Blood and Tissue

Flagellates
College of Medical Laboratory Science
Our Lady of Fatima University-Valenzuela

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Phylum Sarcomastigophora
Subphylum Mastigophora
Hemoflagellates - (arthropod borne)

A. Leishmania spp.
- Leishmania tropica
- Leishmania braziliensis
- Leishmania donovani

B. Trypanosoma spp.
- Trypanosoma gambiense
- Trypanosoma rhodesiense
- Trypanosoma cruzi
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Morphologic forms
1. Amastigote
– round, ovoid
─ usually found in small groups of
cyst-like collection in tissues
─ “binary fission”
2. Promastigote
3. Epimastigote
─ “longitudinal fission”
4. Trypomastigote
─ Long slender or short stumpy
─ C, U, S-shaped

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 Trypanosoma spp.
Etiologic Disease Vector Stages
Agent exhibited
T. cruzi Chaga’s Disease or Assasin bug,
American Kissing bug, ALL
Trypanosomiasis Cone nose bug,
Triatomine bugs
Triatoma,
Rhodnius,
Panstrongylus
T. brucei Gambian or West Tsetse fly, Epimastigote
gambiense African Sleeping Glossina spp. and
Sickness Trypomastigote
T. brucei Rhodesian or East only
rhodesiense African Sleeping
Sickness
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 Trypanosoma cruzi

• Belongs to the group Stercoraria (multiplies within the

mammalian host in a continuous manner)
• Infected cells (intracellular parasite):
– Skin
– Gonads
– Intestinal mucosa
– Placenta
– Myocytes (particularly myocardial tissues)
– Reticuloendothelial system cells

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Life cycle of Trypanosoma cruzi
intracellular parasite

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 Trypanosoma cruzi
Pathology
- “Chagas’ disease” or American trypanosomiasis
- usually serious and fatal in young children
- “Chagoma” – furuncle-like lesions, inflammation
at the site of inoculation
- small, painful, reddish nodule
1. Acute trypanosomiasis
- generalized lymphadenopathy
- focal or diffuse inflammation mainly affecting the myocardium
- “Romaña’s sign” – edema of the eyelid if the parasite
penetrates through the conjunctiva
2. Chronic trypanosomiasis
- no characteristic symptom and may last for 20 years or more
- cardiomyopathy, megaesophagus and megacolon
- these advanced conditions can lead to death
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 Trypanosoma cruzi
Infective stage to vector : trypomastigote
to man : metacyclic trypomastigote

Specimen: blood, CSF, fixed lymph node tissues

and lymph juices

Laboratory tests
1. Stained smear – Giemsa staining : demonstration of trypomastigote
2. Concentration methods (Microhematocrit)
3. Blood Cultures – NNN medium (Novey-McNeal-Nicolle),
4. Xenodiagnosis – use of laboratory animal
4. Serologic test (IFA, CFT, IHA, ELISA)
*Dot-immunobinding: small amounts of sample is used
5. Molecular testing (PCR)- amplify DNA from kinetoplast

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 Trypanosoma cruzi

Treatment:
- Nifurtimox and Benznidazole

Prevention and Control

- vector control (insecticide spraying)
- screening and sterilization of transfusion blood
- health education

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 Trypanosoma brucei complex

• Belong to the trypanosome family Salivaria.

• Collectively the three subspecies represent the Trypanosoma
brucei complex:
• Trypanosoma brucei gambiense
• Trypanosoma brucei rhodesiense
• Trypanosoma brucei brucei

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Life cycle of Trypanosoma brucei complex

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 Trypanosoma brucei complex
Pathogenesis
Chancre – earliest sign of African trypanosomiasis
- hard, painful lesion at the site of inoculation

A. Gambian trypanosomiasis
1. Acute: fever, headache, joint and muscle pain, tachychardia,
dizziness and rashes
“ Winterbottom’s sign” – enlargement of the posterior cervical
lymph node and have a ripe plum consistency.

