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Zookeys 918: 41-63 (2020)

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Cryptic diversity and range extension in the big-eyed
bat genus Chiroderma (Chiroptera, Phyllostomidae)

Burton K. Lim!, Livia O. Loureiro?, Guilherme S.T. Garbino?

| Department of Natural History, Royal Ontario Museum, 100 Queen's Park, Toronto, Ontario M5S 2C6, Ca-
nada 2 Department of Ecology and Evolutionary Biology, University of Toronto, Toronto, Ontario M5S 3B2,
Canada 3 PPG-Zoologia, Departamento de Zoologia, Instituto de Ciéncias Biolégicas, Universidade Federal
de Minas Gerais, Avenida Antonio Carlos 6627, Pampulha, 31270-901, Belo Horizonte, Minas Gerais, Brazil

Corresponding author: Burton K. Lim (burtonl@rom.on.ca)

Academic editor: DeeAnn Reeder | Received 26 November 2019 | Accepted 28 January 2020 | Published 12 March 2020
http://zoobank. org/8C780A 96-9353-4F 34-88 92-64822D8E2932

Citation: Lim BK, Loureiro LO, Garbino GST (2020) Cryptic diversity and range extension in the big-eyed bat genus
Chiroderma (Chiroptera, Phyllostomidae). ZooKeys 918: 41-63. https://doi.org/10.3897/zookeys.918.48786

Abstract

Since the last systematic review of Chiroderma (big-eyed bats) more than two decades ago, we report on
biodiversity surveys that expand the distribution and species diversity of this Neotropical genus. The
Caribbean endemic species Chiroderma improvisum is documented for the first time from Nevis in the
northern Lesser Antilles. A broader geographic sampling for a molecular analysis identifies a paraphyletic
relationship in Chiroderma trinitatum with respect to Chiroderma doriae. Cis-Andean populations of C.
trinitatum are most closely related to the morphologically distinctive and allopatrically distributed C. do-
riae in the Cerrado and Atlantic Forest of Brazil and Paraguay. The sister taxon to this grouping includes
trans-Andean populations of C. trinitatum, which we recommend to elevate to species status as C. gorgasi.
This is an example of a cryptic species because C. gorgasi was previously considered morphologically
similar to C. trinitatum, but more detailed examination revealed that it lacks a posterolabial accessory
cusp on the lower second premolar and has a narrower breadth of the braincase. We provide an amended

description of Chiroderma gorgasi.

Keywords

Chiroderma gorgasi, Chiroderma improvisum, cryptic species, cytochrome c oxidase subunit 1, Lesser Antilles

Copyright Burton K. Lim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC
BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

42 Burton K. Lim et al. / ZooKeys 918: 41-63 (2020)

Introduction

Cryptic species, phenotypically similar organisms that are classified as a single species
but are genetically divergent lineages, are being discovered at a greater rate due to the
increasing prevalence of molecular methods, such as DNA barcoding (e.g., Hebert et
al. 2004). It has been estimated that Neotropical mammalian biodiversity is underesti-
mated by one-third (Lim 2012). At typical lowland tropical forest sites, bats comprise
the majority of mammal species diversity (Voss and Emmons 1996), so more species
are expected to be recognized in this group as traditional taxonomic hypotheses are
tested by genetic techniques. In addition, new surveying methods such as the use of
triple-high netting systems to catch higher flying aerial insectivorous bats, and harp
traps to target species that may be able to better detect mist nets, is decreasing the sam-
pling bias associated with traditional mist nets set just above ground level.

The big-eyed bats in the genus Chiroderma Peters (Phyllostomidae) are character-
ized by greatly reduced nasal bones in the skull and a combination of external fea-
tures including a white dorsal stripe that does not extend onto the head; legs and
interfemoral membrane conspicuously hairy; and relatively large eyes (Straney 1984;
Gardner 2008). The genus currently comprises six species (Simmons 2005, Taddei and
Lim 2010): C. doriae Thomas, 1891 occurs in central-eastern Brazil and Paraguay; C.
improvisum Baker & Genoways, 1976 is endemic to the Lesser Antillean islands of
Guadeloupe, Montserrat, and Saint Kitts (Beck et al. 2016); C. salvini Dobson, 1878
is found from Mexico to Bolivia (recent records from Brazil are misidentifications of
C. villosum Peters, 1860 — see Brandao et al. 2019); C. trinitatum Goodwin, 1958 is
distributed from Honduras (Turcios-Casco et al. 2020) and Costa Rica to Amazonian
Brazil and Trinidad; C. villosum ranges from Mexico to southeastern Brazil and Trini-
dad; and C. vizottoi Taddei & Lim, 2010 is found only in northeastern Brazil.

The systematics of Chiroderma was last reviewed by Baker et al. (1994) based on
a phylogenetic study of the mitochondrial DNA cytochrome 6 (Cytb) gene; however,
each of the five species known at the time was represented by a single specimen. With
broader geographic coverage, we re-assess the distributional range, genetic diversity,
and morphological differences in the genus.

Material and methods

Fieldwork

We conducted a survey of bats on the Caribbean island of Nevis from 24-29 April
2016. Live traps used included a harp trap and 6 m or 12 m mist nets set singly in
the forest understory or on a triple-high telescoping pole system. Traps were regu-
larly monitored for the first 2-3 hours after sunset when bat activity is the highest
after they leave their roosts to feed. Individuals not kept as part of the representa-

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 43

tive collection documenting the species diversity were released at point of capture.
A combined scientific research and export permit (F002) was issued through the
authority of the Nevis Historical and Conservation Society. An Animal Use Protocol
(2016-01) was obtained from the Royal Ontario Museum Animal Care Committee.
An import permit (4#2016-02101-4) was authorized by the Canadian Food Inspec-
tion Agency. Use of wild mammals followed the guidelines of the American Society
of Mammalogists (Sikes et al. 2016).

