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We present the results of herpetological surveys in two adjacent mountains where the EcoMinga Foundation protects the cloud forest in the Upper Rio Pastaza watershed, in the Llanganates Sangay Ecological Corridor in Ecuador. A rapid assessment of the amphibian communities of the study sites reveals a diverse and heterogeneous composition, dominated by terrestrial frogs from the genus Pristimantis. We also identify a cryptic diversity with a significant number of candidate new species. We describe two new species of terrestrial frogs of the genus Pristimantis. Pristimantis maryanneaesp. nov. is characterised by not having tympanum externally visible and having 2–3 subconical tubercles in the upper eyelid; and Pristimantis burtoniorumsp. nov. is characterised by the presence of red colouration in hidden surfaces of the hind-limbs, tubercles on the upper eyelid, interorbital tubercle and a row of rounded tubercles along the snout to the tip and a pale red venter with dark brown mottled pattern. Our samples from the two Reserves do not share species between them, so the proportion of shared species seems to be relatively low. In addition, we highlight the importance of updating the knowledge of amphibians that are restricted to this important conservation region and comment about the threats and composition of the amphibian communities on the eastern slopes of the Upper Rio Pastaza watershed.

The Neotropical frog genus Cryptobatrachus includes six currently recognized species distributed throughout the northern Andes in Colombia and Venezuela. Cryptobatrachus conditus, C. pedroruizi, and C. remotus were described from the Colombian and Venezuelan slopes of the Serranía del Perijá. Due to the great morphological similarity among these species, we re-assess their taxonomic status based on morphological and molecular data from types, topotypes, and specimens from localities referred to in the original descriptions of these species. Morphometric comparisons showed that all these species are virtually indistinguishable, although some subtle qualitative differences in morphological traits distinguish C. conditus from the other species. Phylogenetic analyses of DNA sequences found that C. remotus and C. pedroruizi are not reciprocally monophyletic and exhibit low genetic divergence (< 1 %). Therefore, C. remotus should be considered a junior synonym of C. pedroruizi. This work stresses the importance of comprehensive taxon sampling in poorly explored areas, especially between neighbouring countries.

[EN] The genus Gastrotheca (Anura: Hemiphractidae) is a group of marsupial frogs particularly diverse in Andean regions. Several taxonomic studies of this genus have been conducted in the humid cloud forests—or Yungas—of the Andean eastern slopes of central Bolivia (departments of Cochabamba and Santa Cruz). Yet, the distinction among three species that occur sympatrically in these forests, G. lauzuricae (proposed as a junior synonym of G. coeruleomaculatus in 2015), G. piperata, and G. splendens, remains unclear since the morphological characters that purportedly support their differentiation are variable and partly shared among them. We have carried out external morphological studies, including multivariate morphometric analyses, to assess how they support the taxonomic status of these three species. We also evaluated characters of the cranial osteology of a sample of six individuals using micro CT-scanning. Principal component and linear discriminant analyses resulted in a great overlap among the putative species. Cranial osteological comparisons did not reveal highly significant differences among them, but suggested that different degrees of hyperossification could be related to the developmental state of individuals. Our results indicate that most morphological and osteological reported differences between the three species likely represent intraspecific variation. Thus, we propose that the three nominal species belong to a single biological entity, for which the name Gastrotheca splendens (Schmidt, 1857) has priority. We also restrict the name Gastrotheca coeruleomaculatus (Werner, 1899) to externally similar congeneric populations from the Yungas forests of department of La Paz, but highlighting the need of a detailed evaluation of their taxonomic identity., [ES] El género Gastrotheca (Anura: Hemiphractidae) es un grupo de ranas marsupiales altamente diverso, especialmente en las regiones andinas. Varios estudios taxonómicos de este género se han llevado a cabo en los bosques húmedos montañosos—o Yungas—de la vertiente oriental de los Andes del centro de Bolivia (departamentos de Cochabamba y Santa Cruz). La distinción entre tres especies que se encuentran en simpatría en estos bosques,G. lauzuricae (propuesta como sinónimo posterior de G. coeruleomaculatus), G. piperata y G. splendens, no está clara, ya que los caracteres morfológicos que supuestamente apoyan su diferenciación son variables y están parcialmente compartidos entre ellas. Mediante estudios de morfología externa—incluyendo análisis morfométricos multivariantes—evaluamos la validez taxonómica de estas tres especies. Además, examinamos la osteología craneal de una muestra de seis individuos con microtomografía computarizada. Los análisis de componentes principales y discriminante lineal mostraron un gran solapamiento entre las supuestas especies, y las comparaciones osteológicas no revelaron diferencias sustanciales entre ellas, pero sí sugieren que los diferentes grados de hiperosificación podrían estar relacionados con el estado de desarrollo de los individuos. Nuestros resultados indican que la mayoría de las diferencias morfológicas y osteológicas descritas para las tres especies probablemente representan variación intraespecífica, por lo que proponemos la existencia de una única entidad biológica, para la que el nombre Gastrotheca splendens (Schmidt, 1857) tiene prioridad. Se establece que el nombre aplicable a las poblaciones similares de Gastrotheca de los Yungas del departamento de La Paz sería Gastrotheca coeruleomaculatus (Werner, 1899), pendiente de posteriores estudios taxonómicos., This paper was financed by projects CGL2014-53523-P and PGC2018-097421-B-I00 (PI, Ignacio De la Riva) of the Spanish Ministry of Science and Innovation, partially with funds from the European Union.

We describe a new species of marsupial frog, genus Gastrotheca, using morphological characters and molecular data as lines of evidence. The new species was discovered in the páramo and the ecotone between páramo and humid montane forest of Cordillera de Colán, at elevations between 3136 and 3179 m a.s.l., in northeastern Peru. The new species is distinguished from all its congeners by the combination of the following characters: coarsely granular skin on dorsum, a green dorsal coloration without pattern, finger I shorter than finger II, turquoise iris, and a venter without blotches, flecks or dots. Furthermore, we include a detailed osteological description of the new Gastrotheca species based on Micro-CT scanning. Based on our phylogenetic analyses, the new species belongs to the Gastrotheca marsupiata species group, is sister to G. oresbios and closely related to G. psychrophila, G. spectabilis, G. stictopleura and one undescribed species. Additionally, we test for the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). No Bd infection was detected for G. gemma sp. nov. specimens but Bd prevalence was detected among syntopic frogs.

We extend by 300 m the known upper elevational range of the Marsupial frog, Gastrotheca marsupiata (Dumeril & Bibron, 1841), to 4,660 m a.s.l. This record makes G. marsupiata the highest occurring frog in its genus, which is already characterized by high-elevation distributions. We suggest that this record may represent a case of climate-induced range shifting and discuss this record in the context of our still limited understanding of how amphibian distributions are being affected by climate change.

This article provides a brief account of the career of Eugenia M. del Pino. Casual events and serendipity played important roles in modeling her career as a developmental biologist. In collaboration with colleagues and students, she analyzed the biology and development of the marsupial frog Gastrotheca riobambae (family: Hemiphractidae) in comparison with Xenopus laevis and tropical frogs. The emphasis was placed on oogenesis and the early stages of development. Topics include the mono- and multi-nucleated modes of oogenesis. She described two modes of gastrulation in frogs, gastrulation modes one and two, according to the timing of notochord elongation. She was able to establish a pioneer laboratory for the comparative analysis of frog development in Ibero America at the Pontifical Catholic University of Ecuador, in Quito. Her contributions to society include her influence in the establishment of the National Academy of Sciences of Ecuador, and efforts toward the conservation of the Galápagos Archipelago. She is part of a pioneer group of professors that placed Biology as an academic discipline in Ecuador. The experiences of her career reveal that we all face difficulties in our jobs. However, nothing is impossible when we follow a passion. Her work reveals that the key to success is to turn obstacles into opportunities.

Dollo's law of irreversibility states that once a complex structure is lost, it cannot be regained in the same form. Several putative exceptions to Dollo's law have been identified using phylogenetic comparative methods, but the anatomy and development of these traits are often poorly understood. Gastrotheca guentheri is renowned as the only frog with teeth on the lower jaw. Mandibular teeth were lost in the ancestor of frogs more than 200 million years ago and subsequently regained in G. guentheri. Little is known about the teeth in this species despite being a frequent example of trait "re-evolution," leaving open the possibility that it may have mandibular pseudoteeth. We assessed the dental anatomy of G. guentheri using micro-computed tomography and histology and confirmed the longstanding assumption that true mandibular teeth are present. Remarkably, the mandibular teeth of G. guentheri are nearly identical in gross morphology and development to upper jaw teeth in closely related species. The developmental genetics of tooth formation are unknown in this possibly extinct species. Our results suggest that an ancestral odontogenic pathway has been conserved but suppressed in the lower jaw since the origin of frogs, providing a possible mechanism underlying the re-evolution of lost mandibular teeth.

Aspectos da dieta, reproducao e interacoes com outros taxons foram reportados para a perereca marsupial Fritziana goeldii, uma especie bromeligena endemica da regiao sudeste do Brasil. Um total de 56 individuos (48 machos; 8 femeas) foram coletados entre dezembro de 2018 e novembro de 2019. Um total de 218 itens alimentares, representando pelo menos 28 taxons, foram identificados. Hymenoptera e Coleoptera foram os taxons mais abundantes, principalmente nos primeiros meses do ano (janeiro-abril), com Formicidae sendo o grupo mais representativo. Interacoes com ostracodes e helmintos foram observadas em 12 e 13 individuos, respectivamente, com a maior parte desses organismos sendo encontrados no intestino dos especimes de F. goeldii. Fritziana goeldii apresenta dimorfismo sexual, com as femeas (media 36.0 mm) sendo maiores que os machos (media 28.3 mm). Todas as femeas coletadas apresentavam ovocitos, e duas tambem tinham ovos aderidos no dorso. Nenhuma relacao foi encontrada entre o tamanho das femeas e o tamanho e numero de ovocitos. A reproducao de F. goeldii aparenta ser do tipo prolongada, com machos vocalizando em todos os meses ao longo do ano.

Made available in DSpace on 2021-06-25T10:49:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Aplectana longa n. sp. (Ascaridida: Cosmocercidae) from the small intestine of Gastrotheca microdiscus (Amphibia: Hemiphractidae) is described and illustrated. The new species is characterized by the combination of a unique set of morphological characters: 1) Large body size in both sexes; 2) Lateral alae absent; 3) Gubernaculum present, small, thin, weakly sclerotized, pointed at the distal end and curved proximally at the end, with a small and punctiform papilla-like dilation, and 4) Caudal papillae arrangement (9+1:0:6). The distribution pattern of caudal papillae is similar only to Aplectana chamaeleonsis. Nevertheless, Aplectana longa n. sp. is easily differentiated from this species by the arrangement of precloacal papillae. This is the 57th species of the Aplectana and the 16th species reported from Brazil. Universidade Estadual Paulista (UNESP) Campus de Botucatu Instituto de Biociências Setor de Parasitologia, R. Prof. Dr. Antônio Celso Wagner Zanin, 250-Distrito de Rubião Junior Universidade Federal da Bahia Instituto de Biologia Campus de Ondina Universidade Federal de Uberlândia (UFU) Instituto de Ciências Agrárias, LMG-746, Km 1, Monte Carmelo Universidade Estadual Paulista (UNESP) Campus de Botucatu Instituto de Biociências Setor de Parasitologia, R. Prof. Dr. Antônio Celso Wagner Zanin, 250-Distrito de Rubião Junior CNPq: 140871/2017-9 CNPq: 141322/2018-7 CNPq: 150041/2017-9 CNPq: 309125/2017-0 CNPq: 313241/2018-0

Marsupial frogs (Hemiphractidae) evolved exceptional mechanisms for the conquest of terrestrial life. These adaptations include very large eggs. In some species eggs reach 10 mm in diameter, and are considered to be the largest in frogs. Females have reproductive modifications for the incubation of embryos in their bodies. Modifications of embryos include adaptations for development inside the body of the mother, and changes in the developmental pattern. Moreover, in some species, oocytes are multinucleated instead of having a single germinal vesicle as in most vertebrates. This chapter provides an overview of the adaptations of marsupial frogs associated with terrestrial life, with a discussion of gastrulation and multinucleated oogenesis.

Natural history of the marsupial frog Gastrotheca albolineata (Anura: Hemiphractidae) in lowland Brazilian Atlantic Forest. Gastrotheca albolineata is a marsupial frog endemic to the Atlantic Forest in southeastern Brazil. It remains poorly studied in nature and is uncommon in herpetological collections. We studied the natural history of G. albolineata during a four-year period (2015 to 2019), in Ubatuba, São Paulo state, Brazil, at its southernmost distribution. Our results show that G. albolineata is arboreal, perches from low to medium heights, and breeds during the dry season without chorus aggregation. Calling activity occurs during the day but is more intense during the first half of the night. We used dorsal body markings to identify individuals. Six individuals were recaptured during the study, indicating site fidelity during the active season. The defensive repertory of G. albolineata contains seven different behaviors, including a high-pitched distress call. Egg development in the female’s dorsal pouch took at least 87 days, and fully formed froglets were born with a snout–vent length of 16 mm. Our data substantially add to the knowledge of the natural history of Brazilian marsupial frogs and can be helpful to delineate conservation strategies for elusive species such as G. albolineata.

We study the phylogenetic relationships of egg-brooding frogs, a group of 118 neotropical species, unique among anurans by having embryos with large bell-shaped gills and females carrying their eggs on the dorsum, exposed or inside a pouch. We assembled a total evidence dataset of published and newly generated data containing 51 phenotypic characters and DNA sequences of 20 loci for 143 hemiphractids and 127 outgroup terminals. We performed six analytical strategies combining different optimality criteria (parsimony and maximum likelihood), alignment methods (tree- and similarity-alignment), and three different indel coding schemes (fifth character state, unknown nucleotide, and presence/absence characters matrix). Furthermore, we analyzed a subset of the total evidence dataset to evaluate the impact of phenotypic characters on hemiphractid phylogenetic relationships. Our main results include: (i) monophyly of Hemiphractidae and its six genera for all our analyses, novel relationships among hemiphractid genera, and non-monophyly of Hemiphractinae according to our preferred phylogenetic hypothesis; (ii) non-monophyly of current supraspecific taxonomies of Gastrotheca, an updated taxonomy is provided; (iii) previous differences among studies were mainly caused by differences in analytical factors, not by differences in character/taxon sampling; (iv) optimality criteria, alignment method, and indel coding caused differences among optimal topologies, in that order of degree; (v) in most cases, parsimony analyses are more sensitive to the addition of phenotypic data than maximum likelihood analyses; (vi) adding phenotypic data resulted in an increase of shared clades for most analyses., Funding for this study was provided by Conselho Nacional de Desenvolvimento Cientıfico e Tecnologico (CNPq), Brazil to LYE and SC-F (scholarship numbers 830432/1999-0 and 312744/2017-0), Coordenacao de Aperfeicoamento de Pessoal de Nıvel Superior (CAPES), Brazil to LYE (scholarship number 88887.179352/2018-00) and projects CGL2014-53523-P and PGC2018-097421-B-100 of the Spanish Government (PI, Ignacio De la Riva).

We studied the amphibian community of the Parque Nacional da Serra dos Órgãos (PARNASO) for over thirty years. The area of 20,024 hectares has a steep altitudinal gradient (200-2,263 m a.s.l.), and it is located in the municipalities of Guapimirim, Magé, Petrópolis and Teresópolis, middle of the state of Rio de Janeiro, Brazil. Most data were obtained from sampling sites in the municipality of Teresópolis, with additional data from zoological collections and bibliography. We recorded 83 amphibian species distributed in two orders, Anura, 13 families: Aromobatidae (1), Brachycephalidae (11), Bufonidae (5), Centrolenidae (2), Craugastoridae (2), Cycloramphidae (8), Hemiphractidae (7), Hylidae (28), Hylodidae (6), Leptodactylidae (5), Microhylidae (1), Odontophrynidae (3), Phyllomedusidae (3) and Gymnophiona, one family: Siphonopidae (1). In addition, we present six species that occurs in the buffer zone. Ten of these species are endemic of the park, 18 have PARNASO as its type locality, and five the type locality is at the buffer zone. Resumo: Estudamos a comunidade de anfíbios do Parque Nacional da Serra dos Órgãos (PARNASO) por mais de 30 anos. A área de 20.024 hectares possui um gradiente altitudinal de 200 a 2.263m acima do nível do mar, e está localizada nos municípios de Guapimirim, Magé, Petrópolis e Teresópolis, que ficam no meio do estado do Rio de Janeiro, Brasil. A maioria das coletas foram feitas no município de Teresópolis, com dados adicionais de outras coleções zoológicas e de bibliografia. Foram registradas ao todo 83 espécies de anfíbios distribuídos em duas ordens, Anura, 13 famílias: Aromobatidae (1), Brachycephalidae (11), Bufonidae (5), Centrolenidae (2), Craugastoridae (2), Cycloramphidae (8), Hemiphractidae (7), Hylidae (28), Hylodidae (6), Leptodactylidae (5), Microhylidae (1), Odontophrynidae (3), Phyllomedusidae (3) e Gymnophiona, uma família: Siphonopidae (1). Foram registradas, também, seis espécies que ocorrem na zona de amortecimento. Destas espécies 10 são endêmicas, 18 têm o PARNASO como localidade tipo e cinco têm a localidade tipo na zona de amortecimento.

Mountainous areas host a disproportionately large fraction of Earth’s biodiversity, suggesting a causal relationship between mountain building and biological diversification. Mountain clade radiations are generally associated with changes in environment, climate, and the increase in heterogeneity therein during mountain building. However, examining the causal relationship between mountain building and diversification is a complex challenge, because isolating the effects of surface uplift from other abiotic (climate) or biotic variables is not straightforward. In this study, we investigate the relative contributions of abiotic climate-driven (temperature) and geology-driven (elevation) drivers on evolutionary rates of ancient groups of organisms in the South American Andes. We present regional curves of Andean elevation based on a recent compilation of paleo-elevational data back to the Late Cretaceous, and analyse the diversification history of six Andean frog and lizard families that originated equally far back in time. For two clades (Aromobatidae and Leptodactylidae), we find that they diversified most rapidly during the early phase of mountain building (Late Cretaceous - Paleogene), when the first high-elevation habitats emerged in South America. The diversification of one clade (Centrolenidae) is correlated with Cenozoic temperature variations, with higher speciation rates during warm periods. The last three clades (Dendrobatidae, Hemiphractidae and Liolaemidae) are best explained by environment-independent diversification, although for Liolaemidae, an almost equally strong positive correlation was found between speciation and Andean elevation since the late Eocene. Our findings imply that throughout the long-lived history of surface uplift in the Andes, mountain building drove the diversification of different clades at different times, while not directly affecting other clades. Our study illustrates the importance of paleogeographic reconstructions that capture the complexity and heterogeneity of mountain building in our understanding of the effects that a changing environment plays in shaping biodiversity patterns observed today.HighlightsWe provide novel regional paleoelevation curves for the Andes back to the Late CretaceousThe diversification history of six Andean-centered clades is studiedWe find clade-specific responses to environmental changesThe impact of Andean uplift could reach further back in time than previously thought

We describe a new species of the genus Fritziana, historically identified as Fritziana fissilis or F. aff. fissilis. Using adult and tadpole morphology, as well as molecular analyses, we found the new species to be characterized by having the following traits: a mucronate snout tip, a black stripe on the wrist, round subarticular tubercles on the fingers and toes, a closed dorsal egg-pouch in females, tadpoles deposited in bromeliads, and a different mitochondrial transfer RNA gene order, when compared to the general orders already recognized for anurans. This new species occurs in the Atlantic Forest of Brazil, in the states of Sao Paulo, Parana, Santa Catarina, and Rio Grande do Sul, and has the largest geographic distribution recorded for a species of Fritziana.

