**Tadpole:** Not described.

**Distribution** (Figure 40): *Centrolene heloderma* occurs on the Pacific slopes of the Cordillera Occidental in Colombia (departments of Antioquia, Cauca, Valle del Cauca, and Risaralda) south to the Tandayapa and Saloya Valleys, in Ecuador (provinces of Imbabura, Pichincha, and Santo Domingo de los Tsáchilas) at elevations of 1850–2575 m ([87,120,121], this work). In Ecuador, *Centrolene heloderma* has been recorded from seven localities at elevations between 1960 and 2575 m ([87,120,121]) in this work and has a potential distribution of 1067 km2, within the Western Montane Forest ecoregion.

**Figure 40.** Distribution of *Centrolene heloderma* in Ecuador (yellow dots).

**Conservation status:** Globally, *Centrolene heloderma* is currently listed as *Vulnerable* by the IUCN [122]. In Ecuador, major threats include habitat loss, introduction of exotic predatory fish (Trout), climate change, and emerging diseases [87,123]. The type locality (Quebrada Zapadores), as well as nearby localities, have been visited numerous times in the last years (2000–2015; wet and dry seasons) with no new records of the species [91] (JMG, pers. obs.; DFCH, pers. obs.); in this area, the last record of the species dates from March 1979, when three individuals where collected at Quebrada Zapadores. In 2006, an individual apparently assignable to *C*. *heloderma* was photographed in the western versant of the Pichincha volcano (M. H. Yánez-Muñoz, pers. com.). On March 2009, *C*. *heloderma* was discovered at Reserva Las Gralarias (0.00806 S, 78.72433 W, 1852 m), Pichincha Province, where it maintains three nearby, reproductive populations. During September 2015, a population of *C*. *heloderma* was found at Cordillera de Toisán. Krynak et al. [121] report on a relatively large population (20+ individuals) at Río Alambi observed on 24 April 2017. The potential distribution of the species in Ecuador is 1067 km2, 34% of which is affected by human activities. The amphibian chytrid fungus *Batrachochytrium dendrobatidis* has been found infecting *C*. *heloderma* at Reserva Las Gralarias, but no recent declines have been observed [92]. Given the current information of the species, we sugges<sup>t</sup> that it should be considered as *Endangered* in Ecuador, following IUCN criteria B2, Ba, Bb(iii).

**Evolutionary relationships** (Figure 24): *Centrolene heloderma* is sister to a clade formed by several *Centrolene* species.

**Specimens examined:** *Centrolene heloderma:* Ecuador: *Provincia de Imbabura:* Cordillera de Toisán (0.50276◦ N, 78.5515◦ W; 2575 m), MZUTI 4234. *Provincia de Pichincha:* Quebrada Zapadores, 5 km ESE of Chiriboga on Chiriboga–Quito road (0.2453◦ S, 78.726◦ W, 2010 m), KU 164714–15; 9 km SE Tandayapa (0.01667◦ S, 78.683◦ W, 2160 m); 8.6 km SE Tandayapa (0.0333◦ S, 78.7◦ W, 2000 m), USNM 211218; 13.1 km SW Nono (0.0025◦ S, 78.659◦ W, 2140 m), MCZ 97834, USNM 211216–17; Reserva Las Gralarias (0.00806◦ S, 78.72433◦ W, 1852 m), QCAZ 40200. *Provincia de Santo Domingo de los Tsáchilas:* 14 km west of Chiriboga on Chiriboga–Santo Domingo road (0.2653◦ S, 78.848◦ W, 1960 m), KU 164716–21.

*Centrolene huilensis* (Ruiz-Carranza and Lynch, 1995 [26]) (Figure 41).

*Centrolene huilense* Ruiz-Carranza and Lynch, 1995 [26]. Holotype: ICN 7462.

Type locality: "Colombia, Departamento de Huila, Municipio San José de Isnos, 1 km NW Isnos, vertienteorientaldelaCordilleraCentral, 1◦57-LatitudN,76◦15-WGreenwich,2190m".

