BLOOD AND TISSUE

FLAGELLATES
 Flagellates that are found in the
blood and other fluids
(CSF) and in tissues
 Vector borne parasites
 Medically important genera:
o Trypanosoma
o Leishmania
GENERALITES
 Only Trypanosoma and
Leishmania infect humans
 Transmitted by a bite of an
infected vector
 All forms are found in
Trypanosoma cruzi infections
 Only the epimastigote and
trypomastigote are seen in
 Trypanosoma brucei infections
 Only the amastigote and
promastigote are seen in
 Leishmania infections
 Diagnostic Stages (found in
humans) include amastigote
and or trypomastigote
2) Promastigote
- common morphology in the
insect form
-  flagellum is found anterior of
nucleus and flagellum not attached to the
cell body
- kinetoplast is located in front of
the nucleus, near the anterior end of the
body
3) Epimastigote(metacylic from vector)
- a common form in the insect
host
- flagellum exits the cell anterior
of nucleus and is connected to the cell
body for part of its length by an
undulating membrane
-kinetoplast is located between
the nucleus and the anterior end.

4 Morphological Forms:

1) Amastigote
-occurs in all trypanosomal
genera
-leishmanial stage
-no protruding flagellum
- it divides by longitudinal binary
fission at 37C
4) Trypomastigote
- trypanosomal
-in the mammalian host
bloodstream as well as infective
metacyclic stages in the fly vector
-the kinetoplast is near the
posterior end of the body, and the
flagellum lies attached to the cell body
for most of its length by an undulating
membrane.
 Pathogenesis
 Acute inflammatory reaction on
bite
 Uses lectinlike carbohydrates for
binding
 Direct Inflammatory response
 Chaga-toxin
 damage to infected cells
 Destruction of autonomic
AMERICAN TRYPANOSOMIAS nerve ganglions
 Target cells: cells of RES,
Trypanosoma cruzi cardiac cells, skeletal and
 causes Chagas’ Disease or smooth muscles, neuroglia cells
American
 Final Host: Humans
 Reservoir Hosts: Armadillo,
opossum, raccoon, dog, cat, and
 other animals
 Intermediate host Vector:
Reduviid Bug (major),
 Triatomine bugs, kissing bug
(Triatoma, Rhodrius and
Panstrongylus)
 MOT: Feces of vector entering
bite wound; blood
 transfusion, organ transplants;
transplacentally
 Infective Stage to Humans:
Metacyclic Trypomastigote
Disease Manifestation
 Chagoma – local inflammation;
reddish nodule
 Romaña’s sign(1 week after) –
periorbital swelling (edema of
eyelid
 and conjunctiva)
 Acute Phase: fever and
lymphadenopathy
 Intermediate Phase: no
characteristic symptoms
 Chronic Phase: Mega esophagus,
mega colon, cardiomegaly,
cardiac arrhythmia Diagnosis
 Demonstration of trypanosomes
in blood, CSF, tissues,
 lymph (staining using Giemsa)
o Wet smear-living
state(motile)
o Stained smear-
morphology
o Xenodiagnoses- other
species
 Serology: IFAT, Complement
Fixation, ELISA
 Culture: Chang’s , NNN
Treatment and control
 Nifurtimox and Benznidazole
 Vector control, screening of
blood, health education
AFRICAN TRYPANOMIASIS
 Vector: Tsetse Fly (Glossina
spp.)
o T. b. rhodesiense – G.
pallidipes, G. morsitans
o T. b. gambiense – G.
palparis
 Mechanical:
o mother-child infection
(perinatal death)
o blood transfusion and
accidental pricking
 Infective Stage to Humans:
Metacyclic Trypomastigote
 Agent of sleeping sickness
 Trypanosoma brucei brucei
o In cattles
Trypanosoma brucei complex
 Trypanosoma rhodesiense
o Causes Rhodesian or East
African Sleeping
Sickness
o Endemic in East and
South Africa
 Trypanosoma gambiense
 Causes Gambian or West
African Sleeping
Sickness
 Endemic in West and
Central Africa
Morphological forms
 Epimastigote
o Crithidial form in insect
 Trypomastigote
o Trypanosomal form in
mammal
Disease Manifestation:
T. gambiense
 Causes Gambian or West Afrcian
Sleeping Sickness
 Earliest Sign(1st stage or
Hemolymphatic stage):
Trypanosomal Chancre
o Patients still appear
healthy
o Blood Smear: (+)
Trypomastigotes
o Patients experience fever
once the lymph nodes are
Affected(hemolympatic
stage). Other
manifestations include
malaise, weakness,
night sweats, dizziness
and nausea.
o Winterbottom’s
Sign(swelling of lymph
nodes in posterior neck)
 Chronic stage
o CNS Invasion
o Sleeping Sickness Stage
initiated
o Signs and Symptoms:
Severe Headache,
increasing mental
o deterioration and apathy,
meningoencephalitis
o Manifestation of
Kerandel’s Sign
o Terminal Phase: Coma
leading to Death
T. rhodisiense
 causes Rhodesian and East
African Sleeping Sickness
  Similar to Gambian Sleeping  Physical Findings and Patient
Sickness History
 but Acute and rapidly o Card Agglutination test
progressing  direct, Ab test
 CNS stage takes place in the with IGG test for
early stages detection of
 Glomerulonephritis may also be IgM(history) and
seen IgG(acute)
Pathogenesis o Card Indirect
 Generalized Lymphoid Agglutination test
Hyperplasia  test for antigen
 Anemia  high specifity and
 Thrombocytopenia sensitivity
 Hypergammaglobulinemia  Demonstration of
 Immune Evasion through: Trypomastigotes in Blood,
Variant Surface Glycoproteins CSF(lumbar puncture), Lymph
 Acute Infection seen in Node Aspirate
Rhodesian Sleeping Sickness  Concentration of Buffy Coat
 Chronic Infection seen in  Giemsa Stain
 Serology – IHAT, ELISA, Rapid

