia
species
Giardia agilis
Giardia ardeae
Giardia lamblia
Giardia muris
Giardia microti
Giardia psittaci
Protozoan parasites
 Giardiasis : mild diarrhea to severe malabsorption syndrome;
sudden onset of watery diarrhea, often foul-
smelling, with abdominal cramps, flatulence, and
stearorrhea. Blood and pus are rarely present.
 Parasite : Giardia lamblia
 Trophozoite - flagellate protozoan( 10 x 10 um) having two
large nuclei, a large sucking disc, and many
flagella; face - like appearance
 Cyst - slightly smaller the trophozoite with four nuclei and
parabasal body
Pathogenic Protozoa - Giardia lamblia
 Pathogenesis : gastric acid stimulates excystation and releases
trophozoite into small intestine where it attaches via
the sucking disc to the intestinal villi; the alteration of the villi
and inflammation of the mucous causes maladsorption, but
not obvious tissue necrosis; spontaneous recovery occurs in
10 -14 days, but relapse may occur; chronic infection is
especially a problem in patients with IgA deficiency and
diverticula
 Diagnosis : cyts and trophozoites in the feces with the onset of
symptoms; cyts occur in”showers” and are not
present every day, so stoo should be examined
every three days
 in the absence of observable cysts, new tests for fecal antigen are now
available
 Epidemiology : sylvatic and domestic reservoirs, both human
and animals(mostly those around lakes
and streams;
 fecal - oral transmission via contaminated water is most
common mode of transmission; in U.S. domestic setting,
fecal- oral fomites transmission is very common in day care
settings; also uncooked contaminated vegetable and fruits
may serve as a source of the cysts
 cyst are resistant to traditional chlorination methods
 other than day-care centers, most recent outbreaks have
involved contaminated water from reservoirs
Pathogenic Protozoa - Giardia lamblia
Trichomonas vaginalis
Scientific classification
Domain: Eukaryota
(unranked):Excavata
Phylum: Metamonada
Class: Parabasalia
Order: Trichomonadida
Family: Trichomonadidae
Genus: Trichomonas
Protozoan parasites
Incubation Period is typically 5 to 28 days
Trichomonas vaginalis is pathogenic in the
genitourinary tract
In women, it lives in the vagina and the
Skene's glands (of the urethra)
In men, it lives in the urethra
Multiplies when vaginal conditions become
more basic than usual (normal pH is 3.8 to
4.2)
No known cystic form
May survive in a host for 2+ years
 Vaginitis and urethritis : itching, burning, and painful
urination sometime accompanied by scant watery
vaginal discharge
 Parasite : Trichomomas vaginalis
 only a flagellate trophozoite with four flagella and
undulating membrane
 this flagellate has not cyst stage
 Pathogenesis : extensive inflammation and erosion of the
vaginal or urethral epithelium causing itching, burning,
and painful urination; also sometimes a scanty watery
vaginal discharge
Pathogenic Protozoa - Trichomonas vaginalis
 Epidemiology : human reservoir with many asymptomatic
carriers especially in males- females may also be
asymtomatic
 trophozoite is sexually transmitted
 Diagnosis : microscope examination of vaginal or urethral
discharge
 stained smear show trophozoites
 also parasite is detected in fluorescent antibody stains and
nucleic acid probes
Cryptosporidium parvum
Scientific classification
Domain: Eukaryota
Kingdom:
Chromalveolata
Superphylum: Alveolata
Phylum: Apicomplexa
Class: Conoidasida
Subclass: Coccidiasina
Order: Eucoccidiorida
Suborder: Eimeriorina
Family: Cryptosporidiidae
Genus: Cryptosporidium
Species
Cryptosporidium andersoni
Cryptosporidium bailey
Cryptosporidium bovis
Cryptosporidium canis
Cryptosporidium cichlidis
Cryptosporidium cuniculus
Cryptosporidium felis
Cryptosporidium galli
Cryptosporidium hominis
Cryptosporidium meleagridis
Cryptosporidium muris
Cryptosporidium nasoris
Cryptosporidium parvum
Cryptosporidium pestis
Cryptosporidium reichenbachklinkei
Cryptosporidium saurophilum
Cryptosporidium scophthalmi
Cryptosporidium serpentis
Cryptosporidium suis
Cryptosporidium varanii
Cryptosporidium wrairi
Protozoan parasites
Protozoan parasites
 Cryptosporidosis enterocolitis characterized by water
diarrhea without blood - self limiting in immunocompetent
people
 Parasite : Cryptosporidium parvum
 Sporozoan(coccidian) parasite exhibits both
asexual(schizogony) and sexual(gametogony) in the brush
border of the intestinal epithelium
 Oocyts are released from the cells and transmitted
 Pathogenesis : In immunocompromised patients, an
unremitting enterocolitis with as many as 50 water stools per
day caused by the intracellular multiplication of the parasite
Pathogenic Protozoa - Cryptosporidium
parvum
 Epidemiology : reservoir is a variety of mammals, fish, reptiles
 this host seed the environment especially water with oocyts
 a-p, indirect, fecal-oral, contaminated water/fecal droplets
 p-p, indirect, fecal-oral/oral-anal - fomite/direct contact
 high risk groups = homosexuals, day care children/adults,
animal handlers
 Diagnosis : oocysts in the feces: concentrated and stained
 also assay for fecal antigen
Cyclospora cayetanensis
Protozoan parasites
 Cyclosporiasis -water diarrhea accompanied by nausea,anorexia,
abdominal cramping: sometime bloating
and flatulence
 Parasite Cyclospora cayetanensis
 a sporozoan(coccidian) parasite of the small intestine
 asexual and sexual cycle similar to Cryptosporidium but
oocysts are larger
 Pathogenesis inflammatory changes in the jejenum
resulting from the intracellular multiplication of the parasite =
villous atrophy
Pathogenic Protozoan -Cyclospora
cayetanensis
 Epidemiology : reservoir is a variety of mammals, birds, reptile
 source to humans is fecally contaminated water and some
fresh fruits like strawberries
 Diagnosis : detection of oocysts in the stool: concentrated
and stained by a variety of methods
including immunofluorescence
Blood andTissue
Protozoa
 Sporozoan(coccidian)
 malaria Plasmodium spp
 toxoplasmosis Toxoplasma gondii
 Flagellates
 trypanosomiasis Trypanosoma cruzi
 African Sleeping SicknessTyrpanosoma brucei
 leishmaniasis Leishmania spp
Blood and Tissue Protozoa
Malarial Species
 Malaria - influenza-like symptoms( fever, chills) with headache,
muscle pain, photophobia, anorexia,
nausea, vomiting
 Parasites
 Plasmodium vivax - benign tertian malaria
 Plasmodium falciparum - malignant tertian malaria
 Plasmodium malariae - quartan malaria
 Pathogenesis :infection begins in the liver(exoerythocytic)
but becomes a disease of RBC’s
(erythrocytic)
 merozoites released every 48 hrs in benign tertian malaria
 merozoites released every 36-48 hrs in malignant tertian
malaria
 merozoite released ever 72 hrs in quartan malaria
Pathogenic Protozoa – Plasmodium spp
 Epidemiology : reservior in infecte humans and wild primates
 arthopod-borne transmission, via Anopheline mosquitos
where sporogamy produces infectious sporozoites
 Diagnosis : trophozoites(schizonts) in blood smears
Protozoan parasites
Liver stage
Sporozoites
Mosquito Salivary
Gland
Malaria Life Cycle
Gametocytes
Oocyst
Red Blood Cell
Cycle
Zygote
RING FORMS GAMETOCYTE
RING FORMS SCHIZONT
GAMETOCYTE
Species Stages found in the circulating blood
P. vivax All stages. Wide range of stages may be seen
on given film
P. malariae All stages. Wide variety of stages usually not
seen. Relatively few rings, gametocytes
generally present
P. ovale All stages
P.