An inventory of the amphibians of the Reserva Ecológica Michelin – REM in southern Bahia, Brazil is presented. Sixty-nine species were recorded during a ten-year sampling period. Amphibians were distributed in two orders (Gymnophiona and Anura), belonging to twelve families [Aromobatidae (1), Bufonidae (3), Centrolenidae (1), Craugastoridae (5), Eleutherodactylidae (3), Hemiphractidae (2), Hylidae (34), Phyllomedusidae (5) Leptodactylidae (7), Microhylidae (4), Odontophrynidae (3) and Caeciliidae (1)]. Fifty per cent of the reproductive modes known for Atlantic forest anurans were recorded. While no threatened species were found at REM, six species are classified as data deficient (DD) by the Brazilian Red List of threatened species and deserve additional attention.PhasmahylatimboandVitreoranaeurygnataare listed as endangered in Bahia according to the list of threatened species of the state. Despite a higher diversity of amphibians in the Atlantic forest having been reported for mountainous regions, our results revealed that amphibian richness for lowland forests is also high.

Se analizaron y compararon las características de la neurulación, morfología interna de los embriones desde la néurula hasta la eclosión del renacuajo y el modo de miogénesis de dos especies de ranas de la familia Dendrobatidae (Hyloxalus vertebralis y Dendrobates auratus) con otras especies de la misma familia y Xenopus laevis. Se realizó este análisis debido a que los huevos de H. vertebralis y D. auratus tienen el tamaño más grande detectado hasta ahora en la familia Dendrobatidae (2.6 y 3.5 mm de diámetro, respectivamente). La morfología interna de los embriones y la neurulación de H. vertebralis y D. auratus fueron similares a lo observado en X. laevis y otros Dendrobatidae. Se observó retraso en la diferenciación del cerebro en estas especies en comparación con X. laevis. La miogénesis de H. vertebralis y D. auratus ocurre por interdigitación celular como en Bombina variegata (Bombinatoridae), cuatro especies adicionales de Dendrobatidae, dos especies de Leiuperidae y una especie de Hemiphractidae. El estudio comparativo permite documentar la diversidad de programas del desarrollo entre los anuros y contribuir al conocimiento de la fauna ecuatoriana.

We present new information on the distribution of the marsupial frog Gastrotheca testudinea (Jiménez de la Espada,1870) in Ecuador. We provide the first record from the province of Cañar, and the southernmost locality for Ecuador(which also corresponds to the third known report from the province of Zamora-Chinchipe). In addition, we review theelevation range of the species, and propose to change the lowest elevation limit of G. testudinea from 1100 to 700 m.

Report the first records of Gastrotheca longipes, Cochranella resplendens and Teratohyla midas in the Allpahuayo-Mishana National Reserve, Loreto region, Peru. We update the distribution maps of these three species within the Amazon basin, provide information about their natural history, and describe puffing-up body and mouth gapping behaviours in Gastrotheca longipes. Revisado por pares

Gastrotheca yacuri sp. nov. urn:lsid:zoobank.org:act: 6261932E-02AB-4C9E-B2BD-BD213BBDC771 Holotype. CJ 7822 (Figs. 21–22), an adult male, from El Salado de Jimbura, at about 2.5 km (by road) west from the entrance in the Parque Nacional Yacuri, 2914 m (04° 42' 08.37" S, 79° 27' 00.09" W), Loja Province, Ecuador, collected on 9 April 2016 by Paul Székely and Diana Székely. Paratype. Ecuador: Loja: QCAZ 21105, adult male, from El Salado de Jimbura, 2712 m, on 26 June 2002 by Fernando Nogales Sornoza. Referred specimen. Ecuador: Loja: Not collected (Fig. 11O), subadult, 43 mm SVL, from nearby of Lagunas Negras, Parque Nacional Yacuri, 3492 m (04° 42' 31.81" S, 79° 25' 49.61" W). Diagnosis. Included in the genus Gastrotheca by molecular evidence and general morphological similarity with species of Gastrotheca. Although females are unknown, we infer presence of dorsal brood pouch because all its closely related species have it. A moderately large species (57.4–58.9 mm SVL in males, n = 2) with tibia length 47¯49% SVL, larger than foot; (2) interorbital distance greater than width of upper eyelid; (3) skin on dorsum weakly granular to smooth, co-ossified with skull, having a transverse ridge at occipital region; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus smooth; (7) Finger I shorter than Finger II, width of discs much wider than digits; (8) fingers unwebbed; (9) toes one-half webbed, webbing extending to antepenultimate subarticular tubercle on Toe IV, to penultimate subarticular tubercle on Toe V; (10) in life, dorsum uniform green or brown with brown paravertebral marks; (11) head markings consisting of brown labial stripe with cream glandular areas at posterior end, canthal and supraciliary stripe of black, brown, and bronze; (12) dorsolateral stripe absent or present, when present above consisting in a series of bronze and cream warts, bordered below with a black and brown line, (13) flanks brown and green with a blue and black reticulum towards the lower flanks, axillae, groin, anterior and posterior surfaces of thighs, shanks, and tarsus; brown pelvic patch; (14) venter cream with nearly uniformly distributed, small, dark brown marks or brown with cream marks; gular region dark brown in males. Gastrotheca yacuri differs from all other species of Gastrotheca in southern Ecuador by having a blue and black reticulated pattern on flanks, axilla, groin, thighs, and shanks. It most closely resembles two other species, G. pseustes and G. psychrophila by having the blue colors, but they lack a blue and black reticulum (compare these species in Fig. 10). Gastrotheca yacuri differs from its sister species, G. aguaruna, from the Cordillera Central in northern Peru, by lacking blue colors, having the skin co-ossified with skull, and being smaller (SVL of males of G. yacuri 57.4–58.9 mm vs 41.6–46.8 mm in G. aguaruna). The genetic distance between them is of at least 2.8 % (in a DNA dataset of 438 bp, 16S gene). Description of the holotype. An adult male (Figs. 21–22); body moderately robust; SVL 57.4 mm; head wider that long; snout slightly acuminate in dorsal view, bluntly rounded in profile; canthus rostralis angular in section; loreal region concave; lips rounded; top of head flat; interorbital distance 92% of width of upper eyelid; internarial area flat; nostrils not protuberant, directed anterolaterally, at about level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from the eye by distance about equal to diameter of tympanum; tympanic evident; supratympanic fold elevated, extending from behind the tympanum to the insertion of the forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing four teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 28% of SVL); fingers unwebbed bearing fringes; discs large and rounded, width of disc of Finger III lesser that diameter of tympanum; relative lengths of fingers I Skin on dorsum smooth to shagreen; smooth occipital ridge present; U-shaped ridges (vestigial) in sacrum region; skin on flanks rugose and coarsely areolate; skin on throat loose bearing folds of smooth vocal sac; skin on ventral surfaces of thighs, arms, and belly heavely areolate; skin on venter surface of shanks smooth; rugose surfaces around cloacal opening. Tongue broad, suboval, not notched posteriorly, fully attached to mouth floor. Coloration in life. The dorsal surfaces of the head, body, and limbs are green; the hidden surfaces of the forearm, thighs, and shanks are reticulated with black and blue markings (Figs. 21–22). Head markings are brown canthal and labial stripes. The canthal stripe is diffuse; the labial stripe varies from brown anteriorly to white posteriorly until the insertion of the forelimb. The dorsolateral stripe is thin and consists of a series of bronze and cream warts, bordered below with a black and brown line. The flanks are brown and green with a blue and black reticulum towards the lower flanks, axillae, groin, anterior and posterior surfaces of thighs, shanks, and tarsus; the pelvic patch is brown. The venter is cream with nearly uniformly distributed, small, dark brown marks; gular sac is nearly uniform grayish-brown with small dark brown marks. Palms and soles are pinkish gray; ventral fingers and toes are fleshy. The iris is copper with abundant black reticulations. Coloration in preservative. Similar to coloration in life, but green and blue surfaces have begun to fade. Measurements (in mm). SVL: 57.4, TIBL: 27.2, FL: 24.8, HL: 17.1, HW: 19.2, IOD: 6.9, EW: 5.2, IND: 2.8, ED: 6.0, EN: 3.0, TD: 3.3, FFL: 8.2, TFL: 16.2, TFD: 2.8. Variation. Morphometric variation of the male paratype is as follows: SVL: 58.9, TIBL: 28.6, FL: 28.3, HL: 17.9, HW: 20.1, IOD: 6.7, EW: 4.3, IND: 2.8, ED: 5.6, EN: 5.0, TD: 3.0, FFL: 10.8, TFL: 19.0, TFD: 2.7. The skin on the dorsum varies in the subadult by lacking a smooth tranverse occipital ridge. Color variation in life. The referred subadult specimen varies from the holotype in that it is mostly brown; it has a light reddish-brown dorsum of body and limbs with darker, well-defined stripes, two paravertebral and one at the coccyx position; scattered black spots are present on dorsum towards the flanks (Fig. 10O). The loreal region is brown, and a thin dark brown canthal line is present; the labial stripe is dark brown and bordered along its upper margin by a thin pale stripe that varies from brown anteriorly to cream posteriorly until the insertion of the forelimb. There is no dorsolateral stripe but the reddish-brown dorsal color contrasts with a broad dark brown band on the flanks. The flanks are brown with black and cream marks (without blue colors). The venter is mostly brown with cream marks that are larger towards the chest; the gular region is slightly darker that the venter. The palms and soles are gray. The iris is cooper with black reticulations and has a shade of red in the lower half. Vocalization. Four individuals of Gastrotheca yacuri were recorded at El Salado, Loja Province (Appendix III). Descriptive statistics of the acoustic variables are provided in Table 5. The advertisement call is a complex call, composed of one to three (usually two) long pulsed notes and followed (or not) by one to three (usually two) short, single-pulsed notes (Fig. 23 A–G). The long note had a mean duration of 0.634 s (SD = 0.091) and consisted on average of 32.24 (SD = 7.504) distinct pulses, partly fused, without silent intervals (amplitude modulation close but less than 100%). The amplitude of the long note increases gradually towards the middle of the note after which it decreases gradually by the end. The short notes had a mean duration of 0.072 s (SD = 0.020) and the inter-note interval was on average of 0.549 s (SD = 0.214). The mean dominant frequency of the call was 1496.7 Hz (SD = 38.643), with a mean 90% bandwidth frequency of 1366.7–1637.9 Hz, having the highest values recorded among the analyzed species. The fundamental frequency is recognizable; when visible, 5 to 7 harmonics are distinguishable. Comparisons. The advertisement call of Gastrotheca yacuri is most similar to that of G. pseustes, but G. yacuri has a much shorter call duration, shorter long notes duration, shorter inter-note interval, a higher dominant frequency, and a higher 90% bandwidth frequency compared with the call of G. pseustes (Table 5). Also, G. yacuri emits usually only two short notes compared with 4–6 short notes of G. pseustes. The call of G. yacuri can be easily distinguished form those of G. lojana and G. testudinea by the amplitude modulation of the longer note, longer call duration, lower short note rate, larger number of pulses, higher pulse rate, a much higher dominant frequency, and higher 90% bandwidth frequency. Also, G. yacuri emits up to three (usually two) long notes per call compared with the only one emitted by G. lojana and G. testudinea (Table 5). Distribution and ecology. Gastrotheca yacuri is only known from three nearby localities 2.8 km apart (maximum distance) at elevations of 2712–3492 m in the Cordillera Oriental de los Andes in southern Ecuador. One locality is within Parque Nacional Yacuri and the others are in El Salado de Jimbura in Loja Province (Fig. 9). This nocturnal, semiarboreal species inhabits paramo and subparamo in the Evergreen Montane Forest of Catamayo-Alamor (Ministerio de Ambiente del Ecuador 2012), where the average annual rainfall is 895–1159 mm and the average annual temperature is 10.4–14.6 °C (Fick & Hijmans 2017). The male holotype was found near the road, calling from the branches of a bush, at about 3.5 m above ground. In this same location, choruses were heard from April to late July 2016; the males were calling usually from the vegetation nearby the road. An additional subadult was found during the day in a large terrrestrial bromeliad (Fig. 24A, B). Females and tadpoles are unknown. In this location, in Parque Nacional Yacuri, Gastrotheca yacuri is syntopic with G. turnerorum. Conservation status. We suggest that Gastrotheca yacuri should be assigned to the Data Deficient category according to guidelines of the IUCN (2001), given the inadecuacy of the current information to establish its status. Further searches and definition of its distribution, area of occupancy, habitat use, and biological information are required. Currently its known area of occurrence is less than 10 km 2, thus it could qualify for a Critically Endangered category; however, we can reasonably predict that if further surveys are conducted within the protected Yacuri National Park, its area will increase to more than 100 km 2. In any case, Lagunas Negras de Jimbura have suffered the synergistic effects of agriculture, cattle and sheep raising, fires, introduced species such as trout, pesticide use, and unregulated tourism activitites. Areas in the Salado de Jimbura that are out of the Yacuri National Park already have been heavily modified by human actions. Etymology. The specific name yacuri is a noun in apposition and refers to the Yacuri National Park, where this species has been found. According to Chamba-Troya (2017), Yacuri takes its name from the Quechua words Yacu meaning water and Quri that means gold. Ecuador's southernmost national park ranges from 2000 to 3700 meters in the Cordillera Oriental. This park of 43,090,60 hectares is unique by sheltering a series of 48 glacial lakes, and many endemic species of flora and fauna. It is a Ramsar site (The Convention on Wetlands) and part of the Podocarpus-El Cóndor Biosphere Reserve declared by UNESCO. Comments. Gastrotheca yacuri belongs to the subgenus Gastrotheca, and is reported as Species D in Duellman (2015: Fig 12.1).

