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Ticks and Tick-borne Diseases

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Original article

Description of the larva of Amblyomma calcaratum Neumann, 1899

(Acari: Ixodidae) by light and scanning electron microscopy
Fábio S. Barbieri a,∗ , Luciana G. Brito a , Marcelo B. Labruna b , Darci M. Barros-Battesti c ,
Luis Marcelo A. Camargo d , Kátia M. Famadas e
a
Embrapa Rondônia, Rodovia BR 364, Km 5,5, caixa postal 127, 76815-800, Porto Velho, RO, Brazil
b
Faculdade de Medicina Veterinária e Zootecnia/USP, São Paulo, SP, Brazil
c
Laboratório Especial de Coleções Zoológicas/Instituto Butantan, São Paulo, SP, Brazil
d
Instituto de Ciências Biomédicas 5/USP, Monte Negro, RO, Brazil
e
Departamento de Parasitologia Animal/UFRRJ, Seropédica, RJ, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: The larval stage of Amblyomma calcaratum Neumann is described using optical and scanning electron
Received 19 March 2013 microscopy. Unfed larvae were obtained from a colony of A. calcaratum originating from engorged females
Received in revised form 10 June 2013 collected on Tamandua tetradactyla in the Jaraguá Mountain (23◦ 40 S, 45◦ 44 W), São Paulo County, Brazil.
Accepted 11 July 2013
Eleven larvae were prepared and mounted on slides and observed under a light microscope equipped with
Available online xxx
a drawing tube. Three specimens were prepared for SEM. Several morphological characters are described,
including the chaetotaxy of the idiosoma, palpi, and Haller’s organ, as well as morphological features of
Keywords:
the idiosoma, gnathosoma, and legs of A. calcaratum larvae. In addition, topographical and numerical
Amblyomma calcaratum
Larva
patterns of integumentary structures on the larval idiosoma are described using a recently proposed
Description nomenclature. On the idiosoma, setaes, lyrifissures, small glands, and large wax glands were found. These
Morphology structures were observed isolated or associated over the entire idiosoma, except on the scutum, which
Chaetotaxy lacks large wax glands. The topographical and numerical patterns of integumentary structures of the
Integumentary structures A. calcaratum larva showed only minor differences when compared with patterns of other Amblyomma
larvae; however, a few key features can be used to differentiate A. calcaratum from other members of this
genus.
© 2013 Elsevier GmbH. All rights reserved.

Introduction
Currently, there are 130 valid species belonging to the genus
Amblyomma in the world (Guglielmone et al., 2010). In Brazil, there
are 29 established species of Amblyomma (Dantas-Torres et al.,
2009), from which the larval stage has been described for only 19
species (Barbieri et al., 2012). Indeed, this low number of descrip-
tions is related to the difficulty of obtaining engorged females (and
resultant larvae) of some species, since the majority of Brazilian
ticks preferentially feed on wild animals and are hard to breed in
the laboratory (Onofrio et al., 2006; Szabó et al., 2009).
Amblyomma calcaratum Neumann, 1899, has been reported
from Mexico to Argentina, where the adult stage is found feed-
ing almost exclusively on anteaters, while nymphs feed preferably
on passerine birds (Jones et al., 1972; Guglielmone et al., 2003;
Onofrio et al., 2006; Guzmán-Cornejo et al., 2006; Ogrzewalska
et al., 2011). Birds seem also to be preferred hosts for the larva,
∗ Corresponding author. Tel.: +55 69 3901 2520; fax: +55 69 3222 0409.
E-mail address: fabio.barbieri@embrapa.br (F.S. Barbieri).
although there have been only a very limited number of host
records (Ogrzewalska et al., 2010). A. calcaratum is of potential med-
ical importance because a spotted fever group bacterium, identified
as a Rickettsia parkeri-like agent, was recently reported infecting
nymphs collected from birds in Brazil (Ogrzewalska et al., 2013).
In the present study, the larva of A. calcaratum is described and
illustrated by optical and scanning electron microscopy (SEM).
