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Copyright © American Museum of Natural History 2012 ISSN 0003-0082
AMERICAN MUSEUM NOVITATES
Number 3739, 17 pp. March 9, 2012
A Second Species of the Family Allophrynidae
(Amphibia: Anura)
SANTIAGO CASTROVIEJOFISHER, PEDRO E. PÉREZPEÑA, JOSE M. PADIAL,
AND JUAN M. GUAYASAMIN
ABSTRACT
We describe Allophryne resplendens, a new species from two localities in the Amazon rain-
forest of Loreto, Peru, of the family Allophrynidae, which was monotypic until this discovery.
e new species can be readily dierentiated from Allophryne ruthveni on the basis of dorsal
and ventral coloration both in life and in preservative, transverse processes of presacral II ori-
ented anterolaterally (oriented laterally in A. ruthveni), 19 nucleotide autapomorphies for 761
base pairs (bp) of the mitochondrial gene 16S, and 16 for 923 bp of 12S. Maximum parsimony
analysis of the mitochondrial gene 12S and a fragment of up to 1060 bp of 16S supports the
new species as sister to A. ruthveni.
INTRODUCTION
Discoveries and new descriptions of amphibian species have increased in recent years, with
the number of nominal species growing of about 48.2% since 1985 (Frost et al., 2006) and
19.4% during the last decade (Köhler et al., 2005). e bulk of these new discoveries arise from
applying recently developed advances in elds such as molecular biology, bioacoustics, and
geography to clades from tropical regions (e.g., Meegaskumbura et al., 2002; Stuart et al., 2006;
Fouquet et al., 2007; Vieites et al., 2009). Discovery of new species is, however, not randomly
1 Division of Vertebrate Zoology (Department of Herpetology), American Museum of Natural History.
2 Wildlife Conservation Society, Programa Loreto, Perú.
3 Centro de Investigación de la Biodiversidad y Cambio Climático, Universidad Tecnológica Indoamérica, Av.
Machala y Sabanilla, Quito, Ecuador.
2 AMERICAN MUSEUM NOVITATES NO. 3739
distributed across clades of amphibians. Some groups account for most of the newly described
diversity (e.g., Ranidae alone includes 25% of the new species of amphibians described since
2004: AmphibiaWeb, 2011) whereas for other groups it may be decades before a second species
within a genus is discovered. For example, Rhinatrema bivittatum (Guérin-Méneville, 1838)
remained as the only known species of Rhinatrema Duméril and Bribon, 1841, for 169 years
until Gower et al. (2010) described a new species, although that discovery was quickly followed
by another (Wilkinson and Gower, 2010).
Allophryne ruthveni Gaige, 1926, has stood as the only member of this monotypic genus
and the family Allophrynidae for 85 years. It is known to occur in the lowland rainforest of
the Amazon and Orinoco basins, from Venezuela and the Guianas to southwestern Brazil and
probably Bolivia (Frost, 2011). Despite its ample distribution, records are rare and sparse.
Phylogenetically, the relationship Centrolenidae + Allophryne ruthveni is the most accepted
hypothesis, supported by molecular phylogenetics (Austin et al., 2002; Faivovich et al., 2005;
Wiens et al., 2005; Frost et al., 2006; Grant et al., 2006; Guayasamin et al., 2008) and morpho-
logical studies (Duellman, 2001; Burton, 2004; Wiens et al., 2005; but see Fabrezi, 2006). In this
work, we describe the second species in the family Allophrynidae, found in the Amazon rain-
forest of Loreto, Peru.
MATERIALS AND METHODS
N
Frost et al. (2006) placed Allophryne ruthveni in the monotypic subfamily Allophryninae,
within Centrolenidae, whereas Guayasamin et al. (2009) argued to maintain the species in the
monotypic family Allophrynidae. We followed the later arrangement (see Guayasamin et al.,
2009, for argumentation).