Gastrotheca elicioi sp. nov. urn:lsid:zoobank.org:act: 3D26B088-B527-4362-94E8-647FF26C8419 Holotype. CJ 1402 (Fig. 12), an adult male (collected as tadpole in the field and captive raised), from Cajanuma, entrance to Parque Nacional Podocarpus in the Loja-Vilcabamba road, 2456 m (04�� 05' 25.51" S, 79�� 12' 09.97" W), Loja Province, Ecuador, one of a series obtained on 14 June 2011 by Elicio E. Tapia, Sof��a Carvajal-Endara, and Henry Grefa. Paratypes. (Total 18: 11 males, 3 females, 4 juveniles). Ecuador: Loja: CJ 413���4 (juvenile, male), 1398���401 (four males), 1941 (male) collected with the holotype; KU 142603 ���5 (female, subadults), from 5.5 km W Loja, 2330 m (04�� 00' 48.99" S, 79�� 13' 50.99" W), on 17 July 1971 by William E. Duellman and Linda Trueb; KU 142607 ���8 (subadult, female), 148549���51 (males, female), from 5.5 Km W Loja, 2330 m (04�� 00' 48.99" S, 79�� 13' 50.99" W), on 23 July 1971 by William E. Duellman and Linda Trueb; KU 202688 (male), from 5.2 km W Loja, 2310 m (03�� 58' 58.73" S, 79�� 16' 02.21" W), on 10 March 1984 by William E. Duellman; KU 217511 ���2 (males), from 6.8 km E Loja ca. Loja-Zamora line (03�� 59' 19" S, 79�� 09' 21.99" W), on 9 January 1990 by David Kizirian, John J. Wiens, and Luis A. Coloma. Referred specimens. Ecuador: Loja: QCAZ 22370 (male), from Zamora Huayco, Loja, 3018 m (04��0 5' 49.98" S, 79�� 10' 02.5" W), on 2 December 2002 by Diego Almeida-Reinoso; QCAZ 46319���20 (males), from Zamora Huayco, Loja, 3018 m (04�� 05' 49.98" S, 79�� 10' 02.5" W), on 5 December 2009 by Diego Almeida- Reinoso and Samael D. Padilla; CJ 1841���902 (62 juveniles), from Loja, Barrio La Palmera, Parroquia Sucre, on 22���25 April 2013 by Elicio E. Tapia. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (up to 67.5 mm SVL in female, 46.4���67.8 mm SVL in males, n = 13) with tibia length 42��50% SVL, slightly longer than foot; (2) interorbital distance larger than the width of upper eyelid; (3) skin on dorsum finely granular, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus smooth to slightly granular; (7) Fingers I and II about equal in length, width of discs about twice width of the digits proximal to discs; (8) fingers unwebbed; (9) webbing between external toes extending nearly to antepenultimate subarticular tubercle on Toe IV and proximal to penultimate subarticular tubercle on Toe V; (10) in life, dorsum green, tan, brown, or reddish-brown with or without dark paravertebral marks; (11) head markings consisting of a pale labial stripe on the posterior margin of the lip, a black canthal stripe and a dark triangular interorbital mark in some individuals; (12) a fragmented, narrow, pale dorsolateral stripe present; (13) flanks bronze-brown, tan, or green with or without small pale spots; groin and anterior and posterior surfaces of thighs lightly mottled with tinges of green or blue; (14) venter cream with small brown flecks or spots; (15) brood pouch single, dorsal. In comparison with similar species Gastrotheca elicioi is most like G. lojana, G. cuencana, G. litonedis, G. turnerorum, and G. pseustes in Ecuador and G. monticola in Peru. Gastrotheca elicioi differs from G. lojana, G. cuencana, and G. litonedis in color pattern as follows (compare these species in Fig. 10). Gastrotheca elicioi and G. litonedis have distinct dark canthal stripes, which are absent in G. lojana and inconspicuous or absent in G. cuencana. The groin and anterior and posterior surfaces of the thighs are slightly mottled in G. elicioi, whereas in G. lojana they are heavily mottled; these surface are translucent cream without marks in G. cuencana and usually brown in G. litonedis. If any marks are present on the dorsum in G. elicioi, they usually consist of a triangular interorbital mark, which is connected or not with two narrow and curved paravertebral marks. In contrast, in G. lojana the interorbital mark is a transverse bar usually connected with two broad paravertebral marks; in G. cuencana and G. litonedis an interorbital mark is absent. Gastrotheca lojana and G. cuencana have a conspicuous, elevated row of dorsolateral warts, whereas they are barely raised in G. elicioi and G. litonedis. Additionally, in G. elicioi the dark bars on the limbs, when present, are shorter, thinner and less defined than in G. lojana, whereas in G. cuencana and in some G. litonedis the bars are replaced by irregular blotches. Gastrotheca elicioi has a cream venter with dark flecks or marks, whereas the venter is uniform creamy white in G. cuencana and pale brownish gray in G. litonedis. Gastrotheca cuencana also differs from G. elicioi by having cream dorsal surfaces of the fingers, whereas they are brown, green, or tan in G. elicoi. Gastrotheca turnerorum and G. pseustes differ from G. elicioi (compare these species in Fig. 10) by having the skin on dorsum areolate and weakly areolate respectively, whereas in G. elicioi the skin on dorsum is finely granular. Also, they differ by lacking an interorbital mark, and bars on limbs, which can be present G. elicioi. Gastrotheca pseustes is sympatric with G. elicioi, from which it differs notably by having smaller digital discs (larger in G. elicioi) and lacking a pale dorsolateral and supracloacal stripes, which are fragmented in G. elicioi. Gastrotheca monticola is the sister species of G. elicioi and their minimum genetic divergence is 3.0% (in a DNA dataset of 438 bp, 16S gene). Gastrotheca monticola differs from G. elicioi by having the axilla and groin, concealed surfaces of the thighs, and the dorsal surfaces of the feet green with well-defined black spots on a green or tan background, whereas in G. elicioi the groin and anterior and posterior surfaces of thighs are lightly mottled with tinges of green or blue. Finally, the advertisement call of G. elicioi is unique amongst the calls of Gastrotheca in southern Ecuador, in that the long, pulsed notes are produced after the short ones, whereas the other species tend to produce the long, pulsated notes before the short ones. Description of the holotype. An adult male (Fig. 12) collected as a tadpole and raised in the laboratory; body moderately robust; SVL 46.4 mm; head wider that long; snout rounded in dorsal view, bluntly rounded in profile; canthus rostralis round in section; loreal region concave, lips rounded; top of head flat; interorbital distance 142% of width of upper eyelid; internarial area elevated; nostrils not protuberant, directed anterolaterally, posterior to level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from the eye in a distance approximately one and a half the diameter of tympanum; tympanic annulus and membrane smooth; supratympanic fold moderately weak, extending from corner of eye to tympanum and on to insertion of forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing three teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 31% of SVL), unwebbed, with distinct narrow lateral fringes; discs much wider than digits, slightly truncated, width of disc on Finger III greater that diameter of tympanum; relative lengths of fingers I=IIColoration in life (See also under Comments). The dorsum is uniform brown becoming yellowish brown on the flanks. The dorsal surfaces of limbs have an olive-green tinge. A bronze labial stripe is present at the margin of the lip and continues to the insertion of the forelimb. A bronze inconspicuous and highly fragmented dorsolateral stripe is present; a black canthal stripe is present. The flanks are bronze-brown; the groin and anterior and posterior surfaces of the thighs are pale green with white and black spots. The venter is brownish gray with black and white marks; the gular region is gray dark. The iris is copper colored with a few black reticulations. Coloration in preservative. The dorsum is gray without markings. A narrow pale, fragmented dorsolateral stripe is present. The dorsal surfaces of the limbs are tan with irregular gray blotches. A short pale labial stripe extends from the margin of the lip to the level of the insertion of forelimb; a dark canthal stripe is present. The flanks are dark gray anteriorly and pale gray posteriorly; the groin and anterior and posterior surfaces of the thighs are pale gray with black marks. The venter is cream with dark gray spots; the gular region is gray. Measurements (in mm). SVL: 46.4, TIBL: 19.8, FL: 20.0, HL: 14.1, HW: 16.2, IOD: 4.9, EW: 4.6 IND: 3.3, ED: 5.6, EN: 4.1, TD: 2.2, FFL: 7.9, TFL: 14.6, TFD: 2.6. Variation. Morphometric variation of one female and 13 males is summarized in Table 2. The female is larger than most males (67.5 mm; 56.0�� 8.1 mm). The skin on the dorsum is finely granular. The tympanic annulus usually is smooth but is slightly granular in some individuals. Each dentigerous vomerine process has 6���8 teeth (6.3��1.5, n = 3). Color variation in preservative. Preserved specimens have a bluish gray, pale gray, or brownish gray dorsum. Interorbital and paravertebral marks usually are present but the paravertebral marks may be absent. The dark gray interorbital mark, when present, is triangular in most specimens, but it is divided into two blotches in some specimens. A dark canthal stripe is present in all specimens. A short, pale labial stripe is inconspicuous in most specimens. The flanks are gray, darkest anteriorly, with or without white and dark flecks. The groin and anterior surfaces of thighs are pale gray with slight mottling in some; the posterior surfaces of the thighs are pale gray with black flecks and marks usually only distally. In some specimens, the dorsal surfaces of arms and limbs present short dark gray bars. The venter is white with evenly, or densely distributed dark marks. Color variation in life. (Figs. 10H, 10I, 13���14). In living individuals the dorsum is uniform green (CJ 1843), reddish-brown (CJ 1400), or tan (CJ 1842), with or without contrasting dark gray paravertebral marks. When present, these paravertebral marks usually are narrow, curved, and fragmented; these marks are adherent at the scapular level and may be connected with the interorbital mark. A short cream labial stripe is present on the margin of the lip; the stripe continues posteriorly to the insertion of the forelimb. A dark brown canthal stripe is always present, but it is inconspicuous in some individuals with a dark ground color. The tympanum is brown, tan, or olive green. The iris is usually copper almost without black reticulations. A bronze highly fragmented dorsolateral stripe is present in most individuals. The flanks are brown, bronze-brown, or green. The groin has bluish coloration with slight mottling. The posterior surfaces of the thighs usually are pale green with black flecks and some specimens show black fringes. When present, supracloacal and heel stripes are cream and fragmented. The ventral surfaces usually are cream with dark marks, but some specimens lack dark marks. The gular surface varies from brownish gray to white with black spots. Duellman (1974) provided detailed descriptions of coloration in life of four adult males (KU 148549 ���51 and 142603, under the name Gastrotheca lojana). Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 36 (CJ 4311), from a series of 50 tadpoles (CJ 4311) obtained from a pond at Puntzar�� Alto, near Loja city, 2311 m, Province of Loja, Ecuador, by Luis A. Coloma, Manuel A. Morales-Mite, and Elicio E. Tapia on 28 January 2016. Total length 51.4; body length 19.4 (38% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile, sloping from tip of snout to belly; body width at the level of spiracle 11.9, and height at same position 9.9; head width at level of eyes 11.1. Lateral line system present but barely visible, supraorbital and not evident at level of snout, infraorbital line present at level of the eye, touching the inferior portion of the orbit, and making contact with supraorbital line immediately behind the eye. Inferior oral line visible at the eye level, where it contacts angular line, which descends from eye level. Supraorbital line represented for scattered stiches, which does not make contact with the orbit. Posorbital line forming a circle of stitches, just behind the eye. Anterior oral line and loreal lines not visible; dorsal body and middle bodylines not visible. Nostril medium sized (in proportion to body length), ovoid, protruding, having a fleshy annulus, its opening directed anterolaterally. Snout���nostril distance 3.5; internarial distance 2.9. Eye directed dorsally; eye length 2, eye width 1.8; interorbital distance 5. Spiracle sinistral, located at midbody level, spiracular opening oriented posteriorly; distance from tip of snout to spiracular opening 13.0; end of spiracular tube rounded, attached to body wall, inner wall of spiracular tube not evident; spiracle length 2.9, tube transverse width 2.9. Vent tube dextral, opening oriented posteriorly, tube length 3.4, vent tube transverse width 3. Tail length 32.4; caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 4.4, width 3.4; caudal fins well developed and proportional, arising abruptly near tail-body junction and forming a notorious hump, which makes and arc in the body plane. Dorsal fin height 4.18, ventral fin height 3.7; maximum height of tail 11.9; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body; transverse width 4.9. It is surrounded by an uniserial row of marginal papillae, interrupted medially on upper lip; lower lip papillae alternate in orientation, giving appearance of two rows. Upper lip with 23 papillae on right side and 20 papillae on left side; lower lip bearing 52 marginal papillae; upper jaw sheath medium-sized, forming a smooth arch and finely serrated, transverse width 2.6 (53% of oral disc width) height 0.4; lower jaw sheath V- shaped, open and finely serrated, width 4.1, height 1.3. Labial tooth row formula 2/3(1); tooth rows lengths: A1: 4.15, A2: 4.0, P1 right row 1.75, P1 left row 1.75, P1 gap 0.1, P2: 3.75, P3: 3.70. (Fig. 6D). Color in life. Based on a specimen (CJ 4312a) in Stage 37 from a series (CJ 4312) obtained at Loja (neigborhood Puntzar�� Grande), Loja Province, Ecuador, by Luis A. Coloma, Manuel A. Morales-Mite, and Elicio E. Tapia on 28 January 2016 (Fig. 5D). In dorsal and ventral views, body dark brown. Snout and flanks dark brown; guts gray; red gills visible through the throat. Venter gray. Caudal musculature reddish brown with brown stippling, distal portion lacking pigments; dorsal and ventral fins gray with minute cream stippling. Vent tube translucent. Iris reddish-gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 31���40 (CJ 4306, 4310���13) are shown in Table 6. Total length varies between 15.8 (Stage 31) and 73.2 (Stage 39) and tail length proportion varies from 57% to 70% until Stage 38. Number of marginal papillae varies among specimens and Gosner stages; variation in number of ventral papillae at lower lip is high (43���64). Based on specimens CJ 1950���2 from a series obtained at Loja (neigborhoods La Palmera and Puntzar�� Grande), Loja Province, Ecuador, by Elicio E. Tapia on 22���25 April 2013 (Fig. 15). We documented changes in coloration during ontogenetic development of one, mostly brown individual (CJ 1950) (Figs. 15 A���B). At Stage 40, the dorsum and flanks were cream and brown with a diffuse pattern of brown-gray paravertebral marks and a dark gray stripe bordering the canthus and body dorsolaterally, extending to about level of midbody, bordered dorsally by a diffuse cream area. The caudal musculature is pink proximally, with a gray suffusion distally. The caudal fins are pale gray. The iris is pale red. By Stage 46, the markings on the dorsum of body and limbs are diffuse gray and green on a dark brown background; the fingers and toes are yellowish cream dorsally. The flanks have a wide, brown-gray band. There are well-defined creamy white dorsolateral and labial stripes and supracloacal white marks. Color variation in six additional metamorphs (CJ 1951���2, CJ 4313) from Loja and Loja���Abra de Zamora, in Stage 46 is depicted in Figure 16. They vary from plain dark brown to plain green, and have a complete supracloacal stripe. ������.continued on the next page ������.continued on the next page Comparisons. Tadpoles of Gastrotheca elicioi may occur in sympatry with those of G. lojana, G. psychrophila, G. pseustes, and G. turnerorum in the Loja-Abra de Zamora region. Gastrotheca elicioi differs from all of them by having a dorsal gray-pigmented fin that abruptly arises from the body, whereas is nearly translucent and arises gradually in the other species (compare in Fig 5). The tadpole described as G. psychrophila by Duellman (2015) is G. elicioi. It has a similar dorsal fin and fit well our description of G. elicioi tadpoles. Nonetheless, until tadpoles of G. psychrophila are described and unequivocally assigned to its species, some doubts will remain. Vocalization. Seven individuals of Gastrotheca elicioi were recorded from three locations in Loja Province (one individual from the old Loja-Catamayo road, three from Loja, Parque Universitario, Published as part of Carvajal-Endara, Sof��a, Coloma, Luis A., Morales-Mite, Manuel A., Guayasamin, Juan M., Sz��kely, Paul & Duellman, William E., 2019, Phylogenetic systematics, ecology, and conservation of marsupial frogs (Anura: Hemiphractidae) from the Andes of southern Ecuador, with descriptions of four new biphasic species, pp. 1-102 in Zootaxa 4562 (1) on pages 34-48, DOI: 10.11646/zootaxa.4562.1.1, http://zenodo.org/record/2627982, {"references":["Duellman, W. E. (1974) A systematic review of the marsupial frogs (Hylidae: Gastrotheca) of the Andes Ecuador. Occasional Papers of the Museum of Natural History, The University of Kansas, Lawrence, 22, 1 - 27.","Orton, G. L. (1953) The systematics of vertebrate larvae. Systematic Zoology, 2 (2), 63 - 75. https: // doi. org / 10.2307 / 2411661","Duellman, W. E. (2015) Marsupial Frogs: Gastrotheca and Allied Genera. JHU Press, Baltimore, MD, 408 pp.","Fick, S. E. & Hijmans, R. J. (2017) WorldClim 2: new 1 - km spatial resolution climate surfaces for global land areas. International Journal of Climatology, 37 (12), 4302 - 4315. https: // doi. org / 10.1002 / joc. 5086","Pollet, D. (2003) A la Sombra de Otonga. Serie de divulgacion 4. Centro de Biodiversidad y Ambiente, Pontificia Universidad Catolica del Ecuador, Quito, 159 pp.","Paez-Vacas, M. I., Coloma, L. A. & Santos, J. C. (2010) Systematics of the Hyloxalus bocagei complex (Anura: Dendrobatidae), description of two new cryptic species, and recognition of H. maculosus. Zootaxa, 2711 (1), 1 - 75. https: // doi. org / 10.11646 / zootaxa. 2711.1.1"]}

Gastrotheca cuencana sp. nov. urn:lsid:zoobank.org:act: 82F87E67-2940-424C-AC03- BC751224 D5E4 Holotype. CJ 1391 (Fig. 2), an adult male, from the city of Cuenca, 2579 m (02° 53' 57.91" S, 79° 01' 52.79" W), Azuay Province, Ecuador, one of a series obtained on 8 June 2011 by Elicio E. Tapia, Sofía Carvajal-Endara and Henry Grefa. Paratypes. (Total 48: 10 adult males, 21 adult females, 17 juveniles). Ecuador: Azuay: CJ 1392–7, collected with the holotype; KU 120676 (female), 120683 –4, 120690, 120695 (4 juveniles), 120699 (male), 120705 (male), 120709 –10 (males), 120713 (female), 120718 –9 (female, male), 120721 (female), 120722 (male) from Cuenca, 2600 m, (02° 53' 57.91" S, 79° 01' 52.79" W), on 19 June 1968 by John D. Lynch; KU 138616 (female), 138619– 21 (male, females) from 4 km E: Cuenca 2540 m (02° 52' 59.88" S, 78° 22' 0.11" W), on 10 June 1970 by Thomas H. Fritts; KU 141572 (female) from 2.1 km S Cutchil, 2720 m (03° 06' 00"S, 78° 47' 59.99"W), on 16 May 1971 by Richard R. Montanucci; KU 141579 (female) from 3.5 km S Cutchil, 2785 m (03° 04' 59.87" S, 78° 47' 59.99" W), on 14 May 1971 by Richard R. Montanucci; KU 141582 (juvenile) from 8 km NW Cuenca, 2803 m (02° 51' 47.88" S, 78° 58' 59.88" W), on 16 May 1971 by Richard R. Montanucci; KU 141583 (female) from 8.8 km NW Cuenca, 3820 m (02° 52' 00.12" S, 79° 04' 00.12" W), on 15 May 1971 by Richard R. Montanucci; KU 203441 (male) from Laguna de Zurucuchu, 16 km NW Cuenca, 3200 m (02° 50' 23.99" S, 79° 07' 47.99" W), on 5 March 1984 by William E. Duellman; KU 129779 –82 (4 females), 129783–91 (9 juveniles), 129793–94 (juveniles), 129795 (female), 129796 (juvenile) from Río Matadero, 12 km E Cuenca (02° 52' 59.88"S, 78° 58' 00.12" W), on 17 November 1968 by Craig E. Nelson. Cañar: KU 141571 (male) and KU 141573 (female) from Biblián, 2620 (02° 42' 00" S, 78° 52' 00.12" W), on 17 and 23 May 1971, respectively, by Richard R. Montanucci; KU 142620 –24 (4 females, male), from Biblián, 2620 m (02° 42' 00" S, 78° 52' 00.12" W), on 25 July 1971 by William E. Duellman; KU 147113 (female) from Biblián, 2620 m (02° 42' 00" S, 78° 52' 00.12" W), on 15 January 1972 by John E. Simmons. Referred specimens. (Total 36, 28 adult males, 8 adult females). Ecuador: Azuay: QCAZ 1239 (male), from Sigsig, 2480 m (03° 03' 08.4" S, 78° 48' 10.56" W), on 24 July 1989 by Luis A. Coloma and Luis E. López; QCAZ 26309 (female), from Cumbe, 2740 m (03° 05' 55.64" S, 79° 00' 28.73" W), on 8 August 2003 by Patricio Vargas; QCAZ 26348–50 (males), 26353 (female), 26357–64 (8 males), from Cumbe, 2740 m (03° 05' 55.64" S, 79° 00' 28.73" W), on 9 August 2003 by Luis A. Coloma, Ítalo G. Tapia, Andrés Merino Viteri, Erik Wild and Patricio Vargas Mena; QCAZ 26354 (female), from Cumbe, 2740 m (03° 05' 55.64" S, 79° 00' 28.73" W), on 9 August 2003 by Ítalo G. Tapia; QCAZ 26357–8 (males), from Cumbe, 2740 m (03° 05' 55.64" S, 79° 00' 28.73" W), on 9 August 2003 by Andrés Merino Viteri; QCAZ 31477 (male), from Sigsig, 2424 m (03° 03' 08.4" S, 78° 48' 10.56" W), on 8 March 2006 by Ítalo G. Tapia; QCAZ 31509, 31511 (males), from Sigsig, 2424 m (03° 03' 8.4" S, 78° 48' 10.56" W), on 9 March 2006 by Ítalo G. Tapia and Giovanni Onore; QCAZ 34131 (female), from Cuenca, 2579 m (02° 53' 57.91" S, 79° 01' 52.79" W), on 27 February 1979 by Fernando I. Ortiz; QCAZ 37375–6, 37379–81, QCAZ 37386 (6 males), from unknown locality, on 31 May 2007 by Zoológico Amaru; QCAZ 38232 (male), from Carmen del Guzho, 2666 m (02° 56' 00" S, 79° 03' 00" W), on 16 June 2007 by Hari González Maldonado; QCAZ 39384–5 (female, male), from Tarqui, Patapamba, 2600 m (03° 00' 38.16" S, 79° 01' 53.43" W), on 21 June 2007 by Ernesto Arbeláez Ortiz; QCAZ 42720–1 (males) from Cuenca, 2579 m (02° 53' 57.91" S, 79° 01' 52.79" W), on 3 October 2007 by Sofía Carvajal-Endara and Flavio Jaramillo; QCAZ 47106 (male), from Tarqui, 2741 m (03° 00' 57.17" S, 79° 02' 40.13" W), on 21 June 2007 by Ernesto Arbeláez Ortiz. Cañar: QCAZ 42826, 42835, 42841 (females), from Papaloma de la Nube, 3011 m (02° 40' 21.54" S, 78° 54' 21.28" W), on 12–14 August 2008 by Sofía Carvajal-Endara. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (49.0– 61.6 mm SVL in females, n = 28; 45.2–53.7 mm SVL in males, n = 38), with tibia length 39¯50% SVL, slightly shorter than length of foot; (2) interorbital distance slightly larger than width of upper eyelid; (3) skin on dorsum finely granular, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus weakly granular to smooth; (7) Fingers I and II about equal in length, width of discs greater than digits; (8) fingers unwebbed; (9) webbing between external toes extending to antepenultimate subarticular tubercle on Toe IV, to penultimate subarticular tubercle on Toe V; (10) in life, dorsum green, tan, brown, or reddish-brown with or without dark paravertebral marks; (11) head markings consisting of pale labial stripe and, in some individuals, an inconspicuous canthal stripe; (12) pale white or cream dorsolateral stripe present; (13) flanks tan, brown, or green with cream flecks or spots; groin and anterior and posterior surfaces of thighs translucent cream without marks and with a faint pale blue tinge in some; (14) venter uniform creamy white; (15) in females, brood pouch single, dorsal. In comparison with similar species in Ecuador, G. cuencana is most like G. litonedis, G. lojana, G. elicioi, and G. riobambae. It differs from these species by having a uniform creamy white venter; whereas the venter is pale gray in G. litonedis, and cream with dark flecks, spots or mottling in G. lojana, G. elicioi, and G. riobambae. In G. cuencana the dorsal surfaces of fingers are cream, whereas in the other species dorsal surfaces of fingers are of the same color as the rest of the body (tan, brown, or green). Gastrotheca cuencana also differs from G. litonedis, G. elicioi, and G. riobambae by lacking a dark canthal stripe. Moreover, in G. cuencana the groin and anterior surfaces of thighs are mostly translucent cream without marks; whereas, in G. litonedis the groin and anterior surface of thighs are usually bronze-brown or tan with cream or darks flecks and spots. In G. lojana and G. riobambae the groin and anterior surfaces of the thighs are usually cream with black mottling; in G. elicioi the groin and anterior surfaces of thighs usually have dark and cream flecks and spots. In profile, the snout in G. cuencana is rounded above and inclined anteroventrally, whereas the snout in G. elicioi, G. lojana, and G. litonedis is bluntly rounded. Furthermore, G. cuencana has a conspicuous, elevated row of dorsolateral warts whereas these are barely raised in G. litonedi s; a narrow, cream supracloacal stripe is present in G. litonedis, G. lojana, and G. riobambae; whereas the stripe is fragmented in G. elicioi and absent in most G. cuencana. Gastrotheca lojana and G. riobambae also differ from G. cuencana and G. litonedis by having a complex call, whereas the call in the latter two species is a simple call. Additionally, the call of G. cuencana has a lower note rate, shorter note duration, longer inter-note interval and a lower dominant and 90% bandwidth frequency compared with G. litonedis. Finally, G. cuencana differs from its sister species (G. litonedis) by having a genetic distance of 1.2% (in a DNA dataset of 438 bp, 16S gene). Gastrotheca cuencana occurs in syntopy with G. pseustes (sensu lato) throughout most of its range. Gastrotheca pseustes differs from G. cuencana by having coarsely granular skin on the dorsum, in contrast to the usually finely granular dorsum in G. cuencana. In G. cuencana the venter is uniform creamy white, whereas it is cream or white with dark flecks or spots in G. pseustes; also the call of G. cuencana is simple, whereas in G. pseustes the call is complex. Description of holotype. An adult male (Fig. 2); body moderately robust; SVL 50.6 mm; head wider that long; snout rounded in dorsal view, rounded and inclined anteroventrally in profile; canthus rostralis round in section; loreal region concave, lips rounded; top of head flat; interorbital distance 104% of width of upper eyelid; internarial area slightly elevated; nostrils not protuberant, directed anterolaterally, posterior to level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from eye by distance about equal to diameter of tympanum; tympanic annulus and membrane weakly granular; supratympanic fold moderately weak, extending from behind the tympanum to the insertion of the forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing four teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 35% of SVL), unwebbed, with distinct narrow lateral fringes; discs moderately large and rounded, width of disc of Finger III greater that diameter of tympanum; relative lengths of fingers I=II Skin on dorsum finely granular; skin on flanks coarsely granular; skin on throat, venter surfaces of thighs, and arms granular; skin on belly areolate; skin on venter surface of shanks smooth; numerous small tubercles lateral to cloacal opening. Vocal sac single, median, subgular. Vocal slits present at posterior lingual margins of mandibles. Tongue broad, suboval, not notched posteriorly, fully attached to mouth floor. Coloration in life (Fig. 2). The dorsum is yellowish brown with two darker, black delineated, continuous, paravertebral markings. These markings extend to the sacrum where they bifurcate and become two rows of irregular blotches at each side of the vertebral axis. The dorsal surfaces of the limbs are yellowish brown with irregular gray blotches. Cream dorsolateral and labial stripes are present; the latter continues to the insertion of forelimb; a faint brown canthal stripe is present. The flanks are bronze-brown with small black spots. The venter, groin, and anterior surfaces of the thighs are uniform creamy white; the posterior surfaces of the thighs have a faint bluish coloration. The dorsal surfaces of the fingers are cream. The gular region is creamy gray laterally. The iris is reddish bronze with black reticulations. Coloration in preservative. The dorsum is tan with two well-defined, gray paravertebral markings, which bifurcate at the level of sacrum to become two rows of irregular blotches on each side of the vertebral axis. A white dorsolateral stripe is present. The dorsal surfaces of limbs are tan with irregular gray blotches. A cream labial stripe continues to the insertion of the forelimb; a faint canthal is present. The flanks are gray; the venter, groin, and anterior surfaces of thighs are uniform creamy white; the posterior surfaces of thighs have a faint bluish coloration. The gular region is creamy gray. Measurements (in mm). SVL: 50.4, TIBL: 21.9, FL: 24.9, HL: 16.9, HW: 19.1, IOD: 5.2, EW: 5.0 IND: 3.3, ED: 5.6, EN: 4.1, TD: 2.8, FFL: 10.0, TFL: 17.5, TFD: 3.4. Variation. Morphometric variation of 28 females and 38 males is summarized in Table 3. Females are larger than males (54.2± 2.9 mm; 49.6± 2.4 mm). The skin on the dorsum is finely granular in most, but it varies in some specimens from weakly areolate to coarsely granular. All adults have a supratympanic fold that usually extends from the posterior part of the tympanum to point above the insertion of forelimb. The tympanic annulus is usually smooth, but some individuals (e.g., CJ 1395, 1940) have a slightly granular annulus (Fig. 3A, C). Each dentigerous vomerine process has 2–7 teeth (4.4±1.1, n = 39). Color variation in preservative. Preserved specimens have bluish gray dorsum (tan and reddish-brown in CJ 1397). Faint paravertebral marks are present. A canthal stripe is absent in most specimens; the flanks and posterior surfaces of the thighs are dark gray with white spots evenly distributed in some specimens. In all specimens, a prominent white labial stripe extends from the posterior margin of the lip to the insertion of the forelimb. A narrow white dorsolateral stripe is evident in most specimens. The groin and the anterior surfaces of the thighs are uniform creamy gray. The ventral surfaces are uniform creamy white. The posterior surfaces of thighs usually are creamy gray, but they are pale blue in some specimens. Some males have yellowish brown nuptial pads on the medial surface of the thumb. Color variation in life. (Figs. 3–4). The dorsum is uniform green (CJ 1393 ), reddish brown (CJ 1397), or tan (CJ 1392) with (CJ 1391) or without (CJ 1392) darker paravertebral marks. When present, the paravertebral markings usually are well defined; at the level of the sacrum they become two rows of irregular blotches at each side of the vertebral axis. The ventral surfaces are uniform creamy white. A cream labial stripe is extends from the posterior margin of the lip to the insertion of the forelimb; a dark brown canthal stripe is absent in most individuals. The tympanum is brown, tan, or olive green. The iris is reddish bronze with black reticulations. A creamy white dorsolateral stripe is evident in most individuals. The flanks are cream, bronze brown, or green. The groin and anterior surfaces of the thighs are translucent cream without dark markings (Fig. 3). The posterior surfaces of the thighs are translucent, cream, or pale blue or green. When present, supracloacal and heel stripes are cream or tan. The ventral surfaces of the shanks have a faint tinge of pale blue. The gular surface varies from white to creamy gray. ……continued on the next page ……continued on the next page Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 38 (CJ 1945a), from a series of eight tadpoles (CJ 1945) obtained from a pond at south Cuenca, 2579 m, Azuay Province, Ecuador, by Elicio E. Tapia, Sofía Carvajal-Endara, and Henry Grefa on 8 June 2011 (Fig. 5A). Total length 44.6; body length 17.3 (39% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile, sloping from tip of snout to belly; body width at the level of spiracle 11.5, and height at same position 9.3, head width at level of eyes 8.5. Lateral line system present but barely visible, supraorbital and infraorbital lines both rising at level of tip of snout, extending parallel to the eye and making contact immediately behind eye; angular line descending vertically just porterior of eye to throat, it dorsally contacts with post infraorbital line; anterior oral line descending vertically from oral disc level and behind nares level to throat; it makes a curve that parallels infraorbital line, forming a circuit continuous with angular and loreal lines; dorsal body and middle body lines not visible. Nostril medium sized (in proportion to body length), ovoid, protruding, having a fleshy annulus, its opening directed anterolaterally. Snout–nostril distance 2.3; internarial distance 2.4. Eye directed dorsally; eye length 1.8, eye width 1.8; interorbital distance 4.3. Spiracle sinistral, located at midbody level, spiracular opening oriented posteriorly; distance from tip of snout to spiracular opening 11.0; end of spiracular tube rounded, attached to body wall, inner wall of spiracular tube not evident; tube length 3.1, tube transverse width 2.0. Vent tube dextral, opening oriented posteriorly, tube length 1.7, tube transverse width 1.8. Tail length 27.2; caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 3.7, width 3,2; caudal fins well developed and proportional, arising near tail-body junction, forming a low hump; dorsal fin height 3.0, ventral fin height 2.8; maximum height of tail 9.3; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body; transverse width 4.7. It is surrounded by a uniserial row of marginal papillae, interrupted medially on upper lip; lower lip papillae alternate in orientation, giving appearance of two rows; upper lip with 29 papillae on right side and 27 papillae on left side; lower lip bearing 62 marginal papillae; upper jaw sheath medium-sized, forming a smooth arch and finely serrated, height 0.36, transverse width 2.8 (60% of width of oral width); lower jaw sheath V- shaped, open and finely serrated, width 1.92, height 0.7. Labial tooth row formula 2/3(1); tooth rows lengths: A1: 3.6, A2: 3.5, P1 right row 1.5, P1 left row 1.6, P1 gap 0.2, P2: 3.4, P3: 3.0. (Fig. 6B). Color in preservative. Dorsum dull gray with darker areas on flanks, above eye, and on throat; body contour and snout translucent. Caudal musculature and fins with scattered cream dots, with higher suffusion near tail base, otherwise translucent. Venter pale, translucent in belly region, guts exposed; eyes lavender gray; oral apparatus translucent. Color in life. In dorsal view, body olive-cream; snout and flanks paler olive-cream. In ventral view, guts dark gray in belly area; reddish gills visible throughout throat. Caudal musculature covered by a nearly continuous line of cream marks, proximal half of musculature reddish cream with myomeres and medial line well defined, distal half translucent; dorsal and ventral fins translucent, with scattered cream dots, heavily suffused near tail-body junction. Spiracle, oral apparatus, and legs olive-cream. Iris gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 37–39 (CJ 1945) are shown in Table 4. Total length varies

