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MORPHOLOGICAL AND KARYOTYPIC CONTRIBUTIONS FOR A BETTER
TAXONOMIC DEFINITION OF THE FROG ISCHNOCNEMA RAMAGII
(BOULENGER, 1888) (ANURA, BRACHYCEPHALIDAE)
Marcelo F. Napoli1,4, FerNaNdo aNaNias2, patricia M. FoNseca1, aNd aNa paula Z. silva3
1 Museu de Zoologia, Departamento de Zoologia, Instituto de Biologia, Universidade Federal da Bahia,
Rua Barão de Jeremoabo, Ondina, 40170‑115, Salvador, BA, Brasil. E‑mail: napoli@ufba.br, patriciamfonseca@yahoo.com.br
2 Curso de Ciências Biológicas, Universidade São Francisco (USF), Avenida São Francisco de Assis, 218,
Jardim São José, 12916‑900, Bragança Paulista, SP, Brasil. E‑mail: fernando.ananias@saofrancisco.edu.br
3 Laboratório Especial de Ecologia e Evolução, Instituto Butantan, Avenida Vital Brasil, 1500, 05503‑900,
São Paulo, SP, Brasil. E‑mail: pietri@uol.com.br
4 Corresponding author.
abstract. In this study we describe the morphological variation of Ischnocnema ramagii from a population sample within the
Municipality of Salvador, Bahia State, Brazil, and characterize two extremely distinct morphotypes within this taxon (striped vs.
non-striped dorsum); we describe its karyotype, and compare the karyotypic structure of the distinct morphotypes recognized
herein. Specimens with striped dorsa were less common (13-18%) than those with non-striped dorsa (81-86%), the latter group
comprising a mix of discrete color pattern states, whose frequencies were not different among juveniles, males, and females.
The PCAs on morphometric data resulted in a strong degree of superposition between striped and non-striped dorsa, for both
genders. We encountered a diploid number of 30 chromosomes for both males and females, with chromosome pairs 1 to 15
being telocentric. The karyotypic comparison between specimens with striped and non-striped dorsa did not show any noticeable
difference. Nevertheless the karyotype of I. ramagii from Salvador, Bahia, differed from the karyotype described for I. paulodutrai
from Ilhéus, Bahia, which showed a submetacentric chromosome pair. This result supports the presence of two full “sibling”
species in Bahia State, one from the south (I. paulodutrai) and another from the north (I. ramagii), validating the identity of
I. paulodutrai as a full species, and refuting the recognition of Salvador samples as I. paulodutrai. The high number of telocentric
chromosomes suggests a closer relationship between I. ramagii and “Eleutherodactylinae” species from Central America and the
northern region of Brazil.
Keywords. Ischnocnema paulodutrai, Intra-population variation, Karyotype, Polymorphism, Size and shape variation.
iNtroductioN
The “Eleutherodactylinae” frog Ischnocnema ra‑
magii (Boulenger, 1888) (sensu Heinicke et al., 2007)
inhabits Tropical Atlantic forests of northeastern Bra-
zil, from Paraíba State to Bahia State (Frost, 2008).
The species I. ramagii was described as Hylodes
ramagii, based on a single specimen collected by
G. A. Ramage in the Municipality of Igarassu, State
of Pernambuco, northeastern Brazil. In the last few
years, similar morphotypes have been registered from
the State of Bahia, Brazil, usually being recognized
as I. ramagii (e.g., Munduruca, 2005; Juncá, 2006;
Bastazini et al., 2007) or I. paulodutrai (Bokermann,
1975) (e.g., Napoli and Brandão, 2003). Other closely
related morphotypes have been captured within the
Tropical Atlantic Forest and Caatinga biomes by us
and by other researchers during surveys of anurans,
but probably belong to species not yet described
(pers. obs.).
