Hemoflagellates

Morphological and life cycle notes

 These are blood and tissue flagellates
 Clinical morphological forms that are important are; Amastigote,
Promastigote, Epimastigote and Trypomastigote.
 Transmission of all hemoflagellates is accomplished by bite of arthropod
vector
 Members of hemoflagellates belongs to genera leishmania and
trypanosome
 Primary diagnostic form found in leishmania is amastigote
 Primary diagnostic form found in trypanosome is trypomastigote with
exception of T. cruzi in which amastigote may be found.
 Serological tests are done to confirm hemoflagellate diseases.
Amastigote
 Roundish to oval consisting of nucleus and a kinetoplast
 Large single nuclear located off-center or towards the edge
 Has a dot like blepharoblast attached to axoneme
 Axoneme extend towards the edge of organism
 Has a single parabasal body adjuscent to blepharoblast
Promastigote
 Slightly long with slender body
 Large nucleus located near the center
 Kinetoplast located in anterior end of the organism
 Has single free flagellum extend anteriorly from the axoneme
Epimastigote
 Slightly wider than promastigote
 Has large single nucleus located posterior end of the
organism
 Kinetoplast located anterior to the nucleus
 Undulating membrane measures half of the body length
 Free flagellum at anterior end of the epimastigote
Trypomastigote
 Often assumes letter C or U in a stained blood film
 Long, slender with posteriorly located kinetoplast
 Full body length undulating membrane
 Single large nucleus located anteriorly
 Free flagellum may or may not be present
General Life cycle
 Amastigote and trypomastigote are the two common forms routinely
found in human specimen
 Amastigote primarily found in tissues, muscles and CNS within
macrophages
 Trypomastigote reproduce and is visible in peripheral blood

Henry W. Karoki, MSc.| Hemoflagellates

  Promastigote stage may be seen only if the blood sample is collected
immediately after transmission to healthy individuals
 Promastigote may also be seen when appropriate sample is culture
 Epimastigote is found primarily in arthropod vector as well as human
blood sample but in rare occasion.

Pathogenesis / Clinical symptoms

 Symptoms of infection range from small red papule at infection site with
intense itching, secondary bacterial infection, fever, diarrhea, kidney
involvement, mental retardation, comatose state and even death.
 Initial skin lesion spontaneously heal but others may remain dormant for
months or even years.
Laboratory diagnosis
 Blood, lymph node and ulcer aspiration, tissue biopsy, bone marrow and
CSF are specimen of choice for isolation of hemoflagellates morphological
forms
 Serological tests are available for confirmation.

Trypanosome brucei gambience

Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida

Morphology
Trypomastigote

Measures 12-35um long, often assume letter ‘C’ or ‘U’ in a stained blood film,
long slender and posterior located kinetoplasta, full body length undulating
membrane, single large nucleus located anterior free of flagellum may or may
not be present.

Henry W. Karoki, MSc.| Hemoflagellates

Life cycle notes
Tse tse fly responsible for transmission of parasite during blood meals (glossina
palpalis and glossina tachinoides), trypomastogote migrate through blood
stream into lymphatic system and multiply by binary fission, mutation of the
parasite manages to escape host immune system and eventually invade CNS ,
trypomastigote is transmitted back to tse tse fly vector through a blood meal on
infected human, in the fly, trypomastigote continue to multiply and migrate to
the salivary gland converting to epimastigote along the way, once in the
salivary gland, epimastigote transform back to trypomastigote completing the
cycle.
Epidemiology
Found in tropical west, central Africa along the streams banks where vector
breeds, no known animal reservoir host.
Symptoms
Causes west Africa (Gambian) sleeping sickness, include ulcers on bite site,
fever, headache, generalized weakness, lymph node enlargement “winter
bottom’s sigh”, mental retardation, tremor, meningoencephalitis may occur
when CNS in involved, patients slips to coma and death ensures from CNS
damage.

Winter Bottom’s sign

Laboratory diagnosis
blood, lymph node aspiration and CSF is the specimen of choice, giemsa
stained slides of blood revealed trypomastogote forms, other test on CSF and
serum include presence of IgM and presence of protein.
Prevention and control
Destroy breeding areas for Tse tse fly via chemical treatment or clearing bush,
proper protective clothing, use of repellant and screening, prompt treatment of
infected persons.
Treatment
Suramine, Melarsoprol, pentamidine isethionate and eflornithine are
drugs of choice and are dictated by patient’s age, stage of disease and if
the patient is pregnant at the time.
Toxicity levels of these medication are very high and hence care must be
taken when selecting specific treatment and appropriate dosage.

Henry W. Karoki, MSc.| Hemoflagellates

Trypanosoma brucei rhodesience
Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida
Morphology

Measures 12-35um long, often assume letter c or u in a stained blood film,

long slender and posterior located kinetoplasta, full body length undulating
membrane, single large nucleus located anterior free of flagellum may or may
not be present.

Life cycle notes

Transmitted by tse tse fly vector glossina morsitans and glossina pallipedes.
Rest of the cycle is as same as T.b. gambiense.

Epidemiology
Found in east and central Africa especially in bush areas, Reservoir are cattle,
sheep and wild game.