We review the systematics of the species of Gastrotheca (Anura: Hemiphractidae) in the highlands of the southern Ecuadorian Andes. We analyzed phylogenetic, morphological, ecological, and acoustic data from populations in the region. We provide an updated phylogenetic hypothesis inferred from a database that contains 42 species of Gastrotheca, and sequences from mitochondrial (12S, 16S, and ND1) and nuclear (POMC and RAG-1) genes. We describe four new endemic biphasic species for Ecuador (Gastrotheca cuencana, G. elicioi, G. turnerorum, and G. yacuri). In addition, we redefine G. lojana, recently resurrected from the synonymy of G. monticola, and redescribe G. litonedis. Furthermore, we describe the tadpoles and ontogenetic color changes in six species (G. cuencana, G. elicioi, G. litonedis, G. lojana, G. pseustes, and G. turnerorum). Also, we describe the calls of seven species (G. cuencana, G. elicioi, G. litonedis, G. lojana, G. pseustes, G. testudinea, and G. yacuri). The phylogenetic relationships estimated here are congruent with previous phylogenetic hypotheses for the group, except for the placement of G. galeata, G. plumbea, G. orophylax, G. nicefori, and G. griswoldi. Because most sister species in Gastrotheca are allopatric, the evolutionarly scenarios that likely have intervened in shaping the diversity are the uplift of the Andes and the formation of cross-Andean river valleys, which probably promoted conditions that acted as dispersal barriers that led to speciation. Many species of Gastrotheca (44%) are highly threatened. Most monophasic species have not been seen since the late 1980s, whereas the highland biphasic species have survived the catastrophic events that led to the disappearance of many other sympatric anurans. Research and conservation actions are urgently needed for all surviving species treated herein, five of which are either Critically Endangered or Endangered because of their restricted distributions (10–1600 km2), habitat destruction and fragmentation, climate change, and pathogens. Two of the new species, G. cuencana, and G. elicioi, mostly are known from the cities of Cuenca and Loja, respectively, where urbanization threatens their survival. In order to call attention to conservation issues, we name one new species to refer to the Yacuri National Park, and the other three in honor of the people from Cuenca, the Turner family, and Elicio E. Tapia, whose actions are helping to halt amphibian extinctions.

Gastrotheca turnerorum sp. nov. urn:lsid:zoobank.org:act: B72F71A3-671F-4F89-91D5-2410C1D0B6DF Holotype. CJ 393 (Fig. 18), an adult female (collected as tadpole and raised in captivity), from Laguna Negra de Jimbura, Parque Nacional Yacuri, Amaluza, 3406 m (04° 42' 44.24" S, 79° 25' 42.38" W), Loja Province, Ecuador, one of a series collected on 12 June 2011 by Elicio E. Tapia, Sofía Carvajal-Endara and Henry Grefa. Paratypes. (Total 9: 6 males, 2 females, 1 juvenile). Ecuador: Loja: CJ 394–5 (female, male), 415–7 (males, juvenile), 1386 (female), and KU 335390 (male), collected with the holotype. CJ 7823 (male, collected as tadpole and captive raised), from Parque Nacional Yacuri, Amaluza, 3331 m (04° 43' 25.22" S, 79° 26' 15.47" W), Loja Province, Ecuador, one of a series collected on 7 July 2016 by Dan Cogǎlniceanu, Diana Székely and Paul Székely. Zamora Chinchipe: MUTPL 221, male from Reserva Tapichalaca, 3073 m (04° 28' 55.58" S, 79° 09' 29.76" W), collected on 9 December 2014 by Diego Armijos-Ojeda. Referred specimens. Ecuador: Zamora Chinchipe: QCAZ 9575 (captive raised tadpoles and juveniles) from Parque Nacional Podocarpus, Lagunas del Compadre, 3205 m (04° 10' 29.24" S, 79° 06' 59.54" W), on 1 December 1994 by Jenny Rudston; QCAZ 47299 (tadpole), QCAZ (sc 29154) (female) from Parque Nacional Podocarpus, Lagunas del Compadre, 3205 m (04° 10' 29.24" S, 79° 06' 59.54" W), on 20 October 2009 by Elicio E. Tapia. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (54.0– 58.2 mm SVL in females, n = 3; 50.0– 55.6 mm SVL in males, n = 4), with tibia length 37¯42% SVL, shorter than foot; (2) interorbital distance slightly greater than width of upper eyelid; (3) skin on dorsum areolate, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus smooth; (7) Finger I slightly shorter than Finger II, width of discs much wider than digits; (8) fingers unwebbed; (9) foot webbing between external toes extending to antepenultimate subarticular tubercle on Toe IV, to penultimate subarticular tubercle on Toe V; (10) in life, dorsum uniform green without dark paravertebral marks; (11) head markings consisting of pale labial and canthal stripes formed by a series of small bronze dotes; (12) dorsolateral stripe present, also consisting in a series of small bronze dotes; (13) flanks brown and groin green both with or without a few cream spots; anterior surfaces of thighs dark brown, posterior surfaces of thighs bluish brown with numerous cream tubercles proximal to the vent; (14) venter dark brown with uniformly distributed cream spots; gular region dark brown; (15) brood pouch single, dorsal. Gastrotheca turnerorum most closely resembles three other species in southern Ecuador, G. pseustes, G. elicioi, and G. litonedis. Gastrotheca turnerorum differs from all of them by having a different skin texture on the dorsum, and a distinctive color pattern (compare Figs. 10J, 19–20 vs 10A–I, K, L). The dorsal skin is areolate in G. turnerorum, whereas it is weakly areolate or smooth in G. pseustes, finely granular in G. elicioi, and smooth in G. litonedis. In G. turnerorum the venter is dark brown with uniformly distributed cream spots, whereas it is cream with dark marks in G. pseustes and G. elicioi and pale brown-gray in G. litonedis. The labial stripe consists of a series of small bronze dots in G. turnerorum, and mostly uniform cream in G. pseustes, G. elicioi, and G. litonedis. Gastrotheca pseustes and G. elicioi have a dark brown canthal stripe, whereas G. turnerorum has a canthal stripe formed by a series of small bronze dots. Gastrotheca turnerorum is the sister species of G. aguaruna + G. yacuri, with a genetic distance of at least 2.85 % (in a DNA dataset of 438 bp, 16S gene). The Peruvian G. aguaruna differs from G. turnerorum by having a weakly granular to smooth dorsal skin (areolate in G. turnerorum), Fingers I and II equal in length (Finger I shorter in G. turnerorum), and a cream venter with dark fleck or spots (venter dark brown with uniformly distributed cream spots in G. turnerorum). Description of the holotype. An adult female (Fig. 18) that was collected as a tadpole and raised in the laboratory; body moderately robust; SVL 54.0 mm; head wider that long; snout rounded in dorsal view, bluntly rounded in profile; canthus rostralis round in section; loreal region concave; lips rounded; top of head flat; interorbital distance 89% of width of upper eyelid; internarial area flat; nostrils not protuberant, directed anterolaterally, posterior to level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from the eye by distance about equal to diameter of tympanum; tympanic annulus barely evident;, supratympanic fold moderately weak, extending from behind the tympanum to the insertion of the forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing five teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 34% of SVL); fingers unwebbed; discs large and rounded, width of disc of Finger III greater that diameter of tympanum; relative lengths of fingers I Skin on dorsum weakly areolate; skin on flanks coarsely areolate; skin on throat, venter surfaces of thighs, and arms heavily areolate; skin on belly areolate; skin on venter surface of shanks smooth; numerous white tubercles ventrolateral to cloacal opening. Tongue broad, suboval, not notched posteriorly, fully attached to mouth floor. Pouch opening V- shaped with anterior border at level of posterior edge of sacrum. Coloration in life. The dorsal surfaces of the head, body, and limbs are green; the inner and outer surfaces of the forearm, thighs, and shanks are brown (Fig. 18). Head markings are canthal and labial stripes that consist of a series of small bronze dots. The labial stripe continues to the insertion of the forelimb. The dorsolateral stripe also consists of a series of small bronze dots. The flanks are bronze-brown with some small white stippling; the groin is green. The anterior surfaces of the thighs are dark brown with some cream stippling; the posterior surfaces of the thighs are dark brown with cream tubercles lateral and below the vent. The ventral surfaces are dark brown with uniformly distributed cream spots. The iris is yellow-cream with brown reticulations and a brown horizontal bar across the eye. Coloration in preservative. The dorsum is uniform bluish-gray. The pale labial, canthal, and dorsolateral stripes formed by a series of small bluish gray dots. The labial stripe continues to the insertion of the forelimb. The flanks, groin, and anterior surfaces of thighs are uniform dull gray; the posterior surfaces of thighs are dark gray with numerous white tubercles lateral to, and below, the cloacal opening. The ventral surfaces are uniform dark bluish-gray. Measurements (in mm). SVL: 54.0, TIBL: 20.5, FL: 24.7, HL: 17.8, HW: 20.5, IOD: 4.9, EW: 5.6 IND: 3.4, ED: 5.6, EN: 4.3, TD: 2.3, FFL: 9.9, TFL: 18.5, TFD: 3.4. Variation. Morphometric variation of three females and three males is summarized in Table 3. Females are larger than males (55.9± 2.1 mm; 52.7.± 2.5 mm). The skin on the dorsum varies from weakly to coarsely areolate. All adults have a supratympanic fold, which usually extends from the posterior edge of the tympanum to a point above the insertion of the forelimb. Color variation in preservative. All preserved specimens have a uniformly bluish gray dorsum; labial, canthal, and dorsolateral stripes are formed by a series of small brown dots. The dorsal surfaces of hands and feet also have brown dots. The flanks, groin, and anterior surfaces of the thighs are uniform dull gray, but in some specimens the flanks are covered by small dark gray dots. The posterior surfaces of the thighs are dark gray with numerous white tubercles lateral to, and below the cloacal opening; a short, fragmented supracloacal stripe is present in some individuals. The ventral surfaces are uniform dark bluish-gray. Color variation in life. (Figs. 10J, 18–19). The dorsum is uniform green (e.g., CJ 1386) or brown (e.g., CJ 7823). In CJ 1386, the dorsal surfaces of limbs are green, whereas the inner and outer surfaces of the forearms, thighs, and shanks are brown. Dorsal surfaces of hands and feet have a pattern of brown dots over a lighter background. Head markings are like those of the holotype; a dorsolateral stripe is present in all specimens. The flanks are bronze-brown with or without white stippling; the axillae and groin are green, with a bluish suffusion in some specimens. The anterior surfaces of thighs are dark brown with or without cream stippling and, and the posterior surfaces of thighs are dark brown with cream tubercles lateral to, and below, the vent. The ventral surfaces are dark brown with uniformly distributed cream dots, but some specimens have larger cream spots, thereby resulting in a paler venter. Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 39, from a series (CJ 1959) obtained from a pond at Laguna Negra de Jimbura, Parque Nacional Yacuri, 3406 m, Loja Province, Ecuador, by Elicio E. Tapia, Sofía Carvajal-Endara, and Henry Grefa on 12 June 2011 (Fig. 5F). Total length 61.3, body length 23.5 (38% of total length). Body robust, ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile; body width at level of spiracle 14.7, and height at same position 11.9; head width at level of eyes 12.9. Lateral-line system not evident. Nostril medium sized (in proportion to body length), ovoid, protruding, with a fleshy annulus, its opening directed anterolaterally. Snout–nostril distance 3.7. Eyes positioned and directed dorsally, eye length 2.4, eye width 2.2. Spiracle sinistral, located at midbody level; spiracular opening oriented posteriorly; distance from tip of snout to spiracle opening 15.9; spiracle end rounded, not free, attached to body wall, inner wall of spiracle not evident; tube length 3.4, tube transverse width 1.4. Vent tube dextral, the opening oriented posteriorly, tube length 4.0, tube transverse width 2.6. Tail length 38.8, caudal musculature robust in the two-thirds proximal to body, narrowing gradually until tail terminus; tail muscle height 5.4, tail muscle width 4.8; caudal fins thin at proximal half of tail, getting higher on distal half and raising near tail–body junction, dorsal fin height 3.5, ventral fin height 3.7; maximum height of tail 12.7; tail tip rounded, tail musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body, not visible dorsally; transverse width 5.6. It is surrounded by an uniserial row of marginal papillae, interrupted medially in upper lip; lower lip papillae alternating in and out, giving the appearance of two series; upper lip with 18 papillae on right side and 17 papillae on left side; lower lip with 42 marginal papillae; upper jaw sheath medium-sized, forming a finely serrated, smooth arch, height 0.4, transverse width 3.3 (59% of oral disc width); lower jaw sheath V- shaped, open and finely serrated, width 2.4, height 0.7. Labial tooth row formula 2/3(1), tooth rows lengths: A1: 4.7, A2: 4.0, P1 right row 1.9, P1 left row 2.1, P1 gap 0.1, P2: 3.6, P3: 3.5. (Fig. 6E). Color in preservative. Dorsum dull gray, with darker areas on flanks, above the eyes and on the throat; body contour and snout translucent. Caudal musculature and fins with scattered, white spots, densely arranged near tailbody junction; fins otherwise translucent. Venter dark gray; eyes lavender gray, oral apparatus translucent. Color in life. (CJ 1957, Stage 36). Fig. 5F. In dorsal and lateral views, body tan, speckled with black; areas around the snout are lighter. Venter cream with black markings, semi-translucent; guts visible as a darker area; throat translucent with small cream flecks; gills evident as a red hue. Caudal musculature reddish-pink in proximal half, decreasing its intensity towards distal half; myomeres and nerves barely visible; caudal musculature and fins with cream marks, clustered in dorsal line of caudal musculature, otherwise caudal fins translucent. Legs cream. Oral apparatus light cream. Iris copper-yellow, with small black reticulations. Variation. Variation of 26 meristic characters of tadpoles in Stages 35–39 (CJ 1959) are shown in Table 7. Total length varies between 61.3 (Stage 39) and 75.0 (Stage 35); tail length proportion varied from 63.2 to 67.0 until Stage 39; labial tooth row formula was 2/3(1). Number of marginal papillae varied among specimens and Gosner stages, variation in lower lip papillae is high (42–57). ......continued on the next page We documented changes in coloration during ontogenetic development of CJ 1958–9 (Fig. 20). At Stage 36, the dorsum and flanks are pale brown with dark brown areas on posterior body and flanks. By Stage 42, dorsal surfaces of body and limbs are uniform green; fine, cream, dorsolateral stripes are present, extending to midbody. Cream stripes border the outer dorsal margins of the limbs. At Stage 46, dorsal surfaces of body and limbs have more well-defined cream stripes, and a cream labial stripe; the canthus rostralis is green, and remaining flanks are mostly dark brown with an upper diffuse black-stripe. Dorsal surfaces of fingers and toes are brown except on finger and toe discs, which are yellowish-cream. The iris is copper-yellow, lacking the brown horizontal band that is observed in adults. Comparisons. Tadpoles of Gastrotheca turnerorum may occur in sympatry with those of G. elicioi, G. lojana, G. psychrophila, and G. pseustes, in the Loja-Abra de Zamora region. Gastrotheca turnerorum differs from G. elicioi by lacking a dorsal gray-pigmented fin that abruptly arises from the body; from G. lojana by having a more rounded tail terminus and having bold cream marks in a dorsal line of caudal musculature, and from G. pseustes by having a more rounded tail terminus (compare in Fig. 5). For G. psychrophila, see remarks under G. elicioi tadpole account. Distribution and ecology. Gastrotheca turnerorum only is known from three localities in the Cordillera Oriental of the Andes in southern Ecuador. These localities are in Parque Nacional Yacuri, Parque Nacional Podocarpus, and Reserva Tapichalaca in Zamora Chinchipe and Loja provinces. Its elevational range is 3073–3406 m in an area of extent of occurrence of about 450 km 2. This mostly nocturnal, semiarboreal species inhabits mainly paramos and a few forests in the Evergreen Shrub and Herbazal of Paramo, and the Evergreen Montane Forest of Catamayo-Alamor (Ministerio de Ambiente del Ecuador 2012), where the average annual rainfall is 895–1278 mm and the average annual temperature is 10.4– 13.5 °C (Fick & Hijmans 2017). At the Lagunas del Compadre in the Parque Nacional Podocarpus, a gravid female was collected on 1 December 1994. It deposited tadpoles on 12 December in the laboratory, some of them were raised, and others were preserved. On 20 September 2009, another female with eggs in her pouch was found at the same locality. It was sitting on moss at approximately 2 m from the Río Sabanilla, which is about 1.5 km from Lagunas del Compadre. The holotype, from Laguna Negra de Jimbura in the Parque Nacional Yacuri (Fig. 11C), was found at the edge of a pond about 3 x 2 m at the edge of a lagoon, at 14:00 h. The water temperature was 16.4° C, and water pH was 5.3. Eighteen tadpoles were collected at this pond. Another group of tadpoles was found at 18:00 h; at the same locality in another lagoon, the water temperature was 7.4 ° C. Strong winds blow across the lagoon, and only a small patch of forest remained at one border (Elicio E. Tapia field notes, 12 June 2011). A brooding female is depicted in Figure 10J. Conservation status. We suggest that Gastrotheca turnerorum should be considered as an Endangered species according to criteria B2ab(i,ii,iii) of the IUCN Red List. Although this species occurs at the Yacuri and Podocarpus National Parks, we suggest this conservation status because of its small known area of occurrence (159 km 2) that is vulnerable to improperly regulated tourism activities (Aguirre 2001), introduced species such as trout, climate change, and pathogens. Laguna Negra de Jimbura has suffered the synergistic effects of agriculture, cattle and sheep raising, fires, introduced species, pesticide use, and unregulated tourism activitites. Etymology. The specific name turnerorum is a latinized word that honors the Turner family. As unique as this marsupial frog is to the amphibian world, the Turner family stands out in their exceptional, unwavering and ever- growing commitment to conservation of the world natural resources. From Ted Turner’s pioneering efforts to protect very large land areas and restore them to their original and natural states of clean water and abundant fauna and flora to granddaughter Elizabeth’s awareness-raising book “Our friends the frogs,” there are multiple generations of active conservationists with a true appreciation for the value of all of our natural resources. In 1991, Laura Turner Seydel (with father Ted) co-founded the Captain Planet Foundation and continues to work diligently to inspire the same appreciation for conservation and sustainable living in the next generation of young people who will surely be making big decisions that will affect the health of our planet for themselves and their own grandchildren. Through the camera’s eye, Rhett Turner has managed to put conservation issues such as the pollinator perils and amphibian crisis in front of millions of people. Through the Turner Endangered Species Fund, there is hope for dozens of threatened species of plants and animals in the US and abroad that was not there before. Together, the Turner family has put their hearts, souls, and personal resources into helping make th