Concerning the correct identification of specimens
of Ischnocnema ramagii and I. paulodutrai, four ma-
jor problems can be postulated: (1) the short original
description of I. ramagii; (2) the poor diagnosis of
I. paulodutrai; (3) possible morphological polymor-
phism within populations; and (4) complex variation
of advertisement calls within and among geographic
populations. The current poor knowledge of local
and geographic variation of both species prevents an
accurate assignment of a specimen to a given taxa,
which is usually done by taking into account the geo-
graphic distance between the area of collection and
the respective type-localities (I. paulodutrai, Ilhéus,
southern Bahia State, 14°47’S, 39°02’W; I. rama‑
gii, Igarassu, Pernambuco State, 07°50’S, 34°54’W;
ca. 890 km distance from each other). Because of
the above statements, we did not follow Napoli and
Brandão (2003) in allocating specimens from Salva-
dor to I. paulodutrai.
In this study, we make a first assessment to im-
prove the morphologic and cytogenetic knowledge of
I. ramagii, using population samples from a patch of
Atlantic Forest remnant located in the campus of the
Universidade Federal da Bahia (UFBA), at a neigh-
borhood named Ondina, in the Municipality of Sal-
vador, Bahia State, Brazil. We decided to limit our
analysis to this local population for three reasons:
(1) only one type of advertisement call is found in
South American Journal of Herpetology, 4(2), 2009, 164-172
© 2009 Brazilian Society of Herpetology
this population, which led us to assume that only one
species occurs at the locality; (2) this local population
contains two extremely distinct morphotypes, distin-
guished by the presence or not of longitudinal brown
stripes on the dorsum, which makes their descriptions
very important for an accurate definition of species
limits; (3) there is no doubt about the conspecific-
ity of these morphotypes, as we obtained 13 frogs of
both (UFBA 2914; 54% striped dorsum; 46% non-
striped dorsum) within a single clutch of large-yolked
terrestrial eggs; and (4) many specimens are available
from this locality (n = 151).
The goals of the present study are to (1) charac-
terize the morphological variation of Ischnocnema
ramagii from a population sample within the Munici-
pality of Salvador, Bahia State, Brazil; (2) describe
the karyotype; (3) assess the level of karyotypic poly-
morphisms in the population; and (4) compare the
karyotype of I. ramagii with the karyotype of I. pau‑
lodutrai from Ilhéus, Bahia State, by Siqueira et al.
(2008).
Material aNd Methods
Specimens used for descriptions or examined for
comparisons are deposited in the following Brazilian
collections (acronyms in parenthesis): Comissão Ex-
ecutiva do Plano de Cultivo da Lavoura Cacaueira,
Municipality of Ilhéus, Bahia State (CEPLAC); Cé-
lio Haddad Collection (CFBH), deposited in the De-
partamento de Zoologia, Instituto de Biociências,
Universidade Estadual Paulista, Municipality of Rio
Claro, São Paulo State; Museu de Zoologia, Univer-
sidade Federal da Bahia, Municipality of Salvador,
Bahia State (UFBA).
We analyzed 151 specimens of Ischnocnema ra‑
magii (89 males, 22 females, and 40 juveniles) from
a patch of Atlantic Forest remnant located in the cam-
pus of the Universidade Federal da Bahia, at a neigh-
borhood named Ondina, in the Municipality of Salva-
dor, Bahia State, Brazil (12°59’S; 38°27’W).
We developed outline drawings as a standard
series for color patterns of dorsum, interocular bar,
post-orbital region, and sacral surface, recorded for
each specimen examined. When a new pattern was
encountered, it was added to the standards. We fol-
lowed Heyer (1984) as several aspects of the dorsal
pattern are inherited independently in some species
of Terrarana (Goin, 1950, 1960). The purpose of
this procedure was to analyze (1) variation within
this population and (2) differences among juveniles,
males, and females. Drawings were made using a
Leica MZ6 stereomicroscope with a drawing tube.
Figure 1. Dorsal views of Ischnocnema ramagii from the Municipality of Salvador, Bahia State, Brazil. A, adult female with striped dor-
sum, UFBA 3158, SVL 25.0 mm; B, adult female with non-striped dorsum, UFBA 2271, SVL 25.5 mm. Photographs by Rafael O. Abreu.
165
Napoli, M. F. et al.
Chi-square analyses were performed to test whether
patterns observed differed in occurrence among juve-
niles, males, and females. The α-level of significance
(P ≤ 0.05) was Bonferroni corrected (Bland, 2004).