 *Centrolene huilensis*—Barrio-Amorós, Rojas-Runjaic, and Señaris, 2019 [85].

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

**Etymology:** The specific epithet *huilensis* refers to the type locality of the species, within the Huila Department, Colombia [26].

**Identification:** *Centrolene huilensis*(Figure 41) can be di fferentiated from other glassfrogs by having a green dorsum with a combination of dark green to dark lavender and white spots, white tubercles on the ventrolateral edges of the Finger V, forearm, elbow, Toe V, tarsus, and heel, a humeral spine in adult males, and a relatively large body size (SVL = 23.6–26.7 mm in males; SVL = 28.7 mm in 1 female). Species with a similar dorsal color pattern include *Centrolene peristicta*, *C*. *daidalea*, *C*. *condor*, *C*. *lynchi*, *C*. *muelleri*, *C*. *pipilata*, *C*. *savagei*, *C*. *solitaria*, and *Nymphargus truebae*. Body size of *C*. *huilensis* is larger than *C*. *peristicta*, *C*. *pipilata*, *C*. *savagei*, and *C*. *solitaria* (in *C*. *peristicta*, male SVL = 17.9–22.0 mm, female SVL = 20.8–20.9 mm; in *C*. *pipilata*, male SVL 19.7–22.6 mm, female SVL 22.6–23.6 mm; in *C*. *savagei*, male SVL = 23.3–23.9 mm, female SVL = 19.8–22.6 mm; in *C*. *solitaria*, holotype male SVL = 19.3). Males of *C*. *huilensis* have humeral spines, which are absent in males of *C*. *daidalea*, *C*. *savegei*, *C*. *solitaria*, and *N*. *truebae*. Additionally, some of the species are located in di fferent biogeographic regions; *C*. *peristicta* and *C*. *lynchi* are found on the Pacific slopes of the Andes of Ecuador and Colombia ( *C*. *peristicta* is also found on the western slopes of the Cordillera Central, Colombia), and *C*. *muelleri* and *N*. *truebae* are only known from the Andes of Peru.

**Figure 41.** *Centrolene huilensis* in life. Ecuador, Napo province, Yanayacu Biological Station, QCAZ 45905. Photos by Santiago R. Ron (BioWebEcuador).

**Diagnosis:** (1) Vomers lacking teeth; (2) snout short, round in dorsal aspect, slightly sloping in lateral view; (3) tympanum moderate, oriented almost vertically, with slight lateral and posterior inclinations, its diameter 31%–40% of eye diameter; tympanic annulus completely visible, except for upper border; supratympanic fold evident; tympanic membrane translucent and pigmented as surrounding skin; (4) dorsal skin shagreen, males with low warts and spicules uniformly distributed; (5) pair of enlarged subcloacal warts (Figure 15); (6) anterior 40%–60% of the ventral parietal peritoneum white, posterior portion translucent (condition P3); white pericardium; no iridophores in peritonea covering intestines, stomach, kidneys, gall bladder, and urinary bladder (condition V1); (7) lobed liver, lacking iridophores (condition H0); (8) males with conspicuous humeral spines; (9) webbing absent between Fingers I, II, and III; moderate between outer fingers; webbing formula: III (2<sup>+</sup>–21/2)—(2–2<sup>+</sup>) IV; (10) webbing between toes extensive; foot three/fourths webbed: I (1–1<sup>2</sup>/3)—(13/4–2+) II (1–1<sup>+</sup>)—(2−–21/2) III (1–1<sup>1</sup>/2)—(2<sup>+</sup>–22/3) IV (2–3−)—(1−–11/2) V; (11) ulnar and tarsal folds with white tubercles; (12) concealed prepollex; nuptial pad Type I; (13) Finger I slightly shorter than Finger II (Finger I length 94%–97% of Finger II); (14) disc of Finger III width 51%–63% of eye diameter; (15) in life, dorsum green with small di ffuse dark green to dark lavender spots and smaller white spots; bones green; (16) in preservative, dorsum cream to light lavender with dark lavender spots and smaller white spots; (17) in life, iris cream with a slight yellow hue and thin black reticulations, yellowish–cream circumpupilar ring; (18) melanophores absent from dorsal surfaces of fingers and toes, except for Finger IV, and Toes IV and V; (19) males call from upper side of leaves; call undescribed; (20) fighting behavior unknown; (21) eggs deposition site unknown; parental care unknown; (22) tadpoles unknown; (23) small to medium body size; in Colombian populations, adult males, SVL 23.6–26.7 mm ( X = 25.1 ± 1.703, *n* = 7 [26]); in Ecuador, SVL 23.8 in one adult male and 28.7 mm in one adult female.

