**Tadpole:** Not described.

**Distribution** (Figure 126): *Hyalinobatrachium yaku* is known from three localities on the Amazonian lowlands of Ecuador at elevations between 300–360 m. It is likely that *H*. *yaku* has a broader distribution, including areas in Amazonian Colombia and Peru [241].

**Figure 126.** Distribution of *Hyalinobatrachium yaku* in Ecuador (yellow dots).

**Conservation status:** According to Guayasamin et al. [241], available information is insufficient to sugges<sup>t</sup> a conservation category, thus *H*. *yaku* is a *Data Deficient* species.

**Evolutionary relationships** (Figure 101): *H. yaku* and *H*. *pellucidum* are sister species.

**Specimens examined:** *Hyalinobatrachium yaku*: Ecuador: *Provincia Pastaza:* stream affluent of Kallana river (1.4696◦ S, 77.2784◦ W, 325 m), MZUTI 5001 (holotype), 5002 (paratypes). *Provincia Orellana:* Timburi-Cocha Research Station (0.4800◦ S, 77.2829◦ W, 300 m) near San José de Payamino, QCAZ 55628, 53352 (paratypes). *Provincia Napo:* Ahuano (1.0632◦ S, 77.5265◦ W, 360 m), ZSFQ 2322.

**Genus** *Nymphargus* Cisneros-Heredia and McDiarmid 2007 [17].

**Etymology**: The name *Nymphargus* is formed from the Greek *nymphae* in allusion to the nymphs, beautiful goddesses in Greek mythology that personify the creative and fostering activities of nature, living in mountains, valley, springs, and rivers; and, *argus* in allusion to the mythological Greek Argus, nephew of the nymph Io, a giant with a hundred eyes, whose eyes became the ocelli in the peacock's tail. The name is masculine and alludes to the ocelli found on the dorsum of some of the species of the genus [17].

*Nymphargus anomalus* (Lynch and Duellman, 1973 [22]; Figures 127–130).

*Centrolenella anomala* Lynch and Duellman, 1973 [22]. Holotype: KU 143299. Type locality: "Río Azuela, 1740 m, Quito–Lago Agrio road, Provincia Napo, Ecuador". *Cochranella anomala—*Ruiz-Carranza and Lynch, 1991 [6], 57:21. *Nymphargus anomalus—*Cisneros-Heredia and McDiarmid, 2007 [17].

**Common names:** English: Anomalous Glassfrog. Spanish: Rana de Cristal Anómala.

**Etymology:** The specific name *anomalous* refers to the unusual dorsal coloration of this species [22]. **Identification:** *Nymphargus anomalus* is unique by having, in life, a tan dorsum with black ocelli with orangish–tan centers (Figures 127 and 128), instead of the usual green that characterizes most glassfrogs. Among centrolenids, only one species, *N*. *ignotus*, has a similar dorsal color pattern; however, *N*. *ignotus* is found on the Pacific slopes of the Andes, whereas *N*. *anomalus* is restricted to the Amazonian slopes of the Andes. *Nymphargus anomalus* further differs from other species having ocellated patterns (*N*. *cochranae*, *N*. *ignotus*, *N*. *laurae*) by having scattered black and lavender flecks between ocelli (Figure 127).

**Diagnosis:** (1) Vomers lacking teeth; (2) snout truncated in dorsal and lateral profiles (Figure 129); (3) tympanum relatively small, oriented almost vertically, with evident lateral and posterior inclinations, its diameter about 20% of eye diameter; tympanic annulus mostly visible, with supratympanic fold covering its dorsal margin; tympanic membrane differentiated and translucent, partially pigmented; (4) in males, dorsal skin shagreen with or without minute spiculae; in females, dorsal skin shagreen without spiculae; (5) venter areolate; pair of enlarged subcloacal warts (Figure 15); (6) anterior half of ventral parietal peritoneum white, posterior half transparent (condition P2); white pericardium; translucent peritoneum covering intestines (condition V1); (7) liver with four clearly defined lobes covered by transparent peritoneum (condition H0); (8) humeral spines absent; (9) webbing absent between Fingers I–III, basal or moderate between Fingers III and IV; webbing formula: III (2<sup>1</sup>/3–3<sup>+</sup>)—(2<sup>+</sup>–22/3) IV (Figure 129); (10) webbing between toes moderate; webbing formula on foot I <sup>2</sup>−—(2–2<sup>1</sup>/4) II (1–1<sup>1</sup>/4)—(2<sup>+</sup>–21/3) III (1–1<sup>+</sup>)—(2<sup>+</sup>–21/4) IV (2<sup>+</sup>–22/3)—(1<sup>+</sup>–11/2) V; (11) ulnar fold low; inner and outer tarsal folds present, but low and difficult to distinguish; (12) concealed prepollex; nuptial pad Type I; (13) Finger I slightly shorter than Finger II (Finger I length 91.5%–99.2% of Finger II); (14) disc width of Finger III about 50% of eye diameter; (15) in life, dorsum pale brown or tan with dark brown flecks and black ocelli with brownish–orange centers (Figure 127); color of bones white; (16) in preservative, dorsum cream to tan with black dots and lavender flecks and black and lavender ocelli enclosing whitish centers; (17) iris pale yellow with thin black reticulation; bright yellow circumpupilar ring; (18) dorsal surfaces of all fingers and toes usually with few melanophores, except Finger I that usually lacks melanophores; (19) males call from the upper surfaces of leaves; calls undescribed; (20) fighting behavior unknown; (21) eggs deposited on mossy branches over streams; parental care unknown; (22) tadpoles unknown; (23) small body size; SVL in males 21.2–24.8 mm (X = 23.1 mm; *n* = 16); SVL in three adult females: 25.7, 27.0, and 27.0 mm.

**Figure 127.** *Nymphargus anomalus* in life, ZSFQ 2123, from a tributary of Río San Jacinto, 1672 m, Tungurahua province. Photos by Jose Vieira/Tropical Herping.

**Figure 128.** Ontogenetic color change in *Nymphargus anomalus*, QCAZ 48107. Photos by Luis A. Coloma.

**Figure 129.** *Nymphargus anomalus*, adult male, KU 143299. (**A**) Head in lateral view. (**B**) Hand in ventral view. Illustrations by Juan M. Guayasamin.

**Color in life** (Figures 127 and 128): Dorsum pale brown or tan with slight yellow to pink hue; dorsal surfaces of head, body, and limbs with small flecks and black ocelli that enclose orange spots. Anterior part of venter white, posterior half of venter translucent. Iris pale yellow with black reticulations; color of iris becomes whitish towards margins. *Nymphargus anomalus* exhibits ontogenetic variation in its dorsal coloration, being green in postmetamorphs and then gradually acquiring the tan to pale brown coloration that is characteristic of adults. The spotted pattern also changes ontogenetically; postmetamorphs have yellow spots that progressively change towards ocelli composed of black rings with orange centers.

**Color in ethanol:** Dorsum cream to tan with lavender flecks and black and lavender ocelli enclosing whitish centers. Eyelids bear some iridophore aggregations visible through the skin. White pericardium; hepatic and visceral peritonea are translucent. Peritonea covering urinary and gall bladders, kidneys, and testes without iridophores.

**Biology and ecology:** The holotype of *Nymphargus anomalus* was found on a mossy limb of a bush about 1.5 m above a cascading rivulet at night. The following species were found in the same stream and in other small streams nearby: *Hyloscirtus phyllognathus*, *Centrolene pipilata*, *Nymphargus megacheirus*, *N*. *siren*, and *Hyalinobatrachium pellucidum* [22]. At Río Yana Challuwa Yaku, males where found calling on leaves and branches during the night. Two egg clutches were on a mossy branch above the stream. Parental care is unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 130): *Nymphargus anomalus* is known only from four localities (Volcán Sumaco, Río Azuela, Río Yana Challuwa Yaku, nearby Río Jacinto) on the Amazonian slopes of the Ecuadorian Andes, at elevations between 1668–1795 m. The habitat of the species is within the Eastern Montane Forest region ([22], this work).

**Conservation status:** *Nymphargus anomalus* is listed as *Critically Endangered* by the IUCN [243]. Until recently, the species was known from a single individual collected on 23 October 1971 (Río Azuela). Surveys at the type locality have failed in find this species ([91], RWM and JMG, pers. obs.), but other populations have been discovered (Figure 130; Specimens examined). In Ecuador, because of its restricted distribution and habitat fragmentation, we sugges<sup>t</sup> that it should be considered as *Endangered* (IUCN criteria B1, B2a, B2biii).

**Figure 130.** Distribution of *Nymphargus anomalus* in Ecuador (yellow dots).

**Evolutionary relationships** (Figure 101): Lynch [244] suggested that *N*. *anomalus* is the sister species of *N*. *ignotus*. The hypothesis by Lynch is based on the presence of two putative synapomorphies: Pale brown coloration and small ocelli on elevated warts, with the corresponding plesiomorphic conditions being green coloration and no ocelli. Based on mitochondrial genes and a taxon sampling that does not include *N*. *ignotus*, we found that the sister species of *N*. *anomalus* is *N*. *megacheirus*, which has a coloration that matches the plesiomorphic states.

**Specimens examined:** *Nymphargus anomalus:* Ecuador: *Provincia de Napo:* Río Azuela (0.11667 S, 77.6167 W, 1740 m), KU 143299. Parque Nacional Sumaco, Volcán Sumaco, near the Pavayacu refuge (0.61497 S; 77.59065 W; 1771 m), QCAZ 41312–13. *Provincia de Pastaza*: Reserva Comunitaria Ankaku, zona de amortiguamiento del Parque Nacional Llanganates, cabecera del Río Yana Challuwa Yaku (01.26764 S; 78.04797 W; 1668 m), males: QCAZ 45696–97, 45699, 45701, 45703–11, 45728–29; females: QCAZ 45698, 45700, 45702. *Provincia de Tungurahua:* stream tributary of the San Jacinto River (1.3447 S, 78.1814 W; 1795 m asl), ZSFQ 899.

*Nymphargus balionotus* (Duellman, 1981 [120]; Figures 131–133).

*Centrolenella balionota* Duellman, 1981 [120]. Holotype: KU 164702.

Type locality: "3.5 km (by road) northeast of Mindo, 1540 m, Provincia de Pichincha, Ecuador (00◦01- S, 78◦44- W)".

*Cochranella balionota—*Ruiz-Carranza and Lynch, 1991 [6].

*Centrolene balionotum—*Cisneros-Heredia and McDiarmid, 2006 [17].


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

**Etymology:** The specific epithet *balionotus* combines the Greek words *balios* (spotted, dappled) and *notos* (back), and is used in allusion to the dorsal color pattern of the species [120].

