Corresponding author: Santiago R. Ron(
Academic editor: Franco Andreone
The holotype of
Recently collected specimens of
Records of
Total DNA was extracted from muscle or liver tissue preserved in 95% ethanol and tissue storage buffer using a guanidine tiocyanate protocol. Polymerase chain reaction (PCR) was used to amplify the mitochondrial genes 12S rRNA and ND1. We amplified one DNA fragment for 12S and one or two overlapping fragments for ND1 using primers listed in
A list of the samples included in the phylogenetic analyses is shown in
Specimens used in the phylogenetic analysis. Numbers correspond to those in the Figures 1 and 2.
Museum No. | Species | No. | Genbank Accession No. | Reference | |
---|---|---|---|---|---|
12S | ND1 | ||||
SBH 266458 |
|
DQ380357 | EU034080 | ||
USNM 327241 |
|
DQ380382 | EU034083 | ||
SBH 266457 |
|
DQ380383 | EU034086 | ||
SBH 191985 |
|
AY819436 | EU034087 | ||
QCAZ 15981 |
|
1 | HQ600629 | HQ600596 | This study |
LAC 2216 |
|
30 | DQ380378 | EU034082 | |
QCAZ 14947 |
|
17 | HQ600628 | HQ600595 | This study |
QCAZ 24446 |
|
13 | HQ600633 | HQ600600 | This study |
QCAZ 24447 |
|
14 | HQ600634 | HQ600601 | This study |
QCAZ 28231 |
|
25 | HQ600654 | HQ600621 | This study |
QCAZ 28277 |
|
15 | HQ600639 | HQ600606 | This study |
QCAZ 28395 |
|
16 | HQ600640 | HQ600607 | This study |
QCAZ 32506 |
|
22 | HQ600651 | HQ600618 | This study |
QCAZ 32508 |
|
24 | HQ600652 | HQ600619 | This study |
QCAZ 37175 |
|
23 | HQ600653 | HQ600620 | This study |
QCAZ 25469 |
|
21 | HQ600650 | HQ600617 | This study |
CORBIDI 623 |
|
28 | HQ600649 | HQ600616 | This study |
QCAZ 38420 |
|
31 | HQ600646 | HQ600613 | This study |
QCAZ 39364 |
|
29 | HQ600648 | HQ600615 | This study |
QCAZ 41039 |
|
27 | HQ600647 | HQ600614 | This study |
QCAZ 20785 |
|
4 | HQ600631 | HQ600598 | This study |
QCAZ 25603 |
|
6 | HQ600631 | HQ600598 | This study |
QCAZ 28223 |
|
12 | HQ600638 | HQ600605 | This study |
QCAZ 28646 |
|
7 | HQ600641 | HQ600608 | This study |
QCAZ 28647 |
|
8 | HQ600642 | HQ600609 | This study |
QCAZ 30926 |
|
11 | HQ600643 | HQ600610 | This study |
QCAZ 40253 |
|
10 | HQ600644 | HQ600611 | This study |
QCAZ 42999 |
|
9 | HQ600645 | HQ600612 | This study |
QCAZ 20797 |
|
3 | HQ600632 | HQ600599 | This study |
QCAZ 18230 |
|
2 | HQ600630 | HQ600597 | This study |
QCAZ 15942 |
|
18 | HQ600659 | HQ600626 | This study |
QCAZ 15991 |
|
19 | HQ600656 | HQ600623 | This study |
QCAZ 20544 |
|
33 | HQ600655 | HQ600622 | This study |
QCAZ 32032 |
|
20 | HQ600658 | HQ600625 | This study |
QCAZ 41108 |
|
26 | HQ600660 | HQ600627 | This study |
QCAZ 27816 |
|
34 | HQ600636 | HQ600603 | This study |
QCAZ 27998 |
|
5 | HQ600637 | HQ600604 | This study |
For ease of comparison, we generally follow the format of
Examined specimens (listed in the type-series and Appendix I) are housed at Museo de Zoología, Pontificia Universidad Católica del Ecuador (QCAZ), the Herpetology Collection, Escuela Politécnica Nacional (EPN-H), and the collection of the División de Herpetología, Centro de Ornitología y Biodiversidad (CORBIDI).