Molecular analyses

The cytochrome c oxidase subunit 1 (CO1) gene is the best represented molecular
marker for Chiroderma on the genetic sequence database GenBank (www.ncbi.nih.gov/
genbank). There are 117 samples from nine countries in Central and South America
(Brazil, Ecuador, El Salvador, French Guiana, Guatemala, Guyana, Mexico, Panama,
and Suriname). We add 26 new sequences to bring the sample size to 143 sequences
representing 12 countries in the Neotropics, including Venezuela, Peru, and Nevis,
and five species in the genus (Appendix 1). There are no tissue samples or nucleotide
sequences on GenBank of any genes for the recently described Chiroderma vizottoi
(Taddei and Lim 2010). Outgroup taxa were other genera in the subtribe Vampyressi-
na Baker et al., 2016 (Platyrrhinus incarum Thomas, 1912 and Uroderma bilobatum
Peters, 1866) of the New World leaf-nosed bats, for direct comparison to Baker et al.
(1994) in their analysis of Cytb. Alternative phylogenetic relationships within the sub-
tribe are given by Baker et al. (2016) and Rojas et al. (2016). We also analyzed Cytb,
but there are only 11 sequences on GenBank, although we did add one new sequence
of Chiroderma trinitatum gorgasi from Panama (Appendix 2).

Molecular methods for new sequences of CO1 follow the protocol for DNA ex-
traction, PCR amplification, and automated nucleotide sequencing outlined in Lim
(2017). For Cytb, extraction, amplification, and sequencing followed Lim et al. (2008).
Base calls were confirmed with bidirectional sequences and aligned using Sequencher
version 4.8 (Gene Code Corporation, Ann Arbor, Michigan). Phylogenetic and mo-
lecular evolutionary analyses were conducted using MEGA version 6 (Tamura et al.
2013). For a robust comparison of phylogeny, we used parsimony as a method that
minimizes evolutionary change without an explicit model of evolution and maximum
likelihood as a probabilistic method with an explicit model of evolution. Maximum
parsimony used the subtree pruning regrafting inference method with 500 bootstrap
replicates to test branch supports. Maximum likelihood used the Tamura 3-parameter
substitution model and gamma distributed rates with invariant sites for COI as deter-
mined by the best fit test. For Cytb, the Tamura Nei model and gamma rates were the
best fit. Tree inference used nearest neighbor interchange heuristic inference with 500
bootstrap replicates. Genetic distances were calculated with the Tamura 3-parameter
model with gamma distributed rates among sites for the larger COI dataset.

44 Burton K. Lim et al. / ZooKeys 918: 41-63 (2020)

Morphological analyses

Morphological and morphometric comparisons included 138 specimens from five species
of Chiroderma, including two C. improvisum, four C. doriae, seven C. salvini, 58 C. trini-
tatum, and 66 C. villosum (Appendix 3). We also analyzed the holotypes of C. trinitatum
gorgasi Handley, 1960 and C. trinitatum trinitatum Goodwin, 1958, but did not have
specimens of the most recently described C. vizottoi. Only adults (defined as having closed
cranial sutures and complete epiphyseal ossification of metacarpal and phalanx joints) of
both sexes were examined. Specimens are deposited in the following institutions; Royal On-
tario Museum (ROM, Toronto, Canada); National Museum of Natural History (USNM,
Washington, DC, USA); American Museum of Natural History (New York, USA); Texas
Tech University (Lubbock, USA); and Field Museum of Natural History (Chicago, USA).
Measurements defined below were taken with digital calipers accurate to 0.01 mm
following the descriptions of Handley (1960): forearm length (FA); greatest length of skull
(GSL); interorbital width (TOW); postorbital width (POW); braincase width (BCW); con-
dyloincisive length (CIL); zygomatic breadth (ZB); width across upper molars (M-M);
width across upper canines (C-C); and length of maxillary toothrow (C-M). An analysis of
variance (ANOVA) for each measurement and a multivariate analysis of variance (MANO-
VA) were performed to examine the significance of morphometric divergence among spe-
cies of Chiroderma. The level of significance was p = 0.05 for all statistical tests. The homo-
scedasticity of each variable was tested using Bartlett’s test with the R package mvoutliers.
Statistical analyzes were performed using R 3.1.0 (R Core Team 2005) and PAST 2.17.
Variables were log-transformed and a correlation matrix was used in a Principal Compo-
nents Analysis (PCA) to assess phenetic differences in multivariate morphological space.

Results

We report the first occurrence of Chiroderma improvisum (Fig. 1) from Nevis in the
northern Leeward Islands of the Lesser Antilles in the Caribbean. An adult male was
caught at Barnes Ghaut on April 28, 2016, in a harp trap set across a dry ravine in for-
est bisected by a road and surrounded by residential homes (Fig. 2). Other equipment
deployed included 6 m mist nets set on a triple-high telescoping pole system, a single
6 m mist net, and a single 12 m mist net from 1900-2100 h. In addition to the new
distributional record for the island, one Ardops nichollsi, one Noctilio leporinus, and 12
Artibeus jamaicensis were captured.

Molecular analyses

For COI, the 657 basepairs (bp) at the 5’ end were available for most (82%) of the speci-
mens analyzed. ‘The complete 1140 bp of Cytb were available, including the newly gener-
ated sequence, for most (75%) of the specimens analyzed. ‘The topology of the Chiroderma
COI maximum likelihood tree identified six primary terminal clades with (1) C. salvini as

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 45

Figure |. First record of the big-eyed bat Chiroderma improvisum from Nevis (ROM 126002).

Figure 2. Collecting locality of the first record of the big-eyed bat Chiroderma improvisum from Nevis

caught in a harp trap on April 28, 2016. The habitat is a dry ravine within forest that is bisected by a road
in the residential area of Barnes Ghaut.