**Identification:** *Nymphargus balionotus* is unique by having, in life, a pale green dorsum with reddish–brown dorsolateral stripes, small reddish–brown spots, and larger yellow spots (Figure 131). Additionally, males are small (SVL < 23 mm) and possess a blade-like ventral crest on the humerus that can be confused with a humeral spine. See Remarks.

**Figure 131.** *Nymphargus balionotus* in life, ZSFQ 533. Ecuador, Reserva Río Manduriacu, 1254 m, Tungurahua province. Photos by Jose Vieira/Tropical Herping.

**Diagnosis:** (1) Vomers lacking teeth; (2) snout truncated in dorsal view, protruding lateral profile (Figure 132); (3) tympanum relatively small, oriented almost vertically, with evident lateral and posterior inclinations, its diameter 23%–30% of eye diameter; tympanic annulus mostly visible, with supratympanic fold covering its dorsal margin; tympanic membrane differentiated and translucent, not pigmented; (4) in males, dorsal skin smooth to shagreen, lacking spicules; females unknown; (5) venter slightly granular; pair of enlarged subcloacal warts; (6) anterior half of ventral parietal peritoneum with iridophores, posterior half transparent (condition P2); pericardium covered by iridophores; peritoneum covering digestive tract lacking iridophores (condition V1); (7) liver with four or five lobes, lacking iridophores (condition H0); (8) males with a bladelike ventral crest on the humerus that resembles a small humeral spine; (9) webbing absent between inner fingers; webbing formulae for outer fingers IV (2–2<sup>1</sup>/2)—(2−–2) V (Figure 132); (10) webbing between toes moderate; webbing formula on foot: I (1<sup>1</sup>/3–11/2)—(2–2+) II (1<sup>+</sup>–11/2)—(1–11/2) III (1<sup>1</sup>/3–2)—(2−–3) IV (2−–3)—1<sup>1</sup>/<sup>2</sup> V; (11) ulnar fold low; inner and outer tarsal folds present, but low and di fficult to distinguish; (12) concealed prepollex; nuptial pad in males variable, ranging from Type I to Type III; (13) Finger I slightly longer than Finger II (Finger II length 91.8%–97.4% Finger I); (14) disc of Finger III width about 36%–44% of eye diameter; (15) in life, dorsum pale green with reddish–brown dorsolateral stripes, small reddish–brown spots, and larger yellow spots (Figure 131); color of bones pale green; (16) in preservative, dorsum cream with lavender and white dorsolateral stripes, small dark lavender spots, and larger white spots; (17) iris greyish white with numerous minute brown spots; (18) dorsal surfaces of fingers and toes lacking melanophores; (19) males call from the upper sides of leaves overhanging streams; call undescribed; (20) fighting behavior unknown; (21) egg deposition site unknown; parental care unknown; (22) tadpoles unknown; (23) minute body size; in adult males, SVL 20.1–21.8 mm ( X = 20.8 ± 0.534, *n* = 11); females unknown.

**Figure 132.** *Nymphargus balionotus*, adult male, KU 164708. ( **A**) Head in lateral view. (**B**) Hand in ventral view. Illustrations Juan M. Guayasamin.

**Color in life** (Figure 131): Pale green dorsum with reddish–brown dorsolateral stripes, small reddish–brown spots, and elevated larger yellow spots. Head with reddish–brown interorbital bar and postorbital stripe that is continuous with dorsolateral stripe. Large, elevated yellow spot on anteromedial edge of each eyelid. Pale green bones ([120], this work).

**Color in ethanol:** Dorsal surfaces cream with lavender and white dorsolateral stripes, small dark lavender spots, and larger white spots. White lining covers about anterior half of ventral parietal peritoneum. White pericardium; digestive tract, liver and kidneys covered by translucent peritonea.

**Biology and ecology:** During the night, all individuals at the type locality were calling less than 1 m above the water from the upper surfaces of leaves of herbs and ferns overhanging a trickling stream. Other stream-breeding frogs at the type locality included *Nymphargus grandisonae*, *N*. *gri*ffi*thsi*, and *Hyloscirtus alytolylax* [120]. *Nymphargus balionotus* is nocturnal, arboreal, epiphyllous, and usually found on vegetation along clear-water streams in pristine or in moderately disturbed foothill and cloud forests [87]. Parental care unknown.

**Call:** Not described. **Tadpole:** Not described.

**Distribution** (Figure 133): *Nymphargus balionotus* occurs between 400 and 1540 m along the western slope of the Cordillera Occidental of Colombia, from El Tambito in the Departamento de Cauca, south to Ecuador [21,87,115,116,120,245]. In Ecuador, it is known from localities in the provinces of Carchi, Imbabura, Pichincha, and Cotopaxi at elevations between 1400 and 1540 m (Specimens Examined). It is found within the Western Foothill and Montane Forest ecoregions.

**Figure 133.** Distribution of *Nymphargus balionotus* in Ecuador (yellow dots).

**Conservation status:** *Nymphargus balionotus* is listed by the IUCN as *Vulnerable* at a global level [245]. The species was abundant at the type locality (3.5 km NE of Mindo), where 13 individuals were observed in two nights (7–8 April 1975) [120]. A single reproductive population is known in Ecuador, Reserva Río Manduriacu, which is threatened by mining activities [21]. Thus, in Ecuador, we sugges<sup>t</sup> the category of *Critically Endangered*, following IUCN criteria B2a + B2b(iii).

**Evolutionary relationships:** Lynch and Ruiz-Carranza [246] suggested that *Nymphargus balionotus* is the nearest relative of *Nymphargus armatus* and *N*. *gri*ffi*thsi*, based on the following characters: (i) Snout short and truncate, (ii) slender habitus, (iii) similar webbing and dentition, and (iv) a large bladelike ventral crest present on the humerus. Based on mitochondrial data, a recent study found that *N*. *balionotus* is sister to *N*. *manduriacu* [21].

**Remarks:** Ecuadorian individuals of *Nymphargus balionotus* have a small, pointed humeral spine. The Colombian populations have a variable *crista ventralis* that, in some cases, takes the form of a pointed humeral spine. The polymorphism in this characteristic has created confusion in the generic placement of the species [6,21,139]. It also has been suggested that at least two species are currently confused under the name *Nymphargus balionotus* (M. Rada, pers. com.)

**Specimens examined:** *Nymphargus balionotus:* Ecuador: *Provincia de Carchi:* Cabeceras del Río Baboso (00◦53" N, 78◦27" W; 1400 m), DH-MECN 0865. *Provincia de Pichincha:* 3.5 km NE Mindo (0.0322 S, 78.761 W, 1540 m), KU 164702 (holotype), 164701, 164703–11. *Provincia de Imbabura:* Reserva Río Manduriacu, 1240–1254 m, ZSFQ 0531–33.

**Photographic record:** Ecuador: *Provincia de Cotopaxi:* Río Lomapi, NE of La Maná, Reserva Ecológica Illinizas (0.8270275 S, 79.0843359 W, ca. 1300 m); photo by Martín Bustamante.

*Nymphargus buenaventura* Cisneros-Heredia and Yánez-Muñoz, 2007 [115] (Figures 134 and 135).

*Cochranella buenaventura* Cisneros-Heredia and Yánez-Muñoz, 2007 [115]. Holotype: DHMECN 3563.

Type locality: "Reserva Buenaventura (03◦38- S, 79◦45- W, 1200 m elevation), canton Piñas, Provincia de El Oro, Rep ública del Ecuador".

*Nymphargus buenaventura—*Cisneros-Heredia and McDiarmid, 2007 [17].

**Common names:** English: Buenaventura Glassfrog. Spanish: Rana de Cristal de Buenaventura. **Etymology:** The specific epithet refers to the type locality, Reserva Buenaventura [115].

**Identification:** *Nymphargus buenaventura* can be distinguished from most centrolenids by its green dorsum with small pale yellow to cream spots, reduced webbing between fingers, absence of humeral spine, and absence of iridophores on the digestive visceral peritonea, but with iridophores covering the renal capsules. The last character is shared with at least three other species, *N*. *cariticommatus*, *N*. *gri*ffi*thsi*, and *N*. *wileyi*. *Nymphargus cariticommatus* occurs only on the Amazonian Andean slopes of Ecuador, whereas *N*. *buenaventura* is restricted to the Pacific Andean slopes. *Nymphargus gri*ffi*thsi* di ffers by having dark dorsal spots. *Nymphargus wileyi* di ffers by having a uniform green dorsum and inhabiting the Amazonian slopes of the Ecuadorian Andes. Another species that could be confused with *N*. *buenaventura* is *N*. *lasgralarias*, which lacks the yellow spots characteristic of the former.

**Figure 134.** *Nymphargus buenaventura* in life from Reserva Buenaventura, 1200 m, El Oro province, Ecuador. Photo by Mario Yánez-Muñoz.

**Diagnosis:** *Nymphargus buenaventura* has the following traits: (1) Vomerine teeth absent; (2) snout truncated in dorsal view and profile; nostrils elevated, slightly concave internarial area; (3) lower half of tympanic annulus evident, oriented dorsolaterally with dorsoventral inclination; supratympanic fold absent or very weak; (4) dorsal skin slightly shagreen with scattered flat tubercles corresponding to light spots; (5) ventral skin areolate; pair of large, flat subcloacal warts; subcloacal skin granular and enameled; (6) upper half of parietal peritoneum covered by iridophores (condition P2), all other peritonea transparent, except for renal capsules and pericardium covered by iridophores (condition V1); (7) liver tetralobed, uncovered by iridophores (condition H0); (8) humeral spine absent; (9) webbing absent between Fingers I, II, and III; basal between Fingers III and IV; webbing formulae: III 22/3—21/<sup>2</sup> IV; (10) webbing on feet moderate; webbing formulae: I 2—2<sup>+</sup> II 1<sup>+</sup>—21/<sup>2</sup> III 1—21/<sup>2</sup> IV 21/2—12/<sup>3</sup> V; (11) outer ventral edges of forearms and tarsi with low folds; (12) unpigmented nuptial pad Type I, concealed prepollex; (13) first finger shorter than second, (14) eye diameter larger than width of disc on Finger III; (15) color in life: Dorsum green with scattered pale yellow spots; bones green; (16) color in preservative: Dorsal surfaces pale lavender with scattered cream spots; (17) in life, iris yellowish silver with thin dark maroon reticulations; (18) yellow–green hands with bright yellow discs, melanophores present on outer fingers and outer toes; (19) males call from upper sides of leaves along streams; call undescribed; (20) fighting behavior unknown; (21) egg clutches placed on upper sides of leaves along streams; parental care unknown; (22) tadpoles unknown; (23) small body size; SVL of adult males 20.9–22.4 mm (*n* = 4); SVL in one adult females 23.5 mm.