T S M
Specimens were collected in the eld according to approved animal use and care protocols
(Heyer et al., 1994), and euthanized in Xylocaïne solution prior to xation and preservation in
ethanol 70%. Specimens studied are listed in appendix 1. Color characteristics are described
according to digital photographs taken from life specimens in the eld. External morphological
characters were examined under a dissecting microscope. Measurements were taken with digi-
tal calipers on preserved specimens to the nearest 0.01 mm and rounded to 0.1 mm. Variables
measured, as described in Duellman (2001), are as follows: snout-vent length (SVL); head
length; head width; interorbital distance; eye diameter; upper eyelid; tympanum diameter; eye-
nostril distance; distance from anterior margin of eye to tip of snout; internarial distance; eye-
tympanum distance; length of forearm; hand length; nger I length; nger II length; width of
disc of nger III; width of nger III; thigh length; shank length; foot length. Ontogenetic status
was determined by examination of the development of gonads and presence of secondary
sexual characteristics (e.g., vocal slits and sacs). Terminology for webbing is that described by
Savage and Heyer (1967), as modied by Guayasamin et al. (2006). e osteological description
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 3
FIG. 1. Photographs of preserved specimens of Allophryne. A–C, Allophryne resplendens, new species, adult
female holotype (SVL = 28.4 mm); D–E, Allophryne ruthveni, adult female, Surinam, AMNH 87687 (SVL =
27.0 mm). Photographs by JMP (A–C) and SCF (D–E).
4 AMERICAN MUSEUM NOVITATES NO. 3739
A
C
B
D
E F
FIG. 2. Photographs of live specimens of Allophryne. A, Allophryne resplendens, n. sp., adult female holotype
(SVL = 28.4 mm); B, A. resplendens, n. sp., specimen not collected, sex and size unknown, topotype; C–D, A.
resplendens, new species, adult male, Quebrada Ungurahue (SVL = 25.0 mm); E–F, A. ruthveni, adult male,
Venezuela, MHNLS 20231 (SVL = 23.5 mm). Photographs by Mark Bowler (A), WCS–Perú (B), William
Lamar (C–D), and SCF (E–F).
was based on X-rays of the holotype; osteological terminology is that of Duellman and Trueb
(1986), Fabrezi (1992, 1993), and Trueb (1973, 1993). Museum acronyms are: American
Museum of Natural History (AMNH), Museo Historia Natural La Salle (MHNLS), and Museo
de Zoología de la Universidad Nacional de la Amazonía Peruana (MZUNAP).
M A
We sequenced the complete mitochondrial gene 12S (962 bp) and a fragment of the gene
16S (1060 bp) from the specimen of Allophryne collected in Loreto, Peru, and three specimens
of A. ruthveni from Delta Amacuro, Venezuela. Genomic DNA was isolated using a standard
phenol-chloroform extraction protocol (Sambrook et al., 1989). e selected markers were
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 5
amplied and sequenced using the same primers and PCR conditions as in Guayasamin et al.
(2008) with the addition of primers 16L19 and 16H24 (Heinicke et al., 2007). Sequences from
heavy and light strands were compared to generate a consensus sequence for each specimen
using Sequencher 4.6 (Gene Codes Corp., 2006). We compared these sequences with those
from GenBank for Allophryne ruthveni, as well as representative species of all other Neotropical
families within Hyloidea, according to the taxonomy used in Frost (2011). We used sequences
of Mantella madagascariensis to root phylogenetic trees. GenBank accession codes of all the
sequences studied are provided in appendix 2.
Sequences of each marker were independently aligned in MAFFT (Katoh et al., 2005)
under the E-INS-i option. e resulting multiple sequence alignments (Dryad Repository:
doi:10.5061/dryad.2kp7q7vb) were concatenated and analyzed under the maximum parsimony
criterion (MP) using TNT 1.1, Willi Hennig Society Edition (Golobo et al., 2008) with gaps
as a h character and using the “New Technology search” option at level 100, including secto-
rial searches, ratchet (see Nixon, 1999), dri, and tree fusing (see Golobo, 1999). Bootstrap
support (BS, Felsenstein, 1985) was evaluated through 1000 pseudoreplicates, with 10 RAS
using TBR and 10 trees saved per replication. Uncorrected genetic distances were calculated
from 761 bp of the 16S and 923 of the 12S (maximum length overlapping for all the Allophryne
sequences, Dryad Repository: doi:10.5061/dryad.2kp7q7vb) in PAUP* 4.0b10 (Swoord, 1998).