46 Burton K. Lim et al. / ZooKeys 918: 41-63 (2020)

Chiroderma trinitatum gorgasi ®

Chiroderma doriae @

Chiroderma improvisum ©

Chiroderma villosum ©

Chiroderma salvini ®
Platyrrhinus incarum

Uroderma bilobatum

0.05

Figure 3. Maximum likelihood tree of cytochrome c oxidase subunit 1 gene for big-eyed bats Chiroderma

and general localities of molecular sampling sites. Bootstrap percentages show support at each node.

sister species to all other taxa; (2) C. improvisum and (3) C. villosum as sister species; and
(4) C. doriae sister to (5) C. trinitatum trinitatum with (6) C. trinitatum gorgasi sister to
these taxa (Fig. 3; Suppl. material 1: Fig. S1). These phylogenetic relationships were sup-
ported by bootstrap values >85 and were congruent with the maximum parsimony tree
(Suppl. material 2: Fig. S2), which had bootstrap values >73. Not surprisingly for linked
mtDNA loci, the same interspecific topology was recovered by the smaller Cytb dataset
analyzed by maximum likelihood (Suppl. material 3: Fig. $3) and maximum parsimony
(Suppl. material 4: Fig. S4), except for lower bootstrap supports. The unexpected result
was the paraphyly of C. trinitatum in relation to C. doriae. The sister-group relationship of
C. t. trinitatum and C. doriae was well supported by values >73 in all molecular analyses.

Interspecific genetic distances of the larger COI dataset ranged from 11.3% be-
tween C. doriae and C. salvini to 2.5% between C. doriae and C. t. trinitatum (Table 1).
The sequence divergence between C. ¢. trinitatum and C. t. gorgasi was 3.9%. Intraspe-
cific distances were 1% within C. villosum, 0.9% within C. t. trinitatum, and 0.2%
within C. doriae, but three taxa were represented by only one sample.

Morphological analyses

Cranial and body measurements for the six taxa of Chiroderma identified in the mo-
lecular analyses are shown in Table 2. Chiroderma trinitatum gorgasi and C. trinitatum
trinitatum are the smallest members of the genus, whereas C. improvisum is the largest
for most measurements. In the PCA, there are three main groups of species (Fig. 4). The

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 47

Table |. Genetic divergence of cytochrome c oxidase subunit 1 for the big-eyed bat Chiroderma and out-
group taxa Uroderma and Platyrrhinus. Interspecific distances shown in the lower left matrix; intraspecific
distances shown in bold in the diagonal.

(1) (2) (3) (4) (5) (6) (7) (8)
U. bilobatum (1) -
P incarum (2) 0.203 -
C. villosum (3) 0.223 0.178 0.010

C. improvisum (4) 0.231 0.194 0.047 -
C. t. trinitatum (5) 0.222 0.184 0.067 0.075 0.009

C. salvini (6) 0.205 0.185 0.093 0.101 0.110 -
C. t. gorgasi (7) 0.195 0.149 0.059 0.070 0.039 0.101 -
C. doriae (8) 0.213 0.173 0.066 0.077 0.025 Ol oe 0.039 0.002

Table 2. Cranial and body measurements of six taxa of the big-eyed bat Chiroderma. See Material and
methods for variable abbreviations.

C. trinitatum C. trinitatum C. villosum C. salvini (N C. doriae(N = 4) C. improvisum
gorgasi(N = 11) trinitatum(N = 47) (N = 66) = 6) (N = 2)

FA 37.7 (37.0-40.5) 38.9 (37.1-42.6) 47.9 (44.6-51.0) 49.6 (49.3—50.0) 54 (53.0-55.0) 58.2 (56.2-60)

GLS 21.2 (20.6-21.7) 21.1 (20.0—22.7) 24.5 (23.2-25.7) 26.1 (24.2-26.5) 28.2 (27.6-28.9) 29.3 (28.7-29.9)

CLI 17.3 (16.3-18.1) 17.4 (16.1-18.8) 20.3 (18.7-21.58) 21.6 (21.5-22.3) 23.9 (23.1-24.5) 26.3 (25.3—27.8)

ZB 13.0 (12.5-13.5) 12.9 (11.7-14.2) 15.5 (14.4-16.7) 16.2 (15.8-16.7) 17.84 (17.7-18.1) 18.75 (18.5-19)

POW 5.3 (4.9-5.6) 5.3 (5.8—5.8) 5.9 (5.3-6.3) 6.2 (5.9-6.3) 6.4 (6.1-6.6) 6.6 (6.5—6.6)
IOW 5.6 (5.2—5.9) 5.5 (5.0-6.2) 6.0 (5.56.8) 6.8 (6.1-7.3) 7.6 (7.1-7.8) 7.4 (7.4-7.4)
BCW 9.4 (8.9-9.8) 9.6 (9.2-10.4) 10.7 (10.1-11.5) 11.21 (11.0-11.5) 11.9 (11.3-12.1) 12.0 (11.5-12.5)
C-M 7.0 (6.5-7.3) 7.1 (6.7-7.8) 8.7 (8.1—9.4) 9.4 (9.1-9.4) 10.3 (10.0-11.1) 11.0 (10.9-11.1)
M-M 9.6 (9.2—10.0) 9.5 (8.7—10.3) 11.3 (10.3-12.4) 12.0 (11.5-12.3) 13.3 (13.0-13.7) 13.8 (13.6-13.9)
C-C 4.7 (4.45.0) 4.6 (4.1-5.0) 5.8 (5.3-6.3) 6.1 (6.0-6.2) 6.5 (6.3-6.8) 7.4 (7.4-7.4)
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Component 1

Figure 4. Principal Component Analysis (PCA) carried out using the correlation matrix of ten measure-
ments for six taxa of the big-eyed bat Chiroderma. C. trinitatum gorgasi (W) C. trinitatum trinitatum (Q),
C. salvini (x), C. villosum (+), C. doriae (@), and C. improvisum (0).