Gastrotheca yacuri sp. nov. urn:lsid:zoobank.org:act: 6261932E-02AB-4C9E-B2BD-BD213BBDC771 Holotype. CJ 7822 (Figs. 21���22), an adult male, from El Salado de Jimbura, at about 2.5 km (by road) west from the entrance in the Parque Nacional Yacuri, 2914 m (04�� 42' 08.37" S, 79�� 27' 00.09" W), Loja Province, Ecuador, collected on 9 April 2016 by Paul Sz��kely and Diana Sz��kely. Paratype. Ecuador: Loja: QCAZ 21105, adult male, from El Salado de Jimbura, 2712 m, on 26 June 2002 by Fernando Nogales Sornoza. Referred specimen. Ecuador: Loja: Not collected (Fig. 11O), subadult, 43 mm SVL, from nearby of Lagunas Negras, Parque Nacional Yacuri, 3492 m (04�� 42' 31.81" S, 79�� 25' 49.61" W). Diagnosis. Included in the genus Gastrotheca by molecular evidence and general morphological similarity with species of Gastrotheca. Although females are unknown, we infer presence of dorsal brood pouch because all its closely related species have it. A moderately large species (57.4���58.9 mm SVL in males, n = 2) with tibia length 47��49% SVL, larger than foot; (2) interorbital distance greater than width of upper eyelid; (3) skin on dorsum weakly granular to smooth, co-ossified with skull, having a transverse ridge at occipital region; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus smooth; (7) Finger I shorter than Finger II, width of discs much wider than digits; (8) fingers unwebbed; (9) toes one-half webbed, webbing extending to antepenultimate subarticular tubercle on Toe IV, to penultimate subarticular tubercle on Toe V; (10) in life, dorsum uniform green or brown with brown paravertebral marks; (11) head markings consisting of brown labial stripe with cream glandular areas at posterior end, canthal and supraciliary stripe of black, brown, and bronze; (12) dorsolateral stripe absent or present, when present above consisting in a series of bronze and cream warts, bordered below with a black and brown line, (13) flanks brown and green with a blue and black reticulum towards the lower flanks, axillae, groin, anterior and posterior surfaces of thighs, shanks, and tarsus; brown pelvic patch; (14) venter cream with nearly uniformly distributed, small, dark brown marks or brown with cream marks; gular region dark brown in males. Gastrotheca yacuri differs from all other species of Gastrotheca in southern Ecuador by having a blue and black reticulated pattern on flanks, axilla, groin, thighs, and shanks. It most closely resembles two other species, G. pseustes and G. psychrophila by having the blue colors, but they lack a blue and black reticulum (compare these species in Fig. 10). Gastrotheca yacuri differs from its sister species, G. aguaruna, from the Cordillera Central in northern Peru, by lacking blue colors, having the skin co-ossified with skull, and being smaller (SVL of males of G. yacuri 57.4���58.9 mm vs 41.6���46.8 mm in G. aguaruna). The genetic distance between them is of at least 2.8 % (in a DNA dataset of 438 bp, 16S gene). Description of the holotype. An adult male (Figs. 21���22); body moderately robust; SVL 57.4 mm; head wider that long; snout slightly acuminate in dorsal view, bluntly rounded in profile; canthus rostralis angular in section; loreal region concave; lips rounded; top of head flat; interorbital distance 92% of width of upper eyelid; internarial area flat; nostrils not protuberant, directed anterolaterally, at about level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from the eye by distance about equal to diameter of tympanum; tympanic evident; supratympanic fold elevated, extending from behind the tympanum to the insertion of the forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing four teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 28% of SVL); fingers unwebbed bearing fringes; discs large and rounded, width of disc of Finger III lesser that diameter of tympanum; relative lengths of fingers IColoration in life. The dorsal surfaces of the head, body, and limbs are green; the hidden surfaces of the forearm, thighs, and shanks are reticulated with black and blue markings (Figs. 21���22). Head markings are brown canthal and labial stripes. The canthal stripe is diffuse; the labial stripe varies from brown anteriorly to white posteriorly until the insertion of the forelimb. The dorsolateral stripe is thin and consists of a series of bronze and cream warts, bordered below with a black and brown line. The flanks are brown and green with a blue and black reticulum towards the lower flanks, axillae, groin, anterior and posterior surfaces of thighs, shanks, and tarsus; the pelvic patch is brown. The venter is cream with nearly uniformly distributed, small, dark brown marks; gular sac is nearly uniform grayish-brown with small dark brown marks. Palms and soles are pinkish gray; ventral fingers and toes are fleshy. The iris is copper with abundant black reticulations. Coloration in preservative. Similar to coloration in life, but green and blue surfaces have begun to fade. Measurements (in mm). SVL: 57.4, TIBL: 27.2, FL: 24.8, HL: 17.1, HW: 19.2, IOD: 6.9, EW: 5.2, IND: 2.8, ED: 6.0, EN: 3.0, TD: 3.3, FFL: 8.2, TFL: 16.2, TFD: 2.8. Variation. Morphometric variation of the male paratype is as follows: SVL: 58.9, TIBL: 28.6, FL: 28.3, HL: 17.9, HW: 20.1, IOD: 6.7, EW: 4.3, IND: 2.8, ED: 5.6, EN: 5.0, TD: 3.0, FFL: 10.8, TFL: 19.0, TFD: 2.7. The skin on the dorsum varies in the subadult by lacking a smooth tranverse occipital ridge. Color variation in life. The referred subadult specimen varies from the holotype in that it is mostly brown; it has a light reddish-brown dorsum of body and limbs with darker, well-defined stripes, two paravertebral and one at the coccyx position; scattered black spots are present on dorsum towards the flanks (Fig. 10O). The loreal region is brown, and a thin dark brown canthal line is present; the labial stripe is dark brown and bordered along its upper margin by a thin pale stripe that varies from brown anteriorly to cream posteriorly until the insertion of the forelimb. There is no dorsolateral stripe but the reddish-brown dorsal color contrasts with a broad dark brown band on the flanks. The flanks are brown with black and cream marks (without blue colors). The venter is mostly brown with cream marks that are larger towards the chest; the gular region is slightly darker that the venter. The palms and soles are gray. The iris is cooper with black reticulations and has a shade of red in the lower half. Vocalization. Four individuals of Gastrotheca yacuri were recorded at El Salado, Loja Province (Appendix III). Descriptive statistics of the acoustic variables are provided in Table 5. The advertisement call is a complex call, composed of one to three (usually two) long pulsed notes and followed (or not) by one to three (usually two) short, single-pulsed notes (Fig. 23 A���G). The long note had a mean duration of 0.634 s (SD = 0.091) and consisted on average of 32.24 (SD = 7.504) distinct pulses, partly fused, without silent intervals (amplitude modulation close but less than 100%). The amplitude of the long note increases gradually towards the middle of the note after which it decreases gradually by the end. The short notes had a mean duration of 0.072 s (SD = 0.020) and the inter-note interval was on average of 0.549 s (SD = 0.214). The mean dominant frequency of the call was 1496.7 Hz (SD = 38.643), with a mean 90% bandwidth frequency of 1366.7���1637.9 Hz, having the highest values recorded among the analyzed species. The fundamental frequency is recognizable; when visible, 5 to 7 harmonics are distinguishable. Comparisons. The advertisement call of Gastrotheca yacuri is most similar to that of G. pseustes, but G. yacuri has a much shorter call duration, shorter long notes duration, shorter inter-note interval, a higher dominant frequency, and a higher 90% bandwidth frequency compared with the call of G. pseustes (Table 5). Also, G. yacuri emits usually only two short notes compared with 4���6 short notes of G. pseustes. The call of G. yacuri can be easily distinguished form those of G. lojana and G. testudinea by the amplitude modulation of the longer note, longer call duration, lower short note rate, larger number of pulses, higher pulse rate, a much higher dominant frequency, and higher 90% bandwidth frequency. Also, G. yacuri emits up to three (usually two) long notes per call compared with the only one emitted by G. lojana and G. testudinea (Table 5). Distribution and ecology. Gastrotheca yacuri is only known from three nearby localities 2.8 km apart (maximum distance) at elevations of 2712���3492 m in the Cordillera Oriental de los Andes in southern Ecuador. One locality is within Parque Nacional Yacuri and the others are in El Salado de Jimbura in Loja Province (Fig. 9). This nocturnal, semiarboreal species inhabits paramo and subparamo in the Evergreen Montane Forest of Catamayo-Alamor (Ministerio de Ambiente del Ecuador 2012), where the average annual rainfall is 895���1159 mm and the average annual temperature is 10.4���14.6 ��C (Fick & Hijmans 2017). The male holotype was found near the road, calling from the branches of a bush, at about 3.5 m above ground. In this same location, choruses were heard from April to late July 2016; the males were calling usually from the vegetation nearby the road. An additional subadult was found during the day in a large terrrestrial bromeliad (Fig. 24A, B). Females and tadpoles are unknown. In this location, in Parque Nacional Yacuri, Gastrotheca yacuri is syntopic with G. turnerorum. Conservation status. We suggest that Gastrotheca yacuri should be assigned to the Data Deficient category according to guidelines of the IUCN (2001), given the inadecuacy of the current information to establish its status. Further searches and definition of its distribution, area of occupancy, habitat use, and biological information are required. Currently its known area of occurrence is less than 10 km 2, thus it could qualify for a Critically Endangered category; however, we can reasonably predict that if further surveys are conducted within the protected Yacuri National Park, its area will increase to more than 100 km 2. In any case, Lagunas Negras de Jimbura have suffered the synergistic effects of agriculture, cattle and sheep raising, fires, introduced species such as trout, pesticide use, and unregulated tourism activitites. Areas in the Salado de Jimbura that are out of the Yacuri National Park already have been heavily modified by human actions. Etymology. The specific name yacuri is a noun in apposition and refers to the Yacuri National Park, where this species has been found. According to Chamba-Troya (2017), Yacuri takes its name from the Quechua words Yacu meaning water and Quri that means gold. Ecuador's southernmost national park ranges from 2000 to 3700 meters in the Cordillera Oriental. This park of 43,090,60 hectares is unique by sheltering a series of 48 glacial lakes, and many endemic species of flora and fauna. It is a Ramsar site (The Convention on Wetlands) and part of the Podocarpus-El C��ndor Biosphere Reserve declared by UNESCO. Comments. Gastrotheca yacuri belongs to the subgenus Gastrotheca, and is reported as Species D in Duellman (2015: Fig 12.1)., Published as part of Carvajal-Endara, Sof��a, Coloma, Luis A., Morales-Mite, Manuel A., Guayasamin, Juan M., Sz��kely, Paul & Duellman, William E., 2019, Phylogenetic systematics, ecology, and conservation of marsupial frogs (Anura: Hemiphractidae) from the Andes of southern Ecuador, with descriptions of four new biphasic species, pp. 1-102 in Zootaxa 4562 (1) on pages 56-61, DOI: 10.11646/zootaxa.4562.1.1, http://zenodo.org/record/2627982, {"references":["Fick, S. E. & Hijmans, R. J. (2017) WorldClim 2: new 1 - km spatial resolution climate surfaces for global land areas. International Journal of Climatology, 37 (12), 4302 - 4315. https: // doi. org / 10.1002 / joc. 5086","IUCN (2001) Red list categories. Fersion 3.1. IUCN Species Survival Comission, Gland, 32 pp","Chamba-Troya, C. A. (2017) Faloracion del servicio ecosistemico de recreacion y belleza escenica del Parque Nacional Yacuri, 2015. Tesis de Economista, Universidad Tecnica Particular de Loja, Loja, 80 pp.","Duellman, W. E. (2015) Marsupial Frogs: Gastrotheca and Allied Genera. JHU Press, Baltimore, MD, 408 pp."]}

