Leucocytozoon atkinsoni n. sp. (Apicomplexa: Leucocytozoidae) from the
avian family Timaliidae
Amy F. Savage1,2,*, Frederic Ariey3& Ellis C. Greiner1
1Department of Pathobiology, College of Veterinary Medicine, University of Florida, Box 110880, Gaines-
ville, Florida 32611, USA
2Department of Epidemiology and Public Health, School of Medicine, Yale University, 60 College Street, New
Haven, Connecticut 06520, USA
3Groupe de Recherche sur le Paludisme, Institut Pasteur de Madagascar, BP 1274, Antananarivo 101,
Accepted for publication 28th August, 2005
Investigators of haematozoa of the Timaliidae have reported the presence of two species of Leucocytozoon
Berestneff, 1904, i.e. L. liothricis Laveran & Marullaz, 1914 and L. timaliae Bennett, Earle ´ & Pierce, 1993.
Blood films collected from 42 wild-caught babblers in Madagascar were stained and examined for the
presence of haematozoa using a compound microscope. To date, no species of avian haematozoa have been
reported from babblers in Madagascar, although haematozoa have been observed. In the present study, we
report a new species of Leucocytozoon, L. atkinsoni n. sp., whose morphometrics fall between those
reported for the two previously described species from timaliids. The parasite is capped by the host cell
nucleus covering 38% of its perimeter. L. atkinsoni n. sp. was found to have a marked, intensely staining,
nucleolus as well as vacuoles in the parasite cytoplasm, in contrast to both L. liothricis and L. timaliae.
Remnants of the host cell cytoplasm are commonly observed in cells infected with L. atkinsoni, a char-
acteristic not reported in association with either of the previously described species from these hosts.
The family Timaliidae (babblers) includes 265
species of birds occurring in Africa, Asia and the
Middle East (Clements, 2000). Of these, seven
species, and many more subspecies, occur in
Madagascar. To date, three published studies have
surveyed birds from Madagascar for haematozoa
(Bennett & Blancou, 1974; Greiner et al., 1996;
Raharimanga et al., 2002). Raharimanga et al.
(2002) reported one of 25 babblers to be infected
with a species of Leucocytozoon Berestneff, 1904,
but the authors did not identify it to the species
level. As a result of a fourth, ongoing survey, we
have reported 10 new species of haematozoa of the
genera Plasmodium Marchiafava & Celli, 1885,
Haemoproteus Kruse, 1890 and Leucocytozoon
from four other avian families in Madagascar
(Savage, 2004; Savage & Greiner, 2004; Savage
et al., 2004; Savage & Greiner, 2005; Savage et al.,
As part of this larger survey, 42 wild-caught
babblers representing five different species on
Madagascar were bled and examined for the
presence of haematozoa. A species of Leucocyto-
zoon, unlike the species previously reported from
timaliids, was observed in one of the 42 birds.
Currently, two species of Leucocytozoon are
known from the babblers, L. liothricis Laveran &
Marullaz, 1914 and L. timaliae Bennett, Earle ´ &
Pierce, 1993 (see Bennett et al., 1993). Several
authors have discussed host-specificity in this
genus, reporting specificity at the host family level
for correspondence(E-mail: haemosporida@
Systematic Parasitology (2006) 64:105–109
? Springer 2006
(Fallis et al., 1954, 1974; Bennett et al., 1991).
Based on our current knowledge regarding host
family specificity, as well as distinct morphometric
characteristics, the new Leucocytozoon species
described here is considered to be specific to the
Materials and methods
All birds were mist-netted at field sites in
Madagascar. Blood smears were made from toe
clippings and heart punctures. Slides were air-
dried, fixed in absolute methanol, and stained with
Giemsa stain. Slides were examined at ? 100,
? 200 and ? 1,000 using a Zeiss compound
microscope for the presence of haematozoa. Par-
asites were photographed on an Olympus XB41
microscope with an Olympus DP70 digital camera.
Images were measured using ImageJ (http://
rsb.info.nih.gov/ij/), a public domain, java-based
program available online (http://rsb.info.nih.gov/
nih-image/) from the National Institutes of Health
(Rasband, 2002). All measurements of erythro-
cytes and parasites were performed as described in
Bennett & Campbell (1972) and Bennett et al.
Leucocytozoon atkinsoni n. sp.
Type-host: Neomixis tenella (Hartlaub), the com-
Type-locality: Ankililoaka-Morombe, Madagascar.
Type-material: Hapantotype: Reference Centre for
Avian Haematozoa, Queensland Museum, South
Brisbane, Australia, G464632, Neomixis tenella,
coll. Willard, Ankilioaka-Morombe, Madagascar,
80 m elevation.
Other hosts: Distribution and additional hosts
currently unknown, although it is presumed that
this parasite can be found throughout the range of
babblers on Madagascar.