48 Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

Table 3. Eigenvalue and loadings for the first and second components in the Principal Component
Analysis (PCA) of big-eyed bats Chiroderma. See Material and methods for variable abbreviations.

PCl PC2
Eigenvalue 57 0:53
% Variance 91.4 ome
IlOW 0.25 0.44
POW 0.21 0.40
C-M 0.34 -0.23
GSL Or 33) -0.05
BCW 0.34 0.35
Gi 0.34 0.07
Ce 0.36 -0.23
M-M 0.36 -0.14
ZB 0.37 -0.12
FA 0.42 0.32

first group is formed by the smaller taxa C. 4. gorgasi and C. t. trinitatum. The second
group has species with medium size, C. villosum and C. salvini, and the third group is
formed by the largest species of the genus, C. doriae and C. improvisum. The first and
second principal components (PC1 and PC2) explained 94.5% of the total variation.
PC1 shows a pattern in general size variation and is explained mostly by C-M, C-C,
and FA. PC2 has positive loadings for most measurements, especially IOW, with the
exception of C-M, C-C, M-M, and ZB that have negative loadings (Table 3). All the
species seem to occupy the entire range of PC2, indicating that the contrast among
measurements is negligible and it is not responsible for the separation of groups.

All variables had p > 0.05 for Bartlett’s test of homoscedasticity, indicating constant
variances (p values: FA = 0.06, GSL = 0.25, IOW = 0.59, POW = 0.31, BCW = 0.06, CIL
= 0.45, ZB = 0.08, M-M = 0.32, C-C = 0.08, and C-M = 0.06). The MANOVA and the
ANOVA demonstrated that C. ¢. trinitatum and C. t. gorgasi are significantly different
from the other taxa of Chiroderma (P < 0.001) for all measured variables (Appendix 3). By
contrast, C. ¢. gorgasi and C. ¢. trinitatum are not significantly different from each other
(p = 0.16, F = 56.0). However, the ANOVA showed that one measurement, BCW (p =
0.01; F = 62.0), was significantly larger for C. ¢. trinitatum than for C. t. gorgasi. All other
cranial measurements had smaller mean values for C. ¢. trinitatum than for C. t. gorgasi.

Although similar in size, C. t. trinitatum has a more robust breadth of the braincase
than C. t. gorgasi. Chiroderma t. trinitatum also has an accessory cusp on the second
lower premolar, which is absent in C. ¢. gorgasi (Fig. 5). In the genetic analyses, C. t.
trinitatum is well supported as the sister species to C. doriae and does not share a most
recent common ancestor with C. ¢. gorgasi. We consider this as a previous example of
a cryptic species and therefore now recognize C. gorgasi as a distinct species from C.
trinitatum. Saez and Lozano (2005: 111) considered cryptic species to be “groups of
organisms that are morphologically indistinguishable from each other, yet found to
belong to different evolutionary lineages”. They also stated that “after detailed com-
parisons of morphological and non-morphological features, we can often establish key

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 49

Figure 5. Lateral view of the second lower premolar on the right mandible of A Chiroderma gorgasi

and B C. trinitatum. The arrow points to the accessory cusp that is absent in C. gorgasi and present in C.

trinitatum. But note the variation in cusp formation in C. trinitatum.

morphological characters for their identification. In those cases, we can then refer to
pseudo-cryptic or pseudo-sibling species”. Because Handley’s original description was
qualitative and univariate, we offer an amended description of this taxon.

Taxonomic account

Chiroderma gorgasi Handley, 1960

Chiroderma gorgasi Handley, 1960:464
Chiroderma trinitatum gorgasi Barriga-Bonilla, 1965:246

Material examined. Holotype. — USNM 309903 (Field number COH 5436), adult
male with skin, skull and partial skeleton. Collected on March 6, 1959, by C. O.
Handley, Jr, and B. R. Feinstein in Tacarcuna Village (8°05'N, 77°17'W), 3200 feet
[975 meters], Rio Pucro, Darién, Panama.

Geographic distribution. Chiroderma gorgasi is distributed west of the Andes in
northwestern Ecuador (Albuja 1989), western Colombia (Gardner 2008), Panama

50 Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

0 500 1000 km

Figure 6. Geographic distribution of Chiroderma gorgasi (&) and C. trinitatum (®) localities analyzed

in our study (See Appendix 3) (@) Represents marginal localities reported for C. trinitatum and (A) C.
gorgasi reported by previous papers (Handley 1967; Pine et al. 1970; Ojasti and Linares 1971; Gardner
1976; Albuja 1989; Timm and LaVal 1998; Lim and Engstrom 2001; Genoways et al. 1981; Webster and
Fugler 1984; Anderson 1997; Ochoa et al. 1988; Simmons and Voss 1998; Gardner 2008).

(Handley 1960), Costa Rica (LaVal and Rodriguez-Herrera 2002), and Honduras
(Turcios-Casco et al. 2020) (Fig. 6).

Description. Chiroderma gorgasi is a small species of Chiroderma (FA 37.0-40.5;
GLS 20.2—22.5) that is similar in size to C. trinitatum (sensu stricto) (Table 2). Overall,
the dorsal pelage is tricolor varying from light to dark brown (Fig. 7). The dorsal hairs
have a dark brown band at the base, a buff coloration in the middle, and brown tips. A
white medial stripe extends from the interscapular region to the base of the rump. Proxi-
mal two-thirds of forearm hairy. Basal third of uropatagium hairy. Conspicuous white
facial stripes extend from the noseleaf to the inner base of the ears, and from the pos-
terior part of the upper lip to the base of the ears. The uropatagium is medium brown.
The skull has an elongated braincase with an undeveloped sagittal and lambdoidal crest.
The nasal aperture is short, not extending beyond the second premolar. The occipital
is rounded in posterior view. The upper incisors are thin and elongated with parallel or
convergent tips, which may or may not touch apically. The second lower premolar lacks
a third cusp (Fig 5). The postorbital processes are undeveloped and rounded (Fig. 8).