Gastrotheca lojana Parker 1932 Gastrotheca marsupiata lojana Parker, 1932:25.��� Holotype: BM 1947.2.31.13 (Fig. 25B) from Loja, Loja Province, Ecuador. Gastrotheca monticola (part)��� Duellman & Hillis 1987:158. Gastrotheca (Duellmania) lojana ��� Dubois, 1987:33. Referred specimens. Ecuador: Azuay: CJ 390, 401���2, from O��a, 2272 m (03�� 27' 41.04" S, 79�� 09' 46.47" W), on 15 June 2011 by Elicio E. Tapia, Sof��a Carvajal-Endara and Henry Grefa; KU 138401���3, from Gir��n, 2310 m (03�� 10' 00.12" S, 79�� 07' 59.88" W), on 0 6 June 1970 by Thomas A. Fritts; QCAZ 2692, from O��a, 2272 m (03�� 27' 41.04" S, 79�� 09' 46.47" W), on 1 December 1999 by Luis A. Coloma and Luis E. L��pez; QCAZ 26314���5, 26318, 26322���3, 26327���8, 26334���5, 26337���8, from O��a, 2272 m (03�� 27' 40.43" S, 79�� 09' 45.11" W), on 8 August 2003 by Luis A. Coloma and ��talo G. Tapia, QCAZ 31521���3, from O��a, 2272 m (03�� 27' 41.04" S, 79�� 09' 46.47" W), on 27 December 2005 by ��talo G. Tapia and Giovanni Onore; QCAZ 32212, from O��a, 2272 m (03�� 27' 41.04" S, 79�� 09' 46.47" W), on 2006 by Luis A. Coloma; QCAZ 32571, from O��a, 2272 m (03�� 27' 40.43" S, 79�� 09' 45.11" W), on 8 August 2003 by Luis A. Coloma, Erik R. Wild, Andr��s Merino-Viteri, ��talo G. Tapia and Edwin Patricio Vargas; QCAZ 42725, from San Fernando, Laguna de Busa, 2834 m (03�� 09' 15.80" S, 79�� 15' 49.03" W), on 22 October 2007 by Sof��a Carvajal-Endara, Amaranta Carvajal-Campos, Andrea Carvajal Endara. Loja: BM 1947.2.31.6���10, BM 1947.2.31.13���18, from Loja, 2150 m (04�� 00' 00" S, 79�� 13' 00.12" W), on 1930���1 by Clodoveo Carri��n Mora. KU 120673 ���4, from Loja, 2150 m (04�� 00' 00" S, 79�� 13' 00.12" W), on 9 June 1968 by John D, Lynch; KU 120675, from 2 km E Loja, 2100 m (03�� 59' 56.4" S, 79�� 13' 00.12" W), on 9 June 1968 by John D, Lynch, KU 138233, from 3 km W Loja, 2150 m (03�� 58' 47.99" S, 79�� 12' 54" W), on 21 June 1970 by Thomas H. Fritts; KU 138234, from 10 km W Loja, 2500 m (03�� 59' 56.4" S, 70�� 14' 43.08" W), on 27 June 1970 by Thomas H. Fritts; KU 138235���6, from 5 km N Loja, 2150 m (03�� 52' 00.12" S, 79�� 13' 00.12" W), on 28 June 1970 by Thomas H. Fritts; KU 138404���9, from Saraguro, 2412 m (03�� 36' 00" S, 79�� 13' 00.12" W), on 19���20 June 1970 by Thomas H. Fritts; KU 148568, from Saraguro, 2412 m (03�� 36' 00" S, 79�� 13' 00.12" W), on 22 May 1971 by Richard M. Montanucci; KU 178482���95, from 2 km SSW Saraguro, 2569 m (03�� 38' 22.92" S, 79�� 13' 59.87" W), on 5 January 1978 by John D. Lynch; KU 178496���7, from 2.1 km N Saraguro, 2575 m (03�� 36' 41.76" S, 79�� 13' 00.12" W), on 7 January 1978 by John D. Lynch; QCAZ 22371, from Loja Zamora Huayco, 3018 m (04�� 05' 49.9806" S, 79�� 10' 02.499" W), on 12 December 2002 by Diego Almeida-Reinoso; QCAZ 30788, from Saraguro, 2412 m (03�� 36' 00" S, 79�� 13' 00.01" W), on 20 September 1978 by MC; QCAZ 34505, from 2 km SW Saraguro, 2569 m (03�� 38' 22.99" S, 79�� 14' 24" W), on 5 January 1978 by John D. Lynch. El Oro: QCAZ 32724, from Guanaz��n, 2984 m (03�� 27' 18.43" S, 79�� 29' 23.64" W), on 3 December 2006 by Silvia Ald��s Alarc��n. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (54.1���76.1 mm SVL in females, n = 12; 40.2���61.0 mm SVL in males; n = 24) with tibia length 41��56% SVL, longer than foot; (2) interorbital distance about 1.6 times width of upper eyelid; (3) skin on dorsum finely granular, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus distinct, smooth; (7) Fingers I and II about equal in length, discs on fingers about twice width of digits proximal to discs; (8) fingers unwebbed; (9) webbing between external toes extending to the antepenultimate subarticular tubercle on Toe IV, to point midway between penultimate and distal subarticular tubercles on Toe V; (10) in life, dorsum green, brown, or tan with dark paravertebral marks; (11) markings on the head consisting of pale labial stripe broader at the posterior half of lip and dark interorbital bar or two blotches connected or not to paravertebral marks; (12) pale cream or white dorsolateral stripe fragmented; (13) flanks dark brown, green or tan with pale cream spots ventrally, anterior and posterior surfaces of thighs, groin, and dorsal surface of foot with contrasting pattern of heavily white mottling on a dark background, with bluish or greenish tinges; dorsal surfaces of limbs with dark brown or green bars; (14) venter creamy white with dark brown dots or marks; (15) brood pouch single, dorsal. Gastrotheca lojana most closely resembles three other species in southern Ecuador (G. elicioi, G. cuencana, and G. litonedis) and one species in northern Peru (G. monticola). Gastrotheca lojana differs from G. elicioi in some coloration patterns (described above; also, compare in Figs. 10L, 29, 30 vs 12���14). A black canthal stripe is absent in G. lojana, but present in G. elicioi. The anterior and posterior surfaces of thighs and groin are heavily mottled in G. lojana, but slightly mottled in G. elicioi. The interorbital mark in G. lojana is a transverse bar that sometimes is divided into two blotches and that is usually connected with two broad paravertebral marks. In G. elicioi dorsal marks, when present, consist of a triangular interorbital mark, which may or may not be connected to two narrow and curved paravertebral marks. Gastrotheca lojana has a conspicuous, elevated row of dorsolateral warts, whereas it is barely raised in G. elicioi. In addition, in G. elicioi the dark bars on limbs, when present, are shorter and less defined than in G. lojana. Gastrotheca lojana differs from its most related species, G. cuencana and G. litonedis (compare Figs. 10L, 29, 30 vs 2���4, 26���27) by having: a cream venter with dark flecks or spots (uniform creamy white in G. cuencana and pale brownish gray in G. litonedis); an interorbital bar or blotches usually connected to paravertebral marks (absent in G. cuencana and G. litonedis); dorsal surfaces of limbs with dark transverse bars (irregular dark blotches in G. cuencana and usually unmarked in G. litonedis); anterior and posterior surfaces of thighs and groin heavily mottled (translucent cream without marks in G. cuencana and usually brown and slightly molted in some G. litonedis); dorsal surfaces of fingers cream or with dark brown spots (cream in G. cuencana and of same color of the rest of the body in G. litonedis); a fragmented dorsolateral stripe (continuous in G. cuencana and G. litonedis); and complex call (simple call in G. cuencana and G. litonedis). The genetic divergence between G. lojana and G. litonedis is 2.5% and between G. lojana and G. cuencana is 2.8% (in a DNA dataset of 438 bp, 16S gene). Gastrotheca lojana differs from G. monticola by having the concealed surfaces of thighs, groin and dorsal surfaces of the feet with white mottling on a dark background, whereas in G. monticola the axilla and groin, concealed surfaces of thighs, and dorsal surfaces of the feet are green with black spots on a green or tan background. A distinctly different species, G. pseustes, occurs sympatrically with G. lojana in part of its range. Gastrotheca lojana differs from G. pseustes by having finely granular skin on the dorsum (smooth to coarsely granular in G. pseustes), larger digital discs, and pale dorsolateral and supracloacal stripes. Also the call of G. lojana can be easily distinguished form the call of G. pseustes by the amplitude modulation of the longer note, shorter call duration, higher short note rate, shorter interval between notes, fewer pulses, lower pulse rate, a much lower dominant frequency, and a lower 90% bandwidth frequency. Variation. Morphometric variation of 12 females and 24 males is summarized in Table 3. Females are larger than males (61.6�� 5.7 mm; 49.4�� 4.5 mm). All adults have a moderate supratympanic fold, which usually extends from the superior part of tympanum to point just above the insertion of forelimb. The dorsolateral row of warts can be less conspicuous in some specimens. Each vomer has four to ten teeth (5.7��1.4, n = 44). Color variation in preservative. In most preserved specimens the dorsum is bluish gray or brown with darker interorbital, paravertebral marks and bars on the limbs. Some specimens also have dark flecks densely distributed on dorsum. All specimens lack a dark canthal stripe and have a cream white supralabial stripe, which is broader at the posterior margin of the lip and that is extended from the posterior margin of the lip to the insertion of the forelimb. Most of the specimens have a dark brown stippling on the supralabial stripe. A fragmented supracloacal and heel stripes are present; also a white mark extends laterally from the vent. The flanks are dark gray with black and white dots; the posterior part of the flanks, groin, concealed surfaces of thighs and dorsal surface of foot are heavily mottled. The venter is white and varies from having a few dark flecks to having a heavily mottled pattern. Color variation in life. (Figs. 10L, 29, 30). In living individuals the color of the dorsum is green (CJ 408), brown (CJ 405) or tan (CJ 407) with darker interorbital and paravertebral marks. The interorbital mark is usually a bar that connects the two eyelids; however, in some individuals it is divided into two blotches (CJ 411). Paravertebral marks are broad and are usually connected with the interorbital mark (CJ 407) or, in some individuals (CJ 405), they are fused at the level of the scapular region. The venter is white in most specimens, some have dark flecks and spots evenly distributed, and others have a heavily mottled pattern. On the head there is a cream supralabial stripe, which is broader at the posterior margin of the lip and extends from the posterior margin of the lip to the insertion of the forelimb. The tympanum is brown, tan or olive green. The iris is reddish bronze with a few black reticulations. The flanks are brown, tan, or green with black and white spots. The posterior part of the flanks, groin, anterior and posterior surfaces of thighs have a heavily mottled pattern, some individuals have green or purple tinge on the groin and posterior surface of the thighs (Fig. 30). Supracloacal and heel stripes are cream or tan. The ventral surfaces of the shanks have a faint tinge of pale blue. Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 36, from a series (CJ 1948) obtained from a pond at O��a, 2272 m, Azuay Province, Ecuador, by Elicio E. Tapia, Sof��a Carvajal-Endara, and Henry Grefa on 15 June 2011 (Fig. 5C). Total length 55.7; body length 19.8 (36% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat shape concave in lateral profile; body width at the level of spiracle 12.3, and height at same position10.2; head width at level of eyes 10.5. Lateralline system barely visible, supraorbital and infraorbital lines both originating at tip of snout, running in parallel to the eye and making contact immediately behind the eye; angular line descending vertically from just posterior of eye to throat; anterior oral line descending vertically from level of oral disc and just anterior to level of nares to throat there it curve to parallel infraorbital line, thereby forming a continuous circuit with angular and loreal lines; dorsal body and middle body lines present. Nostril medium sized (in proportion to body length), ovoid, protruding, with a fleshy annulus, its opening directed anterolaterally. Snout���nostril distance 2.8; internarial distance 3.2. Eye positioned and directed dorsally, not visible from below; eye length 2.1, eye width 1.9; interorbital distance 5.7. Spiracle sinistral, located ventrolaterally; spiracular opening directed posteriorly; distance from tip of snout to spiracular opening 13.7; spiracle end rounded, attached to body wall, inner wall of spiracle not evident; tube length 4.0, tube transverse width 2.9. Vent tube dextral, its opening directed posteriorly, tube length 3.7, tube transverse width 2.1. Tail length 37.0, caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 5.2, caudal muscle width 4.3; caudal fins well developed and about the same size, originating near tail-body junction; maximum height of tail 13.6; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body, not visible dorsally; transverse width 4.1. It is surrounded by a uniserial row of short, marginal papillae, interrupted medially on upper lip; lower lip papillae alternating in and out, giving the appearance of two series; upper lip with 21 papillae on right side and 19 papillae on left side; lower lip with 63 marginal papillae; upper jaw sheath medium-sized, forming a finely serrated, smooth arch, height 0.4, transverse width 3.0 (48% of oral disc width); lower jaw sheath V- shaped, open and finely serrated, width 2.1, height 0.7. Labial tooth row formula 2/3(1), tooth rows lengths: A1: 4.2, A2: 4.1, P1 right row 1.9, P1 left row 1.6, P1 gap 0.3, P2: 3.2, P3: 3.5. (Fig. 6C). Color in preservative. Dorsum dull gray, with paler (translucent) areas on the flanks and snout. Caudal muscles pale gray with small cream flecks; dorsal and ventral fins translucent with evenly distributed cream flecks, except at tip where flecks are absent. Venter translucent on throat and belly regions, guts not exposed, white pigment partially covers the belly, producing a black and white mottling; eyes lavender gray with white flecks; oral apparatus translucent. Color in life. In dorsal and lateral views, body olive-cream; loreal and snout areas palest. Ventrally, guts not visible, belly white with gray marks; gills visible through the throat, with a reddish hue. Caudal musculature cream-gray; proximal third reddish with myomeres and medial line well defined; distal two thirds with small cream flecks; dorsal and ventral fins translucent, suffused with minute cream flecks, clustering in rounded patches near borders of fins; most dense near tail-body junction. Spiracle, oral apparatus, and legs olive-cream. Iris gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 34���41 (CJ 1948) are shown in Table 9. Total length varies between 40.9 (Stage 34) and 61.2 (Stage 41) and tail length proportion varies from 59.1 to 63.7 until stage 41; labial tooth row formula 2/3(1). Number of marginal papillae varies among specimens and Gosner stages, variation in number of ventral papillae at lower lip is moderate (60 and 69). The color of venter varies from nearly plain gray (CJ 4303) (Fig. 31) to gray flecks in a white background (CJ 1948) (Fig. 5C). Duellman (2015) described a tadpole (KU 203548) from 7.9 Km W Loja. Nonetheless, it may also belong to either G. pseustes, G. psychrophila, or G. turnerorum. We documented changes during ontogenetic development of CJ 4303, 4304 (Figs. 31 A���C, 32). At Stage 46, the dorsum is nearly uniform clear brown; the paraverterbal marks and interorbital marks are green and vary from nearly absent to well defined. Comparisons. Tadpoles of Gastrotheca lojana may occur in sympatry with those of G. elicioi, G. psychrophila (tadpole unknown, see Remarks under G. elicioi tadpole account), G. pseustes 1, and G. turnerorum in the Loja- Abra de Zamora region, with G. litonedis and G. pseustes 2 (tadpole not described), in the Laguna de Busa area and with at least G. pseustes in other localities (compare in Fig. 5). Gastrotheca lojana differs from G. elicioi by lacking a dorsal gray-pigmented fin that abruptly arises from the body, from G. pseustes 1 by lacking a reticulated pattern on flanks, from G. turnerorum by having a more pointed tail terminus and lacking bold cream marks in a dorsal line of caudal musculature, and from G. litonedis by having a highest tail dorsal fin. Vocalization. Five individuals of Gastrotheca lojana were recorded from one location in Azuay Province (one individual from San Fernando, Laguna de Busa) and from one location in Loja Province (four individuals from O��a) (Appendix III). The advertisement call of G. lojana is a complex call, composed of one long pulsed note and followed (or not) by one or two short, single-pulsed notes (Fig. 17 H���N). The long note had a mean duration of 0.562 s (SD = 0.125) and consisted on average of 17.59 (SD = 2.418) distinct pulses (pulse series), separated by silent intervals (amplitude modulation of 100%). The amplitude of the long note increases gradually towards the end, without falling. The short notes have a mean duration of 0.064 s (SD = 0.027) and the inter-note interval is on average of 0.356 s (SD = 0.107). The mean dominant frequency is 1118.1 Hz (SD = 126.872), with a mean 90% bandwidth of 986.1���1348.4 Hz. The fundamental frequency is not clearly recognizable; when visible, 5 to 7 harmonics are distinguishable in the short notes. Comparisons. The advertisement call of Gastrotheca lojana is most similar to that of G. testudinea, but G. lojana has a shorter call duration, higher short note rate, shorter long notes, short notes duration, shorter inter-note intervals, higher pulse rate, a higher dominant frequency and a higher 90% bandwidth frequency compared with the call of G. testudinea (Table 5). The call of G. lojana can be easily distinguished form the calls of G. yacuri and G. pseustes by the amplitude modulation of the longer note, shorter call duration, higher short note rate, shorter inter-note interval, smaller number of pulses, lower pulse rate, a much lower dominant frequency, and a lower 90% bandwidth frequency. Also, G. lojana emits only one long note, whereas G. yacuri emits up to three (usually 2) long notes per call and G. pseustes emits a larger number of long and short notes (Table 5). Distribution and ecology. Gastrotheca lojana is known in basins within Azuay, Loja, and El Oro Provinces. Its elevational range is 1682���3018 m in an area of extent of occurrence of about 1621.2 km 2. This nocturnal, semiarboreal species inhabits mostly disturbed areas and a few forests in the Evergreen Montane Shrub in the south of the, Published as part of Carvajal-Endara, Sof��a, Coloma, Luis A., Morales-Mite, Manuel A., Guayasamin, Juan M., Sz��kely, Paul & Duellman, William E., 2019, Phylogenetic systematics, ecology, and conservation of marsupial frogs (Anura: Hemiphractidae) from the Andes of southern Ecuador, with descriptions of four new biphasic species, pp. 1-102 in Zootaxa 4562 (1) on pages 71-79, DOI: 10.11646/zootaxa.4562.1.1, http://zenodo.org/record/2627982, {"references":["Parker, H. W. (1932) Some new or rare reptiles and amphibians from southern Ecuador. The Annals and Magazine of Natural History, Series 10, 9 (49), 21 - 26. https: // doi. org / 10.1080 / 00222933208673460","Duellman, W. E. & Hillis, D. M. (1987) Marsupial frogs (Anura: Hylidae: Gastrotheca) of the Ecuadorian Andes: resolution of taxonomic problems and phylogenetic relationships. Herpetologica, 43, 141 - 173.","Dubois, A. (1987) Again on the nomenclature of frogs. Alytes, 6 (1 - 2), 27 - 55.","Orton, G. L. (1953) The systematics of vertebrate larvae. Systematic Zoology, 2 (2), 63 - 75. https: // doi. org / 10.2307 / 2411661","Duellman, W. E. (2015) Marsupial Frogs: Gastrotheca and Allied Genera. JHU Press, Baltimore, MD, 408 pp.","Fick, S. E. & Hijmans, R. J. (2017) WorldClim 2: new 1 - km spatial resolution climate surfaces for global land areas. International Journal of Climatology, 37 (12), 4302 - 4315. https: // doi. org / 10.1002 / joc. 5086","Duellman, W. E. (1974) A systematic review of the marsupial frogs (Hylidae: Gastrotheca) of the Andes Ecuador. Occasional Papers of the Museum of Natural History, The University of Kansas, Lawrence, 22, 1 - 27.","Loaiza-S, C. R. (2012) Clodoveo Carrion Mora (1883 - 1957). Cientifico Insigne de la Provincia de Loja. Universidad Tecnica Particular de Loja, Ediloja, Loja, 255 pp.","Duellman, W. E., Barley, A. J. & Venegas, P. J. (2014) Cryptic species diversity in marsupial frogs (Anura: Hemiphractidae: Gastrotheca) in the Andes of northern Peru. Zootaxa, 3768 (2), 159 - 177. https: // doi. org / 10.11646 / zootaxa. 3768.2.4"]}

