Morphological Characteristics of Malaria Parasites

MALARIA DIAGNOSIS
DEPARTMENT OF LABORATORY MEDICINE
Morphological Characteristics of
Malaria Parasites
Morphological Characteristics of Malaria Parasites
Learning objectives
Name various parts of malaria parasites.

Recognize the three stages of malaria parasites.
Recognize key diagnostic features of each specie.
Distinguish between species on thick and thin films.
Species of Interest
The four human Plasmodium species
P. falciparum
P. vivax
P. ovale
P. malariae
Malaria Parasite Stages
Trophozoites
Chromatin
Cytoplasm
Vacuole
Pigment
Host Cell
Chromatin This is the nucleus of the parasite and

is usually round in shape and stains purple/red.
Cytoplasm - Early stages usually ring shaped, but
varies as it matures and also with species. Usually
stains blue but also varies with age and species of
the parasite.
Pigment - Coarse brown/black precipitates on
parasite cytoplasm which are by-products of parasite
metabolism. Colour may vary depending on species.
Schizonts
Merozoites
Pigment
Cytoplasm
Formed as a result of simple division of the

parasite nucleus (schizogony).
Number of merozoites vary between age and
species of the parasite.
Pigment may be scattered or concentrated
depending on age or species.
Gametocytes
Nucleus
Pigment
Cytoplasm
This is the sexual stage where parasites

become male or female.
May be banana-shaped or round depending
on the species.
Size of chromatin may help differentiate
between male and female.
General characteristics of P.falciparum
Size - small, compact, dark staining

Shape - regular in shape
Multiple infection - common even in light
infections.
Early trophs & Growing - common even in
light infections.
Late trophs - rarely seen in smear from
peripheral blood. If seen, it is a sign of
severe infection.
General characteristics of P.falciparum
Morphological forms
Marginal/applique - (at the edge of RBC) quite common
Extruding forms - some parasites may seem to extrude

beyond the periphery of the cell.
NB: These forms can only be seen on well made thin films.
Schizonts rarely seen in peripheral blood except in severe
infections.
Gametocytes common in peripheral blood.
Host cell not enlarged, round, sometimes creanated.
Colour normal, sometimes dark with a purple rim.
Stippling Maurers clefts which appear as red spots on host
cell cytoplasm. May be up to 20 in number but normally fewer.
Pigment clumped as a single or double black/brown mass.
P. falciparum Trophozoites
Early trophozoite (young ring)
Often very numerous in number.
Have delicate cytoplasm and small chromatin dot.
Sometimes may have double chromatin dot.

Growing trophozoite (medium ring)
Common in peripheral blood.
Have regular cytoplasmic ring.
Sometimes cytoplasm may appear broken or comma shaped.
A single chromatin or double chromatin dot may be seen.

Mature trophozoite (late)
Not common in peripheral blood except in very heavy infections.
Have solid distinct chromatin dot which could be irregularly

shaped.
Pigment is concentrated or clumped as single dark mass.
Sometimes pigment may be concentrated at two points.
Illustrations of P. falciparum Trophozoites

Thin Film
Thick Film
Photographic Images of P. falciparum Trophozoites
Late Rings
Early Rings
Growing Ring
Early Trophs
Late Trophs
Growing Trophs
Late Ring
Early Rings
Early Trophs
Growing Troph
Growing Rings
Late Trophs
P. falciparum Schizonts
Early schizont (pre-segmenter)
Rarely seen peripheral blood except in heavy

infections.
If present will be associated with numerous typical ring

forms.
Pigment fused as a single dark mass (clumped

pigment).
Mature schizont (segmenter)
Compact and round.
Usually have 12-24 merozoites, range is between 8-40.
Merozoites are normally grouped around the pigment
Pigment single dark mass.
Illustrations of P. falciparum Trophozoites Schizonts
Thin Film
Thick Film
Photographic Images of P.falciparum Schizonts
Thin
Thick
Pigment
Thin
P. falciparum Gametocytes
Immature Gametocyte
Initially are rounded bodies with no vacuole.
Become spindle shaped as they mature.

Mature Female Gametocyte (Macro)
Crescent, sausage or banana shaped with rounded ends.
Cytoplasm stains blue.
Chromatin centrally placed as purplish mass.
Pigments appear as rod-like dark granules in the center

of the parasite.
Mature MaleGametocyte (Micro)
Similar in shape to macro but less crescent shaped.
Cytoplasm is pinkish purple.
Chromatin is more diffused.
Pigment more scattered than in macro.
Illustrations of P. falciparum Gametocytes

Thin
Thick
Photographic Images of P.falciparum Gametocytes
Young
Micro
Growing
Macro
Mature
Micro
Macro
General Characteristics of P. vivax
Size small to large.