Comparisons. Chiroderma gorgasi is morphologically very similar to C. trinitatum.
Both species have a small cranial and body size for the genus (Table 2, Fig. 4), an

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma

Figure 7. Dorsal view of the skin of the holotype of Chiroderma gorgasi (\USNM 309903).

a1

52, Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

Figure 8. Dorsal, ventral, and lateral views of the skull of the holotype of Chiroderma gorgasi (USNM 309903).

undeveloped sagittal and lambdoidal crest, a rounded occipital complex, a short nasal
aperture, and undeveloped supraorbital region. However, C. trinitatum has a third
posterior cusp on the second lower premolar, which is absent in C. gorgasi (Fig. 5). This
cusp in C. trinitatum may vary from very pointed and developed to rounded and less
marked, but is always present. In addition, C. gorgasi tends to have a broader braincase
(Table 4) and a flatter supraorbital region, which tends to be deeper in C. trinitatum.
Chiroderma gorgasi is easily distinguished from other species of the genus by its
smaller cranial and body size (Table 2). C. villosum shares with C. gorgasi an elongated
braincase, rounded occipital region in dorsal view, and absence of a third cusp on the
second lower premolar. However, C. gorgasi has an undeveloped postorbital processes,
a short nasal aperture, and conspicuous white stripes on the face and back, whereas
C. villosum has a very developed and pointed postorbital processes, a long nasal ap-

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 53

Table 4. ANOVA comparing Chiroderma trinitatum trinitatum to C. doriae, C. improvisum, C. villosum,
C. salvini, and C. trinitatum gorgasi. * indicates significantly distinct variables (p < 5%).

Variables C. doriae C. improvisum C. villosum C. salvini C. t. gorgasi
GSL < 2.2e-16* < 2.2e-16* < 2.2e-16* < 2.2e-16* 0.1541
Ch < 2.2e-16* < 2.2e-16* < 2.2e-16* < 2.2e-16* 0.4423
ZB < 2.2e-16* < 2.2e-16* < 2.2e-16* < 2.2e-16* 0.3094
POW <2.2e-16* 1.373e-06* < 2.2e-16* 1.061e-12* 0.6747
IOW < 2,2e-16* 1.347e-09* 2.366e-14* 1.355e-14* 0.6272
BWC < 2.2e-16* 2.602e-08* < 2.2e-16* < 2.2e-16* 00339"
M-C < 2.2e-16* < 2.2e-16* < 2.2e-16* < 2.2e-16* 0.5257
B-M < 2.2e-16* 1.527e-15* < DE2eA Gr < 2.2e-16* 0.1444
B-C < 2.2e-16* < 2.2e-16* < 2.2e-16* < 2.2e-16* 0.1251
FA < 2.2e-16* < 2.2¢e-16* < 2.2e-16* < 2.2e-16* 04753

erture, which extends beyond the first molar, a conspicuous posterior palatine spine,
and usually incipient white stripes on the face and back. Chiroderma salvini resembles
C. gorgasi in the undeveloped sagittal and lambdoidal crest and by the rounded pos-
torbital processes, but a set of other cranial characters distinguish both species, such
as a triangular occipital complex and a long nasal aperture. In the dentition, C. gorgasi
can be readily distinguished from C. salvini and C. villosum by having a tall first lower
premolar, with a crown height approximately 2/3 the height of the crown of the second
lower premolar, and placed approximately in the middle of the distance between the
canine and the second lower premolar. In C. salvini and C. villosum, this tooth is much
smaller, usually with a low crown, shorter than the mesiodistal length of the tooth, and
is nearer to the canine than to the second lower premolar.

Chiroderma doriae and C. improvisum are the largest species of the genus, and unlike
C. gorgasi have a triangular occipital complex in dorsal view, a pointed and developed
supraorbital region, a relatively more developed sagittal and lambdoidal crest, and a long
nasal aperture. In addition, C. doriae also tends to have a relatively broader braincase than
C. gorgasi and the presence of an undeveloped third cusp in the second lower premolar.
We were not able to examine specimens of the more recently described C. vizottoi, but
it is larger than C. gorgasi and most similar to C. doriae in qualitative craniodental traits.

Discussion

The only big-eyed bat species occurring in the Caribbean is Chiroderma improvisum,
which until recently was known from Guadeloupe (Baker and Genoways 1976) and
Montserrat (Jones and Baker 1979; Pierson et al. 1986) by six individuals (Larsen et
al. 2007). Subsequently, it was caught on Saint Kitts by Beck et al. (2016) and we are
the first to report its occurrence on Nevis. Although this species has been sporadically
documented since its discovery, the distribution has broadened in the northern Lesser
Antilles but this may be ephemeral depending on weather systems such as hurricanes
(Larsen et al. 2007).

54 Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

Chiroderma gorgasi was originally described by Handley (1960) using five speci-
mens from the type locality in Panama. The author distinguished the new species from
C. trinitatum by its smaller size, deeper brain case, shorter rostrum, shaper lacrimal
ridge, bulging forehead, larger upper incisors, and thicker white band in the dorsal
hairs. But at that time, C. trinitatum was only known by the holotype from Trinidad
(Goodwin 1958) so the extent of variation within each species was poorly understood.
Based on a specimen from Mitu in Amazonian Colombia, Barriga-Bonilla (1965) rec-
ognized the taxon as two subspecies and assigned his Colombian specimen to C. ¢. gor-
gasi. The subspecies were considered to be distributed from eastern Panama to western
Venezuela for C. ¢. gorgasi and Trinidad to the Amazon basin for C. ¢. trinitatum (Jones
and Carter 1976). However, with more geographic sampling the initial distinctions
between the two taxa were less obvious due to individual and geographic variation
(Simmons and Voss 1998), as also demonstrated by our morphometric analysis. But
the taxonomy and distributional limits were still contentious with Gardner (2008) rec-
ognizing the Andes as the delineation of the subspecies and reassigning the specimen
of Barriga-Bonilla (1965) from Mitu, Colombia, to C. ¢. trinitatum.