We measured 20 morphometric characters from 89
adult males and 22 adult females of I. ramagii, which
were recorded in millimeters. Fifteen measurements
followed Napoli (2005): SVL (snout-vent length), HL
(head length), HW (head width), ED (eye diameter),
UEW (upper eyelid width), IOD (interorbital dis-
tance), IND (internarial distance), END (eye-nostril
distance), NSD (nostril to tip of snout distance), TD
(tympanum diameter), THL (thigh length), TL (tibia
length), FL (foot length), 3FD (third finger disk di-
ameter), and 4TD (fourth toe disk diameter). Three
measurements followed Heyer et al. (1990): UAR
(upper arm), FAR (forearm), HAL (hand length). The
remaining measurements are LF1 (length of finger I)
and LF2 (length of finger II), both as the straight line
distance between the lower and upper borders of the
finger. We measured SVL, HL, HW, ITD, THL, TL,
FL, UAR, FAR, and HAL with a vernier caliper, and
used an ocular micrometer in an SZ Olympus stereo-
microscope for the remaining variables.
We used the principal component analysis (PCA)
to study multivariate patterns of shape and size varia-
tion. The first principal component (PC1) typically
expresses “size” variation when it has loadings of the
same magnitude and sign for all growth-dependent
variables (Humphries et al., 1981). The remaining
Figure 2. Standards for color patterns of (A) dorsum, (B) interocular bar, (C) post-orbital region, and (D) sacral surface. Additional stan-
dards not figured are: A1, B1, C1, and D1, which represent immaculate surfaces. Patterns D2 and D3 may be comprised by only one or
two transverse stripes.
166 Morphology and Karyotype of Ischnocnema ramagii
axes are uncorrelated with PC1 (orthogonal) and are
usually considered as “shape” variation. We obtained
eigenvectors and associated eigenvalues from a vari-
ance-covariance matrix, and the loadings were the
correlations between the original variables and the
scores. The scores were projected onto reduced space
using the axes that account for the largest amounts of
variation among the sample groups.
For cytogenetic analysis, we captured 19 speci-
mens of Ischnocnema ramagii (striped female, CFBH
09670; non-striped females, CFBH 09653-09654,
09657, 09662-09663; striped males, CFBH 09644,
09665-09669; non-striped males, 09651-09652,
09655, 09658-096661), and transported them alive to
the cytogenetics laboratory of the São Francisco Uni-
versity (São Paulo State, Brazil). Karyotypes were
prepared by two different procedures: (1) intestinal
epithelium – animals pretreated with 1% colchicine
solution for at least 4 h (Schmid, 1978) and (2) cyto-
logical preparations of bone marrow and liver – ani-
mals previously submitted to Phytohemagglutinin-P
(Cultilab) and 0.1% colchicine treatments, in the
table 3. Standardized coefficients and factor loadings (r) from
principal component analysis (PCA) for 20 morphometric char-
acters of Ischnocnema ramagii adult females from the Munici-
pality of Salvador, State of Bahia, Brazil. NS, not significant; *
P ≤ 0.05; ** P ≤ 0.01; ** P ≤ 0.001; Cum. Prop., cumulative
proportion of eigenvalues in percentage.
