**Tadpole:** Not described.

**Distribution** (Figure 169): *Nymphargus mariae* is known from several localities on the Amazonian slopes of the Cordillera Oriental of the Ecuadorian Andes at elevations between 400 and 1078 m ([176,266–268], this work). In Ecuador, the potential distribution of the species is 28,298 km2.

**Figure 169.** Distribution of *Nymphargus mariae* in Ecuador (yellow spots).

**Conservation status:** Globally, *Nymphargus mariae* is listed as *Least Concern* by the IUCN [270]. We agree with this conservation status.

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

**Specimens examined:** *Nymphargus mariae:* Ecuador: *Provincia de Pastaza:* 1 km W Puyo (1.4833 S, 78.0 W; 1000–1050 m), MCZ 91187; Andean foothills in the Upper Bobonaza River Basin (ca. 2.0 S, 77.0 W), USNM 291298; stream tributary of Río Lliquino (1.72553 S, 78.98058 W; 400 m), QCAZ 37932; near Villano (1.47445 S, 77.53529 W; 440 m), QCAZ 39293; Sacha Yacu (1.39519◦ S, 77.72946◦ W; 1078 m), MZUTI 183. *Provincia de Napo:* ca. 45 km E of Narupa (ca. 0.729 S, 77.374 W; ca. 800 m), on the Hollín–Loreto road, DFCH-USFQ D285; *Provincia de Orellana:* Reserva Río Bigal (0.52525 S, 77.41785 W; 930 m), QCAZ 48529; Río Huataracu (ca. 0.729 S, 77.374; ca. 800 m), ca. 70 km E of Hollín, on the Hollín-Loreto road, QCAZ 7104, 7499; *Provincia de Sucumbíos:* Lumbaqui (ca. 0.05 N, 77.333 W; ca. 500 m). Peru: *Departamento de Huánuco:* Serranía de Sira (ca. 9.367 S, 74.75 W; 1550 m), KU 174713.

**Localities from the literature:** *Nymphargus mariae:* Ecuador: *Provincia de Pastaza:* Río Pucayacu (1.942 S, 77.042 W) [253]; Conambo (1.86197 S, 76.906 W; 337 m) [176]; 1 km W of Puyo (1.493 S, 78.026 W; 1000–1050 m) [253]; Río Lliquino (1.41486 S, 77.54047 W; 380 m) [176]. *Provincia de Napo:* Río Putuyacu, 45 km E of Narupa (0.734 S, 77.49 W; 800 m) [176,253]. *Provincia de Orellana:* Río Huataraco (1.46986 S, 77.92477 W; 347 m) [176]. *Provincia de Sucumbíos:* Río Verde (0.23786 S, 77.576 W; 726 m) [176]; Lumbaqui (0.04675 S, 77. 34358 W; 515 m) [176].

*Nymphargus manduriacu* Guayasamin, Cisneros-Heredia, Vieira, Kohn, Gavilanes, Lynch, Hamilton, and Maynard, 2019 [21] (Figures 170–173).

*Nymphargus manduriacu* Guayasamin, Cisneros-Heredia, Vieira, Kohn, Gavilanes, Lynch, Hamilton, and Maynard, 2019 [21]. Holotype: ZSFQ 0466, by original designation. Type locality: "Reserva Río Manduriacu (0.310755◦ N, 78.8569◦ W; 1,215 m), Provincia de

Imbabura, República del Ecuador".

**Common names:** English: Manduriacu glassfrog. Spanish: Rana de Cristal de Manduriacu. **Etymology:** The specific epithet "*manduriacu*" refers to the type locality of the species, Río Manduriacu Reserve, Ecuador, a conservation area managed by Fundación EcoMinga (https: //ecomingafoundation.wordpress.com/) [21].

**Identification:** *Nymphargus manduriacu* is easily differentiated from most glassfrogs by lacking webbing between inner fingers (Figure 171) and having, in life, a greyish–green dorsum with numerous yellow spots, which sometimes are surrounded by an ill-defined black ring (i.e., false ocelli; Figure 170). On the Pacific slopes of the Ecuadorian and Colombian Andes, similar species include the following: *N*. *buenaventura*, *N*. *ignotus*, *N*. *spilotus*, and *N*. *luminosus*. *Nymphargus buenaventura* has a light green dorsum with diffuse pale yellow spots; *N*. *ignotus* exhibits a pale tan to olive–brown dorsum with black ocelli surrounding orange or yellow spots; *N*. *luminosus* has a green dorsum with numerous yellow spots; finally, *N*. *spilotus* has an olive green back with small yellow spots [21].

**Figure 170.** *Nymphargus manduriacu* in life from Reserva Río Manduriacu, Imbabura province. (**A**–**C**) Adult male, ZSFQ 0466. (**D**–**F**) Adult female, ZSFQ 0462. Photos by Jose Vieira/Tropical Herping. Obtained from Guayasamin et al. [194].

**Diagnosis:** *Nymphargus manduriacu* exhibits the following combination of traits: (1) Dentigerous process of vomer low or absent, lacking vomerine teeth; (2) snout truncated in dorsal view, and truncated to slight rounded in lateral view; (3) tympanic annulus barely evident, lower three-fourths visible, tympanic membrane colored as dorsal skin, supratympanic fold present; (4) dorsal skin shagreen, with microspicules in adult males; (5) ventral skin granular, subcloacal area with two large subcloacal warts; (6) parietal peritoneum white, iridophores covering one-third to one-half parietal peritoneum (conditions P2 or P3); pericardium white (i.e., covered by iridophores), all other visceral peritonea

clear (condition V1); (7) liver lobed and hepatic peritoneum clear (lacking iridophore layer, condition H0); (8) adult males lacking humeral spines; (9) webbing between Fingers I, II, and III absent, basal between Fingers III and IV (Figure 171); (10) toe webbing basal between Toes I and II, III 11 2 –(2<sup>1</sup> 2 –3−) III (11/3 –1<sup>1</sup> 2 )–(3–3−) IV (3–3−)–(1<sup>1</sup> 2 –2−) V; (11) lacking dermal ornamentations in the form of tubercles, folds, or fringes on hands, arms, feet, or legs; (12) nuptial excrescences Type I and VI; concealed prepollex; (13) Finger I slightly longer than Finger II; (14) diameter of eye larger than width of disc on Finger III; (15) color in life, greyish green to olive green with yellow spots, which, sometimes, are surrounded an ill-defined black ring (i.e., false ocelli); bones green; (16) color in preservative, lavender dorsum with cream spots; (17) iris coloration in life: Light grey with thin grey reticulations and pale yellow hue around pupil; (18) melanophores present and abundant along Fingers III and IV, less dense on Finger II, and rarely present on Finger I; furthermore, present and abundant along Toes IV and V, less dense on Toe III, only at the base of Toes I and II; (19) males call from upper side of leaves; advertisement call is a high-pitched "chirp", with a single, pulsed note with a duration of 0.093–0.118 s ( X = 0.10 ± 0.007; *n* = 10) and a dominant frequency at 4052–4447 Hz ( X = 4267.7 ± 118.3); (20) fighting behavior unknown; (21) egg masses deposited on upper side of leaves, clutch size 15–32 (*n* = 4); no long-term parental care provided by either males or females; (22) tadpoles undescribed; (23) SVL in adult males 24.0–25.7 mm (*n* = 3), and in an adult female 28.8 mm.

**Figure 171.** Hand webbing of *Nymphargus manduriacu* and similar species. ( **A**) *N*. *manduriacu*, ZSFQ 0463, adult male, paratype. (**B**) *N*. *luminosus*, ICN 15930, adult female, holotype. ( **C**) *N*. *spilotus*, ICN 35255, adult female, holotype. Modified from Guayasamin et al. [194].

**Color in life** (Figure 170): Dorsal surfaces greyish green to olive green with yellow spots, with melanophores concentrated around yellow spots, sometimes looking like false ocelli. Upper lip unpigmented. Inner fingers and toes with yellowish hue. Anterior half of ventral parietal peritoneum white, posterior portion translucent. Green bones. Iris light grey with thin, dark grey reticulations and pale-yellow hue around pupil [21].

**Color in ethanol:** Dorsal surfaces of body and limbs grey lavender with small white spots. Parietal peritoneum white, iridophores covering one-third to one-half parietal peritoneum. Heart white (covered by iridophores); all other visceral peritonea unpigmented [21].

**Biology and ecology:** *Nymphargus manduriacu* has only been found at Río Manduriacu Reserve and, although the reserve has been visited several times, *N*. *manduriacu* was only regularly detected during February 2018, with the site experiencing particularly heavy rains on a daily basis [21].

**Call** (Figure 172): The advertising call was described by Guayasamin et al. [21], as follows. Each call is a high-pitched "chirp" that consists of a single note with a duration of 0.093–0.118 s (X = 0.10 ± 0.007; *n* = 10). Notes are clearly pulsed (8–12 pulses per note; X = 10.33 ± 1.366). In each call, there is a slight increase in the dominant frequency with time; the dominant frequency is at 4052–4447 Hz ( X = 4268 ± 118.3). Time between calls is 3.9–8.6 s ( X = 5.72 ± 1.82).

**Figure 172.** Call of *Nymphargus manduriacu*, LBE-C-042, from Reserva Río Manduriacu. (**A**) Oscillogram. (**B**) Audio-spectrogram. Obtained from Guayasamin et al. [21].