Our morphological review identified the presence of three cusps on the second
lower premolar in cis-Andean populations referable to C. trinitatum and two cusps
in trans-Andean populations referable to C. gorgasi that also match the taxonomic
boundaries of Gardner (2008). Morphometrically, C. trinitatum averages smaller than
C. gorgasi in all cranial measurements except for a proportionately broader braincase.
Furthermore, our genetic analyses recovered C. trinitatum as the well-supported sister
species to the larger and morphologically distinctive C. doriae, and not to the super-
ficially similar C. gorgasi. Based on this morphological and molecular evidence, we
recognize C. gorgasi as a distinct species and divergent lineage that does not share the
most recent common ancestor with C. trinitatum (sensu stricto).

The overall topology of the Cytb tree proposed by Baker et al. (1994) is identical
to our tree except for the recognition of C. gorgasi, which they did not have a sample
of, as the sister species to C. trinitatum and C. doriae. The evolution of Chiroderma
was suggested as occurring primarily by allopatric speciation (Baker et al. 1994).
Mote specifically, C. improvisum arose by peripatric speciation in the Lesser Antilles
after dispersing from its most recent common ancestor with C. villosum in South
America. The Andes is an obvious geographic barrier separating C. gorgasi from the
most recent common ancestor of C. trinitatum and C. doriae. A dated phylogeny is
needed to test whether this is an older sundering event associated with the uplift of
the northern Andes in the Late Miocene or a more recent dispersal event followed
by isolation and the cessation of gene flow. Rojas et al. (2016) date the divergence of
Chiroderma species to the Pliocene-Holocene, but C. gorgasi was not included in their
dataset. The allopatric distribution of C. trinitatum and C. doriae suggests that per-
haps the Cerrado Savanna in Brazil acted as a barrier after colonization of the Atlantic
Forest from the Amazon, but the records of C. doriae for the Cerrado and the discov-
ery of a species of Chiroderma in the dry deciduous forests of the Brazilian Caatinga,
C. vizottoi, indicates that species of the genus can adapt to more harsh habitats. ‘The
speciation event that gave rise to C. salvini and the most recent common ancestor

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 55

of the other species of Chiroderma is speculative without a thorough biogeographic
analysis with a dated phylogeny.

Although not an overly species-rich genus, biodiversity surveys and molecular anal-
yses are finding new distributional and taxonomic discoveries in Chiroderma. However,
there are still large geographic gaps in sampling throughout the Neotropics, such as the
Amazon basin in Brazil and northern South America in Colombia and Venezuela. In
addition, this has hindered detailed study of the biogeography of the genus and more
broadly the evolution of bats in the Neotropics.

Acknowledgments

GSTG is grateful to Valéria da Cunha Tavares for the valuable assistance during his
PhD research. GSTG was funded by the Coordenagao de Aperfeicoamento de Pessoal
de Nivel Superior, Brazil (CAPES; Code 001). Funding for fieldwork in Nevis was
obtained by BKL through a generous contribution from the Collections and Research
Fieldwork Fund of the Royal Ontario Museum. DNA sequencing was done by Kristen
Choffe in the Laboratory of Molecular Systematics at the ROM.

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58

Appendix |

Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

Tissue samples of Chiroderma used in the cytochrome c oxidase subunit 1 analysis.

Sample ID
ROM 111114
ROM 111141
ROM 111149
ROM 111163

ROM 105191
ROM 105230
ROM 105243
ROM 105253
ROM 105581
ROM 105685
ROM 105718
ROM 105766
ROM 106342
ROM F40504
ROM 104448
ROM 104540
ROM 104541
ROM 104549
ROM 105244
ROM 105254
ROM 105267
ROM 105361
ROM 105540
ROM 105587
ROM 105719
ROM 105720
ROM 105721
ROM 105928
ROM 105968
ROM F37400
ROM F37774
ROM 101245

ROM 99703

ROM 103486
ROM 103503
ROM 103504
ROM 103505
ROM 107205
ROM 107419
ROM 107476
ROM 108144
ROM 108244

Species
Chiroderma doriae
Chiroderma doriae
Chiroderma doriae
Chiroderma doriae

Chiroderma villosum
Chiroderma villosum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma salvini
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum

GenBank
JF448016
JF446371
JF446373
JF446372
KT236232
KT236233
JF448017
JF448810
JF448806
JF448805
JF448811
JF448807
JF448809
JF448808
JF448812
JF448813
JF448818
JF448829
JF448814
JF448828
JF448815
JF448816
JF448826
JF448825
JF448824
JF448830
JF448822
JF448821
JF448820
JF448817
JF448819
JF448827
JF448823
JF446499
KU295490
JF446777
JF454560
JF454561
MN714876
JF454562
EF080285
EF080286
MN714877
JF454552
JF454559

Country
Brazil
Brazil
Brazil
Brazil
Brazil
Brazil

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

Ecuador

El Salvador
French Guiana
Guatemala

Guyana

Guyana

Guyana

Guyana

Guyana

Guyana

Guyana

Guyana

Guyana

State/Department
Sao Paulo
Sao Paulo
Sao Paulo
Sao Paulo
Espirito Santo
Espirito Santo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo
Napo

Napo
Ahuachapan

EI Progreso

Upper Demerara-Berbice
Upper Demerara-Berbice
Upper Demerara-Berbice
Upper Demerara-Berbice

