Haemoflagellates

By
Mr Matheus Heita Namidi
Oshakati campus 2020
Life cycle of L.d
 Life cycle of L.d

Hosts: complete its life cycle in two hosts

Vertebrate host: man, dogs, rodents
Insect vectors (female sand fly) of the phlebotomus argentipes
• Infective form is the Promastigote forms present in the midgut of the female
sandfly.
• Most of transmission: Bite by an infected sandfly mainly during the late night
time. Maximum10-1000 promastigote per infective bite are required to initiate
the infection.
 Life cycle of L.d
In vertebrate hosts
1. Promastigotes are regurgitated from the midgut of the sandfly into the skin
of the vertebrate host.
2. Promastigote are phagocytosed by the skin macrophages and inside the
macrophages they transform into the Amastigote form within 12-24 hours.
3. Amastigote multiply inside the macrophages causing cell rupture and
releasing many amastigotes into blood circulation.
4. The amastigotes in the circulation are carried to various organs eg The liver,
spleen, bone marrow and then invade the Reticuloendothelial cells.
 Life cycle of L.d

In sandfly
1. During the blood meal, the amastigotes are ingested and transform into
promastigote in the insect gut.
2. The promastigote multiply by longitudenal fission and pass through various
stages.
3. Amastigote procyclic promastigoteNectompnad promastigote
haptomonad promastigote leptomonad promastigote metacyclic
promastigote.
 Life cycle of L.d
• The metacyclic promastigote multiply in the midgut of the vector by binary
fission.
• A small propotion migrate to the foregut to be able to infect a new host during
the next blood meal.
• The cycle in the sandfly last for about 4-18 days depending on the species.
 Clinical features

Visceral leishmaniasis
 Incubation period is 2-6 months
 Causing splenomegaly is the most consistant sign
 Common symptoms: Fever, Bone marrow dysfunction, leucopenia,
thrombocytopia, retinal hemorrhage, gastrointestinal bleeding.
 Lemphadenopathy: common in most of the African endemic regions
 Mucosal lesions in moutha and nasophaynx seen in Sudan.
 Leishmanoma: Nodular skin lesions seen in African cases only.
 Weight loss
 Secondary infections: measles, pneumonia, TB, amoebic/bacterial dysentery
Leishmania Donovani
Treatment:
• Sodium stibogluconate intravenously
• However resistance has been a serious problem. Drug resistant cases have been
treated Pentamidine or Amphotericine B. Miltefosine is newer oral drug.
• Prevention:
• Early detection and treatment of cases
• Eradication of sand flies
• Personal prophylaxis of insect repellents
• No vaccine is available
Post Kala-azar dermal leishmaniasis (PKDL)
• PKDL develop 1- 2 years after completion of treatment of visceral Leishmaniasis
or Kala-azar caused by L.donovani.
• Caused by reversal of the parasite from Viscerotropic to dermotropic.
• Occurring in 10-20% of the patients in INDIA but rare in Africa and China.
• Also happens in patients with no history of visceral disease BUT only in places
where L. dovovani is prevalent.
• Patients develop: Hypopigmented macules, erythematous patches on the nose,
cheeks, lips, forehead and ears….resembling Lepromatous leprosy.
 Post Kala-azar dermal leishmaniasis (PKDL)

• Stained biopsy materials shows amastigote forms of the parasite.

• Treated with the same drugs used for treatment of Kala-azar.
Cutaneous Leishmaniasis
• Its morphologically similar to L. donovani
• Causes cutaneous leishmaniasis disease classes as “ OLD AND NEW WORLD
TYPE OF LEISHMANIASIS”.
• L. major, L. tropica, L. aethiopica causes the Old world cutaneous leishmaniasis.
• L. braziliansis complex, L. Mexicana complex causes the New world cutaneous
leishmaniasis.
• NB: Conditions are called: Oriental sore, Dheli boil, Bagdad button.
Cutaneous Leishmaniasis clinical features
• Oriental sore begins as raised papule which becomes a raised indurated lesion
and in most cases its ulcerate.
• Healing occurs spontaneously in about 6 months leaving a depressed scar. The
lesions occurs mostly on the hands and face.
Cutaneous Leishmaniasis clinical features
• L. aethiopica may cause diffuse cutaneous leishmaniasis
• The new world group may cause cutaneous lesions and occasionally diffuse
cutaneous lesions.
Cutaneous Leishmaniasis clinical features
• Treatment: lesions heals naturally in a few months leaving the patient immune
for a whole life. However if the lesions are disfiguring, treatment is given by
way of ANTIMINIALS such as SIDIUM STIBOGLUTINATE.
• Lesions should always be disinfected and covered to prevent secondary
infections.
 Mucocutaneous leishmaniasis
• Extensive destructions of soft tissues in nasopharynx leads to painful mutilating
erosions named as ESPUNDIA.
• Fever, anaemia and loss of weight are common.
• Death is mainly caused by bacterial super infection, aspiration pneumonia and
respiratory obstruction.
Trypanosomes
These are haemoflagellates which live in the blood and tissues of their hosts
• They are species of:
1. Typanosoma brucei brucei AKA Trypanosoma brucei
2. Trypanosoma gambiense
3. Trypanosoma rhodesiense
• May produce disease in both animals and humans.
Trypanosomes species