**Color in life** (Figure 134): Pale green dorsum with pale yellow to cream spots on all dorsal surfaces, including arms and legs. Hands and feet yellowish green with pale yellow discs. Venter cream. Iris yellowish silver with fine maroon reticulations.

**Color in ethanol:** Lavender dorsum with cream spots. Anterior two-thirds of parietal peritoneum white, posterior section translucent; pericardium white; most visceral peritonea clear except for white peritoneum covering the renal capsules.

**Biology and ecology:** Very little information is known on the natural history of *Nympharugus buenaventura*. It has been found on leaves of shrubs and low trees, 1–3 m above streams. A male was found near an egg clutch with 38 embryos [115]. Parental care is unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 135): *Nymphargus buenaventura* is known from three localities in El Oro province (Reserva Buenaventura, Marcabelí, Cascadas de Manuel) and one in Azuay province (Luz María), southwestern Ecuador, within the Deciduous Forest ecoregion. The localities are: Reserva Buenaventura (03◦38-43" S, 79◦45-48" W, 1200 m), Luz María (02◦41-02" S, 79◦25-01" W, 770 m), Marcabelí (03◦44-48" S, 79◦53-36" W, 667 m), and Cascadas de Manuel (03◦12-22" S, 79◦43-34" W, 800 m) [115,247].

**Figure 135.** Distribution of *Nymphargus buenaventura* in Ecuador (yellow dots).

**Conservation status:** *Nymphargus buenaventura* is classified as *Data Deficient* by the IUCN [248]. The species seems to be endemic to southwestern Ecuador, an area with severe deforestation. Thus, we sugges<sup>t</sup> placing the species in the *Endangered* conservation category, following IUCN criteria B1, B2a, B2biii. The species is partially protected by a private reserve (Reserva Buenaventura, managed by Fundación Jocotoco).

**Evolutionary relationships** (Figure 136): *Nymphargus buenaventura* has not ye<sup>t</sup> been included in any molecular phylogeny. However, because of its close morphological resemblance to *N*. *gri*ffi*thsi*, *N*. *wileyi,* and *N*. *cariticommatus*, it is considered as part of the *Nymphargus* clade (sensu Guayasamin et al. [1]).

**Specimens examined:** *Nymphargus buenaventura:* Ecuador: *Provincia de El Oro:* Reserva Buenaventura (03◦38- S, 79◦45- W, 1200 m), DHMECN 3563 (holotype), 2524, 3561, 3562 (paratypes).

**Figure 136.** Evolutionary relationships of glassfrogs in the genus *Nymphargus*, inferred using maximum likelihood and Bayesian criteria.

## *Nymphargus cariticommatus* (Wild, 1994 [137]; Figures 137–139).

*Cochranella cariticommata* Wild, 1994 [137]. Holotype: KU 202806.

Type locality: "11.2 km WSW Plan de Milagro (03◦02- S, 78◦35- W, 2350 m), Provincia Morona-Santiago, Ecuador".

*Nymphargus cariticommatus*—Cisneros-Heredia and McDiarmid, 2007 [17].

**Common names:** English: Unadorned Glassfrog. Spanish: Rana de Cristal Escueta.

**Etymology:** The specific epithet, *cariticommatus*, is an adjective formed from the Latin word *caritus*, meaning without, joined with *Kommos*, a Greek work meaning decoration or embellishment. The name refers to the species' plain appearance and lack of distinctive features [137].

**Identification:** *Nymphargus cariticommatus* is likely to be confused with *Nymphargus wileyi*, *N*. *gri*ffi*thsi*, and *N*. *buenaventura*. However, in life, *N*. *cariticommatus* has a green dorsum with small yellow dots (Figure 137; uniform green dorsum in *N*. *wileyi*), an esophageal peritonea covered by iridophores (translucent in *N*. *gri*ffi*thsi* and *N*. *buenaventura*, and with a thin layer of iridophores in *N*. *wileyi;* but see Remarks), and slightly less hand webbing on the outer fingers than *N*. *buenaventura*. Additionally, *N*. *cariticommatus* is found on the Amazonian versant of the Andes, whereas *N*. *gri*ffi*thsi* and *N*. *buenaventura* inhabit the Pacific versant.

**Figure 137.** *Nymphargus cariticommatus* in life from Ecuador, Zamora Chinchipe province, Reserva Tapichalaca, 2200 m. Photos by Mario Yánez (**left**) and Marco Reyes (**right**).

**Diagnosis:** (1) Vomers lacking teeth; (2) snout round in dorsal profile, and truncated in lateral profile (Figure 138); (3) tympanum evident and of moderate size (tympanum diameter 26.7%–27.1% of eye diameter), dorsal border of tympanic annulus covered by supratympanic fold, tympanic membrane differentiated from surrounding skin; (4) dorsal surfaces shagreen, with minute spicules in males; (5) pair of enlarged subcloacal warts; (6) anterior two-thirds of ventral parietal peritoneum white, posterior third transparent (condition P2); silvery white pericardium; translucent peritonea covering the intestines, stomach, testes, gall bladder, and urinary bladder; esophagus white; kidneys white with unpigmented spots (condition V1); (7) liver tetralobed, three ventral lobes partially covering one smaller lobe; hepatic peritoneum transparent (condition H0); (8) humeral spines absent; (9) no webbing between inner fingers; webbing formula as follows: IV 23/4—22/<sup>3</sup> V in one male, IV 3<sup>+</sup>—23/<sup>4</sup> V in one female; (10) foot webbing moderate: I 2—21/<sup>4</sup> II (1–1<sup>1</sup>/2)—(21/3–22/3) III (1<sup>1</sup>/3–11/2)—(21/2–22/3) IV (2<sup>2</sup>/3–23/4)—2 V (Figure 138); (11) low ulnar fold in males, not visible in females; low inner tarsal fold present in both sexes; (12) concealed prepollex; nuptial pad Type I in males; (13) Finger II slightly longer than Finger I (Finger I about 90%–94% length of Finger II); (14) disc of Finger III of moderate size, 50%–56% of eye diameter; (15) in life, dorsum green with small yellow spots (Figure 137); (16) in ethanol, dorsum lavender with small white spots; (17) iris silvery white with thin black reticulation, yellow ring around pupil; (18) dorsal surfaces of fingers and toes lacking melanophores; (19) males call from the upper side of leaves; call described as soft 'zeet, zeet, zeet', but specific characteristics unknown; (20) fighting behavior unknown; (21) eggs deposited on the upper side of leaves; parental care unknown; (22) tadpoles unknown; (23) small body size; one adult male, SVL 23.3 mm; one adult female, SVL 25.5 mm.

**Figure 138.** *Nymphargus cariticommatus*. Drawings of the holotype, adult male, KU 202806. ModifiedfromWild[137].

**Color in life** (Figure 137): Dorsal surfaces of head, body, and limbs green with small yellow spots. Dorsal surfaces of fingers and toes yellow. Iris silvery white with black reticulations.

**Color in ethanol:** Dorsum of body and limbs lavender with few, small, scattered white spots (fewer in females). Cheek with pale white flecks; margin of lip unpigmented; tympanum with few lavender flecks in males, lacking in females. Hands and feet lacking pigment [137]. Liver covered by transparent peritoneum; white parietal peritoneum covering anterior two-thirds of venter; silvery white pericardium; transparent peritonea covering the intestines, stomach, testes, gall bladder, and urinary bladder; peritoneum around kidneys with a thin silvery white lining; iris silvery white, with purple reticulations. In the original description, Wild [137] stressed the white esophagus in *N*. *cariticommatus*, similar to that seen in *Teratohyla midas*. In the preserved type series, only a thin layer of iridophores is evident.

**Biology and ecology:** *Nymphargus cariticommatus* was found on the upper sides of leaves and ferns within 2 m of cascading streams [137]. Parental care is unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 139): *Nymphargus cariticommatus* is endemic to the Amazonian slopes of the southern Andes of Ecuador at elevations between 2200 and 2700 m ([115,116,137,139], this work). The species has been recorded from localities in the provinces of Zamora Chinchipe and Morona Santiago. The habitat of the species is within the Eastern Montane Forest ecoregion.

**Figure 139.** Distribution of *Nymphargus cariticommatus* in Ecuador (yellow dots).

**Conservation status:** Globally, *Nymphargus cariticommatus* is listed as *Data Deficient* by the IUCN [249]. The last reports of the species come from surveys at the Reserva Biológica Tapichalaca on 2004–2005 ([115], Yánez-Muñoz and Meza-Ramos, pers. com.). In Ecuador, the species is threatened by habitat destruction and fragmentation because of agriculture, pasture lands, and mining. We sugges<sup>t</sup> considering the species as *Endangered* (IUCN criteria B1, B2a, B2biii).

**Evolutionary relationships** (Figure 136): Based on mitochondrial data, *Nymphargus cariticommatus* is sister to *N*. *sucre*.

**Taxonomic Remarks:** As mentioned in the diagnosis, subtle morphological differences separate *Nymphargus cariticommatus* from *N*. *wileyi*. We have examined all the available material of the two species (see Specimens Examined), including the type series, and observed that none of the specimens of *N*. *wileyi* have dorsal spots, whereas all individuals of *N*. *cariticommatus* have them. Also, in the preserved material, the esophageal peritoneum of *N*. *cariticommatus* is more pronounced and extended than in *N*. *wileyi*, in which the iridophores are restricted to the anterior portion of the esophageal peritoneum. Considering the data at hand, we maintain the species status of *N*. *wileyi*.

**Specimens examined:** *Nymphargus cariticommatus*: Ecuador: *Provincia de Morona Santiago:* 11.2 km WSW Plan de Milagro (03◦07- S, 78◦30- W; 2350 m), KU 202806 (holotype), KU 202805; El Cruzado (3.050 S, 78.517 W; 2194 m), USNM 288435–36. *Provincia de Zamora Chinchipe:* Reserva Biológica Tapichalaca (4.49208 S, 79.128389 W; 2200 m), DH-MECN 1974, 2429; 18.1 km E of Loja on the old road to Zamora, 2700 m, QCAZ 33977; Shucos, on the old road from Loja to Zamora, MRy 544.

*Nymphargus cochranae* (Goin 1961 [97]; Figures 140–144).