Principal Components Analysis (PCA) and Discriminant Function Analysis (DFA) were used to assess the degree of morphometric differentiation between adult
Twelve morphometric variables were measured in tadpoles, following
The models with the best fit and the estimated parameters for each of four partitions for the Bayesian analyses are shown in
Post burn-in averages for parameters of Bayesian analyses. Abbreviations are:
Rate Matrix | Base Frequency | ||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Partition | Best-fit model | AIC score | I | G | K | AC | AG | AT | CG | CT | GT | A | G | C | T |
12S | GTR + G + I | 15762 | 0.386 | 0.740 | - | 0.079 | 0.281 | 0.081 | 0.007 | 0.522 | 0.026 | 0.334 | 0.183 | 0.238 | 0.243 |
ND1, 1st position | HKY + G + I | 14527 | 0.489 | 1.385 | 14.22 | - | - | - | - | - | - | 0.365 | 0.129 | 0.240 | 0.264 |
ND1, 2nd position | GTR + G | 6472 | - | 0.185 | - | 0.024 | 0.589 | 0.047 | 0.042 | 0.274 | 0.020 | 0.261 | 0.096 | 0.307 | 0.334 |
ND1, 3rd position | HKY + G + I | 3745 | 0.295 | 1.617 | 14.22 | - | - | - | - | - | - | 0.397 | 0.104 | 0.216 | 0.280 |
Phylogenetic analysis shows that both
Throughout this section, coloration refers to preserved specimens unless otherwise noted.
Bayesian consensus phylogram depicting relationships within
The holotype is an adult female with SVL = 78 mm (
Dorsolateral and ventral views of
Ventral views of the right hand and foot of
Variation in dorsal and ventral coloration of preserved specimens is shown in
Ventral surfaces of preserved specimens (
Head shape is rounded in dorsal view and rounded (e.g., QCAZ 39803) to bluntly rounded (e.g., QCAZ 39800–01) in lateral view. Lateral head coloration varies between dark brown (QCAZ 11625) to light brown (QCAZ 39810). Except for QCAZ 39802 and 41039, there is a lighter (brown to cream) subocular mark. A tan (QCAZ 39805) to cream (QCAZ 39364) labial stripe is always present. The tympanic annulus is concealed dorsally and has lighter color than the background. Variation in hand and foot webbing is shown in
Variation in webbing in hand and feet of representative adults of
Hand | Foot | |
---|---|---|
QCAZ 39805 (female) | I basal II2½—3¼III3½—2+IV | I1–—1–II1–—1–III1–—1–IV1–—1–V |
QCAZ 39809 (female) | I basal II2½—3¼III3½—2+IV | I1–—1–II1–—2–III1–—2½IV2½—1–V |
QCAZ 39799 (male) | I basal II2½—3¼III3½—2+IV | I1—1II1—1–III1–—2½IV2½—1–V |
QCAZ 39802 (female) | I basal II2½—3¼III3½—2+IV | I1+—2–II1+—2III1—2IV2—1–V |
QCAZ 26304 (female) | I basal II2½—3¼III3½—2–IV | I1–—1–II1–—1–III1–—1–IV1–—1–V |
QCAZ 26488 (male) | I basal II2½—3¼III3½—2–IV | I1–—1–II1–—1–III1–—1–IV1–—1–V |
Morphometric data pertain to adults and are summarized in
Descriptive statistics for morphometric measurements of adult
Species | SVL | FOOT | HL | HW | ED | TD | TL | FL |
---|---|---|---|---|---|---|---|---|
Males (n = 12) | 47.47 ± | 19.38 ± | 14.51 ± | 15.77 ± | 5.01 ± | 2.79 ± | 25.90 ± | 22 ± |
Females (n = 27) | 67.91 ± | 29.79 ± | 20.31 ± | 22.60 ± | 6.25 ± | 3.72 ± | 39.13 ± | 35.35 ± |
Chonza Alta | ||||||||
Males (n = 4) | 43.72 ± | 17.97 ± | 14.07 ± | 13.82 ± | 4.51 ± | 2.55 ± | 24.03 ± | 22.05 ± |
Females (n = 8) | 68.72 ± | 29.33 ± | 20.62 ± | 22.33 ± | 6.2 ± | 3.26 ± | 39.56 ± | 34.63 ± |
Miasí | ||||||||
Female (n = 1) | 79.81 | 36.65 | 21.93 | 26.12 | 6.98 | 3.8 | 45.73 | 42.19 |
Río Lejia | ||||||||
Males (n = 1) | 48.3 | 20.7 | 14.3 | 16.8 | 4.3 | 2.8 | 26.8 | 24.6 |
Females (n = 1) | 72.6 | 30 | 21.1 | 23.2 | 6.1 | 3.7 | 41.2 | 30.4 |
Río Napinaza | ||||||||
Males (n = 4) | 50.82 ± | 21.08 ± | 14.62 ± | 17.4 ± | 5.66 ± | 2.96 ± | 27.67 ± | 24.88 ± |