Chromatin single or double.
Cytoplasm - regular or fragmented, usually amoeboid in
shape.
Number few to numerous.
Schizont - common with 12 24 merozoites, usually 16.
Gametocyte common, normally round in shape.
Host cell markedly enlarged; size of WBC. May be
round or oval.
Stippling Schffners clefts.
Pigment Golden brown.
P. vivax Trophozoites
Early trophozoite
Fairly numerous.
Measure a third of RBC diameter.
Composed of prominent red chromatin dot and a
fine cycle of blue cytoplasm.
Occasionally there may be 2 chromatin dots.
Growing trophozoite
Have increased cytoplasm.
Usually irregular amoeboid appearance.
P. vivax Trophozoites
Late trophozoite
Become very large and markedly amoeboid.
May fill the entire RBC.
Chromatin is large and compact.
Vacuole is almost always present.
Pigment scattered all over the cytoplasm.
May be difficult to distinguish from
gametocytes.
Schffners stippling sometimes seen.
Illustration of P. vivax Trophozoites

Thin
Thick
Photographic Images of P. vivax Trophozoites

Early Rings
Growing Trophs
Late Ring
Growing Ring
Late Ring
Late Troph
Early Rings
Early Trophs
Growing Trophs
Early Trophs
Growing Ring
Late Troph
P. vivax Schizonts
Early Schizont (presegmenter)
Large and amoeboid.
Chromatin divides into small irregular masses.
Schffners stippling often seen as a pink granular halo.

Mature Schizont (segmenter)
8-24 merozoites, usually 12-16.
Relatively large size.
One or two clumps of pigment may be centered in the

cluster of merozoites.
Each merozoite is composed of a dot of chromatin

surrounded by a small mass of cytoplasm.
Usually fills the enlarged RBC.
Schffners stippling evident in the infected cell.
Illustrations of P. vivax Schizont

Thin
Thick
Photographic Images of P.vivax Schizonts

Early
Growing
Mature
Gravid
Mature
Early
Growing
Gravid
P. vivax gametocytes
Immature
Similar to late trophzoites.
No vacuole.
Large piece of chromatin.

Female gametocyte (Macro)
Nearly fills the enlarged RBC
Have a small compact chromatin.
Vacuole absent.
Pigment granules scattered through out the cytoplasm.

Male gametocyte (Micro)
Similar in size and shape to the female.
Distinguished from female by the diffused chromatin.
Light blue to pink or lavender cytoplasm containing

scattered granules of pigment.
Illustrations of P. vivax Gametocytes

Thin
Thick
Photographic Images of P. vivax Gametocytes

Early
Early
Mature
Micro
Mature
Macro
Micro
Early
Macro
General Characteristics of P. ovale
Trophozoites
Size in between Pm and Pv.
Shape ring to rounded, mature forms more
compact.
Chromatin single and prominent.
Cytoplasm fairly regular and fleshy.
Pigment scattered and coarse.
Schizont
Similar to Pm.
Merozoites - 4 12, usually 8 in a loose cluster.
Pigment concentrated.
General Characteristics of P. ovale
Gametocyte
Difficult to distinguish from mature trophozoites.
Mature forms round and compact.
Chromatin single and well defined.
Pigment coarse and scattered.
Host cell
Size enlarged
Shape round to oval, sometimes fimbriated.
Stippling numerous coarse granules called
James clefts.
P.ovale Trophozoites
Early trophozoite
Chromatin large and irregular in shape.
Cytoplasm small and regular in shape.
Growing trophozoite
Usually regular in shape with a vacuole.
Host cell appreciably enlarged and with quite
abundant stippling.
P.ovale Trophozoites
Late Trophozoite
Host cell slightly enlarged.
May be oval and fimbriated.
Cytoplasm more compact and regular
in shape.
Usually does not occupy more than
two thirds of the enlarged host cell.
Stippling and pigment prominent.
Illustration of P. ovale Trophozoites

Thin
Thick
Photographic Images of P. ovale Trophozoites

Rings
Ring
Early Troph
Early Troph
Growing Troph
Late Troph
Growing Troph
Late Troph
P. ovale Schizonts
Early Schizont
Size -medium
Shape round and compact.
Merozoites few in number.
Pigment - coarse and scattered.
Mature Schizont
Size -larger than P.malariae.
Merozoites -irregular rosette formation, usually
8-12,occasionally up to 18.
Pigment - clumped at the center of merozoites.
Stippling James's clefts usually prominent.
Illustrations of P. ovale Schizonts

Thin
Thick
Photographic Images of P. ovale Schizonts
Early
Early
Growing
Growing
Mature
Mature
Gravid
Gravid
P. ovale Gametocytes
Immature forms
Difficult to differentiate from mature trophozoites.
Mature Gametocytes
Typically fill the host cell.
Round or oval and compact sometimes with
irregular margins.
Chromatin well defined.
Stippling (Jamess clefts) well defined.
P. ovale Gametocytes
Female gametocye (Macro)

