Malaria, worms and leishmaniasis (BSH 2022, CDC website)
Preparing Films
Make 4 thick and 4 thin films --> 2 of each to go to reference lab unstained
Thick film
Geimsa or Field stain
Parasitaemia count made by 2 staff, from examination of 200 high power fields each
Thin film
Giemsa or Leishman stain
For species identification
Negative films should be repeated every 12-24 hours if clinical suspicion remains
malaria morphology summary
image bank
Image bank from the CDC
other tests
Immunochromatographic test (Rapid Diagnostic Test (RDT))
Detects malaria antigen
Confirmatory test only, must never replace microscopy (lots of detail in guideline)
Able to give species identity to falciparum and vivax
?Negative RDT in presence of intracellular red cell parasites - think babesiosis and send samples to reference laboratory for further investigation. CDC Babesiosis pages.
Nucleic-acid detection
Turnaround time 2-72 hours depending on method.
Cannot always distinguish current from past infection
PCR
10x more sensitive than microscopy
Better at determining species in cases of mixed infection
LAMP
Methodoligically far simpler. Originally developed for use in the field.
Drug sensitivity testing
Research tool only at present
Quality assurance
Internal
Dual reporting of all slides
New stain batches tested on known vivax and ovale cases
External
NEQAS
Reference lab confirmation
Malaria Biology
Geographical distribution
Falciparum – Global
Malariae – Global
Ovale – Sub-Saharan Africa
Vivax – Rest of Africa
Knowlesi – SE Asia
Life Cycle
During a blood meal, malaria-infected female anopheles mosquitoes inoculate sporozoites into the human host.
Sporozoites infect liver cells and mature into shizonts
Liver shizonts mature then rupture to release merozoites
Merozoites infect red cells and multiply asexually --> ring form immature trophozoite --> trophozoite --> shizonts --> again rupture (this time from red cells) releasing more merozoites. This is responsible for clinical manifestations of the disease
Some parasites differentiate into a sexual stage (microgametocytes (male) and macrogametocytes (female)
The gametocytes are ingested by mosquito at next meal. Microgametocytes penetrate macrogametocytes to generate a zygote. The zygote becomes motile and elongated (ookinete) and invade the gut wall of the mosquito.
The now oocyst grows, ruptures and release sporozoites, which migrate to the salivary glands and the cycle repeats.
worms
Sheathed Worms
Wucheria bancrofti
Central Africa, Nile, Central/South America & Tropical Asia
Mosquito-borne
Clinical Picture
Lymphatic filariasis, but often asymptomatic for years
Pulmonary eosinophilia syndrome
Hydrocoeles
Brugia Malayi
South and SE Asia
Mosquito-borne
Clinical Picture
Lymphatic filariasis, but often asymptomatic for years
Pulmonary eosinophilia syndrome
Hydrocoeles
Loa Loa
West and Central Africa
Fly-borne
Syn. African Eye worm
Clinical Picture
Itchy calabar swellings
Eye worm
But often asymptomatic (More symptomatic if not a native of endemic area)
No Sheath Worms
Mansonella
Africa and tropical Americas
Midge-borne
Clinical Picture
Usually asymptomatic
Non-specific fever, angioedema, itch, arthralgia, headache
Worm Summary
Leishmaniasis
Tropics, Subtropics and Southern Europe
Sand fly-borne
Cutaneous
Skin sores
Often self-resolving but can take years
Visceral
Fever, weight loss, hepatosplenomegaly, pancytopenia
Often fatal if untreated
Tissue stage of larvae is called an amastigote
Often seen inside macrophages, should have a nucleus + rod-shaped kinetoplast
trypanosomiasis
african trypanosomiasis
Species
Trypanosoma Brucei Gambiense
Trypanosoma Brucei Rhodensiense
Morphology
Pleomorphic trypanosomes, 18-35um in length
Single flagellum extending from kinetoplast and running along outer margin of the membrane
Small dot-like parabasal body of the kinetoplast stains darkly
Cytoplasm is pale with granules
Early stage: long slender forms, often seen dividing
Late stage: intermediate and short forms, some with no flagellum
south american
Species
Trypanosoma Cruzi
Morphology
Usually C-shaped, 12-30um in length
Narrow membrane and free flagellum
Large round/oval dark-red kinetoplast at posterior end
Nucleus dark-red and centrally placed