falciparum
Rings and or gametocytes. Other stages develop in
blood vessels of internal organs but are not seen in
peripheral blood except in severe infections
 Trophozoite or ring forms
young form is globose in shape with a
central vacuole, a red chromatin mass
and a blue cytoplasm
- in stained smear, they appear signet
rings
b) Growing form
- single chromatin
mass with a more
abundant cytoplasm
- cytoplasm may
appear compact or
irregular (ameboid)
c) Mature form
- single
chromatin mass but
with an increase
amount of cytoplasm
that partially fills the
erythrocyte
 Early trophozoites have
the characteristic
signet ring shape.
 Also, unique to P.
falciparum is the
presence of multiple
trophozoites in one
cell.
RING FORMS GAMETOCYTE
 The gametocytes of P.
falciparum have a
crescent or banana
shape.
 Red blood cells
infected by P. vivax are
often larger than
uninfected red blood
cells.
 They are
approximately 1.5
times the size of a
normal cell.
RING FORMS SCHIZONT
GAMETOCYTE
 A schizont showing the
large number of
merozoites typical of
this species (16-24).
 Also note the larger
size compared to a
normal red blood cell.
 Characteristic
trophozoites of P.
malariae showing the
ring shape and the
tendency of infected
cells to be of normal or
smaller size (arrows).
 A schizont containing
merozoites (6 to 12)
giving a coarse
granular appearance.
Species rbc stage Size of rbc Schauffner’s
stippling
P. vivax Young rbc enlarged +
P. malariae Mature rbc normal -
Ziemann’s
dots
P. ovale Young rbc enlarged +
P. falciparum rbc of all
stages
Normal/
Multiple
infection
_
Maurer’s dots
 A red blood cell
showing the
Schauffner's dots
characteristic of cells
infected by
Plasmodium vivax and
Plasmodium ovale.
Schauffner’s dots
Species Cytoplasm Pigment No. of
merozoites
Gametocytes
P. vivax Irregular,
ameboid in
trophozoite,
has spread
out
appearance
Golden
brown,
incons-
picuous
12-24
Average is
16
rounded
Species Cytoplasm Pigment No. of
merozoites
Gametocytes
P.
malariae
Rounded,
compact
trophozoite
with dense
cytoplasm/
band form
troph are
rarely seen
Dark
brown,
cons-
picuous
6-12
Average is 8,
“rosette”
schizonts are
sometimes
seen
rounded
Species Cytoplasm Pigment No. of
merozoites
Gametocytes
P. ovale Rounded,
compact
trophozoite
with dense
cytoplasm/
band form
troph are
rarely seen
Dark
brown,
cons-
picuous
6-12
Average is 8,
“rosette”
schizonts are
sometimes
seen
rounded
Species Cytoplasm Pigment No. of
merozoites
Gametocytes
P.
falciparum
Young rings
are small,
delicate,
often with
double
chromatin
mass
Black
coarse and
cons-
picuous in
gametocyte
6-32
Average is
20-24
Crescent,
sausage-
elongate
shape
Toxoplasma gondii
 Toxoplasmosis : infectious mononucleosis -like symptoms
of fever, chills, headache, myalgia, lymphadenitis,
and fatigue; chronic form = hepatitis,
encephalomyelitis, mycocarditis
 Parasite : Toxoplasma gondii
 intracellular coccidian(sporozoan) parasite infecting various
tissues(systemic) but confined to intestines in felines
 infected tissues release pseudocysts containing bradyozoites
 feline intestines release oocyts containing tachyzoites
 Pathogenesis : cellular death due to intracellular muliplication
of trophozoites especially severe in fetal and cerebral
tissues
Pathogenic Protozoa - Toxoplasma gondii
 Epidemiology : reservoir is a variety of mammals and birds
including humans
 fecal-oral transmission of oocyst from felines
 common source transmission of pseudocyts in poorly cooked meat
esp. beef
 Diagnosis : serological detection of antibody: Elisa test for
IgM antibodies is most reliable
 Risk Groups : Pregnant females and immunocompromised
Protozoan parasites
Protozoan parasites
 Enteroepithelial cycle
asexual schizogony
Sexual gametogony
*immature oocyst passed in the cat’s feces
 Immature oocyst mature to the infective stage
2 sporocysts with 4 sporozoites each (2-21 days)
 Ingestion of inadequately cooked meat, esp
lamb or pork
 Ingestion of infective oocysts from material
contaminated by cat feces
 Drinking contaminated water or unpasturized
goat’s milk
 Organ transplantation
 Blood transfusion
 Transplacental
 When mature oocyst is ingested, sporozoites
are released forming trophozoites
*tachyzoites- actively growing trop
*bradyzoites- latent or slow growing
trophozoites
 Active infection (all nucleated cells)
fever
lymphadenopathy
 Congenital
depends on the age of gestation when the
infection is acquired
 1st half pregnancy
- intrauterine death
- microcephaly
- hydrocephaly with intracranial calcification
 2nd half of pregnancy
-usually assymptomatic at birth though fever,
hepatosplenomegaly and jaundice may
appear
-choriorenitis, psychomotor retardation and
convulsive disorders may appear months or
years later
 Immunocompromised individuals
-present CNS involvement
-pneumonitis, myocarditis, retinitis,
pancreatitis or orchitis
 Examination of tissues, blood, and body fluids
- Demonstration of tachyzoites or tissue cysts
- Animal inoculation to uninfected mice
- PCR
- Sabin-Feldman dye test
- IFA
- EIA
Babesiosis
 Infection of rbc often producing febrile illness
 Transmitted by ticks (Ixodes scapularis, Ixodes
pacificus)
 Caused by Babesia microti
 mulitiply in rbc by schizogony but do not produce
gametocytes
 Appear as delicate ring forms that may easily
confused with malaria
 The organisms
resemble
P.falciparum
 they vary more in
shape and in size;
and they do not
produce pigment
 evidence of
Maltese cross
formation
Protozoan parasites
Protozoan parasites
Protozoan parasites
 TICK vector of
Babesia
 The organism
belongs to the
genus Ixodes
The vector: Dog tick
Protozoan parasites
MALARIA BABESIOSIS
Multiple rings _ +
Large
trophozoite
+ _
Gametocytes + _
Hemozoin
pigment
+ _
Patients with spleen only 25% of RBCs could be parasitized
Patients withoutspleen higher parasitemia fever, muscle aches,
chills, fatigue, headache and loss of appetite Capillary microvascular stasis
RBC fragments Hemolytic anemia liver, spleenic, renal and
Central Nervous System involvement.
 Indirect Fluorescent Antibody
 PCR
 antimicrobial drugs such as clindamycin*
plus quinine or atovaquone* plus
azithromycin are often used.
Prevention/Control
- wearing appropriate light-colored clothing
-using tick repellent in areas where the diseases
are endemic
-if possible test blood to be transfused for possible presence
of Babesia species
Blood and Tissue Flagellates
Trypanosomes
Species Disease Portal of Entry and
Final Site
Vector
Trypanosoma cruzi
Trypanosoma brucei
gambiense
Trypanosoma brucei
rhodesiense
American
Trypanosomiasis or
Chagas’ Disease
Gambian or West
African Sleeping
Sickness
Rhodesian or East
African Sleeping
Sickness
Breaks in skin or intact
mucous membranes,
later any organs and
blood stream; Heart and
GIT in chronic form
Skin for both
Lymph nodes, blood,
and brain
Infected
Triatomine
bug
Infected
tse-tse flies
 African Sleeping Sickness CNS infection progressing to
lethargy, tremors, meningoencephalitis, convulsions,
coma, and death
 Parasites
Trypanosoma gambiense - West/Central Africa
 Trypanosoma rhodesiense - East Africa
 Pathogenesis - trypanomastigote multiplies in the blood,
lymph, and cerebrospinal fluid; deprives the brain of
amino acids
Pathogenic Protozoa - Trypanosoma brucei
 Epidemiology : reservoir is a variety of mammals
and birds including humans
 fecal-oral transmission of oocyst from felines
 common source transmission of pseudocyts in poorly
cooked meat esp. beef
 Diagnosis : serological detection of antibody:
Elisa test for IgM antibodies is most reliable
 Risk Groups : Pregnant females
and immunocompromised
 American Trypanosomiasis/Chagas Disease -systemic infection
beginning with fever, chills, malaise, myalgia, fatigue, and
chagoma; progressing to involve heart and brain
 Parasite = Tryanosoma cruzi
 flagellate tryanosome exhibiting both the intracellular
amastigote(leishmanial) form and the extracellular
trypanomastigote(trypanosmal) form
 Pathogenesis - the intracellular amastigote multiplies in cells
and destroys the cells
Pathogenic Protozoa - Trypanosoma cruzi
 Epidemiology- wild and domestic animal reservoirs
 arthropod-borne via the cone-nosed (reduviid) bugs
which live in the cracks of domestic dwellings
 Diagnosis : trypanosomes in the blood; amastigotes
in biopsy; also xenodiagnosis and gene probes
Protozoan parasites
Protozoan parasites
Local inflammation at the site of
inoculation.
CHAGOMA = reddish painful nodule
Trypanosmes may enter the conjunctiva
= edema of eyelid & conjunctiva =
ROMAÑA’S SIGN
Protozoan parasites
Protozoan parasites
Protozoan parasites
Winterbottom's sign
is the swelling of
lymph nodes
(lymphadenopathy)
along the back of the
neck, in the posterior
cervical chain of
lymph nodes, as
trypanosomes travel
in the lymphatic fluid
and cause
inflammation.
w/ CNS invasion = chronic disease known as sleeping sickness stage is
initiated
(Kerandel’s sign) = delayed sensation to pain indicative ofAfrican
trypanosomiasis
Untreated infection may lead to death
Leishmania
Species Disease Portal of Entry and
Final Site
Vector
Leishmania
donovani
Leishmania
tropica
Leishmania
brazilienses
Visceral Leishmanisis
or KALAAZAR
Cutaneous
Leishmaniasis
Mucocutaneous
Leishmaniasis
or ESPUNDIA
Skin/ Macrophages and
monocytes of spleen,
liver, bone marrow
Skin/ Macrophages of
the skin and mucosa
Infected
Phlebotomus
Fly
 Leishmaniasis;Visceral : gradual onset of fever, chill, sweating,
diarrhea, anemia which progresses to enlargement of
liver and spleen with weight loss and emaciation: later
kidney damage and granulomatous skin lesion = also called
kala-azar
 cutaneous form -ulcerative skin lesions which contain parasite:
become infected with secondary bacteria and scar
formation disfigures
 Parasites : Leishmania donovani - visceral- amastigote stage
multiplies in visceral cells causing lysis
 Leishmania tropica - cutanous: Leishmania braziliensis –
mucocutaneous
 Pathogenesis - cells in infected visceral and cutaneous tissues
are destroyed by intracellular amastigote
multiplication
Pathogenic Protozaa - Leishmania
 Epidemiology : animal reservoirs including dogs and cats
 arthropod-borne via the bite of sandflies: the amastigote stage
becomes a motile promastigote in the sandfly gut
 Diagnosis : amastigote is observed in tissue biopsy, body
fluid aspirates, and in the blood: also serology for antibody
presence
Species Amastigote Promastigote Epimastigote Trypomastigote
Leishmania
Animal Host
Vector
Macrophages
-
-
midgut and
pharynx of
sandfly
-
-
-
-
Trypanosoma
Animal Host
Vector
-
-
-
-
-
salivary gland and
gut of flies/
culture
Blood, lymph
nodes, brain, CSF
-
Trypanosoma cruzi
Animal Host
Vector
Intracellular,
esp. striated
and smooth
muscle, brain
Cell cultures
-
-
-
-
Intestine of vector
bugs
Blood and tissues
of mammals
Rectum of vector
bug/culture and
cell cultures
Protozoan parasites
Protozoan parasites
Protozoan parasites
Protozoan parasites
Kalaazar
Cutaneous Leishmaniasis
1. Skin ulcers, w/ painless lesions = ugly scar
2. Subcutaneous nodules
Mucocutaneous Leishmaniasis
Metastatic spread to the oronasal and pharyngeal mucosa
(ESPUNDIA) = highly disfiguring leprosy like tissue destruction &
swelling (“TAPIR NOSE”)
Espundia
The current standard treatment for first stage
disease is:
 Intravenous pentamidine (for T.b. gambiense);
or
 Intravenous suramin (for T.b. rhodesiense)
 Eflornithine
To HIM we LIFT the HIGHEST PRAISE

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Protozoan parasites

  • 2. At the end of this unit, the student is able to:  Classify the Protozoans  Describe the morphology of each protozoa  Explain the pathophysiology, life cycle, infective stages, modes of transmission, epidemiology, prevention and control  Describe the diagnostic features of each parasite.  Enumerate the different specimens used for identification  Identify correctly the diagnostic features of each parasite.
  • 3.  Perform the laboratory tests  Discuss specimen collection,transport, processing, preservation and disposal
  • 4. General Characteristics  They are single-celled organisms which are generally classified according to their organelles of locomotion.  Cilia, flagella & pseudopodia  Considered as the simplest forms of animals
  • 5. Organelles  Nucleus  Nucleulos  Mitochondria  Locomotory Organelles
  • 6. Kingdom Protista  Phylum Sarcomastigophora  Subphylum Mastigophora  (Superclass Rhizopoda) Flagella (whip-like) = organ of locomotion  Subphylum Sarcodina  (Class Zoomastigophora) Pseudopodia (hyaline foot like)= organ of locomotion  Phylum Ciliophora Cilia (hair like) = organ of locomotion  Balantidiasis (Balantidium coli)
  • 7. Kingdom Protista  Phylum Apicomplexa  w/ apical complex at the anterior end (apicoplast)  Babesiosis (Babesia)  Malaria (Plasmodium)  Coccidian diseases including:  Cryptosporidiosis (Cryptosporidium parvum)  Cyclosporiasis (Cyclospora cayetanensis)  Toxoplasmosis (Toxoplasma gondii)  Phylum Microspora  Spore-forming parasites
  • 11. Giardia lamblia Amoeba Amoeba feeding
  • 12. Kingdom Protista  Subkingdom Protozoa  automonous unicellular eucaryotic organisms possessing various degrees of cellular specialization and some type of motility  Organelles specialize in nutrient storage, excretion, digestion, motility  Phylum = based upon motility Classification of Protozoa and Descriptions
  • 13.  Size 2 - 100 micrometers  Parasitic protozoa are most facultative anaerobes;  Mostly heterotrophic, chemoorganotrophs  Nutrients assimilated via phagocytosis, pinocytosis, diffusion  Digestion may be both extracellular and intracellular  Two life stages  Trophozoiteactive pathogenic vegetative stage  Cyst latent survival form  Sites of pathogenesis  Intestinal, Urogenital, Blood and Tissue Pathogenic Protozoa - Characteristics
  • 14.  Phylum Sarcodina  Motility via Pseudopoda  Extensions of the cell membrane into which the cytoplasm flows  Cyclosis  No cell wall; thus, no uniform shape  Shape changes with movement  Reproduce asexually by mitosis; sexually by meiosis  Trophozoite  Motile vegetative stage  Cyst  Latent survival stage Protozoa - Amoeba
  • 15.  Phylum Ciliata  Motility via cilia  Cilia surround the cell; their co-ordinated movement  Uses energy from the cell membrane  Synchronized ciliary movement  Two nuclei per cell  Small micronucleus governs sexually reproduction  Large macronucleus governs metabolism and growth  Both nuclei divide during asexual mitosis  Intracellular organelles  Function in digestion and excretion  Only one ciliate pathogen affects humans Protozoa - Ciliates
  • 16.  Phylum Mastigophora  Motility via flagella  One or more whip-like filaments move in counter- clockwise fashion creating rapid movement  A flagellum is a complex organelle composed of many smaller tubules  Flagella attach to the cell membrane and derive energy from movement from this membrane  Most numerous of the protozoan types  Several are pathogens of humans  All reproduce asexually; some reproduce sexually Protozoa - Flagellates
  • 17.  Phylum Sporozoa  No established means of movement  Thus, not motile  All are intracellular parasites in a variety of hosts  Several parasitic species affect humans  Most are in the Class Coccidia  All lack intracellular organelles  All have three things in common  No motility  Sexual and Asexual mode of Reproduction  Intracellular parasites Protozoa - Sporozoans
  • 18. Classification of Protozoan Parasites: Sarcomastigophora Sarcodina Acanthamoeba castellani, Endolimax nana Entamoeba coli, Entamoeba dispar Entamoeba gingivalis, Entamoeba histolytica Iodamoeba butschlii, Nagleria fowleri Mastigophora Chilomastix mesnili, Dientamoeba fragilis Giardia lamblia,Trichomonas (homini, tenax ,vaginalis) Leishmania (braziliensis, donovani, tropica) Trypanosoma brucei complex, Trypanosoma cruzi Ciliophora Balantidium coli Apicomplexa Bebesia spp., Cryptosporidum hominis, Cyclospora cayetansis, Isospora belli, Plasmodium spp., Toxoplasma Microspora Enterocytozoon bineusi, Encephalitozoon spp. Vittaforma cornea,Trachipleistophora hominis Pleistophora spp., Brachiola vesicularum, Microspordium spp.
  • 19. Intestinal  Amebiasis .......................... Entamoeba histolytica  Giardiasis .......................... Giardia lamblia  Balantidiasis .......................... Balantidium coli  Crytosporidosis ...................... Cryptosporidium parvum  Urogenital  Trichomoniasis ....................... Trichomonas vaginalis  Blood and Tissue  Malaria ................................... Plasmodium spp  Meningoencephalitis ............... Naegleria fowleri  Toxoplasmosis ........................ Toxoplasma gondii  Trypanosomiasis  African Sleeping Sickness ... Trypanosoma brucei  Chagas Disease .................. Trypanosoma cruzi  Leishmaniasis  visceral leishmaniasis( Kala-azar) ... Leishmania donovani  cutaneous leishmaniasis .......... Leishmania topica/braziliensis Protozoan Diseases
  • 21. Intestinal and Luminal Protozoa  Constantly changing shapes, move by ending out fingerlike extensions of their cell membranes called pseudopods or “false- feet” projections  the name comes from the Greek word for “change” is an example of a sarcodine, found in fresh water or salt water
  • 22.  six species have been established as parasites of man: Entamoeba histolytica Entamoeba coli Entamoeba gingivalis Dientamoeba fragilis Endolimax nana Iodamoeba butschlii
  • 23. Intestinal and Luminal Protozoa  Stages of Development Cyst – infective stage Trophozoite – vegetative stage
  • 24. General Statements:  All amoebas are capable of producing cyst and trophozoite except for Entamoeba gingivalis and Dientamoeba fragilis.  Found in the colon except Entamoeba gingivalis which is found in the oral cavity  All are commensals except Entamoeba histolytica.
  • 25. Entamoeba histolytica A) Trophozoite Stage (I & H preparation)  shape: irregular  shape: 10– 60 micra  structures: - cytoplasm is grayish violet with a clear demarcation between the ectoplasm and endoplasm
  • 30. Entamoeba histolytica B. Cystic Stage  shape: spherical  size: 5 – 20 micra  structures: - smooth, definite, relatively thin cystic wall
  • 31. Entamoeba histolytica B. Cystic Stage the number of nuclei range from 1-4, rarely more, with a centrally located karyosome in the younger cyst, the chromatoidal bars with rounded ends and glycogen mass may be present
  • 37. Entamoeba histolytica Diagnosis:  Microscopy  Antigen detection  Serology  PCR  Imaging studies
  • 38. Entamoeba histolytica Treatment:  metronidazole  tetracycline with diiodohydroxyquin
  • 39. Entamoeba coli A. Trophozoite Stage (I & H preparation)  shape: condensed, thickened, rounded mass  size: 15 – 50 micra
  • 40. Entamoeba coli A. Trophozoite Stage (I & H preparation)  structures:  cytoplasm is grayish violet; there is no clear demarcation between the ectoplasm and endoplasm  the endoplasm is highly vacuolated containing bacteria  the nucleus is spherical with thick nuclear membrane lined with coarse, irregular chromatin granules or plaques; the karyosome is moderately large and eccentrically located
  • 43. Entamoeba coli B. Cystic Stage  shape: spherical  size: 10 – 33 micra
  • 44. Entamoeba coli B. Cystic Stage  structures:  smooth, definite, thick cystic wall  number of nuclei range from 1 – 8 rarely more with an eccentric karyosome  in younger cysts, chromatoidal bars with splintered ends and glycogen mass
  • 46. Endolimax nana A.Trophozoite Stage (I & H preparation)  shape: irregular  size: small, 6 – 15 micra
  • 47. Endolimax nana A.Trophozoite Stage (I & H preparation)  structures:  cytoplasm is grayish-violet, vacuolated which may contain bacteria  there is small, spherical, single nucleus with a large irregular, central or eccentric karyosome
  • 49. Endolimax nana B. Cystic Stage  shape: oval size: 6 – 10 micra
  • 50. Endolimax nana B. Cystic Stage  structures:  smooth, definite cystic wall  number of nuclei ranges from 1-4  I & H preparation  cytoplasm is grayish, granular  nuclei are visible as darkly stained dots  chromatoidal bodies occasionally seen as deeply stained, small, slightly curved rods
  • 53. Entamoeba gingivalis A.Trophozoite Stage (I & H preparation)  shape: irregular  shape: 5 – 35 micra  structures:  clear demarcation between the clear ectoplasm and granular endoplasm
  • 54. Entamoeba gingivalis  endoplasm contains food vacuoles with bacteria and rarely rbc  possesses a single nucleus which is spherical in shape, karyosome is near the center  have multiple pseudopodia with progressive directional movement similar to E. histolytica
  • 56. Dientamoeba fragilis A.Trophozoite Stage (I & H preparation)  shape: irregular  size 3 – 22 micra
  • 57. A.Trophozoite Stage (I & H preparation)  structures: cytoplasm is granular with vacuoles containing bacteria nuclei, commonly 2 in number, with karyosome consisting of 4-8 chromatin granules Dientamoeba fragilis
  • 59. Iodamoeba buetschlii A.Trophozoite Stage (I & H preparation)  shape: irregular  shape: 6 – 25 micra
  • 60. Iodamoeba buetschlii A.Trophozoite Stage (I & H preparation)  structures:  granular, vacuolated cytoplasm and vacuoles may contain bacteria and yeast cells  single, spherical nucleus with a large central or eccentric karyosome surrounded by an unstained halo
  • 62. Iodamoeba buetschlii B. Cystic Stage  shape: irregular  size: 6 – 15 micra
  • 63. Iodamoeba buetschlii B. Cystic Stage  structures: smooth, definite cystic wall have a single nucleus, rarely with 2 nuclei large glycogen vacuole
  • 64. Iodamoeba buetschlii B. Cystic Stage I & H preparation violet-gray granular cytoplasm nucleus is visible large glycogen vacuole is unstained chromatoidal bodies are usually absent; if present, they are seen as deeply stained small rods
  • 68. A) Trophozoite Stage (I & H preparation) endoplasm is briefly granular with ingested rbc which are bluish black in color single, spherical nucleus; nucleus membrane is lined by fine, regularly arranged chromatin granules; karyosome is relatively small and centrally located
  • 70.  Amebiasis( Amebic dysentery) - colitis with diarrhea(somtimes bloody) accompanied by abdominal pain and cramping: numerous bloody stools per day  extraintestinal amebiasis a complication of intestinal amebiasis in which the trophozoites enter the blood and are carried to other organs; most often the liver  these patients have fever, leukocytosis, rigors  Parasite : Entamoeba histolytica  Trophozoite : sarcodine protozoa which slowly move among the crypts of the large intestine and are shed into the feces  Cyst : latent survival stage which is significant in transmission; spherical containing four characteristic nuclei; infective stage  Pathogenesis the trophozoites bind to colonic epithelial cells and secrete a cytotoxin which alters cell membrane permeability causing the cells to take in calcium and lyse; necrosis and inflammation also accompanies the lysis of neutrophils, lymphocytes and monocytes Pathogenic Protozoa - Entamoeba histolytica
  • 71.  Diagnosis microscopic observation of cyst in the feces  the trophozoite is seen occasionally  multiple concentrated stools specimens may be required since cysts are distributed randomly  pathogenic amoeba must be distinguished from commensal amoeba  new test are designed to detect fecal antigen and trophozoite DNA  extraintestinal complications can be confirmed via biopsy or serological tests for antibody  Epidemiology  reservior is infected humans, both asymptomatic and symptomatic  from 10 - 50 % of world population is infected  prevalence of infection in the U.S. is 1 - 2 %  asymptomatic individuals are chronic carriers  person -to - person, indirect, fecal-oral mode of transmission  food, water, fomites  fecal comtamination of fresh vegetables, water  flies and cockroaches also transmit the cysts mostly to food and fomites Entamoeba histolytica
  • 72.  Disease : Meningoencephalitis  destruction of the brain tissue causeing symptoms of frontal headache, sore throat, fever, stiff neck, blocked nose with alter senses of taste and smell, and Kernig’s sign(a sign of meningitis in which the patient cannot flex the leg)  cerebrospinal fluid is purulent and may contain erythrocytes  leads to rapid death in 4 - 5 days  Acanthomoeba can also enter the eye and cause keratitis and corneal ulcers  Parasite : Naegleria fowleri or Acanthamoeba spp.  trophozoite is a free living amoeba found in soil and water  cyst Naegleria fowleri/ and Acanthamoeba
  • 73.  Pathogenesis most infections occur when trophozoites in water enter the nasal passages when people swim; they then invade the nasal mucosa and extend into the brain  Diagnosis: trophozoites ( in wet mounts or stained smears) in nasal discharge, cerebrospinal fluid, or corneal scrapings  Epidemiology - this organism is an environmental opportunist acquired via contact with water, soil , or dust
  • 74.  major cause of a disease called PAM  has 2 long flagella at one end  most cases are contracted in lakes and swimming pools  probably the flagellated trophozoites are forced deep into the nasal passages when the victim dives in water  found in USA, Czechoslovakia, Mexico, Africa, New Zealand, Australia
  • 76. PAM
  • 77.  cause chronic infection of the skinor CNS in immunocompromised persons  has small spiky acanthopodia  causes keratitis  GranulomatousAmoebic Encephalitis
  • 80. Naegleria Acanthamoeba Trophozoite with broad pseudopods Actively motile Forms the flagellate stage Single-walled cyst Does not encyst in tissues Trophozoite with filamentous pseudopods Sluggishly motile Does not form flagellate stage Double-walled cyst May encyst in tissues
  • 82. Prevention and Control  Avoid swimming or jumping into bodies of warm fresh water  Avoid swimming in thermally polluted water (water around power plants)  Do not swim in areas posted as "no swimming."  Hold the nose shut or use nose clips when jumping or diving into bodies of fresh water.
  • 84.  Balantidiasis : a dysentery (watery stools with blood and pus) accompanied by nausea, anorexia, tenesmus, and abdominal pain and tenderness  Parasite : Balantidium coli  Trophozoite ,an actively motile ciliate; highly specialized for reproduction and food intake (cytostome)  Cyst, a latent survival stage is significant in transmission; infective stage with one large nucleus Pathogenic Protozoa - Balantidium coli
  • 85.  Pathogenesis : ulceration of the intestinal mucosa especially in the large intestine sometimes compounded by bacterial infection  no extraintestinal invasion as seen in amebiasis  Epidemiology : reservoir is swine and humans  fecal-oral transmission involving water and food  Diagnosis : microscopic observation of cysts in the feces  this trophozoite is very large and often seen also
  • 88. Cysts
  • 91. Scientific classification Domain:Eukaryota Phylum: Metamonada Order: Diplomonadida Family: Hexamitidae Genus: Giardia Species: G. lamblia species Giardia agilis Giardia ardeae Giardia lamblia Giardia muris Giardia microti Giardia psittaci
  • 93.  Giardiasis : mild diarrhea to severe malabsorption syndrome; sudden onset of watery diarrhea, often foul- smelling, with abdominal cramps, flatulence, and stearorrhea. Blood and pus are rarely present.  Parasite : Giardia lamblia  Trophozoite - flagellate protozoan( 10 x 10 um) having two large nuclei, a large sucking disc, and many flagella; face - like appearance  Cyst - slightly smaller the trophozoite with four nuclei and parabasal body Pathogenic Protozoa - Giardia lamblia
  • 94.  Pathogenesis : gastric acid stimulates excystation and releases trophozoite into small intestine where it attaches via the sucking disc to the intestinal villi; the alteration of the villi and inflammation of the mucous causes maladsorption, but not obvious tissue necrosis; spontaneous recovery occurs in 10 -14 days, but relapse may occur; chronic infection is especially a problem in patients with IgA deficiency and diverticula
  • 95.  Diagnosis : cyts and trophozoites in the feces with the onset of symptoms; cyts occur in”showers” and are not present every day, so stoo should be examined every three days  in the absence of observable cysts, new tests for fecal antigen are now available  Epidemiology : sylvatic and domestic reservoirs, both human and animals(mostly those around lakes and streams;  fecal - oral transmission via contaminated water is most common mode of transmission; in U.S. domestic setting, fecal- oral fomites transmission is very common in day care settings; also uncooked contaminated vegetable and fruits may serve as a source of the cysts  cyst are resistant to traditional chlorination methods  other than day-care centers, most recent outbreaks have involved contaminated water from reservoirs Pathogenic Protozoa - Giardia lamblia
  • 97. Scientific classification Domain: Eukaryota (unranked):Excavata Phylum: Metamonada Class: Parabasalia Order: Trichomonadida Family: Trichomonadidae Genus: Trichomonas
  • 99. Incubation Period is typically 5 to 28 days Trichomonas vaginalis is pathogenic in the genitourinary tract In women, it lives in the vagina and the Skene's glands (of the urethra) In men, it lives in the urethra Multiplies when vaginal conditions become more basic than usual (normal pH is 3.8 to 4.2) No known cystic form May survive in a host for 2+ years
  • 100.  Vaginitis and urethritis : itching, burning, and painful urination sometime accompanied by scant watery vaginal discharge  Parasite : Trichomomas vaginalis  only a flagellate trophozoite with four flagella and undulating membrane  this flagellate has not cyst stage  Pathogenesis : extensive inflammation and erosion of the vaginal or urethral epithelium causing itching, burning, and painful urination; also sometimes a scanty watery vaginal discharge Pathogenic Protozoa - Trichomonas vaginalis
  • 101.  Epidemiology : human reservoir with many asymptomatic carriers especially in males- females may also be asymtomatic  trophozoite is sexually transmitted  Diagnosis : microscope examination of vaginal or urethral discharge  stained smear show trophozoites  also parasite is detected in fluorescent antibody stains and nucleic acid probes
  • 103. Scientific classification Domain: Eukaryota Kingdom: Chromalveolata Superphylum: Alveolata Phylum: Apicomplexa Class: Conoidasida Subclass: Coccidiasina Order: Eucoccidiorida Suborder: Eimeriorina Family: Cryptosporidiidae Genus: Cryptosporidium Species Cryptosporidium andersoni Cryptosporidium bailey Cryptosporidium bovis Cryptosporidium canis Cryptosporidium cichlidis Cryptosporidium cuniculus Cryptosporidium felis Cryptosporidium galli Cryptosporidium hominis Cryptosporidium meleagridis Cryptosporidium muris Cryptosporidium nasoris Cryptosporidium parvum Cryptosporidium pestis Cryptosporidium reichenbachklinkei Cryptosporidium saurophilum Cryptosporidium scophthalmi Cryptosporidium serpentis Cryptosporidium suis Cryptosporidium varanii Cryptosporidium wrairi
  • 106.  Cryptosporidosis enterocolitis characterized by water diarrhea without blood - self limiting in immunocompetent people  Parasite : Cryptosporidium parvum  Sporozoan(coccidian) parasite exhibits both asexual(schizogony) and sexual(gametogony) in the brush border of the intestinal epithelium  Oocyts are released from the cells and transmitted  Pathogenesis : In immunocompromised patients, an unremitting enterocolitis with as many as 50 water stools per day caused by the intracellular multiplication of the parasite Pathogenic Protozoa - Cryptosporidium parvum
  • 107.  Epidemiology : reservoir is a variety of mammals, fish, reptiles  this host seed the environment especially water with oocyts  a-p, indirect, fecal-oral, contaminated water/fecal droplets  p-p, indirect, fecal-oral/oral-anal - fomite/direct contact  high risk groups = homosexuals, day care children/adults, animal handlers  Diagnosis : oocysts in the feces: concentrated and stained  also assay for fecal antigen
  • 110.  Cyclosporiasis -water diarrhea accompanied by nausea,anorexia, abdominal cramping: sometime bloating and flatulence  Parasite Cyclospora cayetanensis  a sporozoan(coccidian) parasite of the small intestine  asexual and sexual cycle similar to Cryptosporidium but oocysts are larger  Pathogenesis inflammatory changes in the jejenum resulting from the intracellular multiplication of the parasite = villous atrophy Pathogenic Protozoan -Cyclospora cayetanensis
  • 111.  Epidemiology : reservoir is a variety of mammals, birds, reptile  source to humans is fecally contaminated water and some fresh fruits like strawberries  Diagnosis : detection of oocysts in the stool: concentrated and stained by a variety of methods including immunofluorescence
  • 113.  Sporozoan(coccidian)  malaria Plasmodium spp  toxoplasmosis Toxoplasma gondii  Flagellates  trypanosomiasis Trypanosoma cruzi  African Sleeping SicknessTyrpanosoma brucei  leishmaniasis Leishmania spp Blood and Tissue Protozoa
  • 115.  Malaria - influenza-like symptoms( fever, chills) with headache, muscle pain, photophobia, anorexia, nausea, vomiting  Parasites  Plasmodium vivax - benign tertian malaria  Plasmodium falciparum - malignant tertian malaria  Plasmodium malariae - quartan malaria  Pathogenesis :infection begins in the liver(exoerythocytic) but becomes a disease of RBC’s (erythrocytic)  merozoites released every 48 hrs in benign tertian malaria  merozoites released every 36-48 hrs in malignant tertian malaria  merozoite released ever 72 hrs in quartan malaria Pathogenic Protozoa – Plasmodium spp
  • 116.  Epidemiology : reservior in infecte humans and wild primates  arthopod-borne transmission, via Anopheline mosquitos where sporogamy produces infectious sporozoites  Diagnosis : trophozoites(schizonts) in blood smears
  • 118. Liver stage Sporozoites Mosquito Salivary Gland Malaria Life Cycle Gametocytes Oocyst Red Blood Cell Cycle Zygote
  • 121. Species Stages found in the circulating blood P. vivax All stages. Wide range of stages may be seen on given film P. malariae All stages. Wide variety of stages usually not seen. Relatively few rings, gametocytes generally present P. ovale All stages P. falciparum Rings and or gametocytes. Other stages develop in blood vessels of internal organs but are not seen in peripheral blood except in severe infections
  • 122.  Trophozoite or ring forms young form is globose in shape with a central vacuole, a red chromatin mass and a blue cytoplasm - in stained smear, they appear signet rings
  • 123. b) Growing form - single chromatin mass with a more abundant cytoplasm - cytoplasm may appear compact or irregular (ameboid) c) Mature form - single chromatin mass but with an increase amount of cytoplasm that partially fills the erythrocyte
  • 124.  Early trophozoites have the characteristic signet ring shape.  Also, unique to P. falciparum is the presence of multiple trophozoites in one cell.
  • 126.  The gametocytes of P. falciparum have a crescent or banana shape.
  • 127.  Red blood cells infected by P. vivax are often larger than uninfected red blood cells.  They are approximately 1.5 times the size of a normal cell.
  • 129.  A schizont showing the large number of merozoites typical of this species (16-24).  Also note the larger size compared to a normal red blood cell.
  • 130.  Characteristic trophozoites of P. malariae showing the ring shape and the tendency of infected cells to be of normal or smaller size (arrows).
  • 131.  A schizont containing merozoites (6 to 12) giving a coarse granular appearance.
  • 132. Species rbc stage Size of rbc Schauffner’s stippling P. vivax Young rbc enlarged + P. malariae Mature rbc normal - Ziemann’s dots P. ovale Young rbc enlarged + P. falciparum rbc of all stages Normal/ Multiple infection _ Maurer’s dots
  • 133.  A red blood cell showing the Schauffner's dots characteristic of cells infected by Plasmodium vivax and Plasmodium ovale. Schauffner’s dots
  • 134. Species Cytoplasm Pigment No. of merozoites Gametocytes P. vivax Irregular, ameboid in trophozoite, has spread out appearance Golden brown, incons- picuous 12-24 Average is 16 rounded
  • 135. Species Cytoplasm Pigment No. of merozoites Gametocytes P. malariae Rounded, compact trophozoite with dense cytoplasm/ band form troph are rarely seen Dark brown, cons- picuous 6-12 Average is 8, “rosette” schizonts are sometimes seen rounded
  • 136. Species Cytoplasm Pigment No. of merozoites Gametocytes P. ovale Rounded, compact trophozoite with dense cytoplasm/ band form troph are rarely seen Dark brown, cons- picuous 6-12 Average is 8, “rosette” schizonts are sometimes seen rounded
  • 137. Species Cytoplasm Pigment No. of merozoites Gametocytes P. falciparum Young rings are small, delicate, often with double chromatin mass Black coarse and cons- picuous in gametocyte 6-32 Average is 20-24 Crescent, sausage- elongate shape
  • 139.  Toxoplasmosis : infectious mononucleosis -like symptoms of fever, chills, headache, myalgia, lymphadenitis, and fatigue; chronic form = hepatitis, encephalomyelitis, mycocarditis  Parasite : Toxoplasma gondii  intracellular coccidian(sporozoan) parasite infecting various tissues(systemic) but confined to intestines in felines  infected tissues release pseudocysts containing bradyozoites  feline intestines release oocyts containing tachyzoites  Pathogenesis : cellular death due to intracellular muliplication of trophozoites especially severe in fetal and cerebral tissues Pathogenic Protozoa - Toxoplasma gondii
  • 140.  Epidemiology : reservoir is a variety of mammals and birds including humans  fecal-oral transmission of oocyst from felines  common source transmission of pseudocyts in poorly cooked meat esp. beef  Diagnosis : serological detection of antibody: Elisa test for IgM antibodies is most reliable  Risk Groups : Pregnant females and immunocompromised
  • 143.  Enteroepithelial cycle asexual schizogony Sexual gametogony *immature oocyst passed in the cat’s feces  Immature oocyst mature to the infective stage 2 sporocysts with 4 sporozoites each (2-21 days)
  • 144.  Ingestion of inadequately cooked meat, esp lamb or pork  Ingestion of infective oocysts from material contaminated by cat feces  Drinking contaminated water or unpasturized goat’s milk  Organ transplantation  Blood transfusion  Transplacental
  • 145.  When mature oocyst is ingested, sporozoites are released forming trophozoites *tachyzoites- actively growing trop *bradyzoites- latent or slow growing trophozoites
  • 146.  Active infection (all nucleated cells) fever lymphadenopathy  Congenital depends on the age of gestation when the infection is acquired
  • 147.  1st half pregnancy - intrauterine death - microcephaly - hydrocephaly with intracranial calcification
  • 148.  2nd half of pregnancy -usually assymptomatic at birth though fever, hepatosplenomegaly and jaundice may appear -choriorenitis, psychomotor retardation and convulsive disorders may appear months or years later
  • 149.  Immunocompromised individuals -present CNS involvement -pneumonitis, myocarditis, retinitis, pancreatitis or orchitis
  • 150.  Examination of tissues, blood, and body fluids - Demonstration of tachyzoites or tissue cysts - Animal inoculation to uninfected mice - PCR - Sabin-Feldman dye test - IFA - EIA
  • 152.  Infection of rbc often producing febrile illness  Transmitted by ticks (Ixodes scapularis, Ixodes pacificus)  Caused by Babesia microti  mulitiply in rbc by schizogony but do not produce gametocytes  Appear as delicate ring forms that may easily confused with malaria
  • 153.  The organisms resemble P.falciparum  they vary more in shape and in size; and they do not produce pigment  evidence of Maltese cross formation
  • 157.  TICK vector of Babesia  The organism belongs to the genus Ixodes
  • 160. MALARIA BABESIOSIS Multiple rings _ + Large trophozoite + _ Gametocytes + _ Hemozoin pigment + _
  • 161. Patients with spleen only 25% of RBCs could be parasitized Patients withoutspleen higher parasitemia fever, muscle aches, chills, fatigue, headache and loss of appetite Capillary microvascular stasis RBC fragments Hemolytic anemia liver, spleenic, renal and Central Nervous System involvement.
  • 162.  Indirect Fluorescent Antibody  PCR
  • 163.  antimicrobial drugs such as clindamycin* plus quinine or atovaquone* plus azithromycin are often used. Prevention/Control - wearing appropriate light-colored clothing -using tick repellent in areas where the diseases are endemic -if possible test blood to be transfused for possible presence of Babesia species
  • 164. Blood and Tissue Flagellates
  • 166. Species Disease Portal of Entry and Final Site Vector Trypanosoma cruzi Trypanosoma brucei gambiense Trypanosoma brucei rhodesiense American Trypanosomiasis or Chagas’ Disease Gambian or West African Sleeping Sickness Rhodesian or East African Sleeping Sickness Breaks in skin or intact mucous membranes, later any organs and blood stream; Heart and GIT in chronic form Skin for both Lymph nodes, blood, and brain Infected Triatomine bug Infected tse-tse flies
  • 167.  African Sleeping Sickness CNS infection progressing to lethargy, tremors, meningoencephalitis, convulsions, coma, and death  Parasites Trypanosoma gambiense - West/Central Africa  Trypanosoma rhodesiense - East Africa  Pathogenesis - trypanomastigote multiplies in the blood, lymph, and cerebrospinal fluid; deprives the brain of amino acids Pathogenic Protozoa - Trypanosoma brucei
  • 168.  Epidemiology : reservoir is a variety of mammals and birds including humans  fecal-oral transmission of oocyst from felines  common source transmission of pseudocyts in poorly cooked meat esp. beef  Diagnosis : serological detection of antibody: Elisa test for IgM antibodies is most reliable  Risk Groups : Pregnant females and immunocompromised
  • 169.  American Trypanosomiasis/Chagas Disease -systemic infection beginning with fever, chills, malaise, myalgia, fatigue, and chagoma; progressing to involve heart and brain  Parasite = Tryanosoma cruzi  flagellate tryanosome exhibiting both the intracellular amastigote(leishmanial) form and the extracellular trypanomastigote(trypanosmal) form  Pathogenesis - the intracellular amastigote multiplies in cells and destroys the cells Pathogenic Protozoa - Trypanosoma cruzi
  • 170.  Epidemiology- wild and domestic animal reservoirs  arthropod-borne via the cone-nosed (reduviid) bugs which live in the cracks of domestic dwellings  Diagnosis : trypanosomes in the blood; amastigotes in biopsy; also xenodiagnosis and gene probes
  • 173. Local inflammation at the site of inoculation. CHAGOMA = reddish painful nodule Trypanosmes may enter the conjunctiva = edema of eyelid & conjunctiva = ROMAÑA’S SIGN
  • 177. Winterbottom's sign is the swelling of lymph nodes (lymphadenopathy) along the back of the neck, in the posterior cervical chain of lymph nodes, as trypanosomes travel in the lymphatic fluid and cause inflammation.
  • 178. w/ CNS invasion = chronic disease known as sleeping sickness stage is initiated (Kerandel’s sign) = delayed sensation to pain indicative ofAfrican trypanosomiasis Untreated infection may lead to death
  • 180. Species Disease Portal of Entry and Final Site Vector Leishmania donovani Leishmania tropica Leishmania brazilienses Visceral Leishmanisis or KALAAZAR Cutaneous Leishmaniasis Mucocutaneous Leishmaniasis or ESPUNDIA Skin/ Macrophages and monocytes of spleen, liver, bone marrow Skin/ Macrophages of the skin and mucosa Infected Phlebotomus Fly
  • 181.  Leishmaniasis;Visceral : gradual onset of fever, chill, sweating, diarrhea, anemia which progresses to enlargement of liver and spleen with weight loss and emaciation: later kidney damage and granulomatous skin lesion = also called kala-azar  cutaneous form -ulcerative skin lesions which contain parasite: become infected with secondary bacteria and scar formation disfigures  Parasites : Leishmania donovani - visceral- amastigote stage multiplies in visceral cells causing lysis  Leishmania tropica - cutanous: Leishmania braziliensis – mucocutaneous  Pathogenesis - cells in infected visceral and cutaneous tissues are destroyed by intracellular amastigote multiplication Pathogenic Protozaa - Leishmania
  • 182.  Epidemiology : animal reservoirs including dogs and cats  arthropod-borne via the bite of sandflies: the amastigote stage becomes a motile promastigote in the sandfly gut  Diagnosis : amastigote is observed in tissue biopsy, body fluid aspirates, and in the blood: also serology for antibody presence
  • 183. Species Amastigote Promastigote Epimastigote Trypomastigote Leishmania Animal Host Vector Macrophages - - midgut and pharynx of sandfly - - - - Trypanosoma Animal Host Vector - - - - - salivary gland and gut of flies/ culture Blood, lymph nodes, brain, CSF - Trypanosoma cruzi Animal Host Vector Intracellular, esp. striated and smooth muscle, brain Cell cultures - - - - Intestine of vector bugs Blood and tissues of mammals Rectum of vector bug/culture and cell cultures
  • 189. Cutaneous Leishmaniasis 1. Skin ulcers, w/ painless lesions = ugly scar 2. Subcutaneous nodules
  • 190. Mucocutaneous Leishmaniasis Metastatic spread to the oronasal and pharyngeal mucosa (ESPUNDIA) = highly disfiguring leprosy like tissue destruction & swelling (“TAPIR NOSE”)
  • 192. The current standard treatment for first stage disease is:  Intravenous pentamidine (for T.b. gambiense); or  Intravenous suramin (for T.b. rhodesiense)  Eflornithine
  • 193. To HIM we LIFT the HIGHEST PRAISE