**Color in life** (Figure 41): Dorsum green with dark lavender and dark green spots of di fferent sizes, and smaller white spots; upper lip white; region below eye with small white warts; bones green. Upper flanks with same color pattern as dorsum; lower flanks with numerous small white warts. Ulnar and tarsal folds with white tubercles; small white cloacal tubercles. Iris cream with a slight yellow hue and thin black reticulations, yellowish–cream circumpupilar ring.

**Color in ethanol:** Dorsal surfaces of head, body, and limbs cream to light lavender with dark lavender spots and smaller white spots; margin of upper lip white; region below eye with small white warts; white tubercles just posterior to cloaca. White parietal peritoneum covers anterior 40%–60% of venter; white pericardium; iridophores absent from peritonea covering digestive tract, liver, kidneys, and gall and urinary bladders.

**Biology and ecology:** In Ecuador, individuals were found active during the night on upper surfaces of leaves along slow-flowing streams. Parental care is unknown.

**Call:** Not described.

**Distribution** (Figure 42): *Centrolene huilensis* is known from two localities, one in the Amazonian slope of the Andes of Ecuador (Yanayacu Biological Station, 2000 m, Napo Province), and the type locality in the Cordillera Central of Colombia (near Isnos, Huila Department) at elevations between 2000–2190 m ([26], this work).

**Figure 42.** Distribution of *Centrolene huilensis* in Ecuador (yellow dot).

**Conservation status:** Globally, *Centrolene huilensis* is currently listed as *Endangered* by the IUCN [124]. In Ecuador, assessing the conservation status of this species remains challenging because only a single additional locality (Yanayacu Biological Reserve) has been registered since its description in 1995. Yanayacu Biological Reserve is a relatively well-studied site [20,125] and we have been unable to find it during recent surveys (August 2014, June 2016; January 2017); thus, it is possible that *C*. *huilensis* spends most of its time in the canopy and/or is extremely rare. We sugges<sup>t</sup> the that *C*. *huilensis* should be placed in the *Data Deficient* category for Ecuador.

**Evolutionary relationships** (Figure 24): With the current gene and taxon sampling, *Centrolene huilensis* is inferred as sister to *C*. *muelleri*.

**Specimens examined:** *Centrolene huilensis:* Ecuador: *Provincia de Napo:* Yanayacu Biological Station (0◦41- S, 77◦53- W; 2100 m), QCAZ 37230, 45905.

## *Centrolene lynchi* (Duellman, 1980 [126]; Figures 43–46)

*Centrolenella lynchi* Duellman, 1980 [126]. Holotype: KU 164691.

Type locality: "a stream 4 km northeast (by road) of Dos Ríos, Provincia Pichincha, Ecuador, 1140m(0◦21- S,78◦54- W)"(nowinProvinciadeSantoDomingodelosTsáchilas).

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

*Centrolenella grandisonae*—Lynch and Duellman, 1973 [22].

*Centrolenella gemmata* Flores, 1985 [46]. Holotype: MCZ 104073. Type locality: "San Francisco de las Pampas, 1500 m in elevation, Provincia Cotopaxi, Ecuador (00◦25- S, 78◦57- W, just NW of junction of Río Las Juritas and Río Toachi)". **New synonymy.**

*Centrolene gemmatum*—Ruiz-Carranza and Lynch, 1991 [6]. Guayasamin, Castroviejo-Fisher, Trueb, Ayarzagüena, Rada, and Vilà, 2009 [1].