*Cochranella cochranae* Goin, 1961 [97]. Holotype: BM 1912.11.1.68. Type locality: "El Topo, Río Pastaza, Eastern Ecuador, 4200 feet." *Centrolenella cochranae*—Goin, 1964 [187]. *Cochranella cochranae*—Ruiz-Carranza and Lynch, 1991 [6]. *Nymphargus cochranae*—Cisneros-Heredia and McDiarmid, 2007 [17].

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

**Etymology:** The specific epithet honors Doris M. Cochran (1898–1968), herpetologist and curator of the Smithsonian Institution [97].

**Identification:** *Nymphargus cochranae* can be distinguished from all other glassfrogs by its green dorsum with small dark blue to black ocelli enclosing orange dots (Figure 140), relatively large size (adult males, SVL 24.0–26.2 mm; adult females, SVL 27.8–30.3 mm), absence of webbing between fingers (Figure 141), and lacking humeral spines. *Nymphargus cochranae* further di ffers from other species with dorsal ocellated patterns ( *N*. *anomalus*, *N*. *lindae* sp. nov., *N*. *ignotus*, *N*. *laurae*, *N*. *ocellatus*) by having relatively smaller ocelli (Figure 142). Additionally, *N*. *ignotus* is found on the Pacific slopes of the Andes, whereas *N*. *cochranae* is restricted to the Amazonian slopes of the Andes. *N*. *anomalus* further di ffers by having scattered black and lavender flecks between ocelli (absent in *N*. *cochranae*); *N*. *ocellatus* has a green dorsal coloration with large dark rings that, in life, have greenish white centers; *N*. *laurae*, known from a single adult male is smaller (SVL = 19.7 mm), has pointed papillae on Toes I–IV (absent in *N*. *cochranae*), and larger ocelli. *Nymphargus cochranae* is most similar to its sister species, *N*. *lindae* sp. nov., in female body size ( *N*. *cochranae*, SVL = 27.8–30.3 mm; *N*. *lindae* sp. nov., SVL 27.2–27.8 mm) and the relative size of ocelli (minute *in N*. *cochranae* and clearly larger in *N*. *lindae* sp. nov.), which is the most conspicuous di fference between the two species. The only other species similar to *N*. *cochranae* is *N*. *megacheirus*, the dorsal dots of which coincide with spicules and could be confused with ocelli. However, *N*. *megacheirus* is larger (SVL of males = 26.8–31.5 mm; SVL of females = 31.2–32.9 mm) and the males have conspicuous spicules and spiculated warts on the dorsum (minute spicules in males of *N*. *cochranae*).

**Figure 140.** *Nymphargus cochranae* in life. Ecuador, Volcán Sumaco, 1500 m, QCAZ 31113. Photos by Luis A. Coloma.

**Diagnosis:** (1) Vomers usually with teeth, but see Variation; (2) snout truncated in dorsal profile, and truncated to slightly protruding in lateral profile (Figure 141); (3) tympanum relatively small, its diameter 19.5%–27.8% eye diameter, dorsal border of tympanic annulus covered by supratympanic fold, tympanic membrane pigmented as surrounding skin; (4) dorsal skin of males and females shagreen, minute spicules usually present in males; (5) pair of enlarged subcloacal warts; (6) anterior two-thirds of ventral parietal peritoneum white, posterior third transparent (condition P2); white pericardium; translucent peritonea covering intestines, stomach, testes, kidneys, gall bladder, and urinary bladder (condition V1); (7) liver tetralobed, two large ventral lobes partially covering two smaller lobes; hepatic peritoneum transparent (condition H0); (8) humeral spines absent; (9) hand webbing absent; (10) foot about two-thirds webbed: I (2–2+)—(2<sup>1</sup>/3–21/2) II (1<sup>1</sup>/2–2−)—(3−–3) III (1<sup>3</sup>/4–2)—(3−–3<sup>+</sup>) IV (3−–3<sup>+</sup>)—(2–2<sup>+</sup>) V; (11) ulnar and tarsal folds low; (12) concealed prepollex; nuptial pad Type I in males; (13) Finger I about same length as Finger II (Finger II 96.3%–101.8% of length of Finger I); (14) disc of Finger III small, 19.4%–28.3% of eye diameter; (15) in life, dorsum green with small dark blue to black ocelli enclosing orange dots (Figure 140); bones green; (16) in ethanol, dorsal surfaces lavender with small black ocelli enclosing white dots; (17) iris creamy white with thin dark grey reticulation, pupil surrounded by pale brownish–yellow coloration; (18) dorsal surfaces of hands and feet lacking melanophores, except for proximal portion of Finger IV and Toe V; (19) males call from the upper side of leaves or branches overhanging streams; single, high-pitched note; spectral and temporal characteristics of call unknown; (20) fighting behavior unknown; (21) eggs deposition site unknown; parental care unknown; (22) tadpoles unknown; (23) small to medium body size; SVL of adult males 24.0–26.2 mm ( X = 25.0 ± 0.812, *n* = 6); SVL of adult females 27.8–30.3 mm ( X = 28.9 ± 1.014, *n* = 7).

**Color in life** (Figure 140): Green dorsum with small dark blue to black ocelli enclosing bright orange spots. Ventral surfaces mostly white, except for posterior third, which is translucent. Bones green. Iris greyish white with a thin black reticulation; area immediately around pupil pale brownish yellow.

**Color in ethanol** (Figure 143): Dorsal surfaces lavender with small black ocelli enclosing white spots. Pericardium and anterior two-thirds of ventral parietal peritoneum white (presence of iridophores). Liver, intestines, stomach, testes, kidneys, gall bladder, and urinary bladder without white lining.

**Figure 141.** *Nymphargus cochranae*. Head in lateral and ventral views, adult female, QCAZ 22197. Hand in ventral view, adult male, QCAZ 31113. Illustrations by Juan M. Guayasamin.

**Figure 142.** Dorsal patterns of ocellated glassfrogs, excluding *Nymphargus laurae* and *N*. *lindae* sp. nov. (**A**) *N*. *anomalus*, KU 143299. (**B**) *N*. *cochranae*, KU 121035. (**C**) *N*. *ocellatus*, LSU 25990. (**D**) *N*. *ignotus*, ICN 14750. Figure modified from Lynch and Duellman [22].

**Figure 143.** *Nymphargus cochranae*, holotype in preservative, BMNH 1912.11.1.68. Ecuador, El Topo, Río Pastaza. Photos by Martín Bustamante.

**Variation:** Goin [97] described *Nymphargus cochranae* as lacking teeth on the dentigerous process of the vomer, but also noted the presence of teeth in six other specimens [187]. Most specimens (*n* = 13) examined by us present teeth (USNM 286634 lacks teeth) and we assume that this is the usual character state in the species.

**Biology and ecology:** *Nymphargus cochranae* is active during the night. In the reproductive season, males call at night from the upper surfaces of leaves or branches near streams. Parental care is unknown.

**Call:** Described by Lynch and Duellman [22] as a single, high-pitched note. The specific spectral and temporal characteristics of the call are unknown.

**Tadpole:** Not described.

**Distribution** (Figure 144): *Nymphargus cochranae* occurs in Ecuador and Colombia on the Amazonian slopes of the Andes at elevations between 1170 m and 1960 m. The species has been reported from localities in the provinces of Napo, Orellana, Pastaza, Tungurahua, Sucumbíos, and Zamora Chinchipe (Specimens Examined). There is an unconfirmed record from the Ecuadorian Amazonia (Garza 1, at 300 m; QCAZ 1216). The potential distribution of the species is 21,723 km2, within the Eastern Foothill Forest and Eastern Montane Forest regions. The only record of the species in Colombia comes from the Serranía de Churumbelos [250].

**Figure 144.** Distribution of *Nymphargus cochranae* in Ecuador (yellow dots).

**Conservation status:** Globally, *Nymphargus cochranae*is considered as *Vulnerable* by the IUCN [251]. In Ecuador, the species remains abundant at several localities, including the Sumaco and Cayambe Coca National Parks. However, part of the species' range is fragmented; thus, we sugges<sup>t</sup> considering *N*. *cochranae* as *Near Threatened*.

**Evolutionary relationships** (Figure 136): *Nymphargus cochranae* is sister to *N*. *lindae* sp. nov. However, one sequenced individual (QCAZ 31340) is inferred as sister to a clade formed by *N*. *gri*ffi*thsi* + *N*. *lasgralarias*.

**Taxonomic Remarks:** *Nymphargus cochranae* might be a species complex (see Evolutionary relationships). Also, see taxonomic remarks of *N*. *laurae* and *N*. *lindae* sp. nov.

**Specimens examined:** *Nymphargus cochranae:* Ecuador: *Provincia de Orellana:* km 13 on Loreto–Coca road (0.5836 S, 77.234 W), QCAZ 22196–97; *Provincia Napo:* Cascada San Rafael (0.1127 S, 77.596 W, 1280–1372 m), QCAZ 002, USNM 286632–37, 284304–10; 14.7 km (by road) NE of Río Salado (0.1289 S, 77.608 W, 1310 m), USNM 286638; 14 km by road SW of Reventador (ca. 0.11278 S, 77.596 W; 1400–1500 m), USNM 284304–06; Pacto Sumaco (0.726 S, 77.566 W; 1400 m), QCAZ 31113; Río Salado, 1 km upstream from Rio Coca (0.1916 S, 77.6997 W, 1420 m), KU 164517–18. *Provincia de Pastaza:* Abitagua, 8 km NW Mera (1.41667 S, 78.1667 W, 1300 m), KU 121033–35; *Provincia de Tungurahua:* Río El Topo (1.4166 S, 78.1667 W, 1220 m), BMNH 1912.11.1.68; 11 km E Río Negro (1.433 S, 78.13 W, 1170 m), KU 146605; *Provincia de Sucumbíos:* S slope Cordillera del Due above Río Coca (0.0833 N, 77.66 W, 1150 m), KU 123216–18. *Provincia de Zamora Chinchipe:* Estación Científica San Francisco (3.967 S, 79.066 W, 1960 m), QCAZ 31340–41.

**Localities from the literature:** *Nymphargus cochranae:* Ecuador: *Provincia de Zamora Chinchipe:* Alto Nangaritza (4.25026 S, 78.61746 W; 1256–1430 m) [252]; Contrafuerte de Tzunantza (4.046 S, 78.922 W) [253]; Alto Machinaza 1 (3.8976 S, 78.482 W) [254], Alto Machinaza 2 (3.7708 S, 78.554 W) [254]. *Nymphargus colomai* new species Guayasamin and Hutter (Figure 145).

http://zoobank.org/urn:lsid:zoobank.org:act:46978D55-22DF-409E-870C-F3C2203A5E6E

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

**Etymology:** The specific name is a patronym for Luis A. Coloma in recognition of his pioneer and continual efforts in studying and protecting amphibians, as well as mentoring numerous students, including JMG. Luis Coloma is the Director of the Jambatu Center for Research and Conservation of Amphibians (Centro Jambatu de Investigación y Conservación de Anfibios; see http://www. anfibiosecuador.ec/). The Jambatu Center houses dozens of critically endangered amphibians, in an outstanding effort to conserve frogs and toads. Luis received the *Sabin Award* for Amphibian Conservation in 2007 (http://www.amphibians.org/grants/sabin-award/).