Materials and methods
Unfed larvae were obtained from engorged females of A. calcara-
tum collected on Tamandua tetradactyla at the Jaraguá Mountain
(23◦ 40 S, 45◦ 44 W), São Paulo County, Brazil. Eggs laid by these
females under laboratory conditions (27 ◦ C, 90 ± 5% RH) were
pooled and placed in modified disposable syringes whose open
ends were covered with hydrophilic cotton. Eleven larvae were
individually prepared for light microscopy, as previously described
(Barbieri et al., 2007) while another 5 specimens were processed
for SEM according to Keirans et al. (1976).
The A. calcaratum engorged females were determined to species
using the keys and redescriptions of Guimarães et al. (2001). The

1877-959X/$ – see front matter © 2013 Elsevier GmbH. All rights reserved.
http://dx.doi.org/10.1016/j.ttbdis.2013.07.004

Please cite this article in press as: Barbieri, F.S., et al., Description of the larva of Amblyomma calcaratum Neumann, 1899 (Acari: Ixodidae) by
light and scanning electron microscopy. Ticks Tick-borne Dis. (2013), http://dx.doi.org/10.1016/j.ttbdis.2013.07.004
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Fig. 1. Amblyomma calcaratum, larva. Gnathosoma, dorsal (right) and ventral (left) views. Abbreviations: d, dorsal; v, ventral; a, antiaxial; p, paraxial; t, terminal; F, femur; G,
genu; Tt, tibiotarsus.

chaetotaxy terminology of Clifford and Anastos (1960), Hess and width 0.044 ± 0.003 (0.039–0.047); femur length 0.051 ± 0.003
Vlimant (1983), and Woolley (1988), and the porotaxy terminol- (0.046–0.056), width 0.053 ± 0.002 (0.050–0.056); genu length
ogy of Barbieri et al. (2007) were used in our larval description. The 0.052 ± 0.002 (0.049–0.058), width 0.047 ± 0.003 (0.042–0.053).
frequency of integumentary structures was determined by inde- Femur with sensillum near seta Fa1 (Fig. 5).
pendently analyzing each idiosomal side (left and right), according Ventral: Basis capituli as illustrated (Figs. 1 and 6). Hypostome
to Klompen et al. (1996). Thus, a total of 22 idiosomal sides were compact, spatulate, length from apices to posthypostomal seta
analyzed. All measurements were made with a Wild M11 light 0.092 ± 0.002 (0.087–0.096), dental formula 2/2 in file teeth, api-
microscope and are reported in millimeters. The mean is followed cal corona usually with 9 small denticles; 1 pair of posthypostomal
by the standard error, with the range in parentheses. Four larval setae (Ph1) (Fig. 6). Palpal setae (Fig. 1): 10 setae on tibiotarsus, 6
specimens were deposited in the Acari Collection of the Instituto terminal (Ttt1–Ttt6), 2 paraxial (Ttp1, Ttt2), and 2 antiaxial (Tta1,
Butantan (IBSP 5000), São Paulo, Brazil. Tta2) (Fig. 6); 6 genual setae, 1 paraxial (Gp1), 1 antiaxial (Ga1),
3 dorsal (Gd1, Gd2, Gd3), and 1 ventral (Gv1); 6 femoral setae, 1
Description paraxial (Fd1), 2 antiaxial (Fa1, Fa2), 1 dorsal (Fd1), and 2 ventral
(Fv1, Fv2); trochanter 0.
Amblyomma calcaratum Neumann 1899, Larva
(Figs. 1–10)
Idiosoma: Dorsal surface (Figs. 4 and 7). Length from apices of
scapulae to posterior margin of body 0.602 ± 0.015 (0.579–0.628);
greatest width 0.585 ± 0.012 (0.559–0.598); outline oval, with 11
festoons. Setae: 2 central dorsal pairs (Cd1, Cd2); 8 marginal dor-
sal pairs (Md1–Md8), Md1 and Md2 before a large wax gland
present in segment VIII (WdVIII1), and Md3 pair located in the
inner side behind this gland, Md4–Md8 pairs posterior to large wax
gland, each one in one subsequent festoon. Scutum (Figs. 4 and 7):
outline subtriangular; length 0.273 ± 0.012 (0.255–0.296); breadth
0.430 ± 0.015 (0.408–0.455) at level of eyes. Scutum with few punc-
tations. Eyes slightly bulging and shallow; cervical grooves slightly
convergent, parallel to the proximities of setae Sc3. Setae: 3 scutal
pairs (Sc1, Sc2, Sc3) (Fig. 4).