Similar in appearance to P.vivax,
Slightly smaller compared to Pv
Chromatin compact and dark red.
Male gametocyte (Micro)
Similar in size and shape to the females
Have diffuse pink chromatin.
Cytoplasm pink to purple.
Cannot be reliably distinguished from the male
of P. vivax
Illustrations of P. ovale Gametocytes

Thin
Thick
Photographic Images of P. ovale Gametocytes
Young
Macro
Micro
General Characteristics of P. malariae
Trophozoites
Size small.
Number usually few.
Shape ring to rounded and compact.
Chromatin single and large. Double chromatin very rare.
Cytoplasm regular and dense.
Pigment scattered, abundant with greenish yellowish

tinge.
Band forms common in thin films and multiple infections

very rare.
Schizont
Size small and compact.
Merozoites 6-12, usually 8 in loose cluster.
Pigment-scattered in immature forms but concentrated in

mature forms.
Gametocytes
Immature forms difficult to distinguish from mature

trophozoites.
Round and compact.
Chromatin is single and well defined.
Pigment is coarse and scattered.

Host cell
Not enlarged.
Shape not distorted.
No stippling except with intense staining.
Stippling called Ziemans Clefts.
P. malariae Trophozoites
Early Trophozoite
Ring forms fairly large.
Chromatin may be peripheral.
Occasionally chromatin may be at the centre of the

vacuole.
Pigments may form as early as this stage.

Growing trophozoite
May stretch across the host cell producing band forms.
May exhibit large vacuole surrounded by dense cytoplasm

and elongate mass of chromatin producing basket-like
appearance.
Mature trophozoite
Almost fill the entire host cell.
Have prominent chromatin.
Cytoplasm is compact with scattered coarse pigment.
Illustrations of P.malariae Trophozoites

Thin
Thick
Photographic Images of P. malariae Trophozoites

Late Ring
Early Ring
Growing Rings
Early Troph
Growing Troph
Basket Form
Late Troph
P. malariae Schizonts
Early Schizont
Show fewer divisions.
Round and compact.
Pigment scattered.
Mature Schizont
Usually fill the host cell.
Merozoites range 8-12,usually 8.
Merozoites are in rosette formation.
Pigment is concentrated as a single mass.
Illustrations of P. malariae Schizonts

Thin
Thick
Photographic Images of P.malariae Schizonts

Gravid
Growing
Rosette
Early
Growing
Mature
P. malariae Gametocytes
Gametocyte
Difficult to distinguish from mature trophozoites.
Female gametocyte (macro)
Bluer cytoplasm.
Chromatin smaller, redder and more compact.
Male gametocyte (micro)
Cytoplasm light bluish pink.
Chromatin is diffuse.
Illustrations of P. malariae Gametocytes

Thin
Thick
Photographic Images of P. malariae Gametocytes

Young
Macro
Micro
Micro
Macro
Micro
Differential Characteristics
Pf
Pm
Po
Pv
RBC Infection
All RBCs
Normocyte
Mostly
Mostly
Reticulocytes Reticulocytes
& sometimes & sometimes
Normocytes
Normocytes
Cell
enlargement
None
None
Yes +
Yes ++
Stippling
Maurers
Ziemans
James
Schffners
Pigments
Dark
brown/black
Yellowish/
Greenish
tinge
Brown
Golden
Brown
Merozoite
Number
8-24(40)
8-12
8-12(18)
12-18(24)
Early Trophozoite Stages

Pf - have tiny rings
Pm - early pigment formation
Pv thin delicate cytoplasm which may be irregular
Po larger chromatin
Photographic Images of Early Trophozoites
Growing Trophozoites
Pf
RBC
Round, may be
crenated
May have Maurer
clefts
Pf
Cytoplasm
Complete or
Broken
Comma shaped
or marginal
Pf
Pigment
Pigment; may have
or not
Black/brown
Pm
RBC
-Normal
Pm
Cytoplasm
Compact, deep
blue
Pm
Pigment
Scattered large
grains covering
almost entire
cytoplasm
Yellowish brown
Pv
RBC
Slightly enlarged,
Schuffners dot
Pv
Cytoplasm
Fragmented or
amoeboid
Pv
Pigment
Scattered, Golden
brown
Po
RBC
Round to oval
normal or little
enlarged
Jamess Dot
Po
Cytoplasm
Compact
Vacuole around
chromatin
Po
Pigment
Scattered
Dark brown
Thin
Thick
Photographic Images of Growing Trophozoites
Mature Trophozoites
Pf
RBC
-Normal
-Maurers clefts
Pf
Cytoplasm
-Pale
-Regular
Pf
Pigment
-Clumped
-Black brown
Pm
RBC
-Normal
-Zeimans dots
Pm
Cytoplasm
-Compact
-Coarse
-Elongated band
Pm
Pigment
-Scattered
-Yellowish/
Greenish tinge
Pv
RBC
-Enlarged
-Shffners dots
Pv
Cytoplasm
- Slightly fragmented,
-Amoeboid
-More spread out
Pv
Pigment
-Scattered
-Golden brown
Po
RBC
-Enlarged
-Fimbriated
-James dots
Po
Cytoplasm
-Compact
-Less spread out
Po
Pigment
-Scattered
-Brown
Thin
Thick
Photographic images of P. falciparum Trophozoites
Pf
Po
Pm
Pv
Early and Growing Schizonts