**Holotype:** QCAZ 41590, adult male, collected from Miazi Alto (4.25044◦ S, 78.61356◦ W; 1260 m), Cordillera del Cóndor, Provincia de Zamora Chinchipe, Ecuador, by Juan M. Guayasamin and Elicio Tapia on 8 April 2009.

**Paratypes:** QCAZ 41591–92, adult males, and QCAZ 41641, juvenile, collected by Juan M. Guayasamin, Elicio Tapia, Silvia Aldás, and Holger Braun on 8–12 April 2009, at the same locality as holotype.

**Generic placement:** The new species is placed in the clade *Nymphargus* (sensu Guayasamin et al. [1]) based on morphological and molecular data. All species in *Nymphargus* share an absence of webbing between Fingers I–III and an absence or reduced webbing between Fingers III and IV; additionally, males lack humeral spines (except *N*. *grandisonae*). *Nymphargus colomai* sp. nov. has the aforementioned traits. Molecular analyses of mitochondrial genes unambiguously place the new species in the clade *Nymphargus* (Figure 16).

**Identification:** *Nymphargus colomai* sp. nov. is unique by having a white iris with a contrasting black horizontal stripe and a dorsum that varies from dull yellowish green to tangerine yellow, brown, or grey olive, always with numerous small yellow to orange spots (Figure 145). Additionally, *N*. *colomai* sp. nov. lacks humeral spines and hand webbing. Among glassfrogs that inhabit the Amazonian slopes of the Andes, only *N*. *cariticommatus*, *N*. *chancas*, *N*. *humboldti* sp. nov., *N*. *siren*, and *N*. *sucre* could be confused with *N*. *colomai* sp. nov. However, *N*. *cariticommatus*, *N*. *chancas*, *N*. *siren*, and *N*. *humboldti* sp. nov. have a green dorsum with few and well-defined dorsal spots, whereas *N*. *colomai* sp. nov. has a non-green dorsum (see above) with numerous and diffuse yellow dorsal spots. Moreover, *N*. *siren* is smaller than *N*. *colomai* sp. nov. (*N*. *siren*, SVL < 23.3 mm; *N*. *colomai* sp. nov. SVL = 24.7–25.7 mm); and *N*. *cariticommatus* has white esophagus and renal capsules (cream in *N*. *colomai* sp. nov.) Both *N*. *colomai* sp. nov. and the Peruvian *N*. *chancas* share the clearly defined black horizontal stripe on a silver background, but the two species are easily differentiated by their dorsal color patterns (in *N*. *colomai* sp. nov.*:* Dull yellowish green to tangerine yellow, brown, or grey olive, with numerous small yellow to orange spots; in *N*. *chancas:* Yellowish–green dorsum with small yellow dots; compare Figures 145 and 146) and genetics (Figure 136).

**Figure 145.** *Nymphargus colomai* sp. nov. in life from stream nearby Miazi Alto, Zamora Chinchipe province, Ecuador. (**Top left**): QCAZ 41590, holotype; other frogs are part of the type series. Photos by Holger Braun and Juan M. Guayasamin.

**Figure 146.** *Nymphargus chancas* in life from 5.9 km SE from the type locality of Abra Tangarana, San Martín, 6◦19-15.65" S 76◦41-44.16" W, 973 m, CORBIDI 10471. Photos by Jesse Delia (**upper row**) and Evan Twomey (**lower row**).

**Diagnosis:** (1) Dentigerous process of the vomer lacking teeth or with few teeth (up to 3); (2) snout truncated to slightly protruding in lateral profile; truncated in dorsal view; (3) tympanum oriented almost vertically, with slight lateral and posterior inclinations, its diameter about 23%–27% of eye diameter; upper half of tympanic annulus obscured by supratympanic fold and warts; tympanic membrane pigmented as surrounding skin; (4) dorsal skin shagreen, with numerous spiculated warts and spicules in males; females unknown; (5) venter areolate; pair of enlarged subcloacal warts; (6) white parietal peritoneum covering about anterior half of venter (condition P2); white pericardium; translucent peritonea on kidneys, intestines, stomach, gall and urinary bladders (condition V1); (7) liver lobate, covered by translucent peritoneum (condition H0); (8) humeral spines absent; (9) webbing absent between Fingers I, II, and III, absent or basal between Fingers III and IV; webbing formula III (2<sup>3</sup>/4–3)—(21/2–22/3) IV; (10) feet about two-thirds webbed; webbing formula: I 2−—21/<sup>3</sup> II (1<sup>+</sup>–11/3)—21/<sup>2</sup> III 11/2—(22/3–23/4) IV <sup>3</sup>−—(1<sup>1</sup>/<sup>2</sup> – 2−) V; (11) ulnar and tarsal folds present, low; (12) concealed prepollex; in males, nuptial pad Type I; (13) Finger II slightly longer than Finger I; (14) disc of Finger III width about 46%–50% of eye diameter; (15) in life, dorsum varies from dull yellowish green to tangerine yellow, brown, or grey olive with numerous small yellow to orange spots; bones green; (16) in preservative, dorsum grey to greyish lavender with numerous unpigmented spots; (17) in life, iris silvery white with slight yellow hue and a clearly marked horizontal black stripe; (18) melanophores on dorsal surfaces of Fingers III and IV and Toes IV and V; (19) males call from the upper surfaces of leaves; call unknown; (20) fighting behavior unknown; (21) egg deposition site unknown; parental care unknown; (22) tadpole unknown; (23) small body size; in males, SVL 25.0–25.7 mm (*n* = 3); females unknown. **Description of holotype:** Adult male, SVL 25.0 mm. Head slightly wider than long (head length 98% of head width); snout truncated in dorsal and lateral profiles; canthus rostralis indistinct, slightly concave; loreal region slightly concave; lips slightly flared; nostril protuberant, closer to tip of snout than to eye, directed frontolaterally; internarial area barely depressed. Eyes large, directed anterolaterally at an angle ~50◦; transverse diameter of disc of Finger III 48% eye diameter. Supratympanic fold low, obscuring upper portion of tympanic annulus; tympanum oriented mostly vertically, but with slight posterolateral inclination; tympanic membrane translucent, pigmented as surrounding skin. Dentigerous process of vomer low, situated transversely between choanae, with one or two teeth; choanae large, longitudinally rectangular; tongue ovoid, with ventral posterior fifth not attached to floor of mouth and posterior margin slightly notched; vocal slits extending posterolaterally from about the lateral margin of tongue (at about half the length of tongue) to angle of jaws. Humeral spine absent. Low ulnar folds evident on external and internal ventrolateral margins of arm; relative lengths of fingers: III > IV > II > I; webbing absent between Fingers I–III, basal between Fingers III and IV, webbing formula III 3–22/<sup>3</sup> IV; discs expanded, nearly elliptical; disc pads nearly triangular shaped; subarticular tubercles small, round, simple; low supernumerary tubercles present; palmar tubercle elliptical, simple; nuptial pad large (Type I), ovoid, granular, extending from ventrolateral base to dorsal surface of Finger I, covering proximal half of Finger I. Length of tibia 57% SVL; low inner tarsal fold evident; outer tarsal fold absent; foot two-thirds webbed; webbing formula of foot: I 2−—21/<sup>3</sup> II 11/4—21/<sup>2</sup> III 11/2—23/<sup>4</sup> IV 3−—2− V; discs on toes round to elliptical, lacking papillae; disc on Toe IV slightly narrower that disc on Finger III; disc pads triangular; inner metatarsal tubercle large, ovoid; outer metatarsal tubercle small and inconspicuous; subarticular tubercles small, round; supernumerary tubercles low. Skin on dorsal surfaces of head, body, and lateral surface of head and flanks shagreen with numerous spiculated warts and minute spicules; lower flanks show glandular cells; throat smooth; belly and lower flanks areolate; cloacal opening directed posteriorly at upper level of thighs; cloacal warts present, unpigmented. Ventral surface of thighs with pair of enlarged tubercles.

**Coloration of holotype in life** (Figure 145): The holotype has a grey olive dorsal coloration, with numerous yellow to orange spots. The venter is white anteriorly and translucent posteriorly. The iris is silvery white, with a clearly marked black horizontal stripe. Dorsally, at the posterior end of the head, there is a circular, concave area with dark grey collocation.

**Color variation in life** (Figure 145): Dorsum can be dull yellowish green, tangerine yellow, brown, or grey olive, always with numerous small and diffuse yellow to orange spots. Anterior half of venter white, posterior portion transparent. Iris might have a slight yellow hue.

**Coloration of holotype in ethanol:** Dorsal surfaces of head, body, and limbs greyish brown, with small, unpigmented spots. Middorsal dark lavender spot at scapular level. Anterior half of ventral parietal peritoneum white, posterior half cream. White pericardium; translucent peritonea covering digestive tract, liver, kidneys, and gall and urinary bladders.

**Color variation in ethanol:** Dorsal surfaces of head, body, and limbs vary from greyish brown to lavender with small, unpigmented spots.

**Measurements of holotype (mm):** *Nymphargus colomai* sp. nov., adult male, QCAZ 41590: SVL = 25.0, head length = 8.1, head width = 8.3, snout length = 3.3, IOD = 2.6, upper eyelid width = 2.5, eye diameter = 3.3, tympanum diameter = 0.9, tympanum–eye distance = 1.7, femur = 13.1, tibia = 14.2, foot length = 11.5, hand length = 5.7, disc of Finger III width = 1.6, disc of Toe IV width = 1.5.

**Biology and ecology:** *Nymphargus colomai* sp. nov. is active during the night. All individuals were found on vegetation 100–170 cm above fast-flowing streams. Males call from the upper surfaces of leaves. Parental care is unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 147): *Nymphargus colomai* sp. nov. is known only from its type locality at the Cordillera del Cóndor, Zamora Chinchipe province, Ecuador.

**Figure 147.** Distribution of *Nymphargus colomai* sp. nov. in Ecuador (yellow dot).

**Conservation status:** Following UICN criteria, we consider *Nymphargus colomai* sp. nov. as *Endangered*, following IUCN criteria B2a, B2(iii). The main threats for this species are habitat destruction (i.e., cattle, agriculture) and contamination associated with mining.