Females (n = 16) | 66.37 ± | 29.59 ± | 20.04 ± | 22.53 ± | 6.28 ± | 3.96 ± | 38.35 ± | 35.55 ± |
San Francisco | ||||||||
Female (n = 1) | 69.54 | 29.48 | 19.63 | 21.73 | 5.72 | 3.55 | 39.63 | 35.90 |
San Carlos | ||||||||
Males (n = 3) | 47.74 ± | 19.31 ± | 15.00 ± | 15.86 ± | 5.04 ± | 2.87 ± | 25.72 ± | 23.23 ± |
Based on digital photograph of adult female QCAZ 41039 (
There is significant change in coloration between juveniles and adults. The following description is based on a digital photograph of juvenile QCAZ 38081 (
Three components with eigenvalues > 1.0 were extracted from the PCA for males (
Character loadings and eigenvalues for Principal Components (PC) I–III. The analysis was based seven morphometric variables of adult
PCA Males | PCA Females | |||||
---|---|---|---|---|---|---|
Variable | PC I | PC II | PC III | PC I | PC II | PC III |
Foot length | 0.523 | 0.564 | 0.242 | 0.459 | 0.202 | – 0.151 |
Head length | – 0.243 | 0.647 | 0.121 | – 0.243 | 0.394 | 0.537 |
Head width | 0.407 | – 0.430 | – 0.208 | 0.196 | 0.315 | – 0.680 |
Eye diameter | – 0218 | 0.259 | – 0.673 | – 0.061 | 0.648 | – 0.125 |
Tympanum diameter | – 0.224 | 0.004 | 0.655 | – 0.435 | 0.451 | – 0.016 |
Tibia length | 0.455 | 0.407 | – 0.020 | 0.519 | 0.096 | 0.285 |
Femur length | 0.441 | 0.398 | – 0.005 | 0.476 | 0.266 | 0.356 |
Eigenvalue | 2.570 | 1.744 | 1.030 | 2.722 | 1.517 | 1.382 |
Three components with eigenvalues > 1.0 were extracted from the PCA for females (
In the DFA classification procedure, 24 out of 25 specimens of
Letters in parenthesis refer to individual tadpoles on each lot. The following description is based on lot QCAZ 30511 of ten larvae in Stages 25 (A), 26 (B), 31 (C), 32 (D), 33 (E), 34 (F), 35 (G), 39 (H), 40 (I) and 42 (J). Tadpoles were collected at Río Napinaza by E. E. Tapia and I. G. Tapia on October 2003. These larvae belong to the exotrophic, benthic guild as defined by
Measurements of tadpoles of
Stage | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|
Variable | A (25) | B (26) | C (31) | D (32) | E (33) | F (34) | G (35) | H (39) | I (40) | J (42) |
TL | 30.05 | 29.49 | 37.22 | 35.57 | 34.75 | 34.68 | 39.81 | 40.39 | 40.63 | 40.94 |
BL | 9.51 | 9.19 | 11.06 | 10.07 | 10.82 | 11.04 | 11.44 | 11.13 | 11.81 | 11.16 |
BW | 5.74 | 5.68 | 7.25 | 7.05 | 7.23 | 7.10 | 8.11 | 7.50 | 8.46 | 7.12 |
BH | 4.83 | 4.98 | 5.95 | 5.46 | 5.87 | 5.88 | 6.10 | 6.32 | 6.27 | 4.89 |
TAL | 20.30 | 20.04 | 26.59 | 24.56 | 24.36 | 23.76 | 27.90 | 28.99 | 28.69 | 29.90 |
ED | 1.02 | 1.30 | 1.25 | 1.28 | 1.47 | 1.10 | 1.50 | 1.41 | 1.70 | 1.47 |
ODW | 3.50 | 3.35 | 3.90 | 4.12 | 3.80 | 4.43 | 4.13 | 4.00 | 3.94 | 3.46 |
IOD | 4.27 | 4.34 | 5.15 | 5.40 | 5.08 | 5.45 | 5.04 | 5.26 | 5.74 | 5.02 |
IND | 3.49 | 3.50 | 4.01 | 4.19 | 3.75 | 3.94 | 3.98 | 4.12 | 4.01 | 1.62 |
MTH | 5.30 | 5.37 | 5.88 | 6.10 | 5.85 | 6.05 | 6.70 | 6.62 | 6.75 | 6.06 |
TMH | 2.06 | 2.00 | 2.57 | 2.50 | 3.26 | 2.79 | 3.07 | 3.07 | 3.45 | 2.44 |
TMW | 1.60 | 1.46 | 2.70 | 2.12 | 2.30 | 2.50 | 2.97 | 2.62 | 2.78 | 2.56 |
Tail musculature robust, decreasing in size towards tip of tail. Dorsal fin not extending onto body, slightly convex and attaining its maximum height at mid length of tail; tail tip rounded; ventral fin convex, beginning at tail-body junction and tapering gradually towards tail tip. Medial vent tube with both walls attached directly to ventral fin, opening directed posteroventrally. Limbs with subarticular patches. Dorsal body, middle body, supraorbital, infraorbital, posterior supraorbital, and posterior infraorbital lateral lines evident. No glands.