Clinical symptoms
Causes East African (Rhodesian) sleeping sickness, more virulent than T. b.
gambience, has short incubation period, acute sleeping sickness characterized
by fever, winter bottoms sign, rapid weight loss CNS involvement, mental
disturbance, lethargy, anorexia, death, kidney damage and myocarditis.

Winter Bottom’s sign

Henry W. Karoki, MSc.| Hemoflagellates

Laboratory diagnosis
Giemsa stained blood slides and CSF sediments is the specimen of choice,
protein and IgM studies on CSF may be performed, and serological tests may
be available.

Prevention and control

Early diagnosis and treatment, treatment of domestic animals and use of
repellant, dusting breeding areas, bush and control of fly population.

Treatment
Suramine, Melarsoprol, pentamidine isethionate and eflornithine are
drugs of choice and are dictated by patient’s age, stage of disease and if
the patient is pregnant at the time.
Toxicity levels of these medication are very high and hence care must be
taken when selecting specific treatment and appropriate dosage.

Trypanosoma cruzi
Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida
Morphology

Measures 12-35um long, Often assume letter C or U in a stained blood film,

Long slender and posterior located kinetoplasta, Full body length undulating
membrane, Single large nucleus located anterior free of flagellum may or may
not be present.

Henry W. Karoki, MSc.| Hemoflagellates

Life cycle notes
Most frequent transferred to human host by a reduviid bug (Triatomine
bug/Kissing bug) vector defecates infective trypomastigote. Presence of bite
produces itching sensation, Parasite is literally rubbed into bite wound.
Trypomastigote invade surround cells, Transform to amastigote, Amastigote
multiply, transform back to trypomastigote, Trypomastigote migrate through
blood transforming back to amastigote. Ares affected are heart musles, liver
brain, Trypomastigote is transmitted back to the reduviid bug when it feed via
blood meal, upon ingestion, trypomastigote transform to epimastigote in the
mid gut. Epimastigote multiply and transform back to trypomastigote in the
hind gut. These trypomastigote are passed in feces when bug defecates
Epidemiology
Found in south and Central America and in Brazil. Isolated from reduviid bug
(panstrongylus mengistus) also called kissing bug or cone nose bug.
Mammalian reservoir host are dogs and cats
Clinical symptoms
Causes chag'as disease. Initial symptoms are the development of
erythromatous nodule (chagoma) especially in the face. Mucosal infections
characterized by conjunctivitis, edema of the eyelids (romana’s sign), chronic
chaga's present as myocarditis, enlargement of the colon and esophagus,
hepatosplenomegally, and CNS involvement. Acute phase characterized by
chill, fever, fatigue, malaise, Acute may lead to recovery or death or
transmission to chronic after attack, Most common disease in children under 5
yrs.

Romana’s sign

Laboratory diagnosis
Giemsa stained blood slides and CSF sediments is the specimen of choice.
Protein and IgM studies on CSF may be performed, Serological tests may be
available such as ELISA. Biopsy of involved lymph node may reveal amastigote
forms. Blood or CSF of the patient is inoculated intra-peritoneally into mice
and trypanosomes appear in blood. Blood and other specimen may be
inoculated into NNN medium and incubated at 22-24OC and subcultured every
1-2wks then material examined for trypanosomes. Other techniques are PCR
techniques and Xenodiagnosis.

Henry W. Karoki, MSc.| Hemoflagellates

Prevention and control
Eradication of the reduviid bug, Apply residual insecticide (DDT), Education
programmes in endemic areas and housing improvements.

Treatment
Nifurtimox and Beznidazole may be used for treatment Of American
trypanosomiasis

Leishmania branziliensis
Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida

Morphology
Amastigote

Roundish to oval 5-3 um size, consist of a nuclear & a kinetoplasta, large

single nucleus locates off centre or towards the edge. Dot –like blepharoblast
give rise to & attached to axonemes axonemes extends towards edge of
organism, single parabasal body adjacent to blepharoblast

Life cycle notes

Sand fly responsible for transmission of promastigote via blood meal, it quickly
invade reticuloepithalial cell and transform to amastigote which causes tissue
destruction, skin and mucous membrane primarily affected. Diagnostic stage of
L.branziliensis is the amastigote, it is the infective stage for the sand fly, upon
ingestion, amastigote transform back to promastigote in the fly mid gut.
promastigote multiply and the resulting forms move to the salivary gland ready
for the transfer to the humans.

Henry W. Karoki, MSc.| Hemoflagellates

Epidemiology
Found in South America, eg Brazil, Columbia, Venezuela, Bolivia and Peru.
Infection greatest in rain forest areas esp. where harvesting of chicle sap for
chewing gum is done, reservoir hosts are the forest rodents and domestic dogs

Clinical symptoms
Causes mucocutaneous leishmaniasis, large ulcers in oral and nasal mucosa
after invasion of the reticuloendothelial cells. Untreated cases of mucosal lesion
results in the eventual destruction of the nasal septum, Lips, nose, and other
soft parts may be affected. Edema and secondary bacterial infection and
numerous mucosal lesions may disfigure the face and death ensues.

