**Tadpole:** Not described.

**Distribution** (Figure 76): *Cochranella mache* has been reported from few localities in northwestern Ecuador, provinces of Esmeraldas, Imbabura, and Manabí, at elevations between 38 and 800 m ([157–159], this work), and from the Pacific lowlands of Colombia, Departments of Antioquia and Valle del Cauca, at elevations of 750–1030 m [161]. Three localities lie within protected areas (Reserva Biológica Bilsa, Reserva Biológica Canandé, Reserva Jama-Coaque). In Ecuador, the potential distribution of *C*. *mache* is 27,433 km<sup>2</sup> (see also Ortega-Andrade et al. [159]) within the Chocoan Tropical Forest and the Western Foothill Forest regions. When deforestation is taken into account, ~70% of the predicted distribution is reduced [159].

**Figure 76.** Distribution of *Cochranella mache* in Ecuador (yellow dots).

**Conservation status:** Listed as *Endangered* by the IUCN at a global level [162]. Other studies sugges<sup>t</sup> that the species should be classified into the category of *Critically Endangered* [159,163]. The distribution of the species is likely to correspond to the remaining lowland forest in northwestern Ecuador and southwestern Colombia, an area under the constant pressure by wood companies. Although the range of *C*. *mache* is partially within the Mache-Chindul Ecological Reserve and three private reserves (Reserva Biológica Bilsa, Reserva Biológica Canandé, Reserva Jama-Coaque), most conservation measures are ineffective because of institutional and funding restrictions and a lack of law enforcement. Some of the larger fragments are preserved by private organizations but many remain unprotected. Habitat degradation is mainly caused by unsustainable timber extraction, uncontrolled expansion of the agricultural frontier, and replacement by non-native plantations [159,163]. Records from Colombia, however, expand considerably the distribution of *C*. *mache*. More than half of this distribution area of the species is affected by human activities [159]. Under scenarios of climate changes, the potential distribution of *C*. *mache* suffers a reduction of 13%–21% of its predicted range [159].

**Evolutionary relationships** (Figure 69): *Cochranella mache* is the sister species of *C*. *euknemos*.

**Specimens examined:** *Cochranella mache:* Ecuador: *Provincia de Esmeraldas:* Montañas de Mache, Reserva Biológica Bilsa, Riachuelo La Ducha (0◦20-41" N, 79◦42-36" W; 510 m), tributary of Río Aguacatal, 27.4 km W (airline distance) of the town of Quinindé, QCAZ 22412 (holotype), QCAZ 22413, KU 291176; Río Balthazar (0.9745 N, 78.61675 W; 645 m), QCAZ 27747, 31327; Monte Saíno, Punta Galeras region (0.700 N, 80.017 W; 100 m), DHMECN 2611; 3 km NW of Quinindé (0.350 N, 79.483 W; 150 m), DFCH-USFQ LQ23; Reserva Biológica Canandé (0.433 N, 79.133 W; 270 m), DHMECN 3560. *Provincia de Imbabura:* Río La Carolina (0.70449 N, 78.20115 W; 500 m), on the Ibarra–San Lorenzo Road, nearby Jijón y Caamaño, QCAZ 27764. Colombia*: Departamento de Antioquia:* Municipio Dabeiba, Río Amparradó, Quebrada Iotó, 805 m, ICN 10689–90, 8665; Municipio Frontino, Vereda Venados, PNN Las Oriquideas, Quebrada La Miguera, 1030 m, ICN 19638–9.

**Records from the literature:** Ortega-Andrade et al. [159]: Ecuador: Comunidad San Salvador (0.496710 N, 79.85298 W; 38 m); Hacienda Shangrilá (0.18630 N, 79.03019 W; 499 m).

**Photographic record:** Ryan Lynch: Ecuador: *Provincia de Manabí:* Reserva Jama-Coaque (0.0978 S, 80.147 W).

### *Cochranella resplendens* Lynch and Duellman, 1973 [22] (Figures 77–82).

and

*Cochranella resplendens* Lynch and Duellman, 1973 [22]. Holotype: KU 118053. Type locality: "Santa Cecilia, 340 m, Provincia de Napo (Sucumbíos), Ecuador". *Cochranella resplendens*—Ruiz-Carranza and Lynch, 1991a. Guayasamin, Castroviejo-Fisher,

Trueb, Ayarzagüena, Rada, Vilà, 2009 [1]. *Cochranellaphryxa*AguayoandHarvey, 2006[164].Holotype:CBG778.Typelocality:

 "approximately 20 km west of Población de la Cascada (Territorio Comunitario de Origen

y Reserva de la Biósfera Pilón Lajas), La Paz Department, Bolivia (15◦22-37" S, 67◦12-4" W), 1000 m". **New synonymy.**

**Common names:** English: Resplendent Glassfrog. Spanish: Rana de Cristal Resplandeciente. **Etymology:** The specific name *resplendens* is derived from the Latin verb *resplendo*, meaning to glitter, and is used in allusion to the jewel-like appearance of this frog [22].

**Identification:** *Cochranella resplendens* is distinguished from most glassfrogs by having a snout gradually inclined in profile and ventrolateral edges of Finger IV, forearm, elbow, Toe V, tarsus, and heel with dermal folds and enameled tubercles (Figure 77, Figure 78, Figure 81). Additionally, the species lacks humeral spines and has a white venter except for the posterior one-fourth, which is transparent. The following species could be confused with *Cochranella resplendens: Centrolene daidalea*, *Cochranella mache*, *Centrolene savagei*, and *Centrolene solitaria*. However, none of these species is found in the Amazon basin, where *Cochranella resplendens* occurs; *C*. *mache* is distributed in the Pacific lowlands of Ecuador, whereas *C*. *daidalea*, *C*. *savagei*, and *C*. *solitaria* are endemic to the Colombian Andes.

**Figure 77.** *Cochranella resplendens* in life. (**Left**): Amplectant pair. (**Right**): Female brooding clutch. Locality: 2 km N from HW 20 on a dirt road out of Guagua Sumaco, Napo province, Ecuador. Photos by Jesse Delia.

**Diagnosis:** (1) Each vomer with three or four teeth on dentigerous process; (2) snout round in dorsal aspect and gradually inclined in lateral profile (Figure 78); (3) tympanum oriented almost vertically, with slight lateral and posterior inclinations, its diameter about 38% of eye diameter; tympanic annulus partially visible, low supratympanic fold evident, tympanic membrane barely translucent, pigmented as surrounding skin; (4) dorsal skin shagreen with elevated and spiculated warts corresponding to white spots; (5) venter granular; pair of slightly enlarged subcloacal warts; enlarged cloacal fold present; (6) white lining (iridophores) on the anterior three-fourths of the ventral parietal peritoneum, posterior one-fourth transparent (condition P3); iridophores in pericardium and peritonea covering intestines and stomach; renal capsules and gall bladder and urinary bladder lacking iridophores (condition V2); (7) liver lobate, covered by transparent peritoneum (condition H0); (8) humeral spines absent; (9) webbing absent between inner fingers, and extensive between outer fingers (Figure 78); webbing formula III (1<sup>1</sup>/2–2−)—(1–1<sup>1</sup>/2) IV; (10) webbing between toes

extensive (Figure 81); webbing formula on foot I (1–1+)—(1<sup>2</sup>/3–2) II (0–0+)—(2–2+) III (1–1<sup>1</sup>/2)—(2–2+) IV (2−–2<sup>+</sup>)—(1−–1<sup>+</sup>) V; (11) ventrolateral edges of Finger IV, forearm, elbow, Toe V, tarsus, and heel with dermal folds and enameled tubercles (Figures 77, 79 and 81); (12) concealed prepollex; nuptial pad not evident; (13) Finger I about same length as Finger II; (14) disc of Finger III width about 55% of eye diameter; (15) in life, dorsum dark green with numerous small white spots (Figures 77 and 79); bones green; (16) in preservative, dorsum lavender with small white spots (Figure 81); (17) iris whitish cream to beige, with a fine grey to brown reticulation; pale yellow circumpapilar ring; (18) dorsal surfaces of fingers and toes with few small, enameled spots; melanophores only on surfaces of Finger IV and Toe V; (19) calling behavior unknown; call undescribed; (20) fighting behavior unknown; (21) egg clutches placed on upper sides of leaves; short-term maternal care present; parental care provided by males absent; (22) tadpoles with non-emarginate oral apparatus; tooth row formula 2(2)/3; upper jaw slightly curved; (23) medium body size; in two adult males, SVL 26.5–26.6 mm; females unknown.

**Figure 78.** *Cochranella resplendens*, holotype KU 118053. (**A**) Head in dorsal view. (**B**) Head in lateral view. (**C**) Hand in ventral view. Drawings of head not to scale. Illustrations by Juan M. Guayasamin.

**Color in life** (Figures 77 and 79): Dorsum green with numerous, small white to bluish–white spots. Ventral surfaces mostly white, except posterior portion, which is translucent. Upper lip, tubercles on forearm and foot, and cloacal warts white. Bones green. Iris whitish cream to beige, with fine grey to brown reticulation; pale yellow circumpapilar ring ([22,164], this work).