**Evolutionary relationships** (Figure 136): *N. colomai* sp. nov. is inferred as sister to *N*. *mariae*.

**Taxonomic comments:** *Nymphargus colomai* sp. nov. was previously confused with *N*. *chancas*[252], which we restrict to Peru. Preserved specimens are morphologically similar, but in life, several differences are evident (Figures 145 and 146). The two species are not closely related (Figure 136).

*Nymphargus garciae* (Ruiz-Carranza and Lynch, 1995 [255]; Figures 148 and 149).

*Cochranella garciae* Ruiz-Carranza and Lynch, 1995 [255]. Holotype: ICN 11752.

Type locality: "Departamento de Cauca, municipio de Inzá, Km 64–73 carretera Popayán a Inzá, vertiente oriental Cordillera Central, 2◦34- latitud N, 76◦4- W de Greenwich, 2590–2660 m [Colombia]".

*Nymphargus garciae*—Cisneros-Heredia and McDiarmid, 2007 [17].

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

**Etymology:** The specific epithet is a patronym for Dr. Evaristo García, author of the pioneer book *Los ofidios venenosos del Cauca*, published in 1896 [255].

**Identification:** *Nymphargus garciae* can be di fferentiated from other glassfrogs by its moderate body length (SVL 25.9–28.4 mm in adult females, SVL 25.1–29.9 mm in adult males) ([255], this work), green dorsum usually with dark green to blue spots, and absence of humeral spines. In Ecuador, *N*. *garciae* could be confused with *Centrolene buckleyi* and *N*. *megacheirus*. However, *C*. *buckleyi* has basal webbing between Fingers III and IV (absent in *N*. *garciae*), lacks dark spots on the dorsum (usually present in *N*. *garciae*), and males with prominent humeral spines (spines absent in *N*. *garciae*). *Nymphargus megacheirus* has a similar dorsal coloration and hand-webbing pattern as *N*. *garciae*, but the two species have allopatric distributions, *N*. *garciae* inhabiting higher elevations (1900–2700 m) than *N*. *megacheirus* (1300–1740 m). Additionally, *N*. *megacheirus* is slightly larger than *N*. *garciae* (in *N*. *megacheirus*, SVL 26.8–31.5 mm in adult males, 31.2–32.9 mm in adult females). Two species endemic to Colombia, *N*. *nephelophilus* and *N*. *oreonympha*, are extremely similar to *N*. *garciae*. *Nymphargus oreonympha* is reported to have basal webbing between Fingers III and IV (webbing absent in *N*. *garciae*), whereas *N*. *nephelophilus* has a smooth skin (shagreen in *N*. *garciae*) and lacks ulnar and tarsal folds (present in *N*. *garciae*). See Remarks.

**Figure 148.** *Nymphargus garciae* in life from Ecuador, Napo province, trail from Oyacachi to El Chaco, 3012 m, MZUTI 764. Photos by Eduardo Toral.

**Diagnosis:** (1) Vomerine teeth absent; (2) snout rounded in dorsal aspect, truncated to slightly sloping in lateral profile; (3) tympanum oriented almost vertically, with slight posterior inclination, small in size (tympanum diameter 22.9%–30.6% of eye diameter); tympanic annulus visible except for dorsal border covered by supratympanic fold; tympanic membrane partially pigmented but di fferentiated from surrounding skin; (4) dorsal surfaces shagreen, with spicules evident in males; (5) pair of enlarged subcloacal warts (Figure 15); (6) anterior half of parietal peritoneum white (condition P02; white pericardium and renal capsules, translucent visceral peritoneum (condition V1); (7) tetralobed liver covered by transparent peritoneum (condition H0); (8) humeral spines absent; (9) no webbing between fingers, although junction of lateral fringes resemble basal webbing; (10) webbing formula on foot: **I** (2−–2<sup>+</sup>)—(2<sup>+</sup>–22/3) **II** (1–1<sup>3</sup>/4)—(21/2–3−) **III** (1<sup>1</sup>/4–13/4)—(21/3–3−) **IV** (2<sup>1</sup>/3–3+)—(1<sup>3</sup>/4–2+) V; (11) ulnar and tarsal folds present, enameled; (12) nuptial pad Type I, concealed prepollex; (13) Finger II slightly longer than Finger I (FI/FII = 0.87–0.95); (14) disc of Finger III relatively large (3DW/ED = 0.56–0.77); (15) in life, dorsum green with or without green to blue spots; white upper lip and ulnar and tarsal

folds; bones green; (16) in preservative, dorsum lavender with or without dark lavender spots; (17) iris greyish white with thin black reticulation; (18) fingers lacking melanophores dorsally; melanophores restricted to proximal dorsal areas of Toes IV and V; (19) males call from upper side of leaves; call unknown; (20) fighting behavior unknown; (21) eggs deposition site unknown; parental care unknown; (22) tadpoles unknown; (23) medium body size; in Ecuador, males SVL 24.9–29.9 mm (X = 26.7 ± 1.723; *n* = 7); in Colombia, males SVL 25.1–27.7 mm (X = 26.3 ± 0.745; *n* = 15), females SVL 25.9–28.4 mm (*n* = 3).

**Color in life** (Figure 148): Dorsum dark green with cream tubercles and dark green to blue spots; hands and feet yellowish green. Glandular pericloacal region white. Pericardium and anterior half of ventral parietal peritoneum white; translucent visceral peritonea. Based on observations on preserved specimens, we assume that part of the type series of *Nymphargus garciae* (ICN 11743, 11759) was uniform green in life, corresponding to the uniform lavender coloration in ethanol. Some of the photographs of Ecuadorian individuals of *N*. *garciae* show a uniform green dorsal coloration.

**Color in ethanol:** Dorsal surfaces lavender with or without small dark spots; spicules resemble minute white spots in males. White upper lip; tympanic membrane pigmented with purple specks. Cloacal region with white warts. Dorsally, fingers and toes mostly unpigmented, but some pigmentation visible on proximal half of Toes IV and V. White parietal peritoneum covering anterior half of venter; silvery–white pericardium; transparent peritoneum covering liver and urinary bladder; gastrointestinal peritoneum translucent; renal capsules white with minute unpigmented spots, as those in *N. wileyi* ([20]: Figure 12). Internal traits based on a dissected male (ICN 11759).

**Biology and ecology:** Adults of *Nymphargus garciae* were found on vegetation and rocks along streams; one gravid female (ICN 7495) had numerous eggs, which had a dark brown animal and a cream vegetal pole [255]. Parental care is unknown.

**Call:** Not described.

**Tadpole:** Not described.

**Distribution** (Figure 149): *Nymphargus garciae* is known from localities on the eastern slope of the Cordillera Central at elevations of 1900–2700 m in Colombia [101,255], and from the Cordillera Oriental of the Ecuadorian Andes at 2550–3012 m. In Ecuador, this species is found in the Eastern Montane Forest ecoregion.

**Figure 149.** Distribution of *Nymphargus garciae* in Ecuador (yellow dots).

**Conservation status:** *Nymphargus garciae* is classified as *Vulnerable* by the IUCN [256]. We consider the current conservation status justified.

**Evolutionary relationships** (Figure 136): *N. garciae* is the sister species to *N*. *vicenteruedai*.

**Remarks:** Given the morphological similarities between *Nymphargus garciae*, *N*. *nephelophilus*, and *N*. *oreonympha*, it is possible that they are actually one species with geographic variation.

**Specimens examined:** *Nymphargus garciae:* Ecuador: *Provincia de Napo*: 11 km ESE Papallacta (0.38694◦ S, 78.05694◦ W; 2660 m), KU 164658–62; 60 km E San Miguel de Salcedo (0.96667◦ S, 78.21667◦; 2550 m), KU 202793; trail from Oyacachi to El Chaco (0.21891◦ S, 78.04442◦; 3012 m), MZUTI 764. *Provincia de Sucumbíos*: 18 km E Santa Bárbara (0.575◦ N, 77.51138◦ W), KU 202796.

*Nymphargus grandisonae* (Cochran and Goin, 1970 [96]; Figures 150–153).

*Centrolenella grandisonae* Cochran and Goin, 1970 [96]. Holotype: BM 1910.7.11.68. Type locality: "Pueblo Rico, (Departamento) Caldas, southwestern Colombia, 5000 feet." *Centrolene grandisonae*—Ruiz-Carranza and Lynch, 1991 [6].

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

**Common names:** English: Red-spotted Glassfrog, Measles' Glassfrog, Grandison's Glassfrog. Spanish: Rana de Cristal Sarampiona, Rana de Cristal de Grandison.

**Etymology:** The specific epithet is a patronym for Alice G. C. Grandison, former curator of the British Museum [96].

**Identification:** *Nymphargus grandisonae* is unique by having a green dorsum with small bright red spots (Figure 150). Another diagnostic trait is the presence of a white urinary bladder, a characteristic otherwise known only in glassfrogs from the Atlantic forest of Brazil and Argentina.

**Figure 150.** *Nymphargus grandisonae* in life. (**A**) Adult male, QCAZ 40001. (**B**) Adult female, ventral view, QCAZ 32282. (**C**) Metamorph. (**D**) Egg clutch, QCAZ 40004. (**E**) Fight between males. Photos by: (**A**,**C**,**D**) Luis A. Coloma, (**B**) Martín Bustamante, (**E**) Carl R. Hutter.

**Diagnosis:** (1) Dentigerous process of vomer usually bearing teeth, each process with zero to seven teeth; (2) snout round in dorsal view, truncated in lateral profile (Figure 151); (3) tympanum partially hidden under skin, oriented almost vertically, its diameter 27.0%–34.5% of eye diameter; supratympanic fold moderate; tympanic membrane pigmented, barely differentiated from surrounding skin; (4) dorsal surfaces of males and females finely pustular, minute spicules evident only in males; (5) pair of enlarged subcloacal warts; (6) white iridophores covering all or most of ventral parietal peritoneum (condition P4); white pericardium; translucent peritoneum covering intestines, stomach, testes, and gall bladder; kidneys dorsally and laterally covered with white lining; urinary bladder completely covered with white iridophores (condition V4); (7) liver tetralobed, covered by transparent peritoneum (condition H0); (8) in males, humeral spines present, small; (9) webbing absent between Fingers I, II, and III; moderate webbing between Fingers III and IV; webbing formula III (2<sup>1</sup>/3–21/2)—(2–21/4) IV (Figure 151); (10) webbing formula on foot, I (1–1<sup>3</sup>/4)—(2–21/4) II (1–1<sup>1</sup>/2)—(2–21/2) III (1–1<sup>1</sup>/2)—(2–21/4) IV (2–2<sup>1</sup>/3)—(1–11/2) V; (11) ulnar fold low, white; inner tarsal fold low, short; outer tarsal fold absent or low and inconspicuous; (12) concealed prepollex; in males, nuptial pad Type I; (13) Finger I about same length as Finger II (Finger I 92.0%–102.2% of Finger II); (14) disc of Finger III of moderate width, about 55.2%–64.0% of eye diameter; (15) in life, dorsum green with small red spots; upper lip white; bones green; (16) in preservative, dorsal surfaces lavender with cream white spots; (19) males call from upper sides of leaves near streams; advertisement call formed by a single note, note duration 0.056–0.158 s, 18–22 pulses per note, dominant frequency at 3100–4048 Hz; (20) fighting behavior variable (see Biology and ecology section); (21) females deposit eggs on upper side of leaves near streams; short-term maternal care present; parental care by males absent; (22) tadpoles with M-shaped upper jaw sheath, small gap in row A-2, and non-emarginate oral disc; (23) medium body size; males, SVL 25.1–29.3 mm (X = 27.2, *n* = 44); females, SVL 28.9–30.7 mm (X = 29.8, *n* = 4).