Laboratory diagnosis
Specimen of choice for the isolation of the amastogote is biopsy of the infected
ulcer, examination of the giemsa stained slides. Other methods are culture of
infected materials to demonstrate promastigote and also serological tests.

Prevention and control

Public awareness through education programme, exercise personal protection
through use of repellants, protective clothing and screening, prompt treatment
and eradication of the infected ulcer control sand fly population and reservoir.
Host vaccine is on trial.

Treatment
Pentostam (Sodium stibogluconate), camolar (cycloguanil pamoate),
Amphotericin B (given IV), Meglumine antimoniate, Pentamidine, Allopurinol,
Monomycin and Paromomycin are effective medication.

Henry W. Karoki, MSc.| Hemoflagellates

Leishmania donovani complex
Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida

Has three members: L. donovani infantum, L.donovani donovani, L. chagasi.

Morphology
Amastigote

Roundish to oval 5-3 um size, consist of a nuclear & a kinetoplasta, large

single nucleus locates off centre or towards the edge. Dot –like blepharoblast
give rise to & attached to axonemes axonemes extends towards edge of
organism, single parabasal body adjacent to blepharoblast

Life cycle notes

Sand fly responsible for transmission of promastigote via blood meal , it quickly
invade reticuloepithalial cell and transform to amastigote which causes tissue
destruction , skin and mucous membrane primarily affected, diagnostic stage
of L.donovani is the amastigote it is the infective stage for the sand fly , upon
ingestion , amastigote transform back to promastigote in the fly mid gut.
Promastigote multiply and the resulting forms move to the salivary gland ready
for the transfer to the humans.

Epidemiology
Endemic in the Middle East, Yemen, Kuwait, Iraq, Saudi Arabia

Clinical symptoms
Causes visceral leishmaniasis (kala azar or dum dum fever), present with
abdominal illness hepatosplenomegally, fever and chill , onset gradual, and
incubation period is 2wks to 18 months diarrhea and anemia are often present,
weight loss, emaciation, invasion of liver and spleen, kidney damage,
granulomatous area of skin.

Henry W. Karoki, MSc.| Hemoflagellates

Laboratory diagnosis
Giemsa stains blood slides, bone marrow, lymph node aspirate and biopsies of
infected areas are specimen of choice. Culture of the bone marrow, blood and
tissue and may be performed and often shows amastigote forms, giemsa stain
burry coat coat film preparation from venous blood. Serological test is available

Prevention and control

Public awareness through education programme, exercise personal protection
through use of repellants, protective clothing and screening, prompt treatment
and eradication of the infected ulcer control sand fly population and reservoir
host.

Treatment
Pentostam (Sodium stibogluconate), camolar (cycloguanil pamoate),
Amphotericin B (given IV), Meglumine antimoniate, Pentamidine, Allopurinol,
Monomycin and Paromomycin are effective medication.

Leishmania tropica complex

Phylum – kinetoplasta
Sub-phylum - mastigophora
Class - zoomastigophora
Order – trypanosoatida
Has three members: L.major, L. tropica, L. aethiopica.

Morphology
Amastigote

Henry W. Karoki, MSc.| Hemoflagellates

Roundish to oval 5-3 um size, Consist of a nuclear & a kinetoplasta, large
single nucleus locates off centre or towards the edge. Dot –like blepharoblast
give rise to & attached to axonemes. Axonemes extends towards edge of
organism, Single parabasal body adjacent to blepharoblast

Life cycle notes

Sand fly responsible for transmission of promastigote via blood meal, It quickly
invade, reticuloepithalial cell and transform to amastigote which causes tissue
destruction , Skin and mucous membrane primarily affected , Diagnostic stage
of L.Tropica is the amastigote , It is the infective stage for the sand fly, Upon
ingestion , amastigote transform back to promastigote in the fly mid gut.
Promastigote multiply and the resulting forms move to the salivary gland ready
for the transfer to the humans.

Epidemiology
Endemic in the Middle East, Yemen, Kuwait, Iraq, Saudi Arabia

Clinical symptoms
Causes cutaneous leishmaniasis (old world leishmaniasis, oriental sores,
Bagdad or delhi boil. Characterized by one or more ulcers containing pus that
self-heal. Patients develop small red papule on bite site and cause intense
itching. Diffuse cutaneous leishmaniasis (DCL) occurs in limb, and face. Thick
plagues on skin along with multiple lesion or nodules usually results.

Laboratory diagnosis
Giemsa stained blood slides, aspiration of the fluid under- Neath ulcer bed for
amastigote demonstration, Culture in NNN medium reveal promastigote forms,
Serological tests e.g. IIF antibody test.

Prevention and control

Public awareness through education programme, Exercise personal protection
through use of repellants, protective clothing and screening, Prompt treatment
and eradication of the infected ulcer

Treatment
Pentostam (Sodium stibogluconate), camolar (cycloguanil pamoate),
Amphotericin B (given IV), Meglumine antimoniate, Pentamidine, Allopurinol,
Monomycin and Paromomycin are effective medication.

Henry W. Karoki, MSc.| Hemoflagellates