Etymology: This parasite is named for Dr Carter
T. Atkinson for his continued dedication to the
conservation of native Hawaiian avifauna through
his research on the effects of introduced avian
malaria and pox. Additionally, his work with
haematozoa of Pacific island fauna in general is
Description (Figure 1)
Immature gametocyte. Displaces the host cell
nucleus almost immediately, seen to develop in
Macrogametocyte (n=29). Round morph only,
commonly distorted. Stains medium to dark blue
with Giemsa stain (Figure 1A–D). Small vacuoles
observed in parasite cytoplasm. Host-parasite
complex 3.3 times larger (197.4 lm2) than unin-
fected erythrocyte (59.7 lm2). Macrogametocyte
area 137.7 lm2, occupying 69.8% of host-parasite
complex. Parasite nucleus 10.2 lm2or 7.4% of
gametocyte area. Ovoid or round nucleus stains
pink with pronounced nucleolus staining darker
pink or purple. Nuclear cap covers area of
18.1 lm2or 38% of circumference of parasite
and is stretched thin, with irregular margins.
Residual host cell cytoplasm commonly evident,
and, when present, has average size of 33.2 lm2or
up 16.8% of host-parasite complex.
Microgametocyte (n=7). Same characteristics as
described above, with usual gender-associated
differences. Parasites stain very light blue with
Giemsa (Figure 1E-H). Host-parasite complex
26.2% smaller than cells infected with macroga-
metocytes,area 145.7 lm2.
(104.7 lm2) occupies 71.9% of host-parasite com-
plex. Average parasite nucleus area 50.4 lm2.
Of the 42 individuals examined for the presence of
haematozoa, only one was found to be infected
with the species of Leucocytozoon described here.
Morphometric characteristics of the parasite were
compared with the known species from the Timal-
iidae, L. liothricis and L. timaliae, and are
presented in Table 1. Morphometric data for
L. liothricis and L. timaliae were taken from that
presented in Bennett et al. (1993). Based on these
comparisons, L. atkinsoni n. sp. is easily distin-
guished from L. liothricis and L. timaliae. Some
morphological characteristics of L. atkinsoni fall in
between the extremes noted for L. liothricis and
L. timaliae. For example, the caps formed by the
host-cell nuclei in cells infected with these parasites
are each very distinct. The cap for L. liothricis is
described as a thin band, covering 80% of the
parasite perimeter and occupying 24% of the
host-parasite complex. Figures show a cap with a
consistent thickness throughout. On the other
hand, L. timaliae has a small, bulbous cap
covering 21% of the parasite circumference and
occupying 13% of the host-parasite complex.
Morphometrics observed for L. atkinsoni are
formed by the host-cell nucleus encloses 38% of
the parasite perimeter and occupies 16.8% of the
host-parasite complex. The nuclei of both known
species are described as being round to ovoid,
having no marked karyosome and occupying 5%
of the parasite area. The nucleus observed in
L. atkinsoni is slightly larger, occupying 7.4% of
the parasite area, and there is a distinct nucleolus
(karyosome) visible. It is felt that the distinct
nucleolus is a true character, rather than the result
of staining, as consistent differences in the nucleo-
lus between two Leucocytozoon species have
been noted previously (Savage et al., 2004).
Vacuoles are typically observed in the cytoplasm
of L. atkinsoni, while they are uncommon in both
L. liothricis and L. timaliae. Finally, in neither of
these previously described species is there residual
host cell cytoplasm associated with infected cells,
whereas this is common in L. atkinsoni.
Babblers are found throughout the Old World,
and have been included in many surveys. The first
species of Leucocytozoon reported from this family
Figure 1. Gametocytes of Leucocytozoon atkinsoni n. sp.: A-D, macrogametocytes; E-H, microgametocytes.
was described in 1914 from birds suspected to be
imported from Vietnam. Nearly 80 years later,
Bennett et al. (1993) described L. timaliae* from a
babbler sampled in South Africa. Both of these
species are easily identified and have distinctive
morphologies, which serve to separate them from
L. atkinsoni n. sp.. Like L. timaliae, L. atkinsoni is
reported from a single babbler on Madagascar.
Additional sampling is necessary to determine the
true range of this parasite on Madagascar, as well
as on the African continent. The effects of L.
atkinsoni on both the host and on the vector or
vectors of this parasite are not currently known.
The authors wish to gratefully acknowledge
Dr Steven M. Goodman of the Field Museum of
Natural History, as well as Dr David E. Willard
for the collection of samples in the field. Addi-
tionally, Dr Vincent Robert and the Institut de
Pasteur de Madagascar are gratefully acknowl-
edged for their continued assistance and partner-
ship. Finally, the authors thank Dr Carter
Atkinson for providing workspace.
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Table 1. Morphometric parameters of the Leucocytozoon spp. from the Timaliidae (means in lm or lm2, followed by the standard
deviations in parentheses, or percentages where indicated).
L. liothricis n=65 L. timaliae n=15L. atkinsoni n. sp. n=36
Macrogametocyte host parasite complex
Microgametocyte host parasite complex
Macrogametocyte nucleus area
Microgametocyte nucleus area
Host cell nucleus area
Residual host cell cytoplasm
Cap coverage 28.7 (80%)11.3 (21%)18.1 (38%)
n/a, not available.
*We consider that this is the correct spelling of this species
name, which was spelled three ways in the original paper
(Bennett et al., 1993).
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