PC1 PC2 PC3 (r) PC1 (r) PC2 (r) PC3
SVL -0.50 0.74 -0.31 -0.91*** -0.40NS 0.11NS
HL -0.23 0.00 0.34 -0.91*** -0.25NS -0.05NS
HW -0.18 0.26 0.49 -0.91*** -0.21NS -0.11NS
ED -0.04 0.05 0.10 -0.49* -0.32NS -0.23NS
END -0.07 -0.02 0.20 -0.73*** -0.10NS -0.27NS
TD -0.03 0.06 0.05 -0.64*** -0.38NS -0.05NS
UEW -0.03 0.06 0.16 -0.49*** -0.57** -0.33NS
IOD -0.04 -0.04 0.06 -0.61*** -0.23NS -0.49*
IND -0.04 0.00 0.05 -0.83*** -0.37NS -0.07NS
NSD -0.02 0.00 0.04 -0.78*** -0.36NS -0.10NS
UAR -0.24 -0.09 0.20 -0.86*** 0.01NS -0.42*
FAR -0.12 0.01 0.13 -0.90*** -0.19NS 0.10NS
HAL -0.16 -0.04 -0.01 -0.87*** 0.21NS 0.01NS
THL -0.30 -0.11 0.35 -0.94*** 0.22** -0.01NS
TL -0.37 -0.42 0.11 -0.81*** 0.52NS 0.02NS
FL -0.52 -0.35 -0.44 -0.94*** 0.31NS 0.08NS
3FD -0.02 0.02 0.03 -0.21NS -0.33NS -0.17NS
4TD -0.02 0.02 0.05 -0.34NS -0.31NS -0.22NS
LF1 -0.10 -0.12 -0.16 -0.72*** 0.41NS -0.10NS
LF2 -0.10 -0.13 -0.11 -0.77*** 0.31NS -0.13NS
Eigenvalues 5.73 0.81 0.17 — — —
Cum. prop.
(%) 78.51 89.72 92.17 — — —
table 2. Standardized coefficients and factor loadings (r) from
principal component analysis (PCA) for 20 morphometric char-
acters of Ischnocnema ramagii adult males from the Municipal-
ity of Salvador, State of Bahia, Brazil. NS, not significant; *
P ≤ 0.05; ** P ≤ 0.01; ** P ≤ 0.001; Cum. Prop., cumulative
proportion of eigenvalues in percentage.
PC1 PC2 PC3 (r) PC1 (r) PC2 (r) PC3
SVL -0.50 0.74 -0.31 -0.90*** 0.42*** -0.09NS
HL -0.23 0.00 0.34 -0.88*** 0.01NS 0.21NS
HW -0.18 0.26 0.49 -0.78*** 0.37*** 0.35***
ED -0.04 0.05 0.10 -0.57*** 0.23* 0.21NS
END -0.07 -0.02 0.20 -0.73*** -0.07NS 0.31**
TD -0.03 0.06 0.05 -0.48*** 0.31** 0.14NS
UEW -0.03 0.06 0.16 -0.41*** 0.23* 0.29**
IOD -0.04 -0.04 0.06 -0.55*** -0.16NS 0.11NS
IND -0.04 0.00 0.05 -0.74*** 0.01NS 0.13NS
NSD -0.02 0.00 0.04 -0.56*** 0.05NS 0.14NS
UAR -0.24 -0.09 0.20 -0.86*** -0.11NS 0.12NS
FAR -0.12 0.01 0.13 -0.79*** 0.04NS 0.13NS
HAL -0.16 -0.04 -0.01 -0.91*** -0.08NS -0.01NS
THL -0.30 -0.11 0.35 -0.94*** -0.12NS 0.18NS
TL -0.37 -0.42 0.11 -0.92*** -0.33** 0.05NS
FL -0.52 -0.35 -0.44 -0.96*** -0.20NS -0.13NS
3FD -0.02 0.02 0.03 -0.50*** 0.12NS 0.11NS
4TD -0.02 0.02 0.05 -0.52*** 0.14NS 0.16NS
LF1 -0.10 -0.12 -0.16 -0.79*** -0.29** -0.19NS
LF2 -0.10 -0.13 -0.11 -0.79*** -0.32** -0.14NS
Eigenvalues 4.21 0.42 0.11 — — —
Cum. prop.
(%) 80.37 88.49 90.59 — — —
table 1. Occurrence of pattern states (Fig. 2) among juveniles, males, and females of Ischnocnema ramagii from Ondina, Municipality of
Salvador, Bahia State, Brazil. Values in parentheses are percentages.