**Color in ethanol** (Figure 81): Dorsal surfaces of head, body, and limbs slate grey to lavender with small white spots, often corresponding to small, flat tubercles. Upper lip white; tympanum pigmented as surrounding skin. White tubercles present on Fingers III and IV and Toes II–V. White on dermal folds of forearm and tarsus and on tips of Finger IV and Toe V. White cloacal tubercles. Pericardium and anterior three-fourths of the ventral parietal peritoneum white. White peritonea covering intestine, stomach, and colon. Transparent peritonea covering kidney, gall bladder, and urinary bladder. Hepatic peritoneum clear in holotype and most known specimens, but with a patch of iridophores in two Ecuadorian specimens and in Bolivian specimen ([22,164], this work).

**Biology and ecology:** *Cochranella resplendens* is a rare species. Since its description in 1973, only eight additional individuals have been reported (Specimens Examined). The scarcity of *Cochranella resplendens* is not related to low sampling effort. At the type locality (Santa Cecilia), fieldwork from 1967 through 1972 resulted in the collection of 7765 specimens of amphibians and

reptiles, but no additional specimens of *Cochranella resplendens* [165]. However, canopy surveys may reveal that its rarity is an artifact of microhabitat sampling [166]. Sixty-six species of amphibians, including *Teratohyla midas* and *Hyalinobatrachium munozorum*, are known to occur at Santa Cecilia [165]. At the Bolivian locality (20 km W of Población La Cascada), *Cochranella resplendens* was found sympatric with *Rulyrana spiculata* and *Cochranella* sp. [164]. At the Tiputini Biodiversity Station, *Cochranella resplendens* was found sympatric with *Teratohyla midas*, *Hyalinobatrachium munozorum*, and *Vitreorana ritae* ([134], this work). Females provide short-term parental care; male parental care is absent [25].

**Figure 79.** Life cycle in *Cochranella resplendens*. (**A**) Juvenile, QCAZ 38088. (**B**) Ontogenetic variation of tadpoles. (**C**) Egg clutch. Photos by Luis A. Coloma. Figure modified from Terán-Valdez et al. [167].

**Tadpole** (Figures 79 and 80): At Río Napinaza, a clutch with 74 embryos was found on the upper side of a leaf [167]. According to Terán-Valdez et al. [167], the tadpole of *Cochranella resplendens* has the following traits (based on tadpole in Gosner Stage 36): Body elongated, oval-depressed (sensu [168]), wider than high; snout rounded in dorsal and lateral views. Eyes located on dorsal surface of head; in early stages, eyes C-shaped; after Stage 35, eyes become round. Short, single, sinistral spiracle, at the posterolateral region of the body; vent tube short and abdominal, free posteriorly, opening directed posteriorly; myotomes visible throughout length of tail; dorsal fin originating at about mid-length of tail; ventral fin originating almost at base of tail muscle and reaching its maximum height posterior to mid-length of tail. Oral disc non-emarginate; marginal papillae uniserial, distributed around oral disc, but larger on the lower labium; upper labium with papillae only on lateral extremes. Upper jaw sheath completely keratinized with serrated edge; slightly curved. Lower jaw sheath keratinized, U-shaped, and with serrated edge. Labial tooth row formula 2(2)/3; A-2 with medial gap. For ontogenetic variation, see Terán-Valdez et al. [167]. Dorsally, the tadpole is mostly red, with some aggregations of iridophores that form an interorbital line and a middorsal band from behind the eyes to nearly the end of the body (based on a tadpole in Gosner Stage 36). Dorsally, the anterior-most part of the body is pale yellow. Laterally, the tadpole is translucent. The iris is mostly black with a yellow ring around the pupil [167].

**Figure 80.** Oral apparatus of different species of Centrolenidae (from Terán-Valdez et al. [167]). *Cochranella granulosa* and *Hyalinobatrachium ibama* are in Gosner Stage 24, *Cochranella resplendens* in Stage 36, *H*. *cappellei* in Stage 25, *H*. *aureoguttatum* in Stage 35. Gosner stages of the remaining species are not provided in the original descriptions. Figures modified from: (**A**,**G**) Rada et al. [169]; (**B**,**D**,**F**,**I**,**J**) Starrett [146]; (**C**,**E**) Terán-Valdez et al. [167], (**H**) Noonan and Bonett [170].

**Taxonomic Remarks:** Aguayo and Harvey [164] described *Cochranella phryxa* from the Amazonian slopes of the Bolivian Andes. These authors mentioned that *C*. *phryxa* can be distinguished from *Cochranella resplendens* (characteristics in parenthesis) by having a hidden tympanum (tympanum visible), a first finger longer than the second (second finger longer than the first), and a straight cloacal fold (U-shaped cloacal fold). After examining the holotype and six additional specimens (QCAZ 38099, MHNSM 19507, DFCH-USFQ D103–4; FHGO 1305, 1324) of *Cochranella resplendens*, we find that the tympanic membrane in *Cochranella resplendens* is only slightly differentiated from the skin surrounding the tympanum. Therefore, one could interpret the tympanum in *Cochranella resplendens* as hidden, which is the character state that Aguayo and Harvey [164] described for *C*. *phryxa*. The relative length of the two innermost fingers has been used for many authors when comparing centrolenid species. Although we agree that the character is useful when the differences are obvious (e.g., comparing species of *Hyalinobatrachium*), it is problematic when differences are minor, as in *C*. *resplendens*. We have tried

to minimize the error in the assessment of this character by measuring the fingers with a digital caliper. Consistently, we found that these fingers have almost the same length in *Cochranella resplendens* (Finger I length 96%–102% of Finger II). The last character used by Aguayo and Harvey [164] is the shape of the cloacal fold, which they described as straight in *C*. *phryxa* and U-shaped in *Cochranella resplendens*. Di fferences in shape of the cloacal ornaments may be either artificial, associated with how the specimens are preserved (i.e., posterior extremities fixed with an anterior, posterior, or parallel orientation in relation to the sagittal axis of body), or due to intraspecific variation. The cloacal ornamentation (i.e., white warts and folds) is the same in the two species (compare Lynch and Duellman [22]: Figure 2c; Aguayo and Harvey [164]: Figure 4). *Cochranella mache*, a species with U-shaped cloacal folds, shows the same intraspecific variation observed between *C*. *resplendens* and *C*. *phryxa*. The only di fference that we find between the holotype of *Cochranella resplendens* and the Bolivian specimen is the presence of a small patch of iridophores on the hepatic peritoneum of the latter. However, it is possible that this patch is either an artifact of preservation (i.e., iridophores from the ventral parietal peritoneum can be attached to the liver) or intraspecific variation (i.e., the presence/absence of patches of iridophores in the peritoneum has been observed in other centrolenids (e.g., *Cochranella mache* [17,158]). For the reasons discussed above, we formally place *Cochranella phryxa* Aguayo and Harvey, 2006 [164], in the synonymy of *Centrolenella resplendens* Lynch and Duellman, 1973 [22].

**Figure 81.** *Cochranella resplendens* in preservative. Holotype (KU 118053) from Santa Cecilia, Sucumbíos province, Ecuador. ( **A**) Dorsal view. (**B**) Ventral view. ( **C**) Lateral view. Photos by Martín Bustamante.

**Distribution** (Figure 82): *Cochranella resplendens* in known from the Amazon basin of Colombia, Ecuador, Peru, and Bolivia, at elevations between 190 and 1100 m ([22,134,164,167,171], this work).

A disjunct population of the species was recently found on the eastern slope of the central Andes in the Departamento Antioquia, Colombia, at much higher elevations (1309–1699 m) [172]. In Ecuador, this species is known from localities in the provinces of Morona Santiago, Napo, Orellana, Pastaza, and Sucumbíos at elevations between 250–1100 m. In Ecuador, the potential distribution of *C*. *resplendens* is 77,793 km<sup>2</sup> within the Amazonian Tropical Rainforest and Eastern Foothill Forest ecoregions.

**Figure 82.** Distribution of *Cochranella resplendens* in Ecuador (yellow dots).

**Conservation status:** Globally, *Cochranella resplendens* is currently listed as *Least Concern* by the IUCN [173]. Although it is evident that *C*. *resplendens* is rare in collections (see above), we presume that this is a consequence of inadequate sampling [166]. The potential distribution of this species in Ecuador is 77,793 km2, 14% of which is affected by human activities. Given the broad distribution of *C*. *resplendens* and the lack of immediate threats, the category of *Least Concern* is justified.

**Evolutionary relationships** (Figure 69): With the current genetic and taxon sampling, the sister species of *C*. *resplendens* is *C*. *granulosa*. These two species are an example of vicariant speciation, mediated by the uplift of the Andes.

**Specimens examined:** *Cochranella resplendens:* Colombia: *Departamento de Putumayo:* Santa María de Sucumbíos (ca. 00◦16- N, 76◦55- W), AMNH 88083 (Lynch and Duellman, 1973). Ecuador: *Provincia de Sucumbíos:* Santa Cecilia (00◦03- N, 76◦58- W; 340 m), KU 118053 (holotype). *Provincia de Morona Santiago:* Río Napinaza (2.927◦ N, 78.407◦ W; 1100 m), QCAZ 38088. *Provincia de Napo*: Reserva Yachana (00◦52-21.71" S, 77◦14-13.43" W; 300–350 m), QCAZ 38099; *Provincia de Orellana*: San José Viejo de Sumaco (0.5333 S, 77.4167 W; ca. 810 m), USNM 288460; Tiputini Biodiversity Station (00◦37- S, 76◦10- W; 190–270 m), DFCH-USFQ D103–04; *Provincia de Pastaza:* Pozo Garza, Oryx (01◦26- S, 77◦03- W; 300 m), FHGO 1305, 1324. Peru: *Departamento de San Martín:* Cainarachi Valley (6◦43-10 S, 76◦29-13 W; 550 m), Km 33, Carretera Tarapoto-Yurimaguas, MHNSM 19507. Bolivia: *Departamento de La Paz:* approximately 20 km west of Población de la Cascada (15◦22-37" S, 67◦12-4" W; 1000 m), CBG 778 [164].