Ventral surface (Figs. 4, 8 and 9): Anal aperture on central portion
of opisthosoma. Setae: 3 sternal pairs (St1, St2, St3), 2 preanal pairs
(Pa1, Pa2), 4 premarginal pairs (Pm1–Pm4), 5 marginal ventral pairs
(Mv1–Mv5), and 1 anal pair (A1) (Figs. 4, 8 and 9).
Gnathosoma: Dorsal (Figs. 1 and 5). Basis capituli triangu-
lar in outline; length from posterior margin of trochanter to
posterior margin of capituli 0.073 ± 0.005 (0.065–0.079), width
0.157 ± 0.004 (0.151–0.163). Posterior margin straight, cornua
absent. Palpal grooves on segments well-defined (Fig. 5). Palp
length from apices of tibiotarsal segment to posterior margin
Figs. 2 and 3. Amblyomma calcaratum, larva. Tarsus I: (Fig. 2) dorsal view; (Fig. 3)
of trochanter 0.122 ± 0.003 (0.116–0.125), width 0.055 ± 0.004 ventral view. Abbreviations: d, dorsal; v, ventral; a, antiaxial; p, paraxial; la, lateral
(0.051–0.061); trochanter length 0.019 ± 0.002 (0.016–0.022), anterior; lp, lateral posterior.

Please cite this article in press as: Barbieri, F.S., et al., Description of the larva of Amblyomma calcaratum Neumann, 1899 (Acari: Ixodidae) by
light and scanning electron microscopy. Ticks Tick-borne Dis. (2013), http://dx.doi.org/10.1016/j.ttbdis.2013.07.004
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Fig. 4. Amblyomma calcaratum, larva. Segmentation model of the idiosoma. Segments are indicated by Roman numerals (III–VI and VIII–XIV) and delimitated by dashed lines
(- - -); series are indicated by Arabic numbers and delimitated by dotted lines (· · ·). Integumentary structures are illustrated.

Legs: Coxa I with 2 triangular spurs, the external slightly longer
than the internal; coxae II and III each with a single, large spur.
Setae: 3 on coxa I, 1 anterior (CIa), 1 posterior (CIp), and 1 parax-
ial (CIpa); coxae II and III each with 2 setae, 1 anterior (CIIa,
CIIIa) and 1 posterior (CIIp, CIIIp) (Fig. 8). Trochanter lacking spur.
Tarsus I length 0.220 ± 0.006 (0.210–0.227), width 0.085 ± 0.004
(0.076–0.090). Setae (Figs. 2, 3 and 10): dorsal, 2 in dorsal I group
(dI1, dI2); 6 dorsal II (dII1–dII6) (Figs. 2 and 10); 2 dorsal III (dIII1,
dIII2); 2 dorsal IV (dIV1, dIV2); 0 dorsal V; 2 dorsal VI (dVI1, dVI2);
ventral, 2 ventral I (vI1, vI2), 2 in II group (vII1, vII2), and 2 in III
(vIII1, vIII2); lateral anterior, 1 in lateral anterior I group (laI1) and
3 in laII group (laII1, laII2, laII3); lateral posterior, 1 in lateral poste-
rior I group (lpI1) and 3 in lpII group (lpII1, lpII2, lpII3). Ambulacrum
as illustrated (Figs. 2 and 3).