General dorsal pattern Interocular bar Post-orbital maculae Sacral marks
Sample A1 A2 A3 A4 B1 B2 B3 C1 C2 C3 C4 D1 D2 D3 D4
Juveniles 11
(27.5)
23
(57.5)
6
(15.0)
1
(3.4)
18
(62.0)
10
(34.4)
1
(3.4)
19
(65.5)
7
(24.1)
2
(6.8)
3
(10.3)
7
(24.1)
13
(44.8)
6
(20.6)
Males 6
(7.6)
12
(15.3)
46
(58.9)
14
(17.9)
45
(93.7)
3
(6.2)
5
(7.5)
41
(62.1)
15
(22.7)
5
(7.5)
9
(13.6)
17
(25.7)
35
(53.0)
5
(7.5)
Females 4
(23.5)
9
(52.9)
4
(23.5)
9
(69.2)
4
(30.7)
8
(61.5)
4
(30.7)
1
(7.6)
1
(7.6)
3
(23.0)
7
(53.8)
2
(15.3)
167
Napoli, M. F. et al.
proportion of 0.1 ml/10 g animal weight, during 48 h
and 6 h, respectively (Baldissera et al., 1993; Silva
et al., 2000). We stained the slides with 10% Giemsa
solution for chromosome morphological studies, and
performed the nucleolus organizer regions (NOR)
by silver nitrate staining (Ag-NORs) (Howell and
Black, 1980). We identified chromosomes according
to Green and Sessions (1991).
results
We found two clearly distinct color patterns with-
in the examined population of Ischnocnema ramagii;
a morphotype with striped dorsum (Fig. 1A) and an-
other with non-striped dorsum (Fig. 1B). Specimens
with non-striped dorsa were more common (males:
n = 79, 86.81%; females: n = 18, 81.8%) than striped
specimens (males: n = 12, 13.18%; females: n = 4,
18.1%); these two morphotypes exhibit a mixture of
discrete pattern states of general dorsal patterns, in-
terocular bar, post-orbital maculae, and sacral marks
(Fig. 2; Table 1), whose frequencies were not differ-
ent among juveniles, males, and females (after Bon-
ferroni correction significant at P < 0.01: X2 = 0.46,
DF = 2, P = 0.79; X2 = 1.45, DF = 2, P = 0.48;
X2 = 0.11, DF = 2, P = 0.94; X2 = 0.46, DF = 2,
P = 0.79; respectively).
Principal component analyses showed a strong de-
gree of overlap between specimens of I. ramagii with
striped and non-striped dorsa, separately for males
and females (Fig. 3; Tables 2 and 3).
We encountered a diploid number of 30 chromo-
somes in mitotic cells for both males and females.
Chromosome pairs 1 to 15 were telocentric. A slight
and continuous size decrease was observed in chro-
mosome length from pairs 1-5 and 6-15 (Fig. 4).
Stained chromosomes presented Ag-NOR at the dis-
tal region of chromosome 7, which coincided with
the site of the secondary constriction (Fig. 5). The
karyotypic comparison between I. ramagii speci-
mens with striped and non-striped dorsa (dorsal
staining polymorphism) did not show any noticeable
difference.
Figure 3. Projection of individual scores resulted from the principal component analysis (PCA) for 20 morphometric characters from adult
males (A, B) and adult females (C, D) of Ischnocnema ramagii in the space of the first with the second axes. Standardized coefficients and
loadings are presented in tables 2 and 3, respectively.
168 Morphology and Karyotype of Ischnocnema ramagii
discussioN
The two morphotypes and the color patterns de-
scribed herein for Ischnocnema ramagii from Salva-
dor (northern Bahia) are also observed within popu-
lation samples from southern Bahia, Municipality of
Ilhéus (type-locality of I. paulodutrai). Comparisons
of specimens of I. paulodutrai with I. ramagii from
Salvador did not reveal any clear differences in exter-
nal morphology. Furthermore, I. ramagii from Salva-
dor showed the same diploid number (2n = 30) and
similar chromosome morphology as I. paulodutrai
from Ilhéus (Siqueira et al., 2008). Nevertheless, the
karyotype of I. paulodutrai lacks the submetacentric
chromosome pair present in I. ramagii from Salvador.
This result supports the presence of two full “sibling”
species in Bahia State: one from the south (I. pau‑
lodutrai) and another from the north (I. ramagii). It
also validates the identity of I. paulodutrai as a full
species, and refutes the recognition of Salvador sam-
ples as I. paulodutrai by Napoli and Brandão (2003).