Species Subspecies Disease

T. brucei • T. brucei  African sleeping sickness

• T. gambiense  Animal pathogen
• T. rhodesiense

T. cruzi • T.cruzi  Chaga`s disease

T. rangeli • T. rangeli  Infects human but doesn’t cause

disease
Trypanosoma brucei

• T. gambiense occurs in West and central Africa

• T. b rhodesiense confined to East and central Africa.
• Both are similar in all aspects except their geographical distribution and clinical
features.
Trypanosoma brucei
Morphology
• The parasite exist in vertebrate hosts as Trypomastigote which occur in three
forms.
1. A long slender form with a flagellum present in the blood extracellularly,
lymph and tissues. Colorless, spindle shapes, blunt posterior end, finely
pointed anterior end. Its motile. Have a large oval centrally positioned
nucleus.
2. A short stumpy form without a free flagellum and an intermediate form. The
portion of the flagella present intracellularly is called exoneme.
NB: The short stumpy form does not contain a free flagella.
Trypanosoma brucei life cycle
• T. gambiensi and rhodesiensi require two hosts to complete their life cycle
1. Definitive hosts: Humans and other vertebrates
2. Intermediate hosts: Insects
• For T. gambiensi, both humans are the only definitive hosts but for T.
rhodesiesnsi humans and wild animals act as the definitive hosts.
• The tsetse fly of the genus Glossina is the intermediate host.
• Both male and female flies bite man.
• The infective form for humans and other vertebrates is the metacyclic
trypomastigote.
Trypanosoma brucei life cycle
1. The infected tsetse fly introduces the metacyclic trypomastigote into the
definitive host.
2. Metacyclic trypomastigotes transform into a long slender form at the site of
inoculation.
3. These then convert into the short stumpy form and then into the
intermediate form with no free flagella. The short stumpy trypomastigotes
are non-dividing form.
4. Parasitaemia occurs due to invasion of blood stream by parasites
5. The Tsetse fly suck the short stumpy form during the blood meal
Trypanosoma brucei life cycle
6. The short stumpy form reach the Midgut of the fly and develop into the long
slender form and multiply.
7. The long slender form reach the SALIVARY GLANDS where they develop into
EPIMASTIGOTES.
8. The epimastigotes also start to multiply and convert non-dividing metacyclic
forms which are highly motile, stumpy and short.
9. On maturation, these metacyclic forms detach from the cells of the salivary
glands and are infective to the hosts which they enter during the blood meal.
Trypanosoma brucei life cycle
T.brucei & T. gambiense Pathogenesis
• Cause West African and East African trypanosomiasis (sleeping sickness)
• African sleeping sickness in fatal unless treated
• Incubation is about one week from the bite of the tsetse fly.
• A hard but tender chancre develop at the site of the bite
• The chancre heals but the injected organisms invade the blood causing
parasitaenia.
 T.brucei & T. gambiense Pathogenesis
• Antibodies develop but the mutant antigens escape the destruction. It is these
mutants that produce a new wave of parasitaemia.
• These mutants and parasitaemia continue to recur for months.
• If its untreated, the CNS gets involved.
• Progressive neurological impairment ends in coma and death
• East African sleeping sickeness is more acute and CNS get involved much earlier.
Death may occur within a year of onset even before CNS involvement.
• West African sleeping sickness is chronic in nature and may last for 4 years.
T.brucei & T. gambiense Pathogenesis

Clinical features
• Fever, Weakness and Rapid loss of weight are usual symptoms.
• CNS involvement symptoms characteristic include Severe headache, inability to
concentrate, behavioral changes from aggressiveness to day time sleeping
status (sleeping sickness).
• NB: The patient may fall asleep while eating, standing or sitting. In the final
stages it ends in coma and death.
T.brucei & T. gambiense