Porotaxy: Dorsal (Fig. 4): Large wax glands – one pair located
on the lateral margin of the idiosoma, in segment VIII – WdVIII1
(22/22 = 22 idiosomal sites containing the structure/22 observed
idiosomal sites). Lyrifissures – 11 pairs distributed in the follow-
ing segments: 1 pair in segment III – LdIII5 (22/22); 1 in IV–LdIV2
(19/22); 2 in V – LdV1 (17/22), LdV6 (19/22); 1 in VI – LdVI1 (20/22);
2 in VIII – LdVIII4 (18/22), LdVIII6 (21/22); 2 in IX – LdIX4 (20/22),
LdIX6 (21/22); 2 in XII – LdXII4 (20/22), LdXII7 (22/22). Small glands
– 40 pairs, 4 on the scutum: 3 pairs in segment III – SdIII1 (21/22),
SdIII2 (19/22), and SdIII5 (22/22); 1 in IV – SdIV6 (22/22); and an
additional asymmetric small gland on the posterior central margin
of the scutum – SdV7 (11/11). On the alloscutum, 36 pairs dis-
tributed in the following segments: 1 in IV – SdIV2 (19/22); 4 in
V – SdV2 (21/22), SdV3 (21/22), SdV4 (22/22), SdV6 (20/22); 4 in
VI – SdVI2 (22/22), SdVI4 (21/22), SdVI5 (22/22), SdVI6 (22/22);
4 in VIII – SdVIII3 (21/22), SdVIII4 (21/22), SdVIII6 (21/22), SdVIII7
(22/22); 3 in IX – SdIX1 (15/22), SdIX2 (22/22), SdIX4 (20/22), SdIX7
(22/22); 5 in X – SdX1 (21/22), SdX3 (15/22), SdX4 (17/22), SdX5
(21/22), SdX6 (22/22), and dorsal fovea (series 7 – 22/22); 3 in XI
– SdXI1 (22/22), SdXI2 (22/22), SdXI7 (22/22); 4 in XII – SdXII1
(21/22), SdXII2 (21/22), SdXII5 (21/22), SdXII7 (18/22); 5 in XIII –
SdXIII1 (12/22), SdXIII2 (22/22), SdXIII3 (21/22), SdXIII5 (22/22),
SdXIII6 (22/22); 2 in XIV – SdXIV1 (22/22), SdXIV2 (22/22).
Ventral (Fig. 4): Large wax glands – 4 pairs: 1 in the outer mar-
gin of coxa I in segment III – WvIII1 (22/22); 1 posterior to coxa
II in segment IV – WvIV4 (22/22); 1 posterior to coxa III in seg-
ment V – WvV5 (22/22); 1 on the 5th festoon in segment XIII –
WvXIII1 (22/22). Lyrifissures – 13 pairs distributed in the follow-
ing segments: 1 pair in segment IV – LvIV6 (22/22); 1 in V – LvV6
(22/22); 1 in VIII – LvVIII5 (19/22); 4 in IX – LvIX1 (22/22), LvIX3
(22/22), LvIX5 (19/22), LvIX6 (22/22); 1 in X – LvX2 (21/22); 1 in XI
– LvXI2 (22/22); 1 in XII – LvXII3 (22/22); 2 in XIII – LvXIII2 (22/22),
LvXIII6 (22/22); 1 in XIV – LvXIV2 (22/22). Small glands – 21 pairs
distributed in the following segments: 1 in III – SvIII6 (22/22); 1 in
IV–SvIV6, (22/22); 2 in V – SvV1 (20/22), SvV5 (22/22); 1 in VI –
SvVI2 (21/22); 1 in VIII – SvVIII1 (22/22); 3 in IX – SvIX2 (22/22),
SvIX4 (22/22), SvIX6 (22/22); 3 in X – SvX1 (22/22), SvX3 (21/22),
SvX4 (22/22); 2 in XI – SvXI1 (22/22), SvXI3 (19/22); 2 in XII – SvXII2
(22/22), SvXII4 (22/22); 2 in XIII – SvXIII3 (19/22), SvXIII6 (22/22); 3
in XIV – SvXIV1 (21/22), SvXIV3 (19/22), SvXIV5 (22/22). Asymmet-
ric small glands found at low frequency. SdVI3 (3/22), SdX7 (1/22),
and SvIX6 (2/22).