Chromosome numbers 2n = 28 and 2n = 30, with a
high number of telocentric pairs, have been described
for species of the Central American genera Euhyas
Fitzinger, 1843, Pelorius Hedges, 1989, and Syrrho‑
phus Cope, 1878 (see Campos and Kasahara, 2006 for
Figure 4. Karyotypes of Ischnocnema ramagii (A) with striped dorsum and (B) with non-striped dorsum after Giemsa staining. The second-
ary constrictions are indicated by arrows in pair 7, corresponding to homomorphic state of nucleolar organizer regions (NOR).
169
Napoli, M. F. et al.
a synthesis), currently placed in the genus Eleuthero‑
dactylus Duméril and Bibron, 1841 and Pristimantis
Jiménez de la Espada, 1870 (Frost, 2008), and also
for the Brazilian species Pristimantis dundeei (Heyer
and Muñoz, 1999), P. aff. dundeei, and I. paulodutrai
(Siqueira et al., 2008).
According to King (1990), Kuramoto (1990), and
Campos and Kasahara (2006), nearly 100 species of
Eleutherodactylus (many of them currently assigned
to other genera; see Heinicke et al., 2007 and Hedges
et al., 2008 for taxonomic arrangement) have, to some
extent, been chromosomally examined. Contrary to
the conservative karyotypic pattern of most anuran
genera, Eleutherodactylus and allied genera show a
high variation in diploid number, ranging from 2n = 18
to 2n = 36, and also a range of the fundamental num-
ber of chromosome arms, from FN = 32 to FN = 58,
even among species sharing the same diploid number.
In Brazil, seven “Eleutherodactylinae” species have
been karyotyped: Haddadus binotatus (Spix, 1824),
I. guentheri (Steindachner, 1864), I. holti (Cochran,
1948), I. juipoca (Sazima and Cardoso, 1978), I. lac‑
tea (Miranda-Ribeiro, 1923), I. parva (Girard, 1853),
and I. paulodutrai. With the exception of I. pau‑
lodutrai all species cytogenetically analyzed showed
2n = 20 or 2n = 22 chromosomes (Beçak, 1968; Beçak
and Beçak, 1974; de Lucca et al., 1974; de Lucca and
Jim, 1974; Siqueira et al., 2004; Siqueira et al., 2008;
Campos et al., 2008). The number of telocentric chro-
mosomes is highest among those species possessing
the largest chromosome numbers, which suggest that
karyotype evolution may have occurred through cen-
tric fusions and/or centric fissions. Sexual chromo-
somes were described for Eleutherodactylus maussi
Stejneger, 1904 (currently Strabomantis biporcatus
Peters, 1863; Schmid et al. 1992, 2002b); E. euphro‑
nides Schwartz, 1967 and E. shrevei Schwartz, 1967
(both currently allocated in the genus Pristimantis;
Schmid et al. 2002a), and for E. riveroi (= Pristiman‑
tis, Schmid et al. 2003).
The high number of telocentric chromosomes
suggests a closer relationship between I. ramagii and
“Eleutherodactylinae” species from Central Amer-
ica and northern Brazil. Molecular studies obtained
by Heinicke et al. (2007) and Hedges et al. (2008)
revealed three large radiations of species with un-
expected geographic isolations: a South American
clade, a Caribbean clade, and a Middle American
clade. According to Heinicke et al. (2007), the great
diversity of species of the South American clade is as-
sociated to the Andes uplift, a relatively recent event.
Mountain building and associated climatic changes
resulted in repeating patterns of habitat isolation,
which, in turn, probably resulted in genetic isolation
and speciation in these amphibians.
The use of other banding techniques, as well as an
increase in the number of specimens and population
samples of Ischnocnema and allied genera are neces-
sary for the understanding of the chromosome evolu-
tion in this group of frogs.
resuMo
No presente estudo descrevemos a variação
morfológica de Ischnocnema ramagii a partir de
uma população presente no Município de Salvador,
Figure 5. Chromosomes of Ischnocnema ramagii treated with the
Ag-NOR method: (A) with striped dorsum and (B) with non-striped
dorsum. Note the chromosome pair 7 bearing de Ag-NOR site.