Potaro-Siparuni

Potaro-Siparuni

Potaro-Siparuni
Cuyuni-Mazaruni

Cuyuni-Mazaruni

Sample ID
ROM 108463
ROM 108554
ROM 108587
ROM 108588
ROM 108714
ROM 108763
ROM 108889
ROM 108950
ROM 108993
ROM 109026
ROM 109195
ROM 109271
ROM 109333
ROM 111627
ROM 111809
ROM 111844
ROM 111884
ROM 111946
ROM 115807
ROM 116630
ROM 118996
ROM 121975
ROM 125124
ROM 103214
ROM 103331
ROM 106644
ROM 107111
ROM 107112
ROM 107394
ROM 108203
ROM 108219
ROM 108764
ROM 108765
ROM 108843
ROM 108998
ROM 109138
ROM 109175
ROM 109221
ROM 109270
ROM 109307
ROM 109308
ROM 109337
ROM 111628
ROM 111629
ROM 111754
ROM 111768
ROM 111769
ROM 111770
ROM 111788
ROM 111836

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma

Species
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum

Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum

Chiroderma villosum

GenBank
JF454544
JF454545
JF454555
JF454554
EF080287
MN714878
EF080288
JF454557
JF454556
MN714879
JF454558
JF454553
JF454542
JF454543
JF454547
MN714880
JF454546
JF454548
JF454550
JF454549
JF454551
MN714881
MN714882
JF454584
JF454585
JF454566
EF080290
EF080291
EF080292
JF454565
JF454564
JF454571
JF454570
EF080289
JF454573
JF454572
JF454569
JF454568
JF454567
JF454583
JF454582
JF454581
EF080293
JF459119
JF454580
JF454579
JF454578
JF454577
JF454576
JF454575

Country
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana

State/Department

Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni

Potaro-Siparuni

Potario-Siparuni

Potaro-Siparuni

Potaro-Siparuni
Potaro-Siparuni

Potaro-Siparuni

Cuyuni-Mazaruni

Cuyuni-Mazaruni

Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni
Potaro-Siparuni

Potaro-Siparuni

59

Upper Takutu-Upper Essequibo

Upper Takutu-Upper Essequibo
Upper Takutu-Upper Essequibo
Upper Takutu-Upper Essequibo

60

Sample ID
ROM 111845
ROM 119167
ROM 119230
ROM 122481
ROM 98850
ROM 125179
ROM F38952
ROM 98702
ROM 96536

ROM FN30654

ROM 104342
ROM 104352
ROM F38210
ROM 122084
ROM 122137
ROM 122149
ROM 122165
ROM 122260
ROM 125567
ROM 126002
ROM 114170
ROM 114213
ROM 114233
ROM 114234
ROM 117003
ROM 117003
ROM 117027
ROM 117083
ROM 117376
ROM 117555
ROM 120098
ROM 120168
ROM 120225
ROM 120384
ROM 114212
ROM 114228
ROM 117119
ROM 117375
ROM 120226
ROM 120239
ROM 120240
ROM 120354
ROM 120364
ROM 121027
ROM 121117
ROM 126174
ROM 113919
ACUNHC 393

Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

Species
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Uroderma bilobatum
Chiroderma villosum
Chiroderma villosum

Chiroderma gorgasi
Chiroderma villosum
Chiroderma villosum

Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma improvisum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma trinitatum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Chiroderma villosum
Platyrrhinus incarum

Chiroderma villosum

GenBank
JF454574
MN714883
JF454586
MN714884
JF454563
MN714885
MN714886
JF435925
JF448018
JP447242
MN714901
JF447405
JF447406
N714887
N714888
N714889
N714890
N714891
N714892
N714893
JF447622
JF447625
JF447623
JF447624
JF447627
MN714894
JF447626
JF447628
EU096695
EU096696
MN714895
HQ545629
HQ545678
HQ919736
JF447630
JF447631
JF447629
EU096697
HQ545679
HQ545445
HQ545446
MN714896
HQ919717

Sots So a ee

MN714897
MN714898
MN714899
JF435616

MN714900

Country
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Guyana
Mexico
Mexico
Panama
Panama
Panama

Peru
Peru
Peru
Peru
Peru
Peru
Nevis

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Suriname

Venezuela

State/Department

Potaro-Siparuni

Upper Takutu-Upper Essequibo
Upper Takutu-Upper Essequibo

Potaro-Siparuni
Barima-Waini
East Berbice-Corentyne
Potaro-Siparuni
Barima-Waini
Campeche
Campeche
Darien
Darien
Darien
Loreto
Loreto
Loreto
Loreto
Loreto

‘Tumbes

Saint Thomas Lowland Parish

Brokopondo
Brokopondo
Brokopondo
Brokopondo
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Brokopondo
Brokopondo
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Sipaliwini
Para
Brokopondo

Amazonas

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 61

Appendix 2

Tissue samples of Chiroderma used in the cytochrome b analysis.

Sample ID Species GenBank Country State/Province
UNESP 16506 Chiroderma doriae 128937 Brazil Sao Paulo
TK 16379 Chiroderma doriae AY169958 Brazil
TRANS 713 Chiroderma improvisum L28938 Montserrat St. Anthony
TK 25052 Chiroderma villosum DQ312414 Trinidad St. George
FMNH 174652 Chiroderma villosum FJ154121 Peru Madre de Dios
TK 17627 Platyrrhinus helleri 128940 Suriname Marowijne
TK 25256 Uroderma bilobatum L28941 Trinidad St. George
TK 22581 Chiroderma salvini 128939 Panama Darien
PRS 2A) Chiroderma trinitatum DQ312413 Trinidad St. George
ASK 7799 Chiroderma villosum JF442196 Ecuador Orellana
ASK 7667 Chiroderma villosum JF442139 Ecuador Napo
MN 36375 Chiroderma villosum DQ903823 Brazil
SK-Bat-61 Chiroderma improvisum JQ915203 Saint Kitts
ROM 104342 Chiroderma gorgasi MN714902 Panama Darien
Appendix 3

Specimens of Chiroderma examined morphologically. Vouchers examined are arranged al-
phabetically by species and country. See “Material and Methods” for collection acronyms.