Pf
RBC
Normal
Pf
Merozoites
-2 or more
-cytoplasm intact
Pf
Pigment
-Single mass
-Dark staining
Pm
RBC
Normal
Pm
Merozoites
-2 or more
-cytoplasm intact
Pm
Pigment
-Scattered
-Yellowish/
Greenish tinge
Pv
RBC
-Larger than Po
Pv
Merozoites
-2 or more
-cytoplasm intact
Pv
Pigment
-Scattered
-Golden brown
Po
RBC
Medium sized compared
to Pv of the same stage
-Fimbriated cell
Po
Merozoites
-2 or more
-cytoplasm intact
Po
Pigment
-Scattered
-Dark brown
Thin
Thick
Photographic Images of Schizonts

Pf
Po
Pm
Pv
Mature Schizonts
Pf
RBC
Normal
Pf
Merozoites
-8-24(40)
Pf
Pigment
-Single mass
-dark brown/black
Pm
RBC
Normal
Pm
Merozoites
-8-12
-Rosette formation
Pm
Pigment
-Clump of granules
-Yellowish/
Greenish tinge
Pv
RBC
Markedly enlarged
Pv
Merozoites
-12-18(24)
Pv
Pigment
-Single mass
-Golden brown
Po
RBC
-Larger than Pm
-Distinct stippling
Po
Merozoites
-Irregular rosette formation
-8-12(18)
Po
Pigment
-Single mass
-Brown
Thin
Thick
Photographic Images of Mature Schizonts

Pf
Po
Pm
Pv
Male Gametocytes (Micro)

Pf
Morphology
-Less crescent
Pf
Chromatin
-Diffuse
-Makes cytoplasm appear
purple- red
Pf
Pigment
-Scattered
-Black and rod like
Pm
Morphology
-Fills host cell
-Round in shape
Pm
Chromatin
-Diffuse
purple- red
Pm
Pigment
-Scattered
-Yellowish/
Greenish tinge
Pv
Morphology
-Resembles Po
-Larger compared to all
Pv
Chromatin
-Diffuse
-makes cytoplasm appear
purple- red
Pv
Pigment
-Scattered
-Golden brown
Po
Morphology
-Resembles Pv
-Fills 2/3 of host cell
Po
Morphology
-Diffuse
purple-red
Po
Pigment
-Scattered
-Brown
Thin
Thick
Photographic Images of Gametocytes

Pf
Po
Pm
Pv
Female Gametocytes (Macro)

Pf
Morphology
-More crescent shaped than
micro
Pf
Chromatin
-Central
-Not diffuse
-Bluish cytoplasm
Pf
Pigment
-Black
-Rod like
Pm
Morphology
-Occupies the whole cell
Resembles mature trophs
Pm
Chromatin
-Compact chromatin
-Located at the periphery
Pm
Pigments
-Coarse & dark
-Scattered
Pv
Morphology
-Larger than other species
- Resembles mature trophs
Pv
Chromatin
-Compact chromatin
-Slightly large chromatin
Pv
Pigments
-Scattered
Golden yellowish
Po
Morphology
-Similar to Pv but smaller
-Fills 2/3 of host cell
-Resembles mature trophs
Po
Chromatin
-Compact
-Large
Po
Pigments
-Scattered
-Dark brown
Thin
Thick
Photographic Images of Female Gametocytes

Pf
Pm
Po
Pv
Key Distinguishing Characteristics
P. falciparum
Crescent gametocyte and pigment rod like
Tiny ring and may have high density
Clumped pigment
P. malariae
Pigment greenish brown and cover almost the whole

cytoplasm\
Band form
Merozoite 8-12
P. ovale
Compact cytoplasm and red zone
Fimbriation of host cell

P.vivax
Amoeboid cytoplasm
QUIZ
References
Basic Malaria Microscopy: Part 1.

Learners Guide.
Bench Aids for the diagnosis of
malaria infections: Second Edition.

Morphological Characteristics of Malaria Parasites

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MALARIA DIAGNOSIS

DEPARTMENT OF LABORATORY MEDICINE