Treatment
• Suramin and Pentamidine are used in early stages where CNS is not involved
because these drugs DO NOT cross the blood-brain barrier
• Melarsoprol that crosses the blood-brain barrier is used in the late stages of
trypanosomiasis where the CNS is involved.
T.brucei & T. gambiense Prophylaxis
Prevention & control/ Prophylaxis
• Protective clothing
• Insect repellants
• Fly traps and screens impregnated with insecticides are the methods mostly
used to reduce the risk of infections.
• Vaccines are presently not available
Trypanosoma cruzi
• Trypanosoma cruzi is confined to central and southern America
• Discovered in 1909 by Carlos Chagas who named it Trypanosoma cruzi.
• T.cruzi causes Chagas` disease
• Morphology: Two morphological forms are seen in human hosts.
1. Trypomastigote form
2. Amastigote form
Trypanosoma cruzi
Trypomastigote form (20 µm in length).
• Appear in peripheral blood of patients
• Large kinetoplast at posterior end and a central nucleus
• In stained preparations, its appears as a C-shaped trypomastigote
• NB: IT DOES NOT MULTIPLY IN HUMANS
Amastigote form: It is the multiplying form.
• It is round in shape( 2-4 µm in diameter).
• Found in the heart and skeletal MUSCLES of patients, neurological cells and R.E cells.
• Another from the EPIMASTIGOTE form is found in insect vectors.
• Has an undurating membrane running along the anterior half of the parasite
• The epimastigote multiply in the hindgut of the vector.
Trypanosoma cruzi

• Can be cultured in NNN medium

• It is the EPIMASTIGOTE that will be present in the media.
Trypanosoma cruzi life cycle
Trypanosoma cruzi life cycle cycle
• T.cruzi passes its life cycle in two hosts.
• Definitive host: Humans
• Intermediate hosts: triatomine bugs
• NB: armadillos, Opossums,woodrats and raccoons are animals which act as
reservoir hosts.
Trypanosoma cruzi life cycle
• Infective form for humans is the METACYCLIC TRYPOMASTIGOTE
• Humans become infected when the REDUVIID BUG and the infected faecal
mater is discharged near the bite wound.
• The faecal mater is then rubbed into the wound by the bitten person because
that area starts itching.
• The infection can also be transmitted to the CONJUNCTIVA by the contaminated
fingers.
 Trypanosoma cruzi life cycle
• Abraded skin, oral and nasal mucosa can also be infected
• The metacyclic trypomastigote then invade the Reticuloendothelial system and
spread to other tissues.
• METACYCLIC TRYPOMASTIGOTE then is converted to AMASTIGOTE forms
• AMASTIGOTE form multiply by binary fission and after passing through
PROMASTIGOTES and EPIMASTIGOTES forms develop into TRYPOMASTIGOTE
forms.
• M. TRYPOMASTIGOTE
AMASTIGOTEPROMASTIGOTESEPIMASTIGOTESTRYPOMASTIGOTE
Trypanosoma cruzi life cycle
• Trypomastigote forms are liberated into the blood.
• When a reduviid bug bites such a person, the trypomastigotes are taken up
along with the blood meal.
• The trypomastigote is converted into Amastigote in the foregut and multiply by
binary fission.
• TrypomastigoteAmastigote (foregut)
• AmastigoteEpimastigote (midgut)
• Epimastigote Metacyclic trypomastigote (hind gut)
• Then the metacyclic trypomastigote are excreted in the faeces of the bug.
• This development of the T.cruzi takes about 10-15 days. The bug bites new
hosts and the cycle is repeated
Trypanosoma cruzi life cycle
• T.cruzi can aslo take place through blood transfusion and via congenital
• Pathogenesis
• T.cuzi causes South American trypanosomiasis AKA Chagas` disease
• Incubation period is 1-2 weeks
• Pathogenesis depents on the intracellular multiplication of Amastigotes and
causes damage to the cells at various sites.
• The MYOCARDIUM, SKELETAL MUSCLES, NEUROLOGICAL CELLS and the
RETICULOENDOTHELIAL CELLS are commonly affected.
 Chagas disease

Clinical features
• Chagas` disease manifests in two forms: Acute ad chronic
• A subcutaneous inflammatory nodule develop at the site of entry of the parasite
known as the CHAGOMA.
• When the entry is via the conjunctiva the patient develop painless unilateral
oedema and conjunctivitis. This is known as ROMANA`S SIGN.
• Acute form is more common in children and the chronic from in adults.
• Cardiac changes are more common and the patients may also develop
DIALATED oesophagus (megaoesophagus). And colon (megacolon).
 Chagas disease
Treatment
• No effective specific treatment is available
• Nifurtimox and Benznidazole have been used in acute infections.

Prevention and control(prophylaxis)

• Elimintio and control of vectors with insecticides
• Since the vectors live in cracks and crannies in walls. The provision of better
housing would contribute to prevention of the infection.
 Trypanosoma Rangeli
• T. rangeli infect humans but its not pathogenic
• T. rangeli infections are present in most areas where T.cruzi is also occur.
Therefore T.rangeli must be differentiated from T.cruzi morphplogically.

Feature Trypanosoma cruzi Trypanosoma rangeli

Trypomastigote
Trypomastigote size 20 30
Trypomastigote shape Often “C” shaped Not “C” shaped
Kinetolplast Large and terminal, situated at Small and subterminal, situated at
terminal end posterior end