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

**Etymology:** The specific epithet *lynchi* honors Dr. John D. Lynch, who has made multiple and meaningful contributions to the field of amphibian systematics.

**Identification:** *Centrolene lynchi* is distinguished from most glassfrogs by having, in life, a green dorsum with minute yellowish–white flecks and diffuse small black spots (Figure 43). Among centrolenids found on the Pacific slopes of the Andes, only *C*. *peristicta* has a similar dorsal color pattern; however, body size is clearly different between the two species (in males, SVL 23.3–26.5 mm in *C*. *lynchi;* 17.9–21.2 mm in *C*. *peristicta)*. The Colombian *C*. *antioquiensis* lacks black spots on the dorsum and has a white gastrointestinal peritoneum. *Centrolene lynchi* is also similar to the Colombian *C*. *quindianum*, which invariably has iridophores on the digestive tract (iridophores absent in *C*. *lynchi*). Other species, such as *Nymphargus truebae* (from the Amazonian slopes of the Peruvian Andes) and *N*. *garciae* (from the Cordillera Central of the Colombian Andes and the Amazonian slope of the Ecuadorian Andes), have a dorsal coloration similar to that found in *C*. *lynchi*, but lack humeral spines in males and webbing between Fingers III and IV.

**Diagnosis:** (1) Vomerine teeth absent; (2) snout round or truncated in dorsal aspect and truncated to slightly inclined in lateral profile; (3) tympanic annulus visible, oriented almost vertically, with slight lateral and posterior inclinations, its diameter about 31.3%–39.4% of eye diameter; supratympanic fold evident; tympanic membrane translucent, partially pigmented, differentiated from surrounding skin; (4) dorsal skin shagreen in males and females, males have low, white warts, and spicules and spiculated warts on sides of head; (5) pair of enlarged subcloacal warts (Figure 15); (6) anterior half to two-thirds of ventral parietal peritoneum white, remaining posterior portion transparent (condition P2–P3); white pericardium; no iridophores in peritonea covering intestines, stomach, and kidneys; transparent peritoneum around gall bladder and urinary bladder (condition V1); (7) liver with four clearly defined lobes, lacking iridophores (condition H0); (8) males with conspicuous humeral spines; (9) webbing absent between Fingers I and II, absent or greatly reduced between Fingers II and III, and moderate between outer fingers; hand webbing formula III (2–2<sup>1</sup>/4)—(2−–2<sup>+</sup>) IV; (10) webbing between toes extensive; foot webbing formula: I (1–1+)—(2–2+) II (1–1+)—(2–2+) III (1–1+)—(2–2+) IV 2<sup>+</sup>—(1–1<sup>+</sup>) V; (11) ulnar fold present, white; external tarsal fold absent, internal tarsal fold short, low; (12) concealed prepollex, except in a few individuals; nuptial pad Type II; (13) Finger I slightly shorter than Finger II or about equal its length (Finger I 94.5%–100.0% of Finger II); (14) disc of Finger III width about 48.4%–57.0% of eye diameter; (15) in life, dorsum dull green with minute yellowish–white warts and small diffuse black spots (Figure 43); green bones; (16) in preservative, dorsum lavender with numerous minute white spots and larger dark lavender spots (Figure 44); (17) in life, iris greyish–white, with a yellow hue around the pupil; (18) melanophores mostly absent from fingers and toes, except for a few on base of Fingers III and IV, and along Toes IV and V; (19) males call from upper sides of leaves; call is relatively short and consists of a tonal note followed by one to three peaked notes; notes separated by 9.0–138.0 ms; mean tonal note dominant frequency 5296 Hz (SD = 58, range = 4995–5599 Hz); mean peaked note dominant frequency 5264 Hz (SD = 72, range = 4995–5513); notes lack frequency modulation; (20) males fight while hanging upside down, grasping one another venter to venter; (21) eggs placed on upper surfaces of leaves; females provide short-term parental care; prolonged parental care absent; (22) tadpoles unknown; (23) small body size; male SVL 23.3–26.5 mm (X = 24.7, *n* = 22); in two females SVL 24.6–25.0 mm.

**Figure 43.** *Centrolene lynchi* in life from Reserva Las Gralarias, Pichincha province, Ecuador. (**A**) Adult male, not collected. (**B**) Adult male in ventral view, QCAZ 40192; photo by Luis A. Coloma. (**C**) Fight between males; photo by Henry Imba and Rebecca Abuza. (**D**) Egg clutch.

**Color in life** (Figure 43): Dorsum yellow green with minute yellowish–white warts (spiculated in males) and small and diffuse black spots; bones green; throat and venter cream white; fingers and toes dull yellow; upper lip white; ulnar and outer tarsal folds with thin white line or low white tubercles; flanks and venter cream white; small white tubercles just posterior to cloaca. White parietal peritoneum covers anterior half to two-thirds of venter; white pericardium; visceral and hepatic peritonea lacking iridophores. Iris greyish white with black reticulation, and a yellow hue surrounding the pupil.

**Color in ethanol** (Figure 44): Dorsal surfaces of head, body, and limbs lavender with minute white warts and small black spots; margin of upper lip white; region below eye with small white warts; small white tubercles just posterior to cloaca. White parietal peritoneum covering the anterior half to two-thirds of the venter; pericardium white; no iridophores in peritonea covering liver, intestines, stomach, kidneys, gall bladder, and urinary bladder.

**Figure 44.** Type material of *Centrolene lynchi and C*. *gemmata* in preservative. (**A**,**B**) *C*. *gemmata*, MCZ 104077, paratype. (**C**) *C*. *lynchi*, KU 164698, paratype, dorsal view. (**D**) *C*. *lynchi*, KU 164691, holotype, ventral view.

**Variation:** The dorsum of most examined specimens except one (KU 164695) have spicules and spiculated warts; it is possible that this variation is a result of the reproductive condition of this particular male individual. Females of *Centrolene lynchi* lack spicules, normal sexual dimorphism in Centrolenidae.

**Biology and ecology:** The following description is from Dautel et al. [50]. At Reserva Las Gralarias (Lucy's Creek), females place clutches of 21–24 neon green eggs on vegetation in males' territories (*n* = 4, mean = 22.25, SD = 1.08); most egg clutches are placed on top of large leaves or ferns above a fast-flowing stream at a height of 165–600 cm (*n* = 14, mean height = 313.3 cm, SD = 78.7). Dautel et al. [50] suggested that males provide long-term parental care; however, a more detailed study by Delia et al. [25] showed that only females provide parental care (short term) and that prolonged parental care is absent in the species.

**Calls** (Figure 45): The following description is from Dautel et al. [50]. The typical advertisement call is relatively short and consists of a tonal note followed by one to three peaked notes. The first note is tonal and generally longer than the following notes, which are pulsed and show clear frequency peaks. Notes are separated by 9.0–138.0 ms. The dominant frequencies of tonal and peaked notes are similar (mean tonal note dominant frequency = 5296 Hz, SD = 58, range = 4996–5599 Hz; mean peaked note dominant frequency = 5264 Hz, SD = 73, range = 4996–5513) and show no frequency modulation. The aggressive call in this species is markedly different in structure from the advertisement call. It consists of a single short note containing two pulses (*n* = 5, mean length duration = 150.4 ms, SD = 6.7, range 140.0–156.0 ms), at a dominant frequency significantly lower than the advertisement call (mean frequency = 4892 Hz, SD = 39, range = 4823–4910 Hz); like the advertisement call, the note of the aggressive call is not frequency modulated.

**Figure 45.** Call of *Centrolene lynchi* (LBE-C-013) recorded at Reserva Las Gralarias, 1822 m, Pichincha province, Ecuador.