2. Chronic : with CNS invasion

- severe headache, alternately morose and excitable, and lack
interest in work.
- tremors and “Kerandel’s sign” or hyperesthesia and inversion
of sleep cycle can be observed
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 Trypanosoma brucei complex
B. Rhodesian trypanosomiasis
- more rapid and fatal than Gambian trypanosomiasis
- CNS involvement appear early
- Neurologic deterioration is rapid

Specimen: blood, CSF and lymph juices

Laboratory tests:
1. Wet smear : presence of live parasite
2. Stained smear – Giemsa staining : demonstration of trypomastigote
3. Serologic test
a. IFA
b. ELISA
c. Mini-anion centrifugation technique
d. Indirect hemeagglutination
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 Trypanosoma brucei complex

Chancre Winterbottom’s Sign

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 Trypanosoma brucei complex
Treatment: effective on earlier stages

- Suramin sodium and Pentamidine

- Melasorprol and tryparsamide (w/ CNS involvement)
• Febrile episode, Jarisch-Herxheimer reaction, due to
trypanosome lysis following melasorprol treatment
- DL-alpha-difluoromethylornithine (DFMO, Eflornithine) for
T. brucei gambiense only

Prevention and Control

- Control of tsetse flies (traps, screens, insecticides)
- reduction of pool of human infection
(diagnose as many as individuals as possible)
- trimming of bushes

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 Leishmania spp.
Etiologic Disease Vector Stages
Agent exhibited
Leishmaina Cutaneous Sandfly vectors V. host:
tropica leishmaniasis (Phlebotomus Amastigote
Leishmania American or spp.) (reticuloendoth
braziliensis Mucocutaneous elial system)
Leishmaniasis
I. host:
Leishmania Visceral
Promastigote (
donovani Leishmaniasis/
midgut and
Kala-azar/
proboscis)
Dumdum fever

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 Leishmania spp.

Etiologic Agent Other name

Leishmania tropica Old world leishmaniasis, Oriental
leishmaniasis, Cutaneous
leishmaniasis, Jericho boil, Baghdad
boil
Leishmania braziliensis New world leishmaniasis
Leishmania donovani Kala-azar/ Dumdum fever

Phlebotomus Lutzomyia
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Life cycle of Leishmania spp.

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 Leishmania spp.
Pathology
A. Cutaneous Leishmaniasis (Leishmaina tropica)
- most common form, skin ulcer (leaving an ugly scar on healing)
- oriental button at inoculation site
- painless lesions
- Diffuse Cutaneous Leishmaniasis resembles
“leprematous leprosy”
- lesions do not heal spontaneously and tend to relapse after
treatment
B. American or Mucocutaneous Leishmaniasis (Leishmaina braziliensis)
- “Espundia” - metastatic spread of lesion to oronasal and
pharyngeal mucosa
- “Tapir nose” – disfiguring leprosy-like tissue destruction and
swelling
- “Chiclero ulcer” – erosion of the pinna of the ear
- “forest yaws”, “pian bois”, ”uta”
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 Leishmania spp.
Pathology
Cutaneous Leishmaniasis (Leishmaina tropica)

American or Mucocutaneous Leishmaniasis (Leishmaina braziliensis)

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 Leishmania spp.
Pathogenesis

C. Visceral Leishmaniasis (Leishmania donovani)

- Kala-azar or Dumdum fever
- twice-daily elevation of fever: prominent finding
- splenomegaly and cachexia
- parasites are numerous in reticuloendothelial cells of the
- spleen, liver, lymph nodes, bone marrow and other organs.
- FATAL

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 Leishmania spp.
Infective Stage to Man : promastigote
to vector : amastigote

Specimen: Blood and Tissue sample

Diagnostic Tests
1. Microscoping determination from tissue scrapings, lesions and biopsy
a. Cutaneous : skin
b. Visceral: spleen or bone marrow

2. Serologic Test
- Complement Fixation Test
- Montenegro’s intradermal test
- IFA
- Counter current electrophoresis techniques
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 Leishmania spp.
Treatment:
*Pentavalent antimonials: sodium stibogluconate and
n- methyl-glucamine antimonite (meglumine)
* Second line drugs
- Amphotericin B (intavenous)
- AmBisome (lipid-based drug for CL, and VL)
- pentamidine (kala-azar, limited use because of drug
resistance)
- Miltefosine (current oral drug for VL)
- Topical paromomucin (for CL)
- metronidazole
- nifurtimox
- Combination therapy
Prevention and Control
- Skin lesions must be protected from insect bites
- Health education
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 REFERENCES
Belizario, V. and De Leon, W. (2015). Philippine Textbook
of Medical Parasitology. Third Edition. University of the
Philippines Manila. Ermita, Manila.

Mikhail A. Valdescona, RMT, MPH. PAR313 Lecture.

Our Lady of Fatima University. Valenzuela City.

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