Allophryne resplendens, new species
Figures 1A–C, 2A–D
Allophryne, sp. nov., Rodríguez and Knell, 2003: 244.
H: MZUNAP-01-605 (eld number PEPP 500), adult female from Lago Preto
(4°27′35.0″S, 71°45′ 3.5″W; 120 m), Río Yavarí, Provincia Ramon Castilla, Departamento
Loreto, Peru, collected by PEPP in November 2009. When collected, the distal section of the
right hind limb of the specimen was missing; we cut a piece of tissue from the stump for
molecular studies purposes.
R: We are aware of three other records of the new species. One is a noncollected
but photographed adult (g. 2B) found in Lago Preto, Yavarí, Loreto, Peru, in July 2008 (pho-
tographs by anonymous volunteers of Earthwatch and deposited at Wildlife Conservation Soci-
ety, WCS–Perú). e second is reported by Rodríguez and Knell (2003) also from Lago Preto
(4°28′S, 71°46′W; 90 m), Yavarí, Loreto, Peru. Rodríguez and Knell (2003) cite this specimen
as deposited at the MUSM. However, in a visit to the MUSM in March 2008, we were unable
to locate the specimen or to get a voucher code for it. e third specimen, an adult male (g.
2C, D), was collected on the south bank of Quebrada Hungurahui (4°14′42.97″S, 74°21′36.02″W;
114 m), an auent of Río Tigre, approximately 2 km north of Comunidad Monteverde, Loreto,
Peru, by William Lamar on 1 August 2006. is specimen was collected and preserved in etha-
nol; unfortunately, the jar did not seal properly and the specimen totally decomposed and
could not be used as a type.
G P: e new species is assigned to the genus Allophryne because of the
following combination of morphological characters: absence of metacarpal III process (its pres-
6 AMERICAN MUSEUM NOVITATES NO. 3739
ence is a synapomorphy of centrolenid frogs), T-shaped terminal phalanges, tibiale and bulare
not fused, absence of neopalatines and quadratojugals, and protruding snout in lateral view;
no phenotypic autoapomorphy is known for the genus. e molecular phylogeny also supports
the placement of the new species in the genus Allophryne. e 10 most parsimonious trees (tree
cost = 11,381 steps) resulting from 100 replicates (each including RAS + TBR + dri and tree
fusing with the best tree hit 100 times) placed Allophryne resplendens, n. sp., as the sister spe-
cies of Allophryne ruthveni (BS = 100; g. 3).
D: Allophryne resplendens, n. sp., is distinguished from A. ruthveni, the only other
known species in the genus and family (character states of the latter are in parentheses), by (1)
color pattern (cf. photos in gs. 1 and 2): dorsolateral surfaces in life black (cream to light
brown) with large and bright glossy spots due to accumulation of iridophores (absent), most
of which containing irregular yellow blotches (absent), dorsolateral surfaces in preservative
dark brown (cream to light brown with dark brown stripes) with large and bright glossy spots
due to accumulation of iridophores (absent), ventral surfaces in life black and opaque (not
pigmented, translucent) with large and bright glossy spots due to accumulation of iridophores
except in palms and plantar surfaces (absent or just a few o-white spots on the throat/chest
and distal parts of legs), same character states for both species for ventral surfaces in preserva-
tive than in life, but coloration dark brown instead of black for A. resplendens; (2) osteology:
transverse processes of presacral II oriented anterolaterally (oriented laterally; observed in two
cleared and stained specimens; see appendix 1, and g. 2C in Fabrezi and Langone, 2000); and
(3) DNA sequences: uncorrected genetic distance > 2.5 % for 761 bp of mitochondrial gene
16S, and ≥ 1.7 % for 923 bp of 12S (table 1), 19 nucleotide autapomorphies for 761 bp of 16S,
and 16 for 923 bp of 12S (table 2).