Discussion
The morphological characters and measurements of A. calcara-
tum were compared with descriptions of other species of the
Amblyomma genus from the Neotropical region. The basis capit-
uli of A. calcaratum is triangular, even as in Amblyomma cajennense
(Fabricius, 1787) (Famadas et al., 1997), Amblyomma longirostre

Please cite this article in press as: Barbieri, F.S., et al., Description of the larva of Amblyomma calcaratum Neumann, 1899 (Acari: Ixodidae) by
light and scanning electron microscopy. Ticks Tick-borne Dis. (2013), http://dx.doi.org/10.1016/j.ttbdis.2013.07.004
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Figs. 5–10. Amblyomma calcaratum, larva: (Fig. 5) Gnathosoma, dorsal view (bar: 30 ␮m); (Fig. 6) Gnathosoma, ventral view (bar: 30 ␮m); (Fig. 7) Larva, dorsal view (bar:
90 ␮m); (Fig. 8) Detail of coxae (bar: 50 ␮m); (Fig. 9) Detail of festoons (bar: 50 ␮m); (Fig. 10) Tarsus I, dorsal view (bar: 15 ␮m).

(Koch, 1844) (Barros-Battesti et al., 2005), Amblyomma ovale Koch,
1844 (Barbieri et al., 2008a), and Amblyomma pacae Aragão, 1911
(Barbieri et al., 2008b). However, in Amblyomma parvum Aragão,
1908, and Amblyomma pseudoparvum Guglielmone, Mangold and
Keirans, 1990 (Guglielmone et al., 1990), the basis capituli is slightly
subtriangular, and in Amblyomma brasiliense Aragão, 1908 (Sanches
et al., 2009) it is rectangular.
The idiosoma of A. calcaratum is oval in outline, as well as
in A. parvum and A. pseudoparvum (Guglielmone et al., 1990), A.
cajennense (Famadas et al., 1997), A. longirostre (Barros-Battesti
et al., 2005), A. ovale (Barbieri et al., 2008a), A. pacae (Barbieri
et al., 2008b), but is rounded in A. brasiliense. The measurements
of the idiosoma of A. calcaratum is similar to those observed in A.
brasiliense and A. ovale (Sanches et al., 2009; Barbieri et al., 2008a).
On the other hand, larvae of A. parvum, A. pseudoparvum, and A.
cajennense (Guglielmone et al., 1990; Famadas et al., 1997) are
smaller than A. calcaratum, and A. longirostre and A. pacae (Barros-
Battesti et al., 2005; Barbieri et al., 2008b) are larger.
The chaetotaxy of the idiosoma, gnathosoma, and tarsus I of the
larvae of A. calcaratum was similar to that found in other species of
this genus, as reported in other descriptions of larval Amblyomma.
Variation in the numbers of setae was observed for the tibiotarsus
of the palpi and for setae in group dII on tarsus I.
The palpal tibiotarsus of A. calcaratum carries 10 setae, 6 termi-
nal (Ttt1), 2 paraxial (Ttp), and 2 antiaxial (Tta). The same number
of setae was reported for Amblyomma varium Koch, 1844, Ambly-
omma dubitatum Neumann, 1899, A. parvum, A. pseudoparvum,
and Amblyomma oblongoguttatum Koch, 1844 (Amorim and Serra-
Freire, 1996, 1999; Guglielmone et al., 1990; Barbieri et al., 2012).
Larvae of Amblyomma rotundatum Koch, 1844, carry 7 setae on
that segment (Amorim and Serra-Freire, 1995), larvae of A. ovale,
A. pacae, and Amblyomma romitii Tonelli-Rondelli, 1939, carry 11
(Barbieri et al., 2008a, 2008b; Barros-Battesti et al., 2013), and
larvae of A. cajennense, A. longirostre, and A. brasiliense carry 12
(Famadas et al., 1997; Barros-Battesti et al., 2005; Sanches et al.,
2009).