170 Morphology and Karyotype of Ischnocnema ramagii
Estado da Bahia, Brasil, e caracterizamos dois
morfótipos extremamente distintos neste táxon
(dorso listrado vs. não-listrado); descrevemos seu
cariótipo e comparamos a estrutura cariotípica dos
distintos morfótipos aqui reconhecidos. Espécimes
com dorso listrado foram menos comuns (13-18%)
do que aqueles com dorso não-listrado (81-86%),
os últimos compostos por uma mistura de padrões
de colorido, cujas freqüências não foram diferentes
entre juvenis, machos e fêmeas. Os PCAs sobre
os dados morfométricos resultaram em alto grau
de sobreposição entre espécimes de dorso listrado
e não-listrado, para ambos os sexos. Encontramos
o número diplóide de 30 cromossomos para
machos e fêmeas, com pares de cromossomos
telocêntricos do 1 ao 15. A comparação cariotípica
entre espécimes de dorso listrado e não-listrado não
demonstrou qualquer diferença notável. Entretanto,
o cariótipo de I. ramagii de Salvador, Bahia,
diferiu do cariótipo descrito para I. paulodutrai
oriundo de Ilhéus, Estado da Bahia, que apresentou
um par de cromossomos submetacêntricos. Este
resultado suporta a presença de duas espécies
“crípticas” plenas no Estado da Bahia, uma no sul
(I. paulodutrai) e outra no norte (I. ramagii), reforça
a identidade de I. paulodutrai como espécie plena
e refuta a identificação da população de Salvador
como pertencendo a I. paulodutrai. O elevado
número de cromossomos telocêntricos sugere um
relacionamento íntimo entre I. ramagii e espécies
de “Eleutherodactylinae” da América Central e da
região norte do Brasil.
acKNowledgMeNts
We acknowledge Milena Camardelli (Post-Graduate student
in Ecology and Biological Monitoring at Universidade Federal
da Bahia) for the measurements taken of Ischnocnema ramagii;
Rafael Oliveira de Abreu (Graduate student in Biological
Sciences at Universidade Federal da Bahia) for the photographs of
I. ramagii; Carlos Alberto Gonçalves da Cruz (Museu Nacional/
UFRJ) and Wilfried Klein (Universidade Federal da Bahia) for
critically reading and improving the manuscript; Antônio Jorge
Suzart Argôlo (Universidade Estadual de Santa Cruz) for allowing
us to examine specimens under his care. The Conselho Nacional
de Desenvolvimento Científico e Tecnológico (CNPq) awarded a
fellowship to Marcelo Felgueiras Napoli (Proc. 302967/2003-6);
the Fundação de Amparo à Pesquisa do Estado de São Paulo
(FAPESP) financially supported Ana Paula Zampieri Silva (Proc.
2006/06634-8). The Instituto Brasileiro do Meio Ambiente
e dos Recursos Naturais Renováveis (IBAMA) permitted
collection of specimens (license numbers 075/03-IBAMA/RAN,
023/05-IBAMA/RAN, 038/05-IBAMA/RAN, 105/05-IBAMA/
RAN, and 106/06-IBAMA/RAN).
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Submitted 03 February 2009
Accepted 23 April 2009
appeNdix 1
Specimens Examined
Ischnocnema paulodutrai – BRASIL: Bahia: Ilhéus, type-locality (CEPLAC 347, 458, 466, 473-476, 533,
541-548, 708-709, 711-712, 714-717, 7119, 1534, 1545-1547, 1597-1599, 1606, 1612-1622, 1619, 1620-1621,
1622, 1715-1717; UFBA 819-823).
Ischnocnema ramagii – BRASIL: Bahia: Salvador (CFBH 9644, 9651-9655, 9657-9663, 9665-9670; UFBA
809-818, 824-842, 844, 846-847, 2158-2168, 2170-2238, 2268-2284, 2914-2915, 3150, 3152, 3158-3160, 5575).
Pernambuco: Igarassu, type-locality (UFBA 7511, 7669-7677). Sergipe: Itaporanga d’Ajuda (UFBA 6809-6815).
172 Morphology and Karyotype of Ischnocnema ramagii