Chiroderma doriae — Brazil: Sao Paulo - ROM 111163, ROM 111141, ROM 111114,
ROM 111149.

Chiroderma improvisum — Montserrat: St. Anthony Parsish — TTU 31403; St. Kitts
and Nevis: Barnes Ghaut - ROM 126002.

Chiroderma salvini — El Salvador: Morazan - ROM 83365, ROM 85948, Santa Ana -
ROM 101526; Guatemala: El Progreso - ROM 99703; Panama: Darien - ROM
78472, ROM 91194.

Chiroderma trinitatum gorgasi — Colombia: Valle del Cauca - USNM 483763, USNM
483765, Antioquia - USNM 499478, USNM 499476; Panama: Bocas Del Toro
— USNM 319498, USNM 335295, Darien - FMNH 128132, ROM 104342,
USNM 309901, USNM 309903-holotype, San Blas - USNM 309905.

Chiroderma trinitatum trinitatum — Colombia: Vaupes - ROM 45276, ROM 45278,
ROM 45280, ROM 45281, ROM 45284, Putumayo — ROM 63236, ROM 63237,
ROM 63238; Ecuador: Napo - ROM 105191, ROM 105243, ROM 105253,
ROM 105685, ROM 105766, ROM 106342; Guyana: Cuyuni-Mazaruni - ROM
108144, Demerara-Berbice - ROM 57392, ROM 103486, ROM 103503, Upper,
Potaro-Siparuni - ROM 107205, ROM 107419, ROM 107476, ROM 108463,
ROM 108554, ROM 108587, ROM 108714, ROM 108763, ROM ROM 108889,

62

Burton K. Lim et al. / ZooKeys 918: 41—63 (2020)

ROM 108950, ROM 108993, ROM 109195, ROM 109333, ROM 111627,
ROM 111809, ROM 111884, ROM 111946, ROM 115807, ROM 116630; Suri-
name: Brokopondo - ROM 114170, ROM 114213, ROM 114233, ROM 114234,
Sipaliwini - ROM 117027, ROM 117376, ROM 120168, ROM 120225, ROM
120384; Trinidad: Saint Andrew County - AMNH 175325-holotype.

Chiroderma villosum — Bolivia: Carrasco - ROM 78471; Colombia: Choco - ROM

85849, Vaupes - ROM 44952, ROM 44953, ROM 44954, ROM 45243, ROM
45245, ROM 45246, ROM 45247, ROM 45249, ROM 45250, ROM 45251,
ROM 45252, ROM 45253, ROM 45254, ROM 45255, ROM 45257; Ecua-
dor: Napo - ROM 104448, ROM 104541, ROM 104549, ROM 105244, ROM
105254, ROM 105361, ROM 105720, ROM 105721); Guyana: Barima-Waini
- ROM 98850, Potaro-Siparuni - ROM 107111, ROM 107112, ROM 107394,
ROM 108219, ROM 108764, ROM 108843, ROM 108998, ROM 109138,
ROM 109175, ROM 109221, ROM 109307, ROM 109308, ROM 109337,
ROM 111628, ROM 111629, ROM 111754, ROM 111768, ROM 111769,
ROM 111770, ROM 111788, ROM 111836, ROM 111845, ROM 122481, Up-
per Demerara-Berbice - ROM 60402, ROM 60423, Upper Takutu-Upper Essequi-
bo - ROM 35614, ROM 103214, ROM 106644, ROM 119167, ROM 119230;
Panama: Darien - ROM 104352; Suriname: Brokopondo - ROM 114212, Sipaliwi-
ni - ROM 117119, ROM 117375, ROM 120226, ROM 120239, ROM 120364,
ROM 121027; Trinidad and Tobago: Nariva - ROM 124684, ROM 124691.

Supplementary material |

Maximum likelihood tree of cytochrome c oxidase subunit 1 gene for big-eyed

bats Chiroderma as presented in Fig. 3, but expanded to show individuals
Authors: Burton K. Lim, Livia O. Loureiro, Guilherme S. T. Garbino

Data type: phylogenetic dendrogram

Copyright notice: This dataset is made available under the Open Database License

(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/zookeys.918.48786.suppl1

Cryptic diversity and range extension in the big-eyed bat genus Chiroderma 63

Supplementary material 2

Maximum parsimony tree of cytochrome c oxidase subunit 1 gene for Chiroder-

ma. Bootstrap percentages show support at each node

Authors: Burton K. Lim, Livia O. Loureiro, Guilherme S. T. Garbino

Data type: phylogenetic dendrogram

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODDbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/zookeys.918.48786.suppl2

Supplementary material 3

Maximum likelihood tree of cytochrome b gene for Chiroderma. Bootstrap per-

centages show support at each node

Authors: Burton K. Lim, Livia O. Loureiro, Guilherme S. T. Garbino

Data type: phylogenetic dendrogram

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODDbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/zookeys.918.48786.supp13

Supplementary material 4

Maximum parsimony tree of cytochrome b gene for Chiroderma. Bootstrap per-

centages show support at each node

Authors: Burton K. Lim, Livia O. Loureiro, Guilherme S. T. Garbino

Data type: phylogenetic dendrogram

Copyright notice: This dataset is made available under the Open Database License
(http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License
(ODDbL) is a license agreement intended to allow users to freely share, modify, and
use this Dataset while maintaining this same freedom for others, provided that the
original source and author(s) are credited.

Link: https://doi.org/10.3897/zookeys.918.48786.suppl4