**Figure 151.** *Nymphargus grandisonae*. (**A**) Head in lateral view, KU 164688, drawing by Juan M. Guayasamin. (**B**) Hand in ventral view, KU 164688. (**C**) Foot in ventral view, KU 118047. (**B**,**C**) Modified from Lynch and Duellman [22], not to scale.

**Color in life** (Figure 150): Dorsum green with small red spots; throat pale green; margin of upper lip white; flanks and venter white; iris yellowish grey; bones green; peritonea on liver and digestive tract lacking iridophores.

**Color in ethanol:** Dorsal surfaces of head and body lavender with creamy–white spots; flanks white. White parietal peritoneum covering all or most of venter; pericardium white; translucent peritoneum covering liver, intestines, and stomach; kidneys dorsally and laterally covered with white iridophores; white urinary bladder.

**Biology and ecology:** *Nymphargus grandisonae*is a nocturnal and epiphyllous glassfrog that prefers riparian vegetation in evergreen and cloud forests. The species is active on vegetation near streams in primary and secondary forest, and on shrubs along the borders of pastures [51,87]. *Nymphargus grandisonae* is a highly territorial species, with males having site fidelity for prolonged periods of time [51]. Also, males have frequent fights and complex combat behaviors (Figure 6), including the following types: (1) Dangling amplexus-like; (2) dangling venter to venter; (3) amplexus-like; (4) a head to vent wrestle; and (5) reverse dangling amplexus-like. As a consequence of fighting, males usually present injuries produced, likely, by humeral spines (see Hutter et al. [51]; Figure 6). Interspecific combat has also been observed between *N*. a ff. *grandisonae* and *Espadarana prosoblepon* [257]. *Nymphargus grandisonae* reproduces in the rainy season (December–April); at this time of the year, males call from the upper sides of leaves. Amplexus occurs within male's territories and egg clutches, containing 30–71 eggs, are deposited on vegetation above streams [51]. At Reserva Las Gralarias, the species has been found at Ballux Creek, Five-frog Creek, Heloderma Creek, Hercules Creek, Chalguayacu River, Kathy's Creek, Lucy's Creek, Waterfall Trail, and Santa Rosa River Trail ([88], this work). Short-term parental care is provided by females; no parental care is provided by males [25].

**Call:** Hutter et al. [51] presented a detailed description of the advertisement, courtship, territorial, encounter, distress, and release calls of *Nymphargus grandisonae*. The information of these calls is summarized in Tables 5–7 and Figure 152.

**Figure 152.** Call types in *Nymphargus grandisonae*. ( **A**) Principal component analyses (PCA) of call variables for di fferent types of calls of all individuals. (**B**) PCA plot showing call variation within an individual. ( **C**–**E**) Oscillogram, spectrogram, and power spectrum of di fferent types of calls. Modified from Hutter et al. [51].


**Table 5.** Call types and associated behaviors described for *Nymphargus grandisonae* [51]. Definitions follow Bogert [258], Duellman and Trueb [259], and McDiarmid and Adler [238].

> **Table 6.** Description of combat types observed for *Nymphargus grandisonae* [51].


**Table 7.** Comparisons of call types recorded for *Nymphargus grandisonae*. Data are the mean ± standard deviation, followed by the range (modified from Hutter et al. [51]).


**Tadpole:** The tadpole of *Nymphargus grandisonae* was described by Ospina-Sarria et al. [260]. Characteristic traits include M-shaped upper jaw sheath, small gap in row A-2, and a non-emarginated

oral disc. In life, tadpoles (Gosner stage 36) exhibit a cream coloration; hyobranchial apparatus and heart are pinkish. Caudal musculature is reddish cream; caudal and ventral fins are translucent, with small darks spots at the distal end of the tail.

**Distribution** (Figure 153): *Nymphargus grandisonae* occurs on the western slopes of the Cordillera Occidental and Cordillera Central in Colombia, from Departamento de Antioquia, south to the Pacific versant of the Andes of north and central Ecuador [51,87,96,127,261,262]. In Ecuador, *N*. *grandisonae* has been found in the provinces of Carchi, Cotopaxi, Pichincha, El Oro, and Santo Domingo de los Tsáchilas, at elevations between 1140–2150 m (Specimens Examined). In Ecuador, the potential distribution of the species is 8087 km2, mostly within the Western Montane Forest ecoregion, but also including the Deciduous and Western Foothill ecoregions.

**Figure 153.** Distribution of *Nymphargus grandisonae* in Ecuador (yellow dots).

**Conservation status:** *Nymphargus grandisonae* is listed as a *Least Concern* species by the IUCN [261] and Arteaga et al. [87]. The species tolerates a certain level of forest disturbance. In Ecuador, there are several recent reports of *N*. *grandisonae* in Chiriboga, Reserva Río Guajalito, Reserva Las Gralarias, Tandayapa, Tandapi, and Reserva Otonga. At Reserva Las Gralarias, this glassfrog is infected by the chytrid fungus *Batrachochytrium dendrobatidis*, but populations have not shown evidence of drastic declines and the species is fairly common in the area [92]. Experimental studies have shown that tadpoles of *N*. *grandisonae* are susceptible to introduced trout, by having a higher mortality and changing its morphology [123]. We consider that the *Least Concern* conservation status is adequate.

**Evolutionary relationships** (Figure 136): *Nymphargus grandisonae* is sister to a clade formed by *N*. *mariae* and *N*. *colomai* sp. nov.

**Taxonomic remarks:** The name "*Centrolenella grandisonae*" was misapplied by Lynch and Duellman [22], who confused *N*. *grandisonae* with *Centrolene lynchi*. Duellman [127] provided a detailed description of these two species. There are reports [257] of *N. grandisonae* at Reserva Buenaventura (El Oro province, Ecuador) that, most likely, represent a new species (Paul Szekely, Diego F. Cisneros-Heredia, pers. comm.)

**Specimens examined:** *Nymphargus grandisonae:* Ecuador: *Provincia de Carchi:* Quebrada Naranjo, near Maldonado (0.9 N, 78.1 W; 1410 m), KU 178168–69. *Provincia de Cotopaxi:* Reserva Otonga (0.4189 S, 79.004 W; 1800 m), QCAZ 20718, 20725, 11683; 18.2 km on the Quillotuña–Pucayacu road (0.6784 S,

79.01564 W), QCAZ 40388. *Provincia de Pichincha:* 3.5 km NE of Mindo (0.0322 S, 78.761 W, 1340 m), KU 164686–90; Quebrada Zapadores, 5 km ESE of Chiriboga on Chiriboga–Quito road (0.2375 S, 78.735278 W; 2010 m), QCAZ 16288; near Tandapi (0.4164 S, 78.7989 W; 1520 m), KU 180319–22; 5 km Won the Tandapi-Atenas road (0.3954 S, 78.7989 W; ca. 1700 m), QCAZ 14292–93, 17744, 17753–58; 25.7 km ENE La Palma on La Palma-Chiriboga road, 1820 m, MCZ 93587; 2 km E Tandapi, (0.4258 S, 78.7853 W; 1550 m), MCZ 93023–26, 97848–51, USNM 211211; 1 km SW Tandayapa (0.033 S, 78.7667; 1640 m), MCZ 97847, USNM 211214; 2.9 km SW Tandayapa on Tandayapa-Mindo road (0.05 S, 78.783 W; 1820 m), USNM 211212–13; 5.1 km SE Tandayapa on Tandayapa-Nono road (0.033 S, 78.7167 W; 1850 m), USNM 211215; Reserva Las Gralarias, Five-frog Creek (0.03098 S, 78.70853 W; 2150 m), MZUTI 430–31. *Provincia de Santo Domingo de los Tsáchilas:* 4 km NE Dos Ríos (0.30278 S, 78.8678 W; 1140 m), KU 164670–85; 1 km W of Río Faisanes and its intersection with Chiriboga-La Palma road (0.3035 S, 78.869 W), QCAZ 15364.

**Localities from the literature:** *Nymphargus grandisonae:* Colombia: Serranía de los Paraguas (Boquerón; 04◦44.2- N, 76◦18.3- W, 2000 m) [260]. Ecuador:

*Nymphargus gri*ffi*thsi* (Goin, 1961 [97]; Figures 154–157).

*Cochranella gri*ffi*thsi* Goin, 1961 [97]. Holotype: BM 1940.2.20.4. Type locality: "Río Saloya, Ecuador, 4000 feet". Centrolenella griffithsi—Goin, 1964 [187]. *Cochranella gri*ffi*thsi—*Ruiz-Carranza and Lynch, 1991 [6]. *Nymphargus gri*ffi*thsi—*Cisneros-Heredia and McDiarmid, 2007 [17].

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

**Etymology:** The specific epithet honors Dr. Ivor Griffiths, of Birkbeck College, London, in recognition of his contributions to the understanding of relationships among frogs [97].

**Identification:** Among glassfrogs found on the Pacific slope of the Andes, *Nymphargus gri*ffi*thsi* can be distinguished by having a uniformly green dorsum with small dark flecks (Figure 154), and by lacking webbing between fingers. The species also lacks humeral spines, although enlarged humeral crests are apparent in some specimens (Figure 155). Only two species, *N*. *buenaventura*, a species restricted to southwestern Ecuador (Reserva Buenaventura, Provincia de El Oro), and *N*. *lasgralarias*, from Reserva Las Gralarias (Pichincha Province), can be confused with *N*. *gri*ffi*thsi*, and no distinctive morphological traits distinguish among these species. However, *N*. *buenaventura* is slightly smaller and has a green dorsum with yellow spots, whereas populations of *N*. *gri*ffi*thsi* lack these yellow spots and have dark flecks on the dorsum (see Taxonomic Remarks). The most reliable traits to differentiate between *N*. *lasgralarias* and *N*. *gri*ffi*thsi* are the calls; *N*. *gri*ffi*thsi* produces a single tonal or multi-pulsed (i.e., two or more pulses) call, while the calls of *N*. *lasgralarias* are always pulsed. Also, *Nymphargus gri*ffi*thsi* emits its advertisement call as a single note, whereas *N*. *lasgralarias* emits its calls singly or in a series. In addition, *N*. *lasgralarias* has shorter call duration than *N*. *gri*ffi*thsi* (call duration in *N*. *lasgralarias* = 0.016–0.044 s; call duration in *N*. *gri*ffi*thsi* = 0.103–0.148 s) [88]

**Diagnosis:** (1) Vomers lacking teeth; (2) snout truncated in dorsal view and truncated to protruding in profile; (3) lower half of the tympanic annulus evident, oriented dorsolaterally with dorsoventral inclination; tympanic membrane pigmented as surrounding skin; supratympanic fold absent or very weak; (4) dorsal skin slightly shagreen; (5) pair of enlarged subcloacal warts; (6) upper half of ventral parietal peritoneum white, posterior half transparent (condition P2); white pericardium, all other peritonea clear (condition V1); (7) liver tetralobed; hepatic peritoneum transparent (condition H0); (8) humeral spines absent, although enlarged humeral crests are apparent in some specimens (Figure 155); (9) hand webbing absent; (10) foot about two-thirds webbed: I (2−–2<sup>+</sup>)—(2<sup>+</sup>–21/4) II

(1<sup>+</sup>–12/3)—(21/2–3−) III (1<sup>1</sup>/2–12/3)—(21/3–3−) IV (2<sup>1</sup>/3–3−)—(1<sup>1</sup>/2–2−) V (Figure 155); (11) ulnar and tarsal folds enameled and low; (12) concealed prepollex; nuptial pad Type I in adult males; (13) Finger I shorter than Finger II; (14) disc of Finger III 20.5%–26.5% of eye diameter; (15) in life, dorsum varies from uniform green to green with small dark flecks; bones green; (16) in ethanol, dorsal surfaces lavender with small dark marks; (17) iris background white to cream beige with brown to black punctuations; (18) fingers and toes lacking melanophores, except some on Toes IV and V; (19) males call from the upper side of leaves overhanging streams; call tonal and composed by single note with a duration of 0.103–0.148 s, and a dominant frequency at 3790–4307 Hz; (20) males show dangling fighting behavior; (21) eggs deposited on tips of leaves overhanging streams; short-term maternal care present; parental care by males absent; (22) tadpoles unknown; (23) small body size; in adult males, SVL 22.5–24.2 mm ( X = 23.0 ± 0.7, *n* = 5); SVL adult females unknown.

**Color in life** (Figure 154): Dorsum varies from uniformly green to green with small dark flecks. Ventral surfaces greenish white. Bones green. Iris from white to cream beige, with brown to black punctuations forming thin reticulation; in some individuals, a yellow hue is visible around pupil.

**Figure 154.** (**A**,**B**) *Nymphargus gri*ffi*thisi* in life, MZUTI 099. ( **C**,**D**) *Nymphargus lasgralarias* in life, MZUTI 094. Photos of both species from Reserva Las Gralarias, 2175–2200 m, Pichincha province, Ecuador. Photos by Carl R. Hutter.

**Figure 155.** *Nymphargus gri*ffi*thisi*. (**A**) Hand in ventral view, KU 118040. (**B**) Foot in ventral view, KU 118040. (**C**) Ventral humeral crests ((**Top**): KU 288992; (**Bottom**): KU 188148). (**A**,**B**) Modified from Lynch and Duellman [22]. (**C**) Modified from Guayasamin et al. [1].

**Color in ethanol:** Dorsal surfaces lavender with small dark marks. Anterior half to two-thirds of parietal peritoneum white, posterior part translucent; pericardium white; most visceral peritonea clear, except for white peritoneum covering renal capsules.

**Biology and ecology:** During the rainy season (December–April), at night, *Nymphargus gri*ffi*thsi* is active on vegetation 50–900 cm above streams, waterfalls, and small rivers in primary and secondary cloud forests [5,87]. Reproductive activity peaks in February–April. Males are highly territorial and engage in aggressive interactions with conspecifics; when fighting, males have been observed dangling upside-down venter to venter holding onto a branch with their hind legs [5]. Females lay clutches of 14–22 eggs on leaves overhanging fast-flowing water. These later expand into a hanging gelatinous mass upon absorption of water; eventually, tadpoles hatch and fall into the stream below [87]. At Reserva Las Gralarias, *N*. *gri*ffi*thsi* has been observed at Five-frog Creek, Heloderma Creek, Hercules Creek, and Kathy's Creek [88,92]. Short-term parental care is provided by females; no parental care is provided by males [25].

**Call** (Figure 156): The information presented below is from Hutter and Guayasamin (2012). The call of *N*. *gri*ffi*thsi* is a tonal call, produced at a rate of 1.1–1.9 calls per minute (mean = 1.6 ± 0.4). The call is composed by a single note with a duration of 0.103–0.148 s (mean = 0.122 ± 0.009) and a dominant frequency at 3790–4307 Hz (mean = 4107 Hz; ± 105.5 Hz).

**Tadpole:** Not described.

**Distribution** (Figure 157): Although *Nymphargus gri*ffi*thsi* has been reported in Ecuador and Colombia [22], herein we restrict its distribution to Ecuador, specifically, to localities in Pichincha, Santo Domingo de los Tsáchilas, and Cotopaxi provinces, at elevations between 1220–2430 m. Other populations that have been previously identified as *N*. *gri*ffi*thsi* require further taxonomic evaluation.

**Figure 156.** Advertisement call of *Nymphargus gri*ffi*thisi* and *N*. *lasgralarias*. (**A**) Oscillogram. (**B**) Audiospectrogram. (**C**) Power spectrum. Modified from Hutter and Guayasamin [88].

**Figure 157.** Distribution of *Nymphargus gri*ffi*thsi* in Ecuador (yellow dots).

**Conservation status:** Globally, *N. gri*ffi*thsi* is listed by the IUCN as *Least Concern* [263]. Recent records in Ecuador come from Reserva Las Gralarias, Reserva Río Guajalito, Reserva Otonga, La Favorita Station, and near the town of Chiriboga. At Reserva Las Gralarias, the species is not infected by the chytrid fungus *Batrachochytrium dendrobatidis* (*n* = 5) [92]. Given the more restricted distribution of the species, *N*. *gri*ffi*thsi* should be considered as *Endangered*, following IUCN criteria B2a, B2(iii).

**Evolutionary relationships** (Figure 136): *Nymphargus gri*ffi*thsi* and *N*. *lasgralarias* are sister taxa.

**Taxonomic remarks:** Several populations previously assigned to *N. gri*ffi*thsi* (see Specimens examined) require further evaluation. Vocalization studies on Colombian populations assigned to *N*. *gri*ffi*thsi* also sugges<sup>t</sup> that several species are concealed under this name [264]. Paratypes of *Centrolenella scirtetes* seem to correspond to *N*. *gri*ffi*thsi* (see Taxonomic remarks of *Centrolene lynchi*).

**Specimens examined:** *Nymphargus griffithsi,* Ecuador*: Provincia de Cotopaxi:* 11.5 km W of Pilaló (0.94815◦ S, 78.989633◦ W; 1500 m), QCAZ 34113; *Provincia de Pichincha:* Río Saloya, 1219 m, BMNH 1940.2.20.4 (holotype), BMNH 1940.2.20.3 (paratype); km 14 on the San Juan de Chillogallo–Chiriboga road (0.275895◦ S, 78.721647◦ W; 2120 m), QCAZ 29531; km 16 on the San Juan de Chillogallo–Chiriboga road (0.278161◦ S, 78.706067◦ W; 2430 m), QCAZ 29524–30; La Victoria (0.16285◦ S, 77.909667◦ W; 2104 m), QCAZ 24801; Tandapi (0.416388◦ S, 78.7988◦ W), QCAZ 351, KU 118009–20; Reserva Las Gralarias "Hercules Giant Tree Frog Creek" (0◦01.529- S, 78◦42.243- W; 2175 m), MZUTI 100, 102, 099; Reserva Las Gralarias "Five Frog Creek" (0◦01.870- S, 78◦42.358- W; 2150 m), MZUTI 101; Reserva Las Gralarias "Heloderma Creek" (0◦01.245- S, 78◦42.370- W; 2200 m), MZUTI 098; 1 km SW San Ignacio (0.44861◦ S, 78.74777◦ W; 1920 m), KU 178108–21; 3.5 NE Mindo (0.03222◦ S, 78.76138◦ W; 1340 m), KU 164564–76; 5 km ESE Chiriboga (0.245277◦ S, 78.7261◦ W; 2010 m), KU 164519–37; 5.6 km SE Tandayapa (0.0333◦ S, 78.7166◦ W; 1910 m), KU 202792. *Provincia de Santo Domingo de los Tsáchilas:* 14 km W of Chiriboga (0.26527778◦ S, 78.847778◦ W; 1960 m), KU 164544–63; 4 km W Chiriboga (0.24277◦ S, 78.7855◦; 2120 m), KU 142649.

*Nymphargus* <sup>a</sup>ff. *gri*ffi*thsi*. Ecuador: *Provincia Imbabura:* 23.2 km W of Apuela, Cordillera de Intag (0.2666◦ N, 78.6◦ W; 2190 m), KU 178122–36; San Antonio de Cuellaje, Finca de Estuardo Ayala (0.4775 N, 78.56263 W), QCAZ 42164. *Provincia de Carchi:* Chilma Bajo (0.86472 N, 78.04972 W; 2071 m), QCAZ 40176–77; 9.9. km E of Maldonado on the Maldonado-Tulcán road (0.83472◦ N, 78.051388◦ W; 2130 m), QCAZ 12572; km 5 on the Chilma Bajo–El Placer road (0.85705◦ N, 78.032476◦ W; 2222 m), QCAZ 39992, 39994; ca. 5 km W La Gruel (0.916667◦ N, 78.1333◦ W; 2340 m), KU 202784–91, 202796, 202801.

*Nymphargus humboldti* new species Guayasamin, Cisneros-Heredia, McDiarmid, Hutter (Figures 158 and 159).