The dorsal setal arrangement of tarsus I for 9 Amblyomma
species considered by Clifford and Anastos (1960) was 2:2:2:2:2.
The larvae of A. calcaratum show the same arrangement, except for
group dII, which includes 6 setae (Fig. 10). This number was also
found in larvae of A. parvum, A. pseudoparvum (Guglielmone et al.,
1990), A. dubitatum (Amorim and Serra-Freire 1999), and A. romitii

Please cite this article in press as: Barbieri, F.S., et al., Description of the larva of Amblyomma calcaratum Neumann, 1899 (Acari: Ixodidae) by
light and scanning electron microscopy. Ticks Tick-borne Dis. (2013), http://dx.doi.org/10.1016/j.ttbdis.2013.07.004
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(Barros-Battesti et al., 2013), but differs from that of A. rotundatum
(Amorim and Serra-Freire, 1995), A. varium (Amorim and Serra-
Freire, 1996), A. ovale (Barbieri et al., 2008a), A. pacae (Barbieri et al.,
2008b), and A. oblongoguttatum (Barbieri et al., 2012), all of which
have 5 setae in group dII, and from A. cajennense (Famadas et al.,
1997), A. longirostre (Barros-Battesti et al., 2005), and A. brasiliense
(Sanches et al., 2009), which have 7 each.
The topographical and numerical patterns of integumentary
structures on the idiosoma of the larvae of A. calcaratum showed the
presence of setae, lyrifissures, small glands, and large wax glands,
each of which easily characterized morphologically (Schulze, 1942;
Dinnik and Zumpt, 1949; Barbieri et al., 2007). As noted by Klompen
et al. (1996), Barbieri et al. (2007, 2008a,b), these structures are
found isolated or associated over the entire idiosoma, except on
the scutum where large wax glands are absent.
The arrangement of the large wax gland on the idiosoma of A.
calcaratum larvae was the same as reported by Clifford and Anastos
(1960) for Amblyomma species: one pair located dorsally on the
lateral margin of the body (segment VIII), and 4 pairs on the ven-
tral surface – one located posterior to each coxa (segments III, IV,
and V) and one pair on the 5th festoon (segment XIII). A similar
arrangement has been reported for Amblyomma glauerti Keirans,
King and Sharrad, 1994, Amblyomma variegatum (Fabricius, 1794),
Amblyomma americanum (Linnaeus, 1758) (Klompen et al., 1996);
A. cajennense (Famadas et al., 1997); A. parvum, A. rotundatum
(Barbieri et al., 2007), A. pacae (Barbieri et al., 2008b), A. brasiliense
(Sanches et al., 2009), and A. oblongoguttatum (Barbieri et al., 2012).
An additional ventral pair of large wax glands on the 4th fes-
toon (segment XII) has been reported for Amblyomma tuberculatum
Marx, 1894, Amblyomma geoemydae (Cantor, 1847), Amblyomma
babirussae Schulze, 1933 (Klompen et al., 1996), Amblyomma aure-
olatum (Pallas, 1772) (Arzua, 2002), A. longirostre (Barros-Battesti
et al., 2005; Barbieri et al., 2007), A. ovale (Barbieri et al., 2008a), and
A. romitii (Barros-Battesti et al., 2013). Further additional pairs have
been reported on segment V for A. geoemydae, segments V and X for
A. babirussae, dorsal on segment V (WdV1) for A. longirostre, and on
segments IV (WdIV3) and V (WdV1) for A. romitii (Barros-Battesti
et al., 2013).
The number of lyrifissures present on the idiosoma of A. cal-
caratum was 24, 11, and 13 on the dorsal and ventral surfaces,
respectively. This number and the pattern is similar to that found
for other Amblyomma species (Klompen et al., 1996; Barbieri et al.,
2007, 2008a,b), but differs from that for larval A. parvum, who carry
only 12 lyrifissures on the ventral surface. Klompen et al. (1996)
noted that the number and pattern of lyrifissures present in all Par-
asitiformes is similar, listing total numbers of lyrifissures of 18, 24,
20, 20 (all larvae) to 28 (adult), and 23–24 (adult) for Prostriata,
Metastriata, Argas, Carios (Argasidae), Zerconidae, and Amblysei-
inae, respectively (the latter 2 are Mesostigmata).