Gastrotheca cuencana sp. nov. urn:lsid:zoobank.org:act: 82F87E67-2940-424C-AC03- BC751224 D5E4 Holotype. CJ 1391 (Fig. 2), an adult male, from the city of Cuenca, 2579 m (02�� 53' 57.91" S, 79�� 01' 52.79" W), Azuay Province, Ecuador, one of a series obtained on 8 June 2011 by Elicio E. Tapia, Sof��a Carvajal-Endara and Henry Grefa. Paratypes. (Total 48: 10 adult males, 21 adult females, 17 juveniles). Ecuador: Azuay: CJ 1392���7, collected with the holotype; KU 120676 (female), 120683 ���4, 120690, 120695 (4 juveniles), 120699 (male), 120705 (male), 120709 ���10 (males), 120713 (female), 120718 ���9 (female, male), 120721 (female), 120722 (male) from Cuenca, 2600 m, (02�� 53' 57.91" S, 79�� 01' 52.79" W), on 19 June 1968 by John D. Lynch; KU 138616 (female), 138619��� 21 (male, females) from 4 km E: Cuenca 2540 m (02�� 52' 59.88" S, 78�� 22' 0.11" W), on 10 June 1970 by Thomas H. Fritts; KU 141572 (female) from 2.1 km S Cutchil, 2720 m (03�� 06' 00"S, 78�� 47' 59.99"W), on 16 May 1971 by Richard R. Montanucci; KU 141579 (female) from 3.5 km S Cutchil, 2785 m (03�� 04' 59.87" S, 78�� 47' 59.99" W), on 14 May 1971 by Richard R. Montanucci; KU 141582 (juvenile) from 8 km NW Cuenca, 2803 m (02�� 51' 47.88" S, 78�� 58' 59.88" W), on 16 May 1971 by Richard R. Montanucci; KU 141583 (female) from 8.8 km NW Cuenca, 3820 m (02�� 52' 00.12" S, 79�� 04' 00.12" W), on 15 May 1971 by Richard R. Montanucci; KU 203441 (male) from Laguna de Zurucuchu, 16 km NW Cuenca, 3200 m (02�� 50' 23.99" S, 79�� 07' 47.99" W), on 5 March 1984 by William E. Duellman; KU 129779 ���82 (4 females), 129783���91 (9 juveniles), 129793���94 (juveniles), 129795 (female), 129796 (juvenile) from R��o Matadero, 12 km E Cuenca (02�� 52' 59.88"S, 78�� 58' 00.12" W), on 17 November 1968 by Craig E. Nelson. Ca��ar: KU 141571 (male) and KU 141573 (female) from Bibli��n, 2620 (02�� 42' 00" S, 78�� 52' 00.12" W), on 17 and 23 May 1971, respectively, by Richard R. Montanucci; KU 142620 ���24 (4 females, male), from Bibli��n, 2620 m (02�� 42' 00" S, 78�� 52' 00.12" W), on 25 July 1971 by William E. Duellman; KU 147113 (female) from Bibli��n, 2620 m (02�� 42' 00" S, 78�� 52' 00.12" W), on 15 January 1972 by John E. Simmons. Referred specimens. (Total 36, 28 adult males, 8 adult females). Ecuador: Azuay: QCAZ 1239 (male), from Sigsig, 2480 m (03�� 03' 08.4" S, 78�� 48' 10.56" W), on 24 July 1989 by Luis A. Coloma and Luis E. L��pez; QCAZ 26309 (female), from Cumbe, 2740 m (03�� 05' 55.64" S, 79�� 00' 28.73" W), on 8 August 2003 by Patricio Vargas; QCAZ 26348���50 (males), 26353 (female), 26357���64 (8 males), from Cumbe, 2740 m (03�� 05' 55.64" S, 79�� 00' 28.73" W), on 9 August 2003 by Luis A. Coloma, ��talo G. Tapia, Andr��s Merino Viteri, Erik Wild and Patricio Vargas Mena; QCAZ 26354 (female), from Cumbe, 2740 m (03�� 05' 55.64" S, 79�� 00' 28.73" W), on 9 August 2003 by ��talo G. Tapia; QCAZ 26357���8 (males), from Cumbe, 2740 m (03�� 05' 55.64" S, 79�� 00' 28.73" W), on 9 August 2003 by Andr��s Merino Viteri; QCAZ 31477 (male), from Sigsig, 2424 m (03�� 03' 08.4" S, 78�� 48' 10.56" W), on 8 March 2006 by ��talo G. Tapia; QCAZ 31509, 31511 (males), from Sigsig, 2424 m (03�� 03' 8.4" S, 78�� 48' 10.56" W), on 9 March 2006 by ��talo G. Tapia and Giovanni Onore; QCAZ 34131 (female), from Cuenca, 2579 m (02�� 53' 57.91" S, 79�� 01' 52.79" W), on 27 February 1979 by Fernando I. Ortiz; QCAZ 37375���6, 37379���81, QCAZ 37386 (6 males), from unknown locality, on 31 May 2007 by Zool��gico Amaru; QCAZ 38232 (male), from Carmen del Guzho, 2666 m (02�� 56' 00" S, 79�� 03' 00" W), on 16 June 2007 by Hari Gonz��lez Maldonado; QCAZ 39384���5 (female, male), from Tarqui, Patapamba, 2600 m (03�� 00' 38.16" S, 79�� 01' 53.43" W), on 21 June 2007 by Ernesto Arbel��ez Ortiz; QCAZ 42720���1 (males) from Cuenca, 2579 m (02�� 53' 57.91" S, 79�� 01' 52.79" W), on 3 October 2007 by Sof��a Carvajal-Endara and Flavio Jaramillo; QCAZ 47106 (male), from Tarqui, 2741 m (03�� 00' 57.17" S, 79�� 02' 40.13" W), on 21 June 2007 by Ernesto Arbel��ez Ortiz. Ca��ar: QCAZ 42826, 42835, 42841 (females), from Papaloma de la Nube, 3011 m (02�� 40' 21.54" S, 78�� 54' 21.28" W), on 12���14 August 2008 by Sof��a Carvajal-Endara. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (49.0��� 61.6 mm SVL in females, n = 28; 45.2���53.7 mm SVL in males, n = 38), with tibia length 39��50% SVL, slightly shorter than length of foot; (2) interorbital distance slightly larger than width of upper eyelid; (3) skin on dorsum finely granular, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus weakly granular to smooth; (7) Fingers I and II about equal in length, width of discs greater than digits; (8) fingers unwebbed; (9) webbing between external toes extending to antepenultimate subarticular tubercle on Toe IV, to penultimate subarticular tubercle on Toe V; (10) in life, dorsum green, tan, brown, or reddish-brown with or without dark paravertebral marks; (11) head markings consisting of pale labial stripe and, in some individuals, an inconspicuous canthal stripe; (12) pale white or cream dorsolateral stripe present; (13) flanks tan, brown, or green with cream flecks or spots; groin and anterior and posterior surfaces of thighs translucent cream without marks and with a faint pale blue tinge in some; (14) venter uniform creamy white; (15) in females, brood pouch single, dorsal. In comparison with similar species in Ecuador, G. cuencana is most like G. litonedis, G. lojana, G. elicioi, and G. riobambae. It differs from these species by having a uniform creamy white venter; whereas the venter is pale gray in G. litonedis, and cream with dark flecks, spots or mottling in G. lojana, G. elicioi, and G. riobambae. In G. cuencana the dorsal surfaces of fingers are cream, whereas in the other species dorsal surfaces of fingers are of the same color as the rest of the body (tan, brown, or green). Gastrotheca cuencana also differs from G. litonedis, G. elicioi, and G. riobambae by lacking a dark canthal stripe. Moreover, in G. cuencana the groin and anterior surfaces of thighs are mostly translucent cream without marks; whereas, in G. litonedis the groin and anterior surface of thighs are usually bronze-brown or tan with cream or darks flecks and spots. In G. lojana and G. riobambae the groin and anterior surfaces of the thighs are usually cream with black mottling; in G. elicioi the groin and anterior surfaces of thighs usually have dark and cream flecks and spots. In profile, the snout in G. cuencana is rounded above and inclined anteroventrally, whereas the snout in G. elicioi, G. lojana, and G. litonedis is bluntly rounded. Furthermore, G. cuencana has a conspicuous, elevated row of dorsolateral warts whereas these are barely raised in G. litonedi s; a narrow, cream supracloacal stripe is present in G. litonedis, G. lojana, and G. riobambae; whereas the stripe is fragmented in G. elicioi and absent in most G. cuencana. Gastrotheca lojana and G. riobambae also differ from G. cuencana and G. litonedis by having a complex call, whereas the call in the latter two species is a simple call. Additionally, the call of G. cuencana has a lower note rate, shorter note duration, longer inter-note interval and a lower dominant and 90% bandwidth frequency compared with G. litonedis. Finally, G. cuencana differs from its sister species (G. litonedis) by having a genetic distance of 1.2% (in a DNA dataset of 438 bp, 16S gene). Gastrotheca cuencana occurs in syntopy with G. pseustes (sensu lato) throughout most of its range. Gastrotheca pseustes differs from G. cuencana by having coarsely granular skin on the dorsum, in contrast to the usually finely granular dorsum in G. cuencana. In G. cuencana the venter is uniform creamy white, whereas it is cream or white with dark flecks or spots in G. pseustes; also the call of G. cuencana is simple, whereas in G. pseustes the call is complex. Description of holotype. An adult male (Fig. 2); body moderately robust; SVL 50.6 mm; head wider that long; snout rounded in dorsal view, rounded and inclined anteroventrally in profile; canthus rostralis round in section; loreal region concave, lips rounded; top of head flat; interorbital distance 104% of width of upper eyelid; internarial area slightly elevated; nostrils not protuberant, directed anterolaterally, posterior to level of anterior margin of lower jaw; diameter of eye greater than its distance from nostril; tympanum round, separated from eye by distance about equal to diameter of tympanum; tympanic annulus and membrane weakly granular; supratympanic fold moderately weak, extending from behind the tympanum to the insertion of the forelimb. Dentigerous vomerine processes narrowly separated medially, each bearing four teeth. Arm robust; ulnar tubercles absent; hand and fingers moderately large (TFL 35% of SVL), unwebbed, with distinct narrow lateral fringes; discs moderately large and rounded, width of disc of Finger III greater that diameter of tympanum; relative lengths of fingers I=IIColoration in life (Fig. 2). The dorsum is yellowish brown with two darker, black delineated, continuous, paravertebral markings. These markings extend to the sacrum where they bifurcate and become two rows of irregular blotches at each side of the vertebral axis. The dorsal surfaces of the limbs are yellowish brown with irregular gray blotches. Cream dorsolateral and labial stripes are present; the latter continues to the insertion of forelimb; a faint brown canthal stripe is present. The flanks are bronze-brown with small black spots. The venter, groin, and anterior surfaces of the thighs are uniform creamy white; the posterior surfaces of the thighs have a faint bluish coloration. The dorsal surfaces of the fingers are cream. The gular region is creamy gray laterally. The iris is reddish bronze with black reticulations. Coloration in preservative. The dorsum is tan with two well-defined, gray paravertebral markings, which bifurcate at the level of sacrum to become two rows of irregular blotches on each side of the vertebral axis. A white dorsolateral stripe is present. The dorsal surfaces of limbs are tan with irregular gray blotches. A cream labial stripe continues to the insertion of the forelimb; a faint canthal is present. The flanks are gray; the venter, groin, and anterior surfaces of thighs are uniform creamy white; the posterior surfaces of thighs have a faint bluish coloration. The gular region is creamy gray. Measurements (in mm). SVL: 50.4, TIBL: 21.9, FL: 24.9, HL: 16.9, HW: 19.1, IOD: 5.2, EW: 5.0 IND: 3.3, ED: 5.6, EN: 4.1, TD: 2.8, FFL: 10.0, TFL: 17.5, TFD: 3.4. Variation. Morphometric variation of 28 females and 38 males is summarized in Table 3. Females are larger than males (54.2�� 2.9 mm; 49.6�� 2.4 mm). The skin on the dorsum is finely granular in most, but it varies in some specimens from weakly areolate to coarsely granular. All adults have a supratympanic fold that usually extends from the posterior part of the tympanum to point above the insertion of forelimb. The tympanic annulus is usually smooth, but some individuals (e.g., CJ 1395, 1940) have a slightly granular annulus (Fig. 3A, C). Each dentigerous vomerine process has 2���7 teeth (4.4��1.1, n = 39). Color variation in preservative. Preserved specimens have bluish gray dorsum (tan and reddish-brown in CJ 1397). Faint paravertebral marks are present. A canthal stripe is absent in most specimens; the flanks and posterior surfaces of the thighs are dark gray with white spots evenly distributed in some specimens. In all specimens, a prominent white labial stripe extends from the posterior margin of the lip to the insertion of the forelimb. A narrow white dorsolateral stripe is evident in most specimens. The groin and the anterior surfaces of the thighs are uniform creamy gray. The ventral surfaces are uniform creamy white. The posterior surfaces of thighs usually are creamy gray, but they are pale blue in some specimens. Some males have yellowish brown nuptial pads on the medial surface of the thumb. Color variation in life. (Figs. 3���4). The dorsum is uniform green (CJ 1393 ), reddish brown (CJ 1397), or tan (CJ 1392) with (CJ 1391) or without (CJ 1392) darker paravertebral marks. When present, the paravertebral markings usually are well defined; at the level of the sacrum they become two rows of irregular blotches at each side of the vertebral axis. The ventral surfaces are uniform creamy white. A cream labial stripe is extends from the posterior margin of the lip to the insertion of the forelimb; a dark brown canthal stripe is absent in most individuals. The tympanum is brown, tan, or olive green. The iris is reddish bronze with black reticulations. A creamy white dorsolateral stripe is evident in most individuals. The flanks are cream, bronze brown, or green. The groin and anterior surfaces of the thighs are translucent cream without dark markings (Fig. 3). The posterior surfaces of the thighs are translucent, cream, or pale blue or green. When present, supracloacal and heel stripes are cream or tan. The ventral surfaces of the shanks have a faint tinge of pale blue. The gular surface varies from white to creamy gray. ������continued on the next page ������continued on the next page Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 38 (CJ 1945a), from a series of eight tadpoles (CJ 1945) obtained from a pond at south Cuenca, 2579 m, Azuay Province, Ecuador, by Elicio E. Tapia, Sof��a Carvajal-Endara, and Henry Grefa on 8 June 2011 (Fig. 5A). Total length 44.6; body length 17.3 (39% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile, sloping from tip of snout to belly; body width at the level of spiracle 11.5, and height at same position 9.3, head width at level of eyes 8.5. Lateral line system present but barely visible, supraorbital and infraorbital lines both rising at level of tip of snout, extending parallel to the eye and making contact immediately behind eye; angular line descending vertically just porterior of eye to throat, it dorsally contacts with post infraorbital line; anterior oral line descending vertically from oral disc level and behind nares level to throat; it makes a curve that parallels infraorbital line, forming a circuit continuous with angular and loreal lines; dorsal body and middle body lines not visible. Nostril medium sized (in proportion to body length), ovoid, protruding, having a fleshy annulus, its opening directed anterolaterally. Snout���nostril distance 2.3; internarial distance 2.4. Eye directed dorsally; eye length 1.8, eye width 1.8; interorbital distance 4.3. Spiracle sinistral, located at midbody level, spiracular opening oriented posteriorly; distance from tip of snout to spiracular opening 11.0; end of spiracular tube rounded, attached to body wall, inner wall of spiracular tube not evident; tube length 3.1, tube transverse width 2.0. Vent tube dextral, opening oriented posteriorly, tube length 1.7, tube transverse width 1.8. Tail length 27.2; caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 3.7, width 3,2; caudal fins well developed and proportional, arising near tail-body junction, forming a low hump; dorsal fin height 3.0, ventral fin height 2.8; maximum height of tail 9.3; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body; transverse width 4.7. It is surrounded by a uniserial row of marginal papillae, interrupted medially on upper lip; lower lip papillae alternate in orientation, giving appearance of two rows; upper lip with 29 papillae on right side and 27 papillae on left side; lower lip bearing 62 marginal papillae; upper jaw sheath medium-sized, forming a smooth arch and finely serrated, height 0.36, transverse width 2.8 (60% of width of oral width); lower jaw sheath V- shaped, open and finely serrated, width 1.92, height 0.7. Labial tooth row formula 2/3(1); tooth rows lengths: A1: 3.6, A2: 3.5, P1 right row 1.5, P1 left row 1.6, P1 gap 0.2, P2: 3.4, P3: 3.0. (Fig. 6B). Color in preservative. Dorsum dull gray with darker areas on flanks, above eye, and on throat; body contour and snout translucent. Caudal musculature and fins with scattered cream dots, with higher suffusion near tail base, otherwise translucent. Venter pale, translucent in belly region, guts exposed; eyes lavender gray; oral apparatus translucent. Color in life. In dorsal view, body olive-cream; snout and flanks paler olive-cream. In ventral view, guts dark gray in belly area; reddish gills visible throughout throat. Caudal musculature covered by a nearly continuous line of cream marks, proximal half of musculature reddish cream with myomeres and medial line well defined, distal half translucent; dorsal and ventral fins translucent, with scattered cream dots, heavily suffused near tail-body junction. Spiracle, oral apparatus, and legs olive-cream. Iris gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 37���39 (CJ 1945) are shown in Table 4. Total length varies, Published as part of Carvajal-Endara, Sof��a, Coloma, Luis A., Morales-Mite, Manuel A., Guayasamin, Juan M., Sz��kely, Paul & Duellman, William E., 2019, Phylogenetic systematics, ecology, and conservation of marsupial frogs (Anura: Hemiphractidae) from the Andes of southern Ecuador, with descriptions of four new biphasic species, pp. 1-102 in Zootaxa 4562 (1) on pages 17-34, DOI: 10.11646/zootaxa.4562.1.1, http://zenodo.org/record/2627982, {"references":["Orton, G. L. (1953) The systematics of vertebrate larvae. Systematic Zoology, 2 (2), 63 - 75. https: // doi. org / 10.2307 / 2411661","Fick, S. E. & Hijmans, R. J. (2017) WorldClim 2: new 1 - km spatial resolution climate surfaces for global land areas. International Journal of Climatology, 37 (12), 4302 - 4315. https: // doi. org / 10.1002 / joc. 5086","Amphibian Ark (2013) El Centro de Conservacion de Anfibios en el Zoologico Amaru, Ecuador. Boletin Informativo Amphibian Ark, 25, 13.","Siavichay-Pesantez, F., Maldonado-Cedeno, G. & Mejia-Coronel, D. (2016) Anfibios Urbanos de Cuenca. Editorial Don Bosco, Centro Grafico Salesiano, Cuenca, 108 pp.","Arbelaez Ortiz, E. & Vega Toral, A. (2008) Guia de Anfibios, Reptiles y Peces del Parque Nacional Cajas / Cajas National Park Amphibian, Reptile and Fish Guide. ETAPA, Municipalidad Cuenca, Grafisum, Cuenca, 106 pp.","Duellman, W. E. (2015) Marsupial Frogs: Gastrotheca and Allied Genera. JHU Press, Baltimore, MD, 408 pp."]}

