Activity 2

Blood and Tissue

Protozoans
Hemoflagellates
 Lumen dwellers in their insect host
 Invaders of tissues and blood stream of
humans.

2 Genera under Trypanosomatidae:

 Trypanosoma spp. and Leishmania spp.
 Four Morphological Forms

1. AMASTIGOTE
• The parasite is more spherical in
Flagellum
shape and has no free flagellum.
Kinetosome

• A basal body and the base of the Kinetoplast

Nucleus
flagellum is still present.
• The kinetoplast is usually
detectable as a darkly staining
body near the nucleus.
• This form is a non-motile
intracellular stage.
Four Morphological Forms

2. PROMASTIGOTE
anterior

• the kinetoplast is towards

Flagellum
the anterior end and a free
Kinetosome
flagellum with no
Kinetoplast
undulating membrane Nucleus
emerges.

posterior
 Four Morphological Forms

3. EPIMASTIGOTE
• The kinetoplast is more anterior

kinetoplast
centrally located, usually just Undulating
anterior to nucleus. membrane

• The flagellum emerges from the

middle of the parasite and
forms a shorter undulating
membrane than observed in
trypomastigotes. posterior

• Epimastigotes are noticeably

less motile than Flagellated forms are
trypomastigotes. adapted for movement
through fluids of their
hosts.
 Four Morphological Forms

4. TRYPOMASTIGOTE
• The kinetoplast is located on anterior

the posterior end of the parasite. Undulating

membrane
• The flagellum emerges from the
posterior end and folds back
along the parasite's body.
kinetoplast

kinetoplast posterior

Flagellated forms are

adapted for movement
through fluids of their
hosts.
 Trypanosomes
Two broad categories:
- Salivarian – anterior station development, trypanosomes
migrate to salivary glands, introduced to host in blood meal.
Eg. African trypanosomes.
- Stercorarian – posterior station development, trypanosomes
in intestines of vector, transmitted in feces. Eg. T. cruzi,
(Chagas disease).
Trypanosoma brucei – has 3 subspecies:
 Trypanosoma b. gambiense African
 Trypanosoma b. rhodesiense sleeping
sickness

 Trypanosoma b. brucei
– Nagana disease (in animals)
Biological vector of
Trypanosoma b. gambiense

Glossina sp. (Tsetse fly)

Developmental cycle of Trypanosoma brucei. Diagram illustrating the development cycle of
Trypanosoma brucei in the mammalian and tsetse fly hosts. The transition between the long
trypomastigote found in the proventriculus and the short epimastigote that invades the salivary glands
is an asymmetric division [1, 2, 4]. Line drawings traced from fixed Giemsa-stained cells.
LIFE CYCLE OF Trypanosoma brucei gambiense
Trypanosoma b. gambiense

• Trypomastigote stage in the blood

of humans.
• The parasites are elongate, and
are numerous in the blood.
• Kinetoplasts are rather small
• Occurs in West and Central
Africa.
African trypanosomiasis
“African Sleeping sickness”

 Gambian and Rhodesian disease

(Trypanosoma b. gambiense and T. b.
rhodensiense)

Stages:
 Bite reaction (chancre)
 Parasitemia (blood and lymphoid tissues)
 Winterbottom’s sign
 CNS stage
 Trypanosoma brucei gambiense

PATHOLOGY - produces a
chronic disease.

1. Bite reaction (chancre)

- skin sore develops at bite site
where trypomastigotes are
inoculated into bloodstream.
 Trypanosoma brucei gambiense

2. Parasitemia and lymph node invasion.

•Trypomastigotes multiply in the blood
until they reach the lymphnodes;
•Marked by attacks of fever which starts 2-
3 weeks after the bite
•Symptoms: malaise, lassitude, insomnia,
headache and lymphadenopathy and edema
•Painful sensitivity of palms and ulnar
region to pressure (Kerandel's sign)
 Trypanosoma brucei gambiense

3. Winterbottom's sign
• Trypomastigotes invade the glands
of the posterior cervical region
where a visible enlargement or
swelling is seen. Very characteristic
of Gambian disease.
 Trypanosoma brucei gambiense

4. CNS stage
• Trypomastigotes invade CNS
• Marked by changes in character and
personality.(lack of interest and disinclination
to work, avoidance of acquaintances, morose
and melancholic attitude alternating with
exaltation, mental retardation and lethargy,
low and tremulous speech, tremors of tongue
and limbs, slow and shuffling gait, altered
reflexes, etc.)
• The later stages are characterized by
drowsiness and uncontrollable urge to
sleep.
• The terminal stage is marked by wasting and
emaciation. Death results from coma,
intercurrent infection or cardiac failure
 Trypanosoma cruzi
 Causes American trypanosomiasis or
“Chagas’ disease”.
 Site of entry of T.cruzi
 Cutaneous - at the bite site of a triatomine
bug – develops a subcutaneous
inflammatory nodule (Chagoma).
 Conjunctiva – develops painless, inflamed,
periophthalmic, unilateral oedema and
conjunctivitis (Romana’s sign).
 Primary lesions accompanied by fever,
acute regional lymphadenitis and
dissemination to blood stream.
 Trypanosoma cruzi
VECTOR -Triatoma infestans and Rhodnius prolixus

Order Hemiptera – family Reduviidae

Genera Triatoma, Panstrongylus, and Rhodnius
- common names: cone-nosed bug, assassin bug,
kissing bug, vinchuca
- adobe huts
- control
LIFE CYCLE OF Trypanosoma cruzi
Acute infections
 Metacyclic trypomastigotes develop into
amastigote forms in tissues.
 Fever, hepatosplenomegaly, generalized
lymphadenopathy, facial or generalized
oedema, rash, vomiting, diarrhea,
anorexia, ECG changes.
 Patients surviving acute infection develop
chronic disease wherein cardiac changes
are most common with arrythmias,
palpitations, chest pains, oedema, dizziness
and dyspnea.
Other chronic manifestations
 hepatosplenomegaly
 Dilatation of oesophagus
 Megaoesophagus
 Megacolon
 Loss of peristalsis
 Regurgitation
 Dysphagia
 Sever constipation

 Leads to DEATH
Trypanosoma cruzi

Trypomastigote stage in blood

stream
 Characteristic C- or ?-shape
 the kinetoplast is relatively
large in proportion to the body.
 Nucleus is central.

An important diagnostic feature.

Apicomplexan/Sporozoan
Invading blood and tissues of
humans
 Plasmodium spp.
 Toxoplasma gondii
 Sarcocystis spp.
LIFE CYCLE OF Plasmodium spp.
 10 mm

Sporozoites of Plasmodium vivax in a squash of an oocyst from an infected

Anopheles mosquito.
Life Cycle: Asexual phase in man

Pre-erythrocytic
schizogony –
inoculation of the
infective sporozoites
to man during a
mosquito blood meal.
Within a few hours,
they are carried with
the blood circulation
into the liver
parenchymal cells.
Erythrocytic phase – Merozoites rapidly and specifically enter
erythrocytes. This specificity is manifested both for
erythrocytes as the preferred host cell type and for a
particular host species, thus implying receptor-ligand
interactions.
Differential diagnostic features of Plasmodium
species (Erythrocytic stages).

P. falciparum P. malariae P. vivax P. ovale

YOUNG
TROPHOZOITE

Fine ring Thick ring

Thick ring Thick ring
multiple infection Often irregular
1 chromatin dot I chromatin dot
1-2 small chromatin dots 1 chromatin dot

MATURE
TROPHOZOITE

Ring enlarged Band forms Irregular Round

Slightly irregular Pigment distinct Amoeboid Compact
 P. falciparum P. malariae P. vivax P. ovale

SCHIZONT

Rarely seen; Merozoites Merozoites arranged as Merozoites arranged Merozoites arranged

arranged irregularly rosette irregularly irregularly
No. of merozoites: 8-24 No. of merozoites: 6-12 No. of merozoites: 12-24 No. of merozoites: 6-14

GAMETOCYTE

Crescent shaped
Oval to round Oval to round Oval to round
or banana-shaped

Size of RBC Unchanged Unchanged Enlarged Enlarged

Sometimes irregular Often irregular with

Shape of RBC and crenated
Unchanged Unchanged
jagged edges

Sometimes present Rarely present Often present

Stippling Always present
(Maurer’s dots – course (Ziemann’s dots – pinkish (Schuffner’s dots – pinkish
(Presence of dots) (Schuffner’s dots)
and irregular) small dots) small and round dots)
Erythrocytic schizogony
During the erythrocytic phase, the organism differen-
tiates into a ring-shaped trophozoite.
The ring form grows into an ameboid form, undergo
a multiplicative process and then differentiates into a
schizont filled with merozoites.
The merozoites are released to infect other RBC.

The simultaneous rupture of the infected

erythrocytes and the concomittent release of
antigens and waste products accounts for the
intermittent fever, chills and sweating
(paroxysms) associated with malaria.
1.Malarial Paroxysm
Cold stage Sweating stage profuse
 feeling of intense cold • sweating
 vigorous shivering
• declining temperature
 lasts 15-60 minutes
• exhausted and weak
Hot stage intense
heat • sleep
 dry burning skin • lasts 2-4 hours
 throbbing headache
 lasts 2-6 hours
The schizogonic cycle in the RBC repeats at regular intervals
typical for each species:
1. Plasmodium malariae – every 72 hours;
Quartan malaria.
2. Plasmodium vivax – every 48 hours;
Benign tertian malaria
3. Plasmodium falciparum – every 36-48
hours; Malignant tertian malaria
4. Plasmodium ovale – every 48 hours;
Ovale tertian malaria
2. Anemia – microcytic or
normocytic type
As a consequence of:
 mechanical destruction of parasitized red
blood cells
 Reduced erythropoeisis in the bone marrow
 Lysis and phagocytosis or uninfected RBC

3. Splenomegaly
 Due to massive proliferation of
macrophages which phagocytize both
parasitized and non parasitized RBC.
LIFE CYCLE OF Toxoplasma gondii
 Enteric cycle – occurs in cat
 Cats are infected by ingestion of tissue cysts
containing tachyzoites and bradyzoites from
intermediate hosts.
 Tachyzoites, bradyzoites and sporozoites from
oocysts can undergo several asexual cycles
 Male and female gametocytes are formed and
give rise to gametes
 Oocysts develop in the cat’s intestinal mucosa
after sexual fusion of male and female
gametes.
 Oocysts sporulate and become infectious to
intermediate hosts.
Exoenteric cycle
 Contaminated food and water containing
sporulated oocysts may cause infection to
intermediate hosts (humans, mice, rats,
and certain birds)
 Ingested oocysts release sporozoites and
develop tissue cysts containing bradyzoites
and tachyzoites in various organs.
 Undercooked meat from intermediate hosts
with tissue cysts can be a source of
infection when ingested by other hosts.
Toxoplasma gondii tissue cysts in
intermediate host
 Infection most
commonly involve
nervous tissues.
 Internal
accumulation as a
result of asexual
reproduction.
Cyst wall  Immune response
by host slows
down the division.
 Tachyzoites within
cysts transform
into bradyzoites.
 SARCOCYSTIS spp.
 S. hominis and S. suihominis
 have humans as their definitive hosts.
 S. hominis – cow is the intermediate host.
 S. suihominis – pig is the intermediate host.
 Other Sarcocystis spp. – have humans as one
of their intermiediate hosts (also as a dead-
end host).
LIFE CYCLE OF Sarcocystis hominis
Sarcocystis sp. tissue cysts/
sarcocysts
 Infection most
commonly involve
muscles.
 Sarcocyst in
skeletal muscle
packed with
bradyzoites.
 Compartmentalized
by internal septa.