**Photographic records:** Ecuador: *Provincia de Napo:* ca. 2 km north from HW 20 on a dirt road out of Guagua Sumaco (0.688◦ S, 77.6◦ W; ca. 900 m); photographs by Jesse Delia (Figure 77).

**Genus** *Espadarana* Guayasamin, Castroviejo-Fisher, Trueb, Ayarzagüena, Rada, & Vilà 2009 [1].

**Etymology:** The name *Espadarana* honors Marcos Jiménez de la Espada, a Spanish zoologist who was part of the Comisión Científica del Pacífico that explored America between 1862 and 1865. Jiménez de la Espada described the first centrolenid frog, *Centrolene geckoideum* in 1872. In Spanish, the word *Espada* means sword, in allusion to the humeral spines present in males of species in this genus. *Espadarana* is a combination of the words *Espada* and *rana* (frog) and is feminine in gender [1].

*Espadarana audax* (Lynch and Duellman, 1973 [22]; Figures 83–87).

*Centrolenella audax* Lynch and Duellman, 1973 [22]. Holotype: KU 146624.

Type locality: "Salto de Agua, 2.5 km NNE of Río Reventador on Quito–Lago Agrio road, 1660 m, Provincia de Napo, Ecuador".

*Centrolene audax*—Ruiz-Carranza and Lynch, 1991 [6].

*"Centrolene" audax*—Guayasamin, Castroviejo-Fisher, Trueb, Ayarzagüena, Rada, Vilà, 2009 [1].

*Centrolene fernandoi*—Duellman and Schulte, 1993 [174]. Synonymy by Cisneros-Heredia and Guayasamin, 2014 [175].

*Espadarana audax*—Twomey, Delia, and Castroviejo-Fisher, 2014 [19].

**Common names:** English: Daring Glassfrog. Spanish: Rana de Cristal Audaz.

**Etymology:** The specific name *audax* is Latin, meaning daring, and is used in allusion to the precipitous regions inhabited by this species [22].

**Identification:** *Espadarana audax* is unique among Ecuadorian glassfrogs by having small yellow spots on the dorsum, short and distally curved humeral spines in males, and extensive webbing only between Fingers III and IV (Figures 83–85). In Ecuador, species with a similar dorsal coloration include *Nymphargus buenaventura*, *N*. *siren*, *N*. *humboldti* sp. nov., *Rulyrana flavopunctata*, and *Teratohyla midas;* however, males of these species lack humeral spines. In addition, *Teratohyla midas* has visceral peritonea covered by iridophores; *N*. *buenaventura*, *N*. *siren*, and *N*. *humboldti* sp. nov. have considerably less hand webbing; and *R*. *flavopunctata* has more webbing between Fingers III and IV. The Colombian *Centrolene notosticta* has a similar dorsal color pattern but lacks vomerine teeth (present in *E*. *audax*) and has less webbing between the outer fingers. *Espadarana audax* is most similar to *E*. *durrellorum* (see Taxonomic Remarks), but the latter differs by lacking yellow dorsal spots. Also, the two species are found at slightly different, although overlapping, elevations; *E*. *audax* inhabits the Amazonian slopes of the Andes (800–1900 m), whereas *E*. *durrellorum* is found mostly in the Amazonian lowlands and foothills (220–1150 m).

**Figure 83.** *Espadarana audax* in life. Adult male, QCAZ 37871, from Gral. Leonidas Plaza Gutiérrez (Limón), Quebrada del Río Napinaza, Morona Santiago, Ecuador. Photos by Luis A. Coloma.

**Diagnosis:** (1) Dentigerous process of the vomer bearing two to four teeth; (2) snout rounded in dorsal profile, rounded to truncated in lateral profile; (3) tympanum of moderate size, tympanum diameter 23%–29% of eye diameter, low supratympanic fold evident, tympanic membrane partially pigmented, clearly di fferentiated from surrounding skin; (4) dorsal surfaces of males and females shagreen, with minute spicules in males; (5) pair of enlarged subcloacal warts, other cloacal ornamentation absent; (6) anterior two-thirds of the ventral parietal peritoneum covered with white iridophores, posterior third transparent (condition P2); white pericardium; no iridophores in peritonea covering intestines, stomach, testes, kidneys, gall bladder, and urinary bladder (condition V1); (7) liver tetralobed, hepatic peritoneum lacking iridophores (condition H0); (8) in adult males, humeral spines present; (9) webbing absent between Fingers I, II, and III; webbing formula for outer fingers III (2−–21/3)—(2−–2<sup>+</sup>) IV; (10) extensively webbed foot: I (1–1+)—(2–2+) II (1−–1<sup>+</sup>)—(2–2<sup>1</sup>/4) III (1–1<sup>1</sup>/4)—(2<sup>+</sup>–21/3) IV (2<sup>+</sup>–21/3)—(1–1<sup>+</sup>) V; (11) ulnar fold present, inner tarsal fold present, outer tarsal fold absent; (12) concealed prepollex; in males, nuptial pad Type I; (13) Finger I about same length as Finger II (Finger II 95.6%–102% length of Finger I); (14) disc of Finger III moderate, its width 43%–56% of eye diameter; (15) in life, dorsal surfaces of head, body, and limbs green with small yellow spots; bones green; (16) in ethanol, dorsal surfaces of head, body, and limbs lavender with small white spots; (17) in life, iris white with yellowish hue and thin black reticulations; (18) melanophores on outer fingers and outer toes; (19) males call from upper sides of leaves; calls are produced in series, with each series having four or five calls; each call is composed by a single, pulsed note; dominant frequency is at 5426–6718 (mean = 6146, SD = 368) Hz; (20) fighting behavior unknown; (21) eggs deposited in moss on branches; short-term maternal care present; parental care by males absent; (22) tadpoles unknown; (23) small body size; in adult males, SVL 21.6–25.5 mm ( X = 23.5 ± 0.722, *n* = 41); in adult females, SVL 24.5–28.8 ( X = 27.0 ± 1.468, *n* = 7).

**Figure 84.** *Espadarana audax*, adult males. ( **A**) Head in lateral view, KU 164503. (**B**) Hand in ventral view, KU 178018. Illustrations by Juan M. Guayasamin.

**Figure 85.** Humeral spines of *Espadarana audax* and *E. durrellorum*. (**A**) *Espadarana audax*, KU 164500. (**B**) *Espadarana durrellorum*, QCAZ 47909. Photos by Juan M. Guayasamin.

**Variation:** In several males, the dorsal spicules are minute and only visible under magnification. A population from Leonidas Plaza Gutiérrez (Limón) has a dorsum with very few and small dorsal yellow spots.

**Color in life** (Figure 83): Dorsum green, with small yellow spots; fingers and toes pale yellow; parietal peritoneum white; heart not visible; visceral peritoneum lacking iridophores. Ventral surfaces of limbs unpigmented; bones green; iris pale bronze to mustard with thin black reticulation ([22], this work).

**Color in ethanol:** Dorsal surfaces of head, body, and limbs lavender with small white dots; Fingers I–III and Toes I–III cream (without melanophores); upper lip white. White lining on the anterior two-thirds of ventral parietal peritoneum; white pericardium; no iridophores in peritonea covering liver, intestines, stomach, testes, kidneys, gall bladder, and urinary bladder. Some preserved individuals (KU 164497–98) have lost their dorsal white spots; however, close examination revealed the presence of collapsed chromatophores (apparently iridophores).

**Biology and ecology:** By day, one individual (KU 146624) was found in a bromeliad. During the night, adults are found on the upper sides of leaves along streams, a site from which males call (W. E. Duellman field notes, 19 March 1975; this work). On 18 July 1981, 12 individuals were found in Río Salado; the males were calling and one female (KU 178023) had dark brown oviductal eggs visible through the body wall. Females provide short-term parental care; male parental care is absent [25].

**Call** (Figure 86): We analyzed 14 notes contained within four calls from two individuals (MZUTI 1492–93). Calls are emitted in series; a typical call series has four or five calls (mean = 4.3, SD = 0.6) and is relatively long (range = 2398–3285 ms, mean = 2711 ms, SD = 497.8 ms). Each call has a single note with a duration of 26–53 (mean = 38, SD = 8.2) ms. Notes have clearly defined pulses with 5–11 (mean = 7.4, SD = 1.7) amplitude peaks throughout the note. Pulses within a note have a rate of 171–208 (mean = 189, SD = 10) pulses per second. Notes have their peak amplitude in the first 50% of the note (relative peak time: Range = 0.0144–0.0722, mean = 0.043, SD = 0.02), whereby the first note usually has the highest amplitude. The dominant frequency of a note measured at peak amplitude is 5426–6718 (mean = 6146, SD = 368) Hz and is contained within the fundamental frequency. The fundamental

frequency has a lower limit of 5254–6288 (mean = 5925, SD = 374) Hz and a higher limit of 6115–7063 (mean = 6608, SD = 297) Hz.

**Figure 86.** Calls of *Espadarana audax*, MZUTI 1492, recorded at Río Hollín, 1038 m, Napo province, Ecuador. (**A**) Call series. (**B**) Single call.

**Tadpole:** Not described.

**Distribution** (Figure 87): *Espadarana audax* occurs through the Eastern Montane region of southeastern Colombia, Ecuador, and northern Peru at elevations between 800 and 1900 m. This species has been recorded from the provinces of Napo, Morona Santiago, Sucumbíos, and Zamora Chinchipe in Ecuador, the departments of Huila (eastern slope of Cordillera Central), Putumayo, Cauca, Caquetá, and Boyacá (eastern slope of Cordillera Oriental) in Colombia, and the Department of San Martín in Peru ([19,22,174–176], this work). In Ecuador, the species is found within the Eastern Foothill Forest and Eastern Montane Forest ecoregions. The potential distribution of *E*. *audax* is 6738 km2, 9% of which is affected by human activities.

**Figure 87.** Distribution of *Espadarana audax* in Ecuador (yellow dots).

**Conservation status:** Globally, *Espadarana audax* is currently listed as *Least Concern* by the IUCN [177]. In Ecuador, *E. audax* has not been found in some of its historical localities (e.g., Río Azuela, Río Salado ([91], JMG, pers. obs.)), but has been discovered recently at several new localities (Miazi Alto, General Leonidas Plaza Gutiérrez, Zamora); see Specimens Examined. The distribution of the species is fragmented because of agriculture, pasture lands, and mining. We sugges<sup>t</sup> that, in Ecuador, the species should be considered as *Near Threatened*.

**Evolutionary relationships** (Figure 88): Given the current gene and taxon sampling, *Espadarana audax* and *E*. *durrellorum* are sister species.

**Figure 88.** Evolutionary relationships among species in the genus *Cochranella*, inferred using maximum likelihood and Bayesian criteria.

 *omma*,

**Taxonomic Remarks:** The morphological di fferences that separate *Espadarana durrellorum* and *E*. *audax* are subtle, but given the available genetic and morphological data, we consider them as valid species. *"Centrolene" fernandoi* was recently placed in the synonymy of *E*. *audax* [175].

**Specimens examined:** *Espadarana audax:* Ecuador: *Provincia Napo:* 2.5 km NNE of Río Reventador on Quito–Lago Agrio road (0.133 S, 77.633 W, 1600 m), KU 146624 (holotype); 16.5 km NNE Santa Rosa (0.2186 S; 77.7319 W, 1700 m), KU 143290, 143292; 2 km SSW of junction between Río Reventador and Baeza-Lumbaqui road (0.1 S, 77.6 W, 1700 m), KU 164497–504; 7 km SW Río Azuela on Quito–Lago Agrio road (0.1667 S, 77.667 W), KU 155502–03; Río Salado (0.19167 S, 77.6997 W, 1420 m), KU 178018–27; 43 km NE Santa Rosa (0.1186 S, 77.6003 W, 1490 m), KU 190015; 8.9 km NE Santa Rosa on Quito–Lago Agrio road (0.1667 S, 77.667 W), KU 190016; Río Hollín (0.72 S, 77.639 W; 1100 m), QCAZ 6898; Reserva Narupa (0.6848 S, 77.742 W, 1164 m), ZSFQ 382–86. *Provincia Sucumbíos:* Río Azuela (0.11667 S, 77.6167 W, 1740 m), KU 164496. *Provincia Morona Santiago:* Quebrada del Río Napinaza, 6.6 km N in the road Limón–Macas (2.92665 S, 78.40701 W; 1100 m), QCAZ 37871, 29439; La Y (3.43236 S, 78.60449 W, 835 m), QCAZ 23910. *Provincia Zamora Chinchipe:* Alto Miazi (4.25026 S, 78.61746 W; 1250 m), QCAZ 41653; Cordillera del Cóndor, Centro Shuar El Tink, ca. 1050 m, QCAZ 48202. Colombia: *Departamento Huila:* Parque Arqueológico San Agustín, 3 km SW of San Agustín, 1750 m. Peru: *Departamento San Martín:* W slope of Abra Tangarana, 7 km (by road) NE of San Juan de Pacaysapa (6.2 S, 76.733 W; 1080 m), KU 211770–75.

#### *Espadarana callistomma* (Guayasamin and Trueb 2007 [9]; Figures 89–92).

 as a

*Centrolene callistommum*—Guayasamin and Trueb, 2007 [9]. Holotype: QCAZ 25832.

Type locality: "stream a ffluent of Río Bogotá (1◦05-13.8" N, 78◦41-25.8" W, 83 m), nearby San Francisco de Bogotá, Provincia de Esmeraldas, Ecuador."

*Espadarana callistomma*—Guayasamin, Castroviejo-Fisher, Trueb, Ayarzagüena, Rada, and Vilà, 2009 [1].

**Common names:** English: Beautiful eyes Glassfrog. Spanish: Rana de cristal de ojos bellos. **Etymology:** The specific name *callistomma* is derived from the Greek *kallistos*–, meaning "most beautiful"andmeaning"eye",reference tothefantasticirispatterninthisspecies[9].

**Identification:** *Espadarana callistomma* is easily distinguished from most glassfrogs by its large size (in males, SVL 26.7–29.6 mm; in females, SVL 29.5–31.8 mm), uniform green dorsal coloration, and silvery white iris with black reticulations (Figure 89). *Espadarana callistomma* closely resembles *Sachatamia ilex;* however, males of *E*. *callistomma* have a conspicuous humeral spine, whereas males of *S*. *ilex* have a small humeral spine that is embedded in the arm musculature (Figure 201). Some populations of *Espadarana prosoblepon* have the same color pattern as *E*. *callistomma*, but they have a significantly smaller body size (in males, SVL 23.2–27.5 mm; in females, SVL 25.3–27.8 mm) and a two-note call (three to four notes in *E*. *callistomma*). See Taxonomic Remarks.

**Figure 89.** *Espadarana callistomma* in life from Durango, Esmeraldas province, Ecuador. Adult male, QCAZ 32055. Photo by Martín Bustamante.

**Diagnosis:** (1) Vomerine teeth present, each vomer with two to seven teeth; (2) snout truncated in dorsal and lateral views (Figure 90); (3) tympanum small (tympanum diameter 20%–31% eye diameter), oriented vertically, with lateral inclination; tympanic annulus visible except for upper border, which is covered by supratympanic fold; tympanic membrane partially pigmented, differentiated from surrounding skin; (4) dorsal surfaces of males and females shagreen, with minute spicules evident in males (visible under magnification); (5) pair of enlarged subcloacal warts (Figure 90); (6) anterior two-thirds of ventral parietal peritoneum covered with white iridophores, posterior third transparent (condition P3); white pericardium; translucent peritoneum covering intestines, stomach, gall bladder, testes, and urinary bladder; kidneys covered with translucent peritoneum (condition V1); (7) liver tetralobed, covered by transparent peritoneum (condition H0); (8) humeral spines present in adult males; (9) no webbing between Fingers I and II, webbing between Fingers II and III reduced, webbing between Fingers III and IV extensive (Figure 90), webbing formula: II (1<sup>2</sup>/3–2)—(3<sup>+</sup>–31/4) III (1<sup>1</sup>/2–12/3)—(1–11/2) IV; (10) webbing on foot extensive (Figure 90), webbing formula: I (0<sup>+</sup>–1)—(2−–2<sup>+</sup>) II (0<sup>+</sup>–1)—(2−–2<sup>+</sup>) III (0<sup>+</sup>–1)—2− IV (2−–21/3)—(1−–1<sup>+</sup>) V; (11) ulnar and inner tarsal folds low; outer tarsal fold absent; (12) concealed prepollex; in males, nuptial pad Type I; (13) Finger II slightly longer than Finger III (Finger III 93.3%–100% length of Finger II); (14) disc of Finger III of moderate size, about 29%–34% eye diameter; (15) in life, dorsum olive green (Figure 89); bones green; (16) in preservative, dorsum dark lavender; (17) iris silvery white with thin black reticulations; (18) melanophores covering dorsal surfaces of Fingers III and IV and Toes IV and V; (19) males call from upper side of leaves; each call consists of three to four pulsed notes, dominant frequency at 5343–5812 Hz; (20) fighting behavior unknown; (21) eggs deposited on upper side of leaves; parental care unknown; (22) tadpoles unknown; (23) in males, SVL 26.7–29.6 mm (X = 27.8 ± 0.762; *n* = 19); in females, SVL 29.5–31.8 mm (X = 30.5 ± 0.796; *n* = 13).

**Figure 90.** *Espadarana callistomma*. (**A**) Head in lateral and dorsal views, paratype, adult male, QCAZ 28803. (**B**) Ventral surfaces of thighs showing enlarged subcloacal tubercles, holotype, adult male, QCAZ 25832. (**C**) Hand and foot in ventral view, paratype, adult male, QCAZ 28803. (**D**) Nuptial pad and thumb in dorsal view, holotype, adult male, QCAZ 25832. Modified from Guayasamin and Trueb [9].

**Osteology:** A detailed osteological description of *Espadarana callistomma* was provided by Guayasamin and Trueb [9]; they mentioned that the frontoparietals were exceedingly slender along the orbital margin of the braincase. After reexamining the cleared-and-stained material, it seems that the frontoparietals extend medially and their medial margins partially delimit the frontal fontanelle (area shown as cartilage in [9]: Figure 8A).

**Color in life** (Figure 89): Dorsum uniform olive green, without spots; upper lip with whitish–cream coloration; iris silvery white with marked black reticulations; flanks white; parietal peritoneum white, covering anterior two-thirds of abdomen (heart not visible); bones green [9].

**Color in preservative:** Dorsal surfaces of head, body, and limbs uniform lavender; upper lip cream; iris white with marked dark lavender reticulations; nuptial pad on Finger II cream; dorsally, Fingers II and III and Toes I–III unpigmented; venter cream [9].

**Biology and ecology:** The natural history of *Espadarana callistomma* is poorly known. The species is active during the night and has been found on leaves along streams. Males call from the upper side of leaves, and females deposit pigmented eggs on the upper side of leaves. It is unknown if males or females provide parental care [9].

**Call** (Figure 91): Description based on a call (LBE-C-022) of a male *E. callistomma* recorded by JMG on 17 May 2010 at Reserva Otokiki, Ecuador. Each call consists of three to four pulsed notes (X = 3.7 ± 0.483, *n* = 10) and lasts 0.28–0.44 s (X= 0.38 ± 0.066, *n* = 10). Notes are produced at a rate of 9.1–11.0 notes/s (X= 9.76 ± 0.596, *n* = 10); each note has a duration of 0.014–0.04 s (X = 0.03 ± 0.007, *n* = 10). The call is not frequency modulated; dominant frequency at 5343–5812 Hz (*n* = 10).

**Figure 91.** Calls of *Espadarana*. (**A**) Call of *Espadarana callistomma* from Reserva Otokiki, 706 m, Esmeraldas province, Ecuador, LBE-C-022. (**B**) Call of *E*. *prosoblepon* from Cordillera de Chontilla, 908 m, Pichincha province, Ecuador, MZUTI 601.

**Tadpole:** Not described.

**Distribution** (Figure 92): *Espadarana callistomma* is known from four localities in the lowland rainforest of northwestern Ecuador (provinces of Esmeraldas and Carchi) at elevations below 500 m ([9], this work) and the Pacific lowlands of Colombia [178]. The habitat of the species is within the Chocoan Tropical Forest and the Western Foothill Forest regions (see Biogeographic Regions).

**Figure 92.** Distribution of *Espadarana callistomma* in Ecuador (yellow dots).

**Conservation status:** Globally, *Espadarana callistomma* is currently listed as *Least Concern* by the IUCN [179]. At present, *E. callistomma* is known from a few localities in the Pacific lowlands of Ecuador and Colombia [9,178]. The most likely scenario, however, is that *E*. *callistomma* is restricted to portions of the Chocó ecoregion, an area with high rates of deforestation and mining; we sugges<sup>t</sup> that the species is considered as *Endangered* in Ecuador, following IUCN criteria B1, B2a, B2b(iii).

**Evolutionary relationships** (Figure 88): *Espadarana prosoblepon* is paraphyletic with respect to *E*. *callistomma*, although *E*. *callistomma* is monophyletic. The most likely explanations to the observed pattern are: (i) Divergence between the two species is recent and reciprocal monophyly was not ye<sup>t</sup> achieved, (ii) *E*. *callistomma* is a synonym *of E*. *prosoblepon*, or (iii) *E*. *prosoblepon* is a species complex that requires further subdivision. Given the morphological and acoustic data at hand, we favor the first and third hypotheses.

**Taxonomic Remarks:** The specific status of *Espadarana callistomma* is uncertain because of the conflict among different sets of data. Genetically, *Espadarana callistomma* and *E*. *prosoblepon* are not reciprocally monophyletic (Figure 88). On the other hand, body size of males and females of *E*. *callistomma* is significantly larger than those of *E*. *prosoblepon* (*t-test*, *P* < 0.001), and the dorsal and iris color patterns of *E*. *callistomma* are different from most *E*. *prosoblepon* (Figure 89 and Figure 95). In terms of vocalization, *E*. *callistomma* has a call with more notes (three to four notes per call) than *E*. *prosoblepon* (two notes per call; Figure 91). Given that we find biologically meaningful differences among the species (i.e., body size, vocalizations), we recognize both species as valid. The lack of molecular divergence could be the product of a recent divergence, but this is a hypothesis that remains untested. Another pending taxonomic problem is that the type of *Hylella parabambae* matches the spotless coloration of *E*. *callistomma* and some populations of *E*. *prosoblepon*.

**Specimens examined:** *Espadarana callistomma:* Ecuador: *Provincia de Esmeraldas:* stream affluent of Río Bogotá (1◦05-13.8" N, 78◦41-25.8" W, 83 m), nearby San Francisco de Bogotá, QCAZ 25832 (holotype), 27776–8, 28555–56, 28557 (C&S), 28558; stream affluent of Río Bogotá (1◦05-9.06" N, 78◦41-8.7" W, 77 m), 2 km E of San Francisco de Bogotá on the San Francisco–Durango road, QCAZ 28803; 2.1 km E of Durango (1.02477 N, 78.61746 W; 284 m), QCAZ 32169. *Provincia de Carchi:* Río La Carolina (0◦42-16.16" N, 78◦12-4.14" W, 500 m), on the Ibarra–Lita road, nearby Jijón y Caamaño, QCAZ 27744–45, 27768.

#### *Espadarana durrellorum* (Cisneros-Heredia, 2007 [180]; Figures 85, 93 and 94).

*Centrolene durrellorum* Cisneros-Heredia, 2007 [180]. Holotype: DFCH-USFQ D131. Type locality: "small rivulet tributary of the Jambue River, ca. 6 km S from Zamora (ca. 04◦03- S, 78◦56- W, 1150 m), on the western slope of Contrafuerte de Tzunantza, Cordillera Oriental, eastern slopes of the Andes, Provincia de Zamora Chinchipe, Rep ública del Ecuador". *"Centrolene" durrellorum—*Guayasamin, Castroviejo-Fisher, Trueb, Ayarzagüena, Rada, and Vilà, 2009 [1].

*Espadarana durrellorum—*Twomey, Delia, and Castroviejo-Fisher, 2014 [19].

**Common names:** English: Durrell's Glassfrog. Spanish: Rana de Cristal de Durrell.

**Etymology:** The specific name *durrellorum* honours Gerald Durrell and Lee Durrell (Durrell Wildlife Conservation Trust) for their contributions to the conservation of biodiversity [180].

**Identification:** *Espadarana durrellorum* can be distinguished from all other glassfrogs by having a green dorsal coloration (Figure 93), humeral spines short and straight in adult males (Figure 85), moderate body size (25.7–26.1 mm adult males), and lacking iridophores on the visceral peritonea. Among Ecuadorian centrolenids, *E*. *durrellorum* is most similar to *E*. *audax*. The main trait to di fferentiate between the two species is the absence of yellow dorsal spots in *E*. *durrellorum* (present in *E*. *audax*). *E*. *audax* inhabits higher elevations (800 to 1900 m) than *E*. *durrellorum* (220–1150 m).

**Figure 93.** *Espadarana durrellorum* in life. Ecuador, Napo province, Comunidad Ñukanchi Allpa, 403 m, QCAZ 47909. Photos by Luis A. Coloma.

**Diagnosis:** (1) Vomerine teeth present; (2) snout rounded in dorsal view, bluntly truncated in lateral view; (3) tympanic annulus evident, oriented dorsolaterally; very weak supratympanic fold above tympanum; (4) dorsal skin shagreen, with minute spicules uniformly distributed; (5) ventral skin granular; cloacal area granular, with two large, rounded, flat subcloacal tubercles, and two folds on the sides of the cloacal opening; distinct cloacal sheath; (6) upper two-thirds of the parietal peritoneum covered by iridophores (condition P3), pericardium white, all other peritonea lacking iridophores (condition V1); (7) liver lobed, lacking iridophores (condition H0); (8) in adult males, humeral spine short and straight, with sharp point; (9) webbing absent between Fingers I and II, basal or absent between II and III, moderate between outer fingers: In males, III 2—2− IV; in one female III <sup>1</sup>2/3—(11/2–2−) IV; (10) webbing formula on feet: In males, I 2—2− II 1—2 III 1<sup>+</sup>—2− IV 21/3—1<sup>+</sup> V; in one female, I 1—13/<sup>4</sup> II 1—2− III 1<sup>+</sup>—2 IV 2—1 V; (11) no dermal folds on hands, forearms, feet, or tarsus; (12) nuptial excrescences present, Type I; concealed prepollex; (13) first finger slightly longer than second; (14) eye diameter larger than width of disc on finger III; (15) in life, dorsal surfaces uniform green; (16) in preservative, dorsal surfaces uniform pale lavender; (17) in life, iris golden with thin dark reticulations; in preservative, lavender with darker reticulation; (18) melanophores widespread on outer fingers and outer toes; (19) males call from upper surfaces of leaves; call undescribed; (20, 21, 22) fighting behavior, egg clutches, tadpoles, and parental care unknown; (23) small body size; SVL 24.8–26.1 mm in adult males (*n* = 3); 27.0 in one female.

**Color in life** (Figure 93): Uniform green dorsum without flecks or spots. Iris golden with dark reticulations. Green bones.

**Color in ethanol:** All dorsal surfaces pale lavender (no light or dark spots). Ventral surfaces cream. Parietal peritoneum covered by iridophores to the level of the lower stomach; pericardium white, all other peritonea lacking iridophores.

**Biology and ecology:** *Espadarana durrellorum* is a recently described species and little information is available on its natural history. It is nocturnal, and males call from the tops of leaves of riverine vegetation in mature Foothill and Lowland Evergreen forests. Parental care unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 94): *Espadarana durrellorum* is known from few localities on the foothills of Cordillera Oriental and the Amazonian lowlands of Ecuador, at elevations of 300–1267 m [176,180].

**Figure 94.** Distribution of *Espadarana durrellorum* in Ecuador (yellow dots).

**Conservation status:** Globally, *E. durrellorum* is *Least Concern* by the IUCN [181]; the species is known from few confirmed records, and a re-evaluation of its conservation status may be required.

**Evolutionary relationships** (Figure 88): *Espadarana durrellorum* and *E*. *audax* are sister species.**TaxonomicRemarks:**SeeTaxonomicRemarks under*Espadaranaaudax*.

**Specimens examined:** *Espadarana durrellorum:* Ecuador: *Provincia de Napo:* ca. 45 km E of Narupa, on the Hollín–Loreto road (ca. 800 m), DFCH-USFQ D291; Parroquia Chontapunta, comunidad Ñukanchi Allpa, cabecera del río Canoayacu (0.99965◦ S, 77.39619◦ W; 403 m), QCAZ 47909; Reserva Yachana (00◦52-21.7" S, 77◦14-13.4" W; 300–350 m), DHMECN 03492, 06790–91; Pungarayacu (00◦42-27.8" N, 77◦44-26.2" W; 1267 m), DHMECN 03476. *Provincia de Sucumbíos:* Zábalo (0.3181333◦ S, 75.76625◦ W; 220 m), QCAZ 27832; Shuara (00◦00-26" N, 76◦33-55.1" W; 300 m), DHMECN 06794–95; Plataforma Shushufindi (00◦05-14.8" S, 76◦40-03.8" W; 330 m), DHMECN 06793. *Provincia de Zamora Chinchipe:* tributary of the Jambue River, ca. 6 km S from Zamora (ca. 04◦03- S, 78◦56- W; 1150 m), on the western slope of Contrafuerte de Tzunantza, Cordillera Oriental, eastern slopes of the Andes, DFCH-USFQ D131.

#### *Espadarana prosoblepon* (Boettger, 1892 [182]; Figures 91 and 95–97).


**Common names:** English: Variable Glassfrog. Spanish: Rana de Cristal Variable.

**Etymology:** The specific name *prosoblepon* is apparently derived from the Greek words *proso* (forward, onward, in front) and *blepo* (look or see), probably referring to the forward orientation of the eyes in centrolenid frogs [84].

**Identification:** *Espadarana prosoblepon* is distinguished from most glassfrogs from the Pacific versant of the Andes by having a large, flat, and projected humeral spine, and green dorsum with small black spots (Figure 95); however, *E*. *prosoblepon* has a highly variable dorsal coloration, including individuals that have both black and yellows spots, only yellow spots, only black spots, or absence of spots all together. In Ecuador, the only species that have a green dorsum with dark spots are *Cochranella litoralis*, *Vitreorana ritae*, *Nymphargus cochranae*, and *N*. *megacheirus*. In three of these species (*V*. *ritae*, *N*. *cochranae*, *N*. *megacheirus*), males lack humeral spines; additionally, they occur on the Amazonian lowlands or Amazonian slope of the Andes and are never in sympatry with *E*. *prosoblepon*. *Cochranella litoralis* has a bright orange iris and is smaller than *E*. *prosoblepon* (in males, SVL < 22.1 mm in *C*. *litoralis;* 23.2–27.5 mm in *E*. *prosoblepon*). Unspotted *E*. *prosoblepon* can be confused with *E*. *callistomma* and *C*. *buckleyi; E*. *callistomma* is distinguished by having a larger body size (in males, SVL 26.7–29.6 mm; in females, SVL 29.5–31.8 mm), contrasting black-and-white iris, and a call with three to four notes (call with two notes in *E*. *prosoblepon;* see Taxonomic remarks). *Centrolene buckleyi* has an inclined snout in lateral view (Figure 26) and is only found at elevations above 2000 m. Individual of *E*. *prosoblepon* with uniform dorsum and white iris with black reticulations resemble *Sachatamia ilex;* however, males of *E*. *prosoblepon* have a conspicuous humeral spine, whereas males of *S*. *ilex* have a small humeral spine that is embedded in the arm musculature (Figure 201). The closely related *E*. *andina* is similar to *E*. *prosoblepon* by having dark spots on the dorsum but has a pointy humeral spine that is almost parallel to the humerus, whereas in *E*. *prosoblepon,* the spine is broad, laminar, and projected at an angle of about 45◦ from the humerus.

**Figure 95.** *Espadarana prosoblepon* in life. Note variation of dorsal patterns. Frogs from Mindo and surroundings, Pichincha province, Ecuador; not collected. Photos by Alejandro Arteaga/Tropical Herping.

**Diagnosis:** (1) Each vomer with two to seven teeth on dentigerous process; (2) snout usually truncated in dorsal and lateral profiles (Figure 96); (3) tympanum small, oriented almost vertically, with slight lateral and posterior inclinations, its diameter 20.5%–30.8% of eye diameter; tympanic annulus mostly visible, with supratympanic fold covering its posterodorsal margin; tympanic membrane translucent, partially pigmented, clearly differentiated from surrounding skin; (4) dorsal skin shagreen; males and females may possess minute spicules on flanks and tympanic region; (5) pair of enlarged subcloacal warts; (6) white iridophores on the anterior 50%–70% of the ventral parietal peritoneum, posterior portion transparent (condition P2–P3); white pericardium; translucent peritonea covering intestines, stomach, kidneys, gall bladder, and urinary bladder (condition V1); (7) liver with four clearly defined lobes covered by transparent peritoneum (condition H0); (8) males with conspicuous humeral spines; (9) webbing absent between Fingers I and II, absent or basal between Fingers II and III; moderate between outer fingers (Figure 96); webbing formula III (1<sup>2</sup>/3–2)—(1–2) IV; (10) webbing between toes moderate; webbing formula on foot I (0<sup>+</sup>–1)—(2−–2<sup>+</sup>) II (0<sup>+</sup>–1)—(2−–2<sup>+</sup>) III (0<sup>+</sup>–1)—(2−–2<sup>+</sup>) IV (2−–21/3)—(1–1<sup>+</sup>) V; (11) ulnar and tarsal folds absent or low and inconspicuous, lacking white coloration; (12) concealed prepollex; nuptial pad Type I; (13) Finger I as long as Finger II or slightly longer (Finger II length 93.8%–101% Finger I); (14) disc of Finger III moderate, width 28.6%–49.0% of eye diameter; (15) in life, dorsum green usually with small dark spots (Figure 95; but see *Color in life* section); bones green; (16) in preservative, dorsum lavender with or without cream and/or dark spots; (17) in life, iris varies from grey with fine dark reticulations to white with thick black reticulations; (18) melanophores covering dorsal surfaces of Fingers III and IV and Toes IV and V; (19) males call from upper side of leaves or small branches; (20) males fight upside down, grasping one another venter to venter; (21) black eggs are deposited on the upper surface of leaves, moss-covered rocks, or branches; short-term maternal care present; males do not provide parental care; (22) tadpoles with non-emarginated oral apparatus; tooth row formula 2/3; upper jaw curved; (23) small to medium body size; in adult males, SVL 23.2–27.5 mm (X = 24.8 ± 0.982, *n* = 53); in adult females, SVL 25.3–27.8 mm (X = 26.6 ± 0.859, *n* = 15).

**Figure 96.** *Espadarana prosoblepon*, KU 132462. (**A**) Head in lateral view. (**B**) Hand in ventral view. Illustrations by Juan M. Guayasamin

**Color in life** (Figure 95): In Costa Rica, dorsal surfaces green dorsum with dark spots [148]. About one-half of Honduran specimens lack small dark dorsal spots [84]. In Ecuador, several color patterns have been reported: (1) Uniform green dorsum; (2) dorsum green with dark spots; (2) dorsum green with yellow spots; (3) dorsum green with dark and yellow spots, separated from each other; and (4) dorsum green with dark spots surrounding yellow spots, forming false ocelli. Iris varies from greyish white to pale bronze with fine dark reticulation, to silvery white with thick black reticulations. Green bones; whitish cream upper lip; white venter that is transparent posteriorly; ventrolateral margins of forearm and tarsus lacking white coloration.

**Color in ethanol:** As described above, but green turns into lavender and yellow spots turn white. Some individuals (e.g., KU 132463–64) present black and smaller white spots on dorsum. White parietal peritoneum covering about anterior 60% of venter. Translucent peritonea cover digestive tract, liver, kidneys, and urinary and gall bladders; white pericardium.

**Biology and ecology:** The following information is based on studies by Jacobson [188], Hayes [189], Savage [148], Hoffmann [147], and Arteaga et al. [87]. At night, during the breeding season, *Espadarana prosoblepon* is found on vegetation along margins of streams in primary to slightly degraded evergreen forests and, sporadically, even in pastures. Reproduction peaks in the rainy season, when males become more vocal and often engage in aggressive territorial combats. Males initiate amplexus by jumping on the back of approaching gravid females. Females lay 20–52 black eggs on a variety of substrates and directly above the water. Females provide short-term parental care; male parental care is absent [25]. After hatching, tadpoles usually hide under leaf litter or sand at the bottom of streams. Males are territorial, and the advertisement call is used to space them along the stream. If an intruder moves into an occupied territory, the resident male produces a rapid series of calls; if the intruder approaches too closely, a fight ensues with both males dangling upside down while holding onto vegetation with their legs and grappling with their arms and humeral spines. The combat concludes when the loser drops from the fight site or signals submission by flattening his body against the leaf surface. *Espadarana prosoblepon* feeds on a variety of small arthropods including beetles, moths, spiders, and mites. As a defense mechanism, this glassfrog produces pungen<sup>t</sup> odors when grabbed. *Espadarana prosoblepon* is the only glassfrog species for which survival information is available; McCaffery and Lips [190] report that mean annual survival probability is 0.46 (0.41–0.52, 95% CI). These authors also provide abundance data on several populations from Panama.

**Call** (Figure 91): The following description is based on a call of a male *Espadarana prosoblepon* (MZUTI 601) recorded by Italo Tapia during the night of 4 July 2012 at Río Sune Chico (908 m, Cordillera Chontilla, Pachijal-Mashpi) Pichincha province, Ecuador. Air temperature at the moment of the recording was 18.8 ◦C. Calls are produced every 4–7 min. Each call consists of two pulsed notes and lasts 0.207–0.223 s (X = 0.212 ± 0.007, *n* = 4). Each note has a duration of 0.032–0.054 notes/s (X = 0.040 ± 0.008, *n* = 8); time between the two notes of each call is 0.130–0.137 s (X = 0.134 ± 0.004, *n* = 4). Calls are frequency modulated; the dominant frequency at the beginning of each note is 5712–5825 Hz (X = 5759 ± 56.1, *n* = 4), whereas at the end of the note is at 6196–6394 Hz (X = 6252 ± 95.7, *n* = 4). Harmonics are also frequency modulated, showing complex spectral patterns.

The call of *E*. *prosoblepon* from Costa Rica shows some differences from the Ecuadorian call described above. The following information is from Jacobson [188] and describes a call from Monteverde, Costa Rica. Males of *Espadarana prosoblepon* call at irregular intervals (17 calls/h) from the upper surfaces of leaves during the night. The advertisement call consists of two to five short "beeps". When encountering another frog within 15 cm, the resident males often give a series of rapid short beeps. Males of *Espadarana prosoblepon* call vigorously while in amplexus and also immediately after egg deposition. Kubicki [24] described the call of *E*. *prosoblepon* as a rapid three-note series, but sometimes males emit four to five notes; each note has an average duration of 0.03 s; the dominant frequency is 5.5–5.8 kHz. Typical call series have a total duration lasting 0.5 s (average three-note series) or 0.75 s (average four-note series). Because males call from exposed sites on the upper surfaces of leaves and twigs, these periodic calling bouts, often synchronized among a few, nearby males, may make it more difficult for predatory bats (i.e., *Trachops cirrhosus*) to locate and eat calling male *E*. *prosoblepon* (RWM, pers. obs.).

**Tadpole:** Females lay egg clutches on the upper side of leaves, moss-covered rocks, or branches; clutch size varies from 20 to 52 eggs that have a dark brown to black coloration; females remain with the eggs for some time (up to 131 min), but once they leave neither parent returns to the clutch [87,147,148,188,189]. Embryos are black but quickly turn yellow. By the time the tadpoles hatch and fall into the stream, they are red colored [148]. At Stage 25 [191], larvae have a sinistral spiracle, a relatively simple oral disc with medium-sized jaw sheaths, 2/3 tooth rows, and a single row of marginal papillae with a wide dorsal gap. The second anterior (upper) tooth row (A-2) also has a wide gap [148]. Drawings of the mouth and body can be found in Starrett [146], Savage [148]), and McCranie and Wilson [84].

**Distribution** (Figure 97): *Espadarana prosoblepon* is found from eastern Honduras, Nicaragua, Costa Rica, and Panama, south along the Pacific slopes of the Andes of Colombia and Ecuador from sea level to about 1600 m elevation. In Colombia, it occurs on the northern and eastern flanks of the Eastern Cordillera south to Caldas, and in the Magdalena Valley [192]. In Ecuador, *Espadarana prosoblepon* has been reported from several localities below 1620 m [87] (Specimens examined). The habitat of the species in Ecuador is mainly within the Chocoan Tropical Forest and the Western Foothill Forest regions, but it is also found in Deciduous Forest and the lower limit of the Western Montane Forest region. In Ecuador, it has a potential distribution of 41,612 km2.

**Figure 97.** Distribution of *Espadarana prosoblepon* in Ecuador (yellow dots).

**Conservation status:** Globally, *Espadarana prosoblepon* is listed as *Least Concern* [192] in view of its large distribution and presumed large populations. In Ecuador, this species is common and is found even in habitats with moderate alterations; the category of *Least Concern* is justified.

**Evolutionary relationships** (Figure 88): *Espadarana prosoblepon* is paraphyletic, containing populations currently assigned to *E*. *callistomma*, a pattern that can be explained, mainly, by recent species divergence or the two species actually representing a single evolutionary lineage. See Taxonomic Remarks below.

**Internal morphology:** A detailed osteological and myological description of *Espadarana prosoblepon* was provided by Eaton [193]. The hyoid apparatus illustrated by Eaton lacks the anterolateral processes that are evident in cleared-and-stained material from Ecuador (KU 178163) and Costa Rica (KU 65178). We assume that the normal condition is the presence of these processes.

**Taxonomic remarks:** While the general morphology of *Espadarana prosoblepon* is mostly conservative, its color pattern is extremely variable, and because of an insufficient understanding of the species' variation, different names have been assigned to distinctive morphs. For example, populations whose individuals have dorsal coloration consisting of black spots surrounding yellow spots and forming ocelli were called *ocellifera*, while others with dorsum lacking spots have been variously named *parambae* and *callistomma*. Several authors, including Lynch and Duellman [22], Savage [42], and Cisneros-Heredia and McDiarmid [17], have described some of the interspecific variation characteristic of *E*. *prosoblepon*, but much remains to be done. For example, most descriptions and color photographs of *E*. *prosoblepon* depict the iris of this species as grey with fine dark reticulations, but Kubicki [24] reported individuals with a white iris with black reticulations. Unfortunately, most centrolenid descriptions have paid little attention to iris patterns; in fact, with the exception of the descriptions of *Sachatamia ilex*, *Hyalinobatrachium ignioculus*, and *H*. *eccentricum* (the last two taxa now under the synonymy of *H*. *cappellei*), iris coloration, if reported at all, usually is based on examination of photographs; thus, the inter and intraspecific variation in iris color is poorly known. Genetic data sugges<sup>t</sup> that *E*. *prosoblepon* and *E*. *callistomma* might represent one evolving lineage (Figure 88). Morphological differences between *E*. *callistomma* and *E*. *prosoblepon* are in body size (*callistomma* being larger), dorsal coloration (without spots in *callistomma*; usually with spots in *prosoblepon*), and iris coloration (white with contrasting black reticulation in *E*. *callistomma;* less contrasting in *E*. *prosoblepon*). In terms of vocalization, *E*. *callistomma* has a call with more notes (three to four notes per call) than *E*. *prosoblepon* (two notes per call; Figure 91). At the moment, we consider that the morphological and acoustic differences justify recognition of *E*. *callistomma* as a valid species.

**Specimens examined:** *Espadarana prosoblepon:* Ecuador: *Provincia de Azuay:* 12.9 km W Luz María (2.6889 S, 79.474 W, 740 m), KU 217502, QCAZ 12603–04; *Provincia de Bolívar:* Balzapamba (1.7667 S, 79.1833 W, 800 m), KU 132555–56, 132462–65; *Provincia de Carchi:* near Maldonado (0.9 N, 78.1 W, 1410 m), KU 178156–57; *Provincia de Cotopaxi:* near La Mana (0.933 S, 79.2167 W, 300 m), QCAZ 8641; near Sigchos (0.7 S, 78.883 W), USNM 288441; km 8 of the Pucayacu-Sigchos road (1.0097 S, 79.23769 W), QCAZ 40674. *Provincia de Esmeraldas:* Reserva Biológica Bilsa (0.3447 S, 79.71 W, 500 m), KU 291165–75, QCAZ 22414–17; Viruela, QCAZ 10267; Reserva Biológica Bilsa (0.3447 S, 79.71 W, 500 m), KU 291165–75; Reserva Ecológica Cotacachi-Cayapas, Charco Vicente (0.792 N, 79.1978 W; 60 m), QCAZ 11364–65; 5 km W of Durango (1.0858 N, 78.74 W), QCAZ 13206, 13212, 13242. *Provincia de El Oro:* near Valle Hermoso (3.50194 S, 79.81722 W; 379 m), QCAZ 37249; Río Chillayacu (3.32834 S, 79.58102 W; 395 m), 16.8 km W of Piñas (3.667 S, 79.667 W, 600 m), USNM 286738–39. *Provincia del Guayas:* Chongon-Colonche hills near Guayaquil, (ca. -2.1 S, -80.15 W), USNM 288438; *Provincia de Imbabura:* near Lita (0.833 N, 78.4667 W, 520 m), KU 133482–83; 6 km E of Lita (0.79472 N, 78.4286 W), QCAZ 4318–19; Zona de amortiguamiento de Reserva Cotacachi Cayapas, near Rio Aguas Verdes (0.331010◦ N, 78.93152◦ W; 670 m), QCAZ 46009; km 5 in the Lita-Ibarra road (0.84773 N, 78.42175 W), QCAZ 39919. *Provincia de Los Ríos:* Estación Biológica Río Palenque, 56 km N of Quevedo (0.55 S, 79.3667 W, 220 m), KU 164616–22. *Provincia de Santo Domingo de los Tsáchilas:* Río Orito, on the Toachi-Chiriboga road (0.30561 S, 78.882 W, 1315 m), QCAZ 15356; Río Faisanes, on the Toachi-Chiriboga road (0.2608 S, 78.845 W; 1400 m), QCAZ 15357, 15360, 15362; La Florida (0.28361 S, 79.0189 W), QCAZ 20726–28, 20730–32; Otongachi, near La Unión del Toachi (0.3167 S, 78.95 W; 900 m), QCAZ 25094; 5 km NE of La Florida (0.25694 S, 79.0539 W), QCAZ 7184–893; 4 km NE of the Dos Ríos–Chiriboga road (0.305139 S, 78.884333 W; 1270 m), QCAZ 31982; Santo Domingo de los Colorados (0.25 S, 79.15 W, 660 m), KU 121054–55; 4 km NE of Dos Ríos (0.30278 S, 78.8678 W, 1140 m), KU 164623–34; 2 km E and 1 km S of Santo Domingo de los Colorados (0.24512 S, 79.15509 W, 600 m), KU 178158–66; La Palma (0.3167 S, 78.9167 W, 920 m), KU 178167. *Provincia de Pichincha:* Reserva Maquipucuna (0.12429 N, 78.62936 W; 1343 m), QCAZ 42179; 6 km NW (by air) of Pedro Vicente Maldonado (0.10421 N, 79.10279 W; 544 m), QCAZ 35430; La Concordia, Bosque Protector La Perla (0.01 N, 79.4 W, 190 m), QCAZ 12602, Reserva Mashpi, 18 km N of San Miguel de Los Bancos (0.15 S, 78.883 W, 1100 m), DFCH-USFQ 293–95.

## **Genus** *Hyalinobatrachium* Ruiz-Carranza and Lynch, 1991 [6].

**Etymology:** The generic name *Hyalinobatrachium* comes from the Greek words *hyalos* (glass) and *batrachion* (frog), alluding to the translucent and delicate appearance of the species in this genus [6].

*Hyalinobatrachium adespinosai* Guayasamin, Vieira, Glor, Hutter 2019 [194] (Figures 98–101).

*Hyalinobatrachium adespinosai* Guayasamin, Vieira, Glor, Hutter, 2019 [194]. Holotype: ZSFQ 1648.

Type locality: "San Jacinto River (1.3447 S, 78.1814 W; 1795 m asl), Tungurahua province, Ecuador".

**Common names:** English: Adela's Glassfrog. Spanish: Rana de Cristal de Adela.

**Etymology:** The specific epithet *adespinosai* honors Adela Espinosa, an Ecuadorian conservationist and board member of the Jocotoco Foundation (http://www.jocotoco.org/wb#/EN/LaFundacion). Adela has concentrated her work to the conservation of species and ecosystems. *Hyalinobatrachium adespinosai* is found only within the limits of a natural reserve owned by Adela and her husband, Antonio Páez [194].

**Identification:** Among glassfrogs, *H*. *adespinosai* (Figure 98) is diagnosable mainly by having a transparent pericardium. However, it is morphologically cryptic with four closely related taxa (*H*. *anachoretus*, *H*. *pellucidum*, *H*. *esmeralda);* these species display a similar size and color pattern (pale green dorsum with yellow dots and a transparent venter and pericardium; red heart visible ventrally). However, calls have diverged noticeably (Figure 99); the major difference is the structure of the call, with two species (*H*. *adespinosai* and *H*. *anachoretus*) having pulsed calls and the others having tonal vocalizations. The call of *H*. *adespinosai* sp. nov. is further differentiated from that of *H*. *anachoretus* by being longer, having more pulses per note, and being produced at a higher rate [194]. Genetic distances for the 16S marker between *H*. *adespinosai* and closely related species (*H*. *pellucidum*, *H*. *esmeralda*, *H*. *yaku)* are >2.5%. However, *H*. *adespinosai* and *H*. *anachoretus* only have a 1% genetic distance, although they are separated by a major biogeographic barrier (i.e., Marañon river valley) [194].

**Diagnosis:** The following characters are found in *Hyalinobatrachium adespinosai:* (1) Dentigerous process of the vomer lacking teeth; (2) snout truncated in dorsal and lateral profiles; (3) tympanum barely visible, hidden under skin, with coloration similar to that of surrounding skin; (4) dorsal skin shagreen, lacking tubercles; (5) ventral skin areolate; cloacal ornamentation absent, paired round subcloacal warts absent; (6) parietal peritoneum transparent; pericardium with thin layer of iridophores (in life, a red heart is mostly visible ventrally); liver, viscera, and testes covered by iridophores; (7) liver white, bulbous; (8) humeral spines absent; (9) hand webbing formula: I (2–3)—(2–2+) II (1–1+)—3<sup>1</sup>/<sup>3</sup> III (2–2<sup>+</sup>)—(2−–2) IV; (10) foot webbing moderate; webbing formula: I 1—(1<sup>2</sup>/3–2−) II (1–1−)—(2−–21/3) III (1–1<sup>+</sup>)—(2<sup>+</sup>–21/3) IV 2<sup>+</sup>—(1<sup>+</sup>–11/3) V; (11) fingers and toes with thin lateral fringes; ulnar and tarsal folds present, but difficult to distinguish, with thin layer of iridophores that extends to ventrolateral edge of Finger IV and Toe V; (12) nuptial excrescence present as a small pad on Finger I (Type V), concealed prepollex; (13) when appressed, Finger I longer than II; (14) diameter of eye about two times wider than disc on Finger III; (15) coloration in life: Dorsal surfaces pale yellowish green with small pale yellow spots and minute grey to black melanophores; bones white; (16) coloration in preservative: Dorsal surfaces pale cream with minute melanophores; (17) iris coloration in life: White with pale yellow hue and minute lavender spots; (18) melanophores absent from most fingers and toes, but present on Finger IV and Toes IV and V; (19) males call from underside of leaves; advertisement call consisting of single note, distinctly pulsed (9–13 pulses per call), with duration of 0.382–0.430 s, and dominant frequency at 4645–5001 Hz; (20) fighting behavior unknown; (21) males attend egg clutches located on the underside of leaves overhanging streams; clutch size of 22 embryos (*n* = 1); (22) tadpoles unknown; (23) SVL in adult males 20.5–22.2 mm (*n* = 3), females unknown.

**Figure 98.** *Hyalinobatrachium adespinosai* in life, holotype, ZSFQ 1648. Photos by Jose Vieira/Tropical Herping.

**Color in life** (Figure 98): Dorsal surfaces apple green to yellowish green with diffuse yellow spots and minute grey to black melanophores. Melanophores absent from fingers and toes, except Finger IV and Toes IV and V. Ventrally, parietal peritoneum and pericardium transparent, with a red heart always visible, even when a very thin layer of iridophores is present on the pericardium of some individuals. Visceral peritoneum of gall bladder and urinary bladder transparent; hepatic and visceral peritonea white; ventral vein red. Iris pale yellowish white, with numerous minute lavender spots. Bones white [194].

**Color in ethanol**: Dorsal surfaces cream dotted with minute dark lavender melanophores; venter uniform cream; visceral peritoneum lacking iridophores; pericardium with a very thin layer of iridophores. Iris silvery white with minute lavender melanophores [194].

**Biology and Ecology:** All individuals were found on the underside of leaves of riverine vegetation along the San Jacinto River. The section of river was fast-flowing and had visible rapids. Although the population is locally abundant, individuals are difficult to observe because they are usually found at the canopy level (4–16 m above ground level). Males were calling in the months of July and August. One male (ZSFQ 1648) was apparently guarding an egg clutch containing 22 embryos; both the adult male and the egg clutch were on the same leaf most of the time, but the male also moved to nearby leaves [194].

**Call** (Figure 99): The description is based on recording from nine individuals. The call of *Hyalinobatrachium adespinosai* has a striking resemblance to the chirp of a cricket. Each call is composed by a single and high-pitched pulsed note and has a duration of 0.38–0.44 s (X = 0.38 ± 0.017). Time between calls varied from 2.0–11.0 s (X = 4.58 ± 2.3). The fundamental frequency, the same as the dominant frequency, is at 4645–5203 Hz (X = 4855 ± 152). There is no frequency modulation. The first harmonic is at 9336–9754 Hz and the second harmonic is at 14,159–14,444 Hz [194].

**Figure 99.** Call of *Hyalinobatrachium adespinosai*, holotype, ZSFQ 1648, recorded in field conditions at the type locality (San Jacinto River, 1795 m asl), Tungurahua province, Ecuador). Air temperature = 18 ◦C. Obtained from Guayasamin et al. [194].