Among the integumentary structures, the small glands are the
most numerous on the idiosoma, but their frequency and pattern
also showed a high degree of variability. This pattern has been
observed previously for larvae of other Amblyomma species, such
as A. cajennense, A. longirostre, A. parvum, A. rotundatum (Barbieri
et al., 2007), A. ovale (Barbieri et al., 2008a), and A. pacae (Barbieri
et al., 2008b), as well as for species within the genera Rhipicephalus
Koch, 1844 (Nawar and Madbouly, 1985), Amblyomma, Dermacen-
tor Koch, 1844, and Ixodes Latreille, 1795 (Klompen et al., 1996).
This variability in the frequency of small glands can be related to
the higher number this structure is present on the idiosoma, since
the lowest variation has been observed for large wax glands, which
had the lowest number on the idiosoma (Barbieri et al., 2007).
In general, the integumentary structures on the idiosoma of the
larvae of A. calcaratum show patterns similar to those observed in
larvae of A. cajennense, A. longirostre, A. parvum, and A. rotunda-
tum (Barbieri et al., 2007), A. ovale (Barbieri et al., 2008a), A. pacae
Please cite this article in press as: Barbieri, F.S., et al., Description of the larva of Amblyomma calcaratum Neumann, 1899 (Acari: Ixodidae) by
light and scanning electron microscopy. Ticks Tick-borne Dis. (2013), http://dx.doi.org/10.1016/j.ttbdis.2013.07.004
5
(Barbieri et al., 2008b), A. oblongoguttatum (Barbieri et al., 2012),
and A. romitii (Barros-Battesti et al., 2013). However, their number,
position, and association patterns may help identifying this stage.
Thus, larvae of A. calcaratum differ from those of A. longirostre, A.
ovale, and A. romitii because these larvae have 2 and one additional
pairs of large wax glands, respectively (Barbieri et al., 2007, 2008a;
Barros-Battesti et al., 2013); from those of A. parvum because this
species has one less pair of lyrifissures (Barbieri et al., 2007). The
association between 2 or more structures can also be auxiliary in
the identification, for example, the association of lyrifissure and
small gland in ventral surface/segment IX/series 6 and lack in A.
cajennense and A. oblongoguttatum (Barbieri et al., 2007, 2012), and
association of the same structures in ventral surface/segment X, XI,
and XII/series 2 in A. pacae and lack in A. calcaratum (Barbieri et al.,
2008b).
This is the first morphological description of the larva of A.
calcaratum. Currently, larval morphological descriptions are avail-
able for only 19 of the 29 Amblyomma species known from Brazil
(Barbieri et al., 2012). Although many of these descriptions provide
general measurements of the body and details on chaetotaxy,
complete analyses of integumental structures (lyrifissures, small
glands, and large wax glands) have been reported for only 8 species
(A. cajennense, A. parvum, A. rotundatum, A. longirostre, A. ovale,
A. pacae, A. oblongoguttatum, and A. romitii) (Barbieri et al., 2007,
2008a,b, 2012; Barros-Battesti et al., 2013). Chaetotaxy is generally
highly conserved among Amblyomma spp. (Barbieri et al., 2007),
but analyses of especially gland distribution patterns in association
with other morphological features and bioecological information
should eventually yield a taxonomic key for identification of the
Amblyomma larvae of the Neotropical Zoogeographic Region.
Acknowledgments
The authors are grateful for financial support from CNPq-
Bolsa de Desenvolvimento Científico Regional (no. 350265/2005-4)
and Scientific Career Scholarships to MBL and DMBB, CNPq (nos.
479877-2004-1 and 304475/2010-6), FAPESP (no. 07/57749-2) and
FAPERJ (no. E-26/170179/2004).
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