D H: Adult female, SVL 28.4 mm. Head wider than long (HL
= 85.7% of HW); snout protruding in lateral prole and broadly rounded in dorsal view (g.
1A, C); canthus rostralis indistinct, straight; loreal region at; lips not ared; nostril small, not
protuberant, closer to tip of snout than to eye. Eye of moderate size (ED = 10.6% of SVL).
Tympanic membrane and tympanic annulus dierentiated; tympanum relatively small (TD =
TABLE 1. Uncorrected genetic distances between specimens of Allophryne inferred from 923 bp of the mitochondrial genes
12S (above the diagonal) and 761 bp of the 16S (below the diagonal). Dryad Repository: doi:10.5061/dryad.2kp7q7vb.
12345678
1 A. resplendens * 2.0% 2.1% 1.7% 2.1% – – 2.1%
2 A. ruthveni 3301 2.6% * 0.1% 0.3% 0.1% – – 0.1%
3 A. ruthveni 3302 2.6% 0.0% * 0.2% 0.0% – – 0.0%
4 A. ruthveni 3303 3.0% 0.7% 0.7% * 0.2% – – 0.2%
5 A. ruthveni AY843564 2.9% 0.5% 0.5% 0.9% * – – 0.0%
6 A. ruthveni EU662973 2.9% 0.4% 0.4% 1.1% 0.4% * – –
7 A. ruthveni AF364512 3.4% 0.9% 0.9% 1.4% 1.5% 1.4% * –
8 A. ruthveni AY819328 – – – – – – – *
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 7
Marker Position A. ruthveni A. resplendens
16S 91 G A
92 T C
101 G A
153 T C
174 C T
240 T C
280 A –
282 A T
333 –/T C
342 T G
464 A G
542 C T
544 T C
558 A G
584 T C
604 A C
624 T C
628 T C
638 A T
12S 59 T C
91 A G
299 G A
341 T C
370 T C
375 T C
390 T C
459 T/- C
583 A G
625 A T
724 G A
748 C T
851 T C
852 T C
861 T C
922 A G
TABLE 2. Autapomorphies for Allophryne resplendens new species with respect to A. ruthveni inferred from a fragment of
761 bp and 923 bp of the mitochondrial genes 16S and 12S respectively. Dryad Repository: doi:10.5061/dryad.2kp7q7vb.
8 AMERICAN MUSEUM NOVITATES NO. 3739
Allophryne ruthveni AF364512
Allophryne ruthveni MHNLS 20233
Allophryne ruthveni MHNLS 20232
Allophryne ruthveni MHNLS 20231
Allophryne ruthveni AY819328
Allophryne ruthveni EU662973
Allophryne ruthveni AY843564
Allophryne resplendens
Mantella madagascariensis
Hypsiboas geographicus
Bro meliohyla bromeliacia
Anotheca spinosa
Duellmanohyla soralia
Hyloscirtus charazani
Osteocephalus taurinus
Ecnomiohyla miotympanum
Isthmohyla rivularis
Bokermannohyla martinsi
Argenteohyla siem ersi
Charadrahyla taeniopus
Aparasphenodon brunoi
Pristimantis galdi
Yunganastes pluvicanorus
Pri stimantis diadematus
Oreobates cruralis
Lynchius avomaculatus
Stefania schuberti
Gastrotheca ssipes
Dendrobates auratus
Ameerega trivittata
Silverstoneia otator
Epipedobates t ricol or
Rheobates palmatus
Allobates ta lamancae
Rhaebo nasicus
Dendrophryniscus minutus
Atelopus peruensis
Rhinella arenarum
Atelognathus patagonicus
Telmatobius sp.
Batrachyla leptopus
Ceratophrys cranwelli
Lepidobatrachus sp.
Eupsophus calcaratus
Alsod es ga rgola
Leptodactylus ocellatus
Lithodytes lineatus
Chimerella mariaelenae
Celsiella revocata
Vitreorana gorzul ae
Hyalinobatrachium taylori
Cochranella nola
Centrolene savagei
Teratohyla pulverata
Rulyrana spiculata
Nymphargus bejaranoi
Sachatamia albomaculata
Espadarana andina
Ikakogi ta yrona
Hyla arborea
Paratelmatobius po ecilogaster
Scythrophrys sawayae
Terrarana
Hemiphractidae
Bufonidae
Dendrobatoidea
Hylidae
Ceratophryidae
Cycloramphidae
Allophrynidae
Centrolenidae
Leptodactylidae
100
99
100
81
97
100
100
99
99 98
74
99
99
98
74
98
99
97
98
76
98 98
95 77
100 87
FIG. 3. Strict consensus of the 10 most parsimonious trees inferred from 12S and 16S sequences (tree cost =
11,381 steps). Bootstrap support values are indicated above branches when ≥ 70.
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 9
FIG. 4. X-ray of the holotype of Allophryne resplendens, n. sp. Arrow points to the anterolaterally oriented
transverse processes of presacral II.
3.9% of SVL). Dentigerous process of vomer indistinguishable. Ulnar tubercles absent; relative
lengths of ngers: III > IV > II > I webbing on hands basal. Discs expanded, truncate; subar-
ticular tubercles round, conspicuous; few supernumerary tubercles; palmar tubercle oval, sim-
ple. Posterior limbs relatively long (FL = 40.8% of SVL; TL = 43.3% of SVL); tarsal tubercles
absent; feet about 3/4 webbed, formula: I 0+–2 II 1––2 III 1–2 IV 3––2– V; discs on toes ellipti-
cal; inner metatarsal tubercle ovoid; outer metatarsal tubercle small, barely evident; subarticu-
lar tubercles round, conspicuous; supernumerary tubercles absent. Skin on dorsal surfaces of
head and body shagreen with hemispherical pustules having a central spicules; venter areolate.
Cloacal opening directed posteriorly at midlevel of thighs, covered by a slightly developed
cloacal sheath; cloacal tubercles absent.
C L: is description is based on photographs of the holotype and the other
three mentioned specimens (g. 2; Rodríguez and Knell, 2003: g. 5C). Dorsolateral surfaces
with large and bright glossy spots due to accumulation of iridophores, most of which contain
yellow irregular blotches, set in a washed black reticulum; reticulum carries smaller and less
bright oval to pentagon-shaped accumulations of iridophores from which brown spicules arise.
10 AMERICAN MUSEUM NOVITATES NO. 3739
Ventral surfaces black and
opaque with large and bright
glossy spots due to accumula-
tion of iridophores except in
palms and plantar surfaces.
Iris dark bronze with dark
reticulations, pupillary ring
absent, pupil black. There
seems to be sexual dichroma-
tism; the female has fewer
spicules and more ventral
glossy spots than the male,
which is also the case in
Allophryne ruthveni.
C P:
This description is based
exclusively on the holotype
(g. 1A–C). Dorsolateral sur-
faces with white large and
bright irregular spots due to
accumulation of iridophores,
yellow irregular blotches
absent, set in a dark brown
reticulum that carries smaller
and less bright oval to penta-
gon-shaped accumulations of
iridophores from which brown spicules arise. Ventral surfaces dark brown with large and bright
glossy spots due to accumulation of iridophores except in palms and plantar surfaces. Iris
brown and pupil white.
O (g. 4): e skull is widest posterior to the orbit and at the level of the articula-
tion of the maxilla with the squamosal. e neopalatines and quadratojugals are absent. e
anterior ramus of the pterygoid articulates with the posterior end of the maxilla. e vertebral
column has eight presacral vertebrae that have transverse processes except presacral I. e
vertebral prole in decreasing order of overall width of bony parts is: sacrum > III > IV > V ≈
VI ≈ VII > II > I. e orientations of the transverse processes of presacral VI are lateral,
whereas those of presacrals II, VII, and VIII are anterolateral, and those of presacral III–V are
posterolateral. e bony sacral diapophysis is broadly expanded distally, and has a slender base
that is approximately a third the width of the distal margin. e leading edge of each diapophy-
sis is slightly concave with an anterolateral orientation, whereas the posterior margin is nearly
straight and oriented posterolaterally. e urostyle is slender and shorter than the length of the
presacral portion of the vertebral column; the urostyle bears a dorsal crest on its anterior half.
FIG. 5. Known distribution of Allophryne resplendens, n. sp. Star rep-
resents type locality.
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 11
e tibiale and bulare are not fused medially. e phalangeal formulae for the hand and foot
are standard, 2-2-3-3 and 2-2-3-4-3, respectively. In increasing order of length, the order of the
digits on the hand is I < II < IV < III and that of the foot is: I < II < III < V < IV. e terminal
phalanges of hands and feet are T-shaped.
M: e morphometric data for the female holotype are (in mm): SVL = 28.4;
head length = 7.8; head width = 9.1; thigh length = 11.6; shank length = 12.3; foot length =
11.5; interorbital distance = 3.6; upper eyelid width = 2.0; internarial distance = 1.7; eye-to-
nostril distance = 2.5; snout-eye distance = 3.3; eye diameter = 3.0; tympanum diameter = 1.1;
eye-tympanum distance = 1.1; forearm length = 5.7; hand length = 8.7; nger I length = 6.1;
nger II length = 6.5; and width of disc of nger III = 1.6.
D: Allophryne resplendens new species is currently known only from two
localities in Peru (g. 5). e type locality is in the Yavarí River drainage. is drainage includes
other rivers, such as the Yavarí Mirín and Quebrada Esperanza. e region is dominated by
the geological formation that covers much of northeastern Peru: the Pebas formation, a thick
slab of clays and sands deposited in ancient lakes and rivers (Räsaänen et al., 1998; Sánchez et
al., 1999; de la Cruz et al., 1999). e southern sector, close to Angamos, is associated with an
uplied geological structure known as the Iquitos Arch, which stretches hundreds of kilometers
across Loreto and into Colombia. e vegetation of the area includes upland (terra rma) forest
concentrated along the Iquitos Arch, ooded (varzea) forest and palm swamps. e second
known locality is the western side of the Amazon River, Quebrada Hungurahui (an auent of
Río Tigre), approximately 2 km north of Comunidad Monteverde, Loreto, Peru. Allophryne
resplendens, n. sp., is likely present in other varzea forests at least around and between the two
known localities. us, it is possible that this species also occurs in Brazil.
N H: is is an arboreal species that has been found perched on leaves and
branches below 2 m. e holotype was found on a Lepidocaryum tenue (Arecaceae). ese
medium-size palm trees form particular forests known locally as irapayales. e specimen from
Quebrada Hungurahui was also found in an irapayal forest. Allophryne resplendens, n. sp., is
very dicult to nd, which could reect very low densities, at least in the lower strata of the
forest outside the breeding season (the sister species is a explosive breeder). Aer 49.45 hours
of search only one specimen was collected (table 3), and Rodríguez and Knell (2003) found
only a single specimen in 200 hours of search. William Lamar (personal commun.) has visited
the locality in Quebrada Hungurahui during the last 10 years in dierent months and has not
TABLE 3. Search eort distributed by forest type and season in the Yavarí river.
Flooding season (November) Dry season (June–July)
Palm swamps Varzea Terra rma Palm swamps Varzea Terra rma
Search hours 2.5 7.42 12.28 8.75 10.50 8.00
Number of people 2 2 1 2 2 2
Specimens found 1 0 0 0 0 0
12 AMERICAN MUSEUM NOVITATES NO. 3739
been able to locate by sight or sound any other specimens. At least 45 species of amphibians
have been reported from the type locality and surroundings, of which 23 have been found in
swamp palm forest.
E: e specic name resplendens is derived from the Latin verb resplendo mean-
ing “to glitter,” which we used in allusion to the bright and ornate coloration of the frog.
DISCUSSION
T
Monotypic supraspecic taxa can result problematic. For example, dierent Linnean ranks
convey the same information (Allophrynidae = Allophryne = Allophryne ruthveni) and phyloge-
netic denitions are repetitive (see Guayasamin et al.’s [2009] attempt to dene Allophrynidae and
Allophryne). Our discovery of a new species of Allophryne partly solves this problem. Allophryne
is no longer equivalent to the species Allophryne ruthveni and we provide a new phylogenetic de-
nition of the family and genus: a clade stemming from the most recent common ancestor of
Allophryne ruthveni Gaige, 1926, and Allophryne resplendens Castroviejo-Fisher et al., 2011.
P R
Our MP tree shows the sequences of A. resplendens, n. sp., as sister to those of A. ruthveni.
However, the MP analysis does not support the relationship (Leptodactylidae, (Allophrynidae,
Centrolenidae)), which has been suggested in other phylogenetic studies (e.g., Frost et al., 2006;
Guayasamin et al., 2008; Heinicke et al., 2009; Pyron and Wiens, 2011) although with low support.
We explain this lack of resolution by the fact that our dataset was primarily designed to evaluate
whether A. resplendens, n. sp., was more closely related to Allophrynidae than to representatives
of all other families of Neotropical Hyloidea. Our results are therefore not comparable to studies
using much larger taxon and character sampling (e.g., Frost et al., 2006; Pyron and Wiens, 2011).
e unique combination of phenotypic characters exhibited by Allophryne ruthveni has long
puzzled herpetologists, resulting in dierent and conicting phylogenetic hypotheses placing
Allophrynidae as the sister taxon of the families Bufonidae (Gaige, 1926; Laurent, 1980, 1986),
Hylidae (Lynch and Freeman, 1966; Hoogmoed, 1969; Duellman, 1975; Duellman and Trueb,
1986), or Centrolenidae (Noble, 1931; Lutz, 1968; Duellman, 2001; Burton, 2004; Wiens et al.,
2005). Although no unique autapomorphy has been described for A. ruthveni, the following
combination of characters has been proposed as diagnostic: intercalary elements absent, biaxial
articulation between last two phalanges, T-shaped terminal phalanges, tibiale and bulare not
fused, absence of neopalatines and quadratojugals, prepollex and prehallux formed by one proxi-
mal element each (Fabrezi and Langone, 2000, 2001; Fabrezi, 2006). Unfortunately, we have been
able to secure only one specimen of A. resplendens, n. sp., so we could investigate only three of
the characters mentioned above (i.e., T-shaped terminal phalanges, tibiale and bulare not fused,
absence of neopalatines and quadratojugals). e next logical step in the study of Allophryne
would be to gather more information on these and other phenotypic characters and study them
in combination with other relevant information and taxa in a cladistic framework.
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 13
ACKNOWLEDGMENTS
We thank Mark Bowler, Miguel Antúnez, Maria Soledad Riveros, Bray Torres, Franco San-
tana, Claudia Rios, Ciro Pinedo, Richard Bodmer, Pablo Puertas, and the volunteers of Earth-
watch Institute for supporting the study in Lago Preto; Juan Carlos Chaparro for his support
and help in the description of this species; and William Lamar for photographs and data on
the new locality. e Wildlife Conservation Society–Perú (WCS–Perú) and AmazonEco pro-
vided nancial support for eldwork. Collection permits in Peru (authorization N° 320-2009)
were issued by AG-DGFFS-DGEFFS. is study is included in the “Contrato Marco de Acceso
a Recursos Genéticos N° 0001, 11 Enero 2007” subscribed between Fundacion La Salle de
Ciencias Naturales and the Ministerio del Ambiente, Venezuela. e work of SCF was nanced
by a Fulbright/Ministry of Education postdoctoral research contract. e Universidad Tec-
nológica Indoamérica supports J.M.G.’s research. J.M.P.’s research is funded by a Gerstner Post-
doctoral Fellowship at the American Museum of Natural History.
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16 AMERICAN MUSEUM NOVITATES NO. 3739
APPENDIX 1
A S S
Allophryne ruthveni: VENEZUELA: D A: Reserva Forestal Rio Grande,
Coderforce (08°16′00.8″N, 61°45′16.3″W; 192 m), MHNLS 20231–6 (adult males). A:
Base camp of Tapirapeco expedition, upper rio Mavaca (150 m), AMNH 131351 (adult female,
cleared and stained). SURINAM: S: Raleigh Cataracts, Coppename river (50 m),
AMNH 87687 (adult female), 876888 (adult male). GUYANA: I: Pakatau creek,
AMNH 163990 (adult female). Marudi creek, AMNH 44749 (male). U L:
AMNH 70108 (adult female), AMNH 70110 (unknown sex, cleared and stained)
.
APPENDIX 2
S, T C S G C, U S.
Sequences newly generated for this study are in boldface.
Family Genus Species 12S 16S
Allophrynidae Allophr yne resplendens JQ436697 JQ436698
Allophryne ruthveni AY819328 —
Allophryne ruthveni JQ436702 JQ436700
Allophryne ruthveni JQ436703 JQ436699
Allophryne ruthveni JQ436704 JQ436701
Allophryne ruthveni AY843564 AY843564
Allophryne ruthveni — AF364512
Allophryne ruthveni — EU662973
Aromobatidae Allobates talamancae 170026605 170026605
Rheobates palmatus 170026602 170026602
Bufonidae Atelopus peruensis 77176491 77176491
Dendrophryniscus minutus 61697193 61697193
Rhaebo nasicus 77176549 77176549
Rhinella arenarum 61697184 61697184
Centrolenidae Celsiella revocata EU663379 EU663019
Centrolene savagei EU663380 187729151
Chimerella mariaelenae EU663350 EU662991
Cochranella nola EU663375 EU663015
Espadarana andina EU663335 EU662976
Hyalinobatrachium taylori EU663420 EU663056
Ikakogi tayrona EU663358 EU662999
Nymphargus bejaranoi EU663422 EU663059
Rulyrana spiculata EU663382 EU663022
Sachatamia albomaculata EU663362 EU663003
Teratohyla pulverata EU663416 EU663053
2012 CASTROVIEJOFISHER ET AL.: NEW SPECIES OF ALLOPHRYNIDAE 17
Family Genus Species 12S 16S
Vitreorana gorzulae EU663348 EU662989
Ceratophryidae Atelognathus patagonicus 61697182 61697182
Batrachyla leptopus 61697183 61697183
Ceratophrys cranwelli 61697186 61697186
Lepidobatrachus sp 37699560 37699560
Telmatobius sp 61697380 61697380
Cycloramphidae Alsodes gargola 61697176 61697176
Eupsophus calcaratus 61697198 61697198
Dendrobatidae Ameerega trivittata 170026722 170026722
Dendrobates auratus 61697192 61697192
Epipedobates tricolor 37624303 37624303
Silverstoneia otator 170026684 170026684
Hemiphractidae Gastrotheca ssipes 61697203 61697203
Stefania schuberti 61697379 61697379
Hylidae Anotheca spinosa 61697177 61697177
Aparasphenodon brunoi 61697178 61697178
Argentohyla siemersi 61697181 61697181
Bokermannohyla martinsi 61697252 61697252
Bromeliohyla bromeliacia 61697223 61697223
Charadrahyla taeniopus 61697290 61697290
Duellmanohyla soralia 61697195 61697195
Ecnomiohyla miotympanum 61697256 61697256
Hyla arborea 61697212 61697212
Hyloscirtus charazani 61697229 61697229
Hypsiboas geographicus 61697239 61697239
Isthmohyla rivularis 61697270 61697270
Osteocephalus taurinus 61697320 61697320
Leptodactylidae Leptodactylus ocellatus 61697299 61697299
Lithodytes lineatus 37699553 37699553
Paratelmatobius poecilogaster 159139110 159139110
Scythrophrys sawayae 159139126 159139126
Mantellidae Mantella madagascariensis 89255435 89255435
Strabomantidae Lync hius avomaculatus 166155972 166155972
Oreobates cruralis 166155971 166155971
Pristimantis diadematus 166155973 166155973
Pristimantis galdi 166155975 166155975
Yunganastes pluvicanorus 61697197 61697197
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