Gambian Sleeping Sickness Tests

 Molecular Methods(PCR)
Diagnosis Treatment:
  Better Prognosis if treatment
started before CNS stage
 Pentamidine and Suramin (blood
and lymphatic stage)
 Melarsoprol (Late Stage)
LEISHMANIA
 Vector Borne Parasitic Disease
o Vector: Sandflies
(Phlebotomus spp.)
 Obligate Intracellular Parasites
 Primarily a zoonotic disease
 Amastigote (leishmania)
o seen in the mammalian
host
 Promastigote(leptomonad)
o seen in sand fly
3 types
1. L. tropica
o Damages subcutaneous
(leishmaniasis)
o Incubation Period: Weeks
to Months
Disease Manifestation
 Elevated Skin
Ulcers (Painless)
 Leaves an Ugly
Scar; Highly
Disfiguring
2. L. braziliensis
o Damages mucocutaneous
lesion(mucocutaneous
leishmaniasis)
o Tapir nose(disfiguration),
Espundia, Chictero’s
ulcer
3. L. donovani
o Damages visceral and
internal organs
o Dum-dum fever or Kala-
azar fever
o Incubation: 1-3 months
o Manifestations
 Fever with twice
daily elevations
(“dromedary fever
peak”),
 Splenomegaly,
Cachexia
 Hepatomegaly
 Darkening of
Skin (forehead,
temples, around
the mouth)
 Dermal
Leishmanoid
Lesions – may be
rarely seen
MOT
 Blood transfusion, contamination
of wounds, contact
 Infective stage: Promastigote
 Target: RE cells
Diagnosis
 Tissue biopsy( braziliense)
 Skin biopsy ( tropica)
 Spleen and lymph node
biopsy( donovani)
Tests
1. Stained smear
2. Culture (Novey McNeal Nicole)
 Non nutrient agar
 Rabbit’s blood( for
enrichment)
3. Intradermal test(Montenegro test)
 Skin test
 Somatic antigen test
4. Formul-gel test
5. Serological test
 CFT, IFAT
6. Total protein: globulin ratio
Treatment
 Antimony compounds
o Amphotericin B
o Pentamidine
o Nifurtimux
o Sodium
Stibogluconate,
o N- methyl
glucamine
antimonite