Oral disc anteroventral (
In preservative, dorsum brown with darker marks between eyes; dark brown dorsolateral stripes extend from mid-body to base of tail; caudal musculature beige with brown spots (
Tadpoles of
Adult
Ventral views of adult
In QCAZ 38074, 26321, 26053, 26498 and 26284 the caudal musculature is cream with brown dots; fins can have dark brown spots without white blotches (e.g., QCAZ 38074, 26321). The LTRF is the same in all stages but in stage 42, rows A2–A4 and P1–P3 have approximately half the number of teeth.
In preservative, ten tadpoles collected in Chonza Alta, Peru, lot CORBIDI-CL-10 in Stages 37 (A-B), 36 (C), 34 (D), 32 (E), 31 (F), 42 (G, H, I) and 44 (J) have dorsum brown with darker marks between the eyes; dark brown dorsolateral stripes from mid-body to base of tail; caudal musculature cream with abundant melanophores; skin of flanks, spiracle, vent tube, and fins transparent; skin around the eyes brown; belly and fins transparent with abundant melanophores. Oral disc with LTRF 5/7; papillae distributed around oral disc; tooth rows complete except for medial gap in row A5.
In life (
Larvae of
Most of our specimens of
All the specimens collected in Las Cataratas de Paraiso (Chonza Alta) and Camñopite were found at night on vegetation 50 to 300 cm above the ground, next to fast running streams. Tadpoles (CORBIDI-CL-10) were found in a rocky stream with average width of 4 m and an average depth of 30 to 40 cm with fast running water, close to the base of a waterfall. At both sites the streams are surrounded by secondary forest, pastures and agricultural lands. The specimens from Bajo Naranjillo and Río Lejia were found at night on branches 150 to 200 cm above the ground (in primary forest at Río Lejia and secondary forest surrounded by pastures at Naranjillo).
At Las Cataratas de Paraiso (Chonza Alta), on 1 December 2007, we found twelve males calling from the low vegetation and six amplectant pairs (
Vegetation types for Ecuadorian localities (according to the classification of
Vegetation types of the Peruvian localities (according to
Principal components from analysis of seven size-corrected morphological variables. See Table 5 for character loadings on each component.
The phylogenetic relationships recovered by this study are consistent with the phylogenies reported by
An unexpected result in our phylogeny is the finding of paraphyly among populations of
This study was supported by grants from the Secretaría Nacional de Ciencia y Tecnología del Ecuador SENACYT (PI-C08-0000470) and Pontificia Universidad Católica del Ecuador. For the loan of specimens we are indebted to A. Almendáriz. William E. Duellman and F. Andreone provided photographs of type material. Ministerio de Ambiente of Ecuador and the Dirección General Forestal y Fauna Silvestre (DGFFS) of Peru issued collection permits. For field assistance we are indebted to P. Peña, D. Salazar-Valenzuela, E. Tapia, I. G. Tapia, and Maik Dobiey. For logistic support in Peru we are indebted to Ronald Wagter. Fieldwork in Peru was supported by the fellowship Koepcke of APECO and Conservation International. Omar Torres-Carvajal helped with translations of relevant literature. Ross MacCulloch and J. J. Wiens provided helpful comments to the manuscript.