Gastrotheca litonedis Duellman & Hillis 1987 Holotype. KU 202690 (Figs. 10A, 25A), an adult female, from 10 km (by road) northeast of Gir��n, 2750 m (03�� 07' 11.8" S, 79�� 06' 28.4'' W), Azuay Province, Ecuador, obtained on 7 March 1984 by William E. Duellman. Referred specimens. (Total 28: 13 males, 15 females). Ecuador: Azuay: CJ 386 ��9 (females), 401��4 (females, male), 1404��8 (males), KU 335388 ���9 (male, female), San Fernando, Laguna de Busa, 2834 m (03�� 09' 15.8" S, 79�� 15' 49.03" W), collected on 9 June 2011 by Elicio E. Tapia, Sof��a Carvajal-Endara and Henry Grefa; QCAZ 42734 (female), San Fernando, Laguna de Busa, 2834 m (03�� 09' 15.8" S, 79�� 15' 49.03" W), collected on 27 October 2007 by Sof��a Carvajal-Endara, Amaranta Carvajal-Campos and Andrea Carvajal-Endara; QCAZ 42855 (female), 42857��9 (females), 42861 (female), 42866 (male), 42871 (male), San Fernando, Laguna de Busa, 2834 m (03�� 09' 15.8" S, 79�� 15' 49.03" W), collected on 18���20 August 2008 by Sof��a Carvajal-Endara; QCAZ 49973 ��4 (female, male), 49976��8 (males), San Gerardo, 2854 m (03�� 08' 00.17" S, 79�� 11' 36.78" W), collected on 1 December 2010 by Sof��a Carvajal-Endara. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (53.5��� 62.4 mm SVL in females, n = 22; 48.9���57.4 mm SVL in males, n = 13) with tibia length 40��54% SVL, slightly longer than foot; (2) interorbital distance about 1.5 times width of upper eyelid; (3) skin on dorsum smooth, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus distinct, smooth; (7) Fingers I and II about equal in length, width of discs notably wider than digits; (8) fingers unwebbed; (9) webbing between external toes extending to the penultimate subarticular tubercle on Toe IV, to the distal subarticular tubercle on Toe V; (10) in life, dorsum green, brown, or tan with or without dark paravertebral marks or dark flecks; (11) head markings consisting of a pale cream labial stripe and narrow dark brown canthal stripe; (12) pale creamy white dorsolateral stripe present; (13) flanks, anterior surfaces of thighs and groin pale brown or tan slightly molted in some, posterior surfaces of thighs dark brown���gray with pale cream warts in some; (14) venter pale brownish gray; (15) brood pouch single, dorsal. Gastrotheca litonedis most closely resembles five other species in southern Ecuador��� G. cuencana, G. plumbea, G. pseustes, G. turnerorum, and G. elicioi. Gastrotheca litonedis differs from all of them by having a pale brownish gray venter; whereas, the venter is uniform creamy white in G. cuencana, cream in G. plumbea, cream with dark flecks or spots in G. pseustes and G. elicioi, and bronze or dark brown in G. turnerorum. Gastrotheca litonedis, like G. plumbea, has smooth skin on the dorsum, whereas it is finely granular in G. elicioi and G. cuencana, weakly areolate to smooth in G. pseustes and areolate in G. turnerorum. A pale supracloacal stripe is present in G. litonedis, G. pseustes, and G. turnerorum, but it is absent in G. cuencana and G. plumbea. Gastrotheca pseustes differs further from G. litonedis by having a complex call structure (long and short pulsed-notes), which is simple in G. litonedis (only short pulsed-notes). The sister species of G. litonedis is the parapatric G. cuencana. Distinctive morphological and coloration features (compare Figs. 26���27, vs 2���4) are (character states for G. cuencana in parentheses): Gastrotheca litonedis is larger than G. cuencana (SVL student���s t-test: t = -6.725, df = 99, p G. litonedis has a higher note rate, longer note duration, shorter inter-note interval, higher dominant frequency, and higher 90% bandwidth frequency compared with G. cuencana. Finally, these two species also differ by having a genetic distance of 1.2 % (in a DNA dataset of 438 bp, 16S gene). Gastrotheca litonedis is syntopic with G. lojana and G. pseustes 2, from which it differs notably by the texture of skin on dorsum, ventral coloration, and call structure. The skin on the dorsum is smooth in G. litonedis, whereas it is finely granular in G. lojana and coarsely granular in G. pseustes 2; furthermore, both species have cream venters with dark flecks, spots, or mottling, and a complex call structure with long and short pulsed-notes (G. lojana call in Fig. 17 H���N). Variation. Morphometric variation of 22 females and 13 males (from Duellman 2015) is summarized in Table 3. Females are larger than males (58.0�� 2.4 mm; 53.8�� 2.8 mm). The skin on the dorsum is mostly smooth in most specimens; however, in some specimens it is weakly areolate. All adults have a moderate supratympanic fold, which usually extends from the upper part of the tympanum to a point above the insertion of the forelimb. Some individuals have several small rounded tubercles anterolateral to the tympanum. Each dentigerous vomerine process has 2���10 teeth (5.4��1.6, n = 34). Color variation in preservative. In most specimens the dorsum is bluish gray without marks; in a few specimens faint paravertebral marks are present. The narrow canthal stripe, flanks, and posterior surfaces of thighs are dark brown. A creamy white supralabial stripe from the posterior margin of the lip to the insertion of the forelimb is prominent in all specimens. Creamy white dorsolateral, supracloacal and heel stripes are present. On the posterior part of the flanks the coloration varies from small white flecks to mottling on a dark brown ground color. The anterior surfaces of the thighs are usually gray with black flecks or spots. The ventral surfaces are uniform dull gray in most specimens; some have a pale gray venter with evenly distributed dark flecks and spots, and in CJ 1412 the venter is pale gray with black markings. Some specimens have a faint tinge of pale blue on the ventral surfaces of the shanks. Males have a cream nuptial pad on the medial surface of the thumb. Color variation in life. (Figs. 26, 27). The dorsum varies from uniform green (QCAZ 42858), to brown (CJ 404); in some individuals dark paravertebral marks and dark brown flecks are present (QCAZ 49977). The ventral surfaces are pale gray in most specimens; some have evenly distributed dark flecks and spots, and some have a faint bluish tinge on the ventral surfaces (QCAZ 49977). A narrow dark brown canthal stripe is present; a cream supralabial stripe extends from the posterior margin of the lip to the insertion of the forelimb. The tympanum is brown, tan, or olive green. The iris is reddish bronze with dense black reticulations. The posterior surfaces of the thighs are dark brown or cream with orange flecks. A cream dorsolateral stripe is evident in most individuals. The flanks, groin, and anterior surface of the thighs usually are bronze-brown or tan with cream or dark flecks and spots. In some individuals a pink-orange tinge is present in the groin (Fig. 27). The supracloacal and heel stripes are cream or tan, and the ventral surfaces of the shanks have a faint pale blue tinge. Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 37 (CJ 6558), from a series of 79 tadpoles (CJ 6557) born and reared in laboratory conditions, from a mother collected when gravid in the field (CJ 6018) at Laguna de Busa, 2780 m, Azuay Province, Ecuador, by Manuel A. Morales-Mite on 0 6 January 2016 (Fig. 5A). Total length 52.8; body length 19.4 (37% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile, sloping from tip of snout to belly; body width at the level of spiracle 12.5, and height at same position 10.3, head width at level of eyes 10.0. Lateral line system present but barely visible, supraorbital and infraorbital lines not evident at level of snout, infraorbital line present at level of the eye, touching the inferior portion of the orbit, and making contact with supraorbital line immediately behind the eye. Postorbital line represented by three stitches arranged in line and forming a 45-degree angle with the body plane. Angular line, anterior oral line and loreal lines not visible; dorsal body and middle body lines not visible. Nostril medium sized (in proportion to body length), ovoid, protruding, having a fleshy annulus, its opening directed anterolaterally. Snout���nostril distance 4.2; internarial distance 2.8. Eyes dorsally positioned; eye length 1.9, eye width 1.8; interorbital distance 5.2. Spiracle sinistral, located at midbody level, spiracular opening oriented posteriorly; distance from tip of snout to spiracular opening 12.9; end of spiracular tube rounded, attached to body wall, inner wall of spiracular tube not evident; spiracle length 2.7, spiracle transverse width 3.5. Vent tube dextral, opening oriented posteriorly, tube length 2.5, tube transverse width 2.2. Tail length 33.3; caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 4.1, width 3.6; caudal fins well developed and proportional, dorsal fin arising abruptly near tail-body junction; dorsal fin height 4.1, ventral fin height 3.7; maximum tail height 9.2; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body; transverse width 5.8. It is surrounded by a uniserial row of marginal papillae, interrupted medially on upper lip. Lower lip papillae alternate in orientation, giving appearance of two rows; upper lip with 23 papillae on right side and 23 papillae on left side; lower lip bearing 57 marginal papillae. Upper jaw sheath medium-sized, forming a smooth arch and finely serrated, transverse width 2.9 (50% of width of oral width) and height 0.3. Lower jaw sheath V- shaped, open and finely serrated, width 2.6, and height 0.7. Labial tooth row formula 2/3(1); tooth rows lengths: A1: 4.5, A2: 4.4, P1 right row 1.65, P1 left row 2.0, P1 gap 0.2, P2: 4.1, P3: 3.95. (See also Fig. 6A of CJ 5292, Stage 35). Color in life. Based on a specimen in Stage 34 from a series (CJ 1947) obtained from San Fernando, Laguna de Busa, Azuay Province, 2834 m, by Elicio E. Tapia, Sof��a Carvajal-Endara, and Henry Grefa on 9 June 2011 (Fig. 6A). In dorsal view, body olive-gray, head and tail suffused with pink and cream. Flanks cream anteriorly to olivegray posteriorly. In ventral view, cream anteriorly in gular region; followed posteriorly by red gills visible troughout the throat, and olive-gray guts. Caudal musculature pink proximally, gradually becoming gray posteriorly; dorsal and ventral fins translucent, with olive stippling distributed regularly along length of tail except distally, where stippling diminishes. Spiracle and oral apparatus nearly translucent. Iris gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 33���40 (CJ 5447, 5449, 5464���67, 6557, 6558) are shown in Table 8. Total length varies between 31.4 (Stage 33) and 61.9 (Stage 40) and tail length proportion varies from 58% to 69% until Stage 40. Number of marginal papillae varies among specimens and Gosner stages; variation in number of ventral papillae at lower lip is high (39���66). We documented changes in coloration during ontogenetic development of CJ 1947 (Fig. 28A���C). At Stage 39, the dorsum and flanks are olive-gray with a diffuse pattern of gray paravertebral marks. By Stage 41, dorsum of body and limbs is green with well-defined dark green paravertebral marks; a cream dorsolateral stripe is bordered below by a fine black line; a creamy white stripe is present on face where upper lip will be formed. The flanks are greenish gray. At Stage 46, the dorsum is nearly uniform green; the paraverterbal marks are barely evident. The flanks are brown, and the iris is red. CJ 6813 is similar to the above described but has scattered cream flecks on a green dorsum. Comparisons. Tadpoles of Gastrotheca litonedis may occur in sympatry with those of G. pseustes 2 and G. lojana. Gastrotheca litonedis differs from them by having less pigmentation on dorsum and venter (compare in Fig. 5). It further differs from G. lojana by having a lowest tail dorsal fin.and from G. pseustes by lacking a reticulated pattern on the flanks. Vocalization. A total of nine individuals of Gastrotheca litonedis were recorded from two locations (6 individuals from San Gerardo and 3 from San Fernando, Laguna de Busa) in Azuay Province (Appendix III). Descriptive statistics of the acoustic variables are provided in Table 5. The advertisement call of G. litonedis is a simple call, composed of a series of short, single-pulsed notes, emitted at regular intervals (Fig. 8 E���H). In our recordings, the calls have between 1 and 21 notes per call. The notes have a mean duration of 0.075 s (SD = 0.025) and a mean inter-note interval of 1.768 s (SD = 0.286). The mean dominant frequency of the call is 1224.0 Hz (SD = 66.415), with mean 90% bandwidth of 1114.7���1308.1 Hz. The fundamental frequency and harmonics are not clearly recognizable. Comparisons. The advertisement call of Gastrotheca litonedis is similar to that of G. cuencana; all the other species of Gastrotheca species in the southern Ecuadorian Andes have complex calls. However, the call of G. litonedis has a higher note rate, longer note duration, shorter inter-note interval, higher dominant frequency, and higher 90% bandwidth frequency compared with G. cuencana (Table 5). Distribution and ecology. Gastrotheca litonedis is known from three localities (ca. Gir��n, San Fernando (Laguna de Busa), San Gerardo) on the eastern slope of the Cordillera Occidental in Azuay Province (Fig. 9). Its elevational range is 2750���2854 m in an area of extent of occurrence of about 10.2 km 2. This nocturnal, semiarboreal species inhabits forest and disturbed areas in the Evergreen Montane Forest of Cordillera Occidental of the Andes (Ministerio de Ambiente del Ecuador 2012), where the average annual rainfall is 611���658 mm and the average annual temperature is 12.2���12.5 ��C (Fick & Hijmans 2017). Most of the specimens collected at Laguna de Busa were found among totora reeds (Schoenoplectus californicus) at the border of the lagoon. Males were calling from totora reeds at approximately 1���2 m above ground. Females were perched on totora reeds approximately 40 cm above ground and along small streams that were close to the lagoon. Some males also were calling from branches of shrubs around the lagoon approximately 2.5 m above ground. During the day a male and a female were basking on leaves of calla lilies (Zantedeschia aethiopica), which were present along irrigation ditches, bordering pastures. At this locality, Gastrotheca litonedis is syntopic with G. pseustes (sensu lato) and G. lojana; all of them have a biphasic mode of development, in which brooding females release tadpoles in ponds, where they complete their development (Figs. 10���11). In all the visits made to this locality (October 2007 and December 2007, August 2008, June 2011), gravid females were found, as well as tadpoles in small streams, temporary ponds, and at the edge of the lagoon. One of the gravid females collected on 9 June 2011 was transported to the laboratory, where it deposited 96 tadpoles. At San Gerardo, most individuals were found on the border of an irrigation ditch in a pasture. Males were calling from the grass at approximately 50 cm above ground. At this locality, G. pseustes (sensu lato) also was found. A brooding female of G. litonedis, showing heliophilic behavior during most of the incubation time, under captive conditions in an outdoor enclosure, is depicted in Figure 10C. Conservation status. IUCN categorizes Gastrotheca litonedis as Endangered (Coloma et al. 2004). However, the current assessment was based on information combining the two species described in this study (G. litonedis and G. cuencana). Thus, we re-categorize this species as Critically Endangered, according with IUCN criteria and sub-criteria B1ab(i,ii,iii,iv). Currently, the known area of occurrence of G. litonedis is extremely small (10.2 km 2), and its area of occupancy is even smaller; although we suspect that its distribution might increase as further searches are conducted. Its populations are in heavily human populated areas. For example, the surroundings of Laguna de Busa have been deforested and modified for human activities (Fig. 11D). The lake contains two introduced, predatory fish (trout and carp) and its surroundings are greatly altered by the introduction of exotic eucalyptus and conifers. Additional threats at this site are unregulated tourisms activities. Comments. In the original description of Gastrotheca litonedis, Duellman and Hillis (1987) included data on nine males and 15 females. As noted by Duellman (2015), of these specimens, only the holotype, KU 202690, is G. litonedis. The other specimens used by Duellman & Hillis (1987) are identified herein as G. cuencana. In addition, the photograph published as the holotype of G. litonedis by Duellman and Hillis (1987:156: Fig. 9) is not the holotype. The frog in the photograph is actually G. lojana (KU 148794, from 2 km SSW of Saraguro, Loja Province, Ecuador). Also, the frog QCAZ 2692 depicted in Duellman (2015: Fig. 11.12B) is G. lojana (not G. litonedis). Genetic sequences generated in this study from tissues of the holotype differ from the sequences of other specimens generated and published by Wiens et al. (2007) (Genbank numbers: DQ679395, DQ679355, DQ679323, DQ679287), which are more similar to sequences of G. cuencana., Published as part of Carvajal-Endara, Sof��a, Coloma, Luis A., Morales-Mite, Manuel A., Guayasamin, Juan M., Sz��kely, Paul & Duellman, William E., 2019, Phylogenetic systematics, ecology, and conservation of marsupial frogs (Anura: Hemiphractidae) from the Andes of southern Ecuador, with descriptions of four new biphasic species, pp. 1-102 in Zootaxa 4562 (1) on pages 61-67, DOI: 10.11646/zootaxa.4562.1.1, http://zenodo.org/record/2627982, {"references":["Duellman, W. E. & Hillis, D. M. (1987) Marsupial frogs (Anura: Hylidae: Gastrotheca) of the Ecuadorian Andes: resolution of taxonomic problems and phylogenetic relationships. Herpetologica, 43, 141 - 173.","Duellman, W. E. (2015) Marsupial Frogs: Gastrotheca and Allied Genera. JHU Press, Baltimore, MD, 408 pp.","Orton, G. L. (1953) The systematics of vertebrate larvae. Systematic Zoology, 2 (2), 63 - 75. https: // doi. org / 10.2307 / 2411661","Fick, S. E. & Hijmans, R. J. (2017) WorldClim 2: new 1 - km spatial resolution climate surfaces for global land areas. International Journal of Climatology, 37 (12), 4302 - 4315. https: // doi. org / 10.1002 / joc. 5086","Coloma, L. A., Ron, S. & Morales, M. (2004) Gastrotheca litonedis. In: IUCN 2013. IUCN Red List of Threatened Species. Fersion 2013.2. Available from: www. iucnredlist. org (accessed 30 October 2018)","Wiens, J. J., Kuczynski, C. A., Duellman, W. E. & Reeder, T. W. (2007) Loss and re-evolution of complex life cycles in marsupial frogs: Does ancestral trait reconstruction mislead? Evolution, 61 (8), 1886 - 1899. https: // doi. org / 10.1111 / j. 1558 - 5646.2007.00159. x"]}

Gastrotheca litonedis Duellman & Hillis 1987 Holotype. KU 202690 (Figs. 10A, 25A), an adult female, from 10 km (by road) northeast of Girón, 2750 m (03° 07' 11.8" S, 79° 06' 28.4'' W), Azuay Province, Ecuador, obtained on 7 March 1984 by William E. Duellman. Referred specimens. (Total 28: 13 males, 15 females). Ecuador: Azuay: CJ 386 ¯9 (females), 401¯4 (females, male), 1404¯8 (males), KU 335388 –9 (male, female), San Fernando, Laguna de Busa, 2834 m (03° 09' 15.8" S, 79° 15' 49.03" W), collected on 9 June 2011 by Elicio E. Tapia, Sofía Carvajal-Endara and Henry Grefa; QCAZ 42734 (female), San Fernando, Laguna de Busa, 2834 m (03° 09' 15.8" S, 79° 15' 49.03" W), collected on 27 October 2007 by Sofía Carvajal-Endara, Amaranta Carvajal-Campos and Andrea Carvajal-Endara; QCAZ 42855 (female), 42857¯9 (females), 42861 (female), 42866 (male), 42871 (male), San Fernando, Laguna de Busa, 2834 m (03° 09' 15.8" S, 79° 15' 49.03" W), collected on 18–20 August 2008 by Sofía Carvajal-Endara; QCAZ 49973 ¯4 (female, male), 49976¯8 (males), San Gerardo, 2854 m (03° 08' 00.17" S, 79° 11' 36.78" W), collected on 1 December 2010 by Sofía Carvajal-Endara. Diagnosis. Included in the genus Gastrotheca by having a closed brood pouch on dorsum of female. A moderately large species (53.5– 62.4 mm SVL in females, n = 22; 48.9–57.4 mm SVL in males, n = 13) with tibia length 40¯54% SVL, slightly longer than foot; (2) interorbital distance about 1.5 times width of upper eyelid; (3) skin on dorsum smooth, not co-ossified with skull, lacking transverse ridges; (4) supraciliary processes absent; (5) heel lacking calcar or tubercle; (6) tympanic annulus distinct, smooth; (7) Fingers I and II about equal in length, width of discs notably wider than digits; (8) fingers unwebbed; (9) webbing between external toes extending to the penultimate subarticular tubercle on Toe IV, to the distal subarticular tubercle on Toe V; (10) in life, dorsum green, brown, or tan with or without dark paravertebral marks or dark flecks; (11) head markings consisting of a pale cream labial stripe and narrow dark brown canthal stripe; (12) pale creamy white dorsolateral stripe present; (13) flanks, anterior surfaces of thighs and groin pale brown or tan slightly molted in some, posterior surfaces of thighs dark brown–gray with pale cream warts in some; (14) venter pale brownish gray; (15) brood pouch single, dorsal. Gastrotheca litonedis most closely resembles five other species in southern Ecuador— G. cuencana, G. plumbea, G. pseustes, G. turnerorum, and G. elicioi. Gastrotheca litonedis differs from all of them by having a pale brownish gray venter; whereas, the venter is uniform creamy white in G. cuencana, cream in G. plumbea, cream with dark flecks or spots in G. pseustes and G. elicioi, and bronze or dark brown in G. turnerorum. Gastrotheca litonedis, like G. plumbea, has smooth skin on the dorsum, whereas it is finely granular in G. elicioi and G. cuencana, weakly areolate to smooth in G. pseustes and areolate in G. turnerorum. A pale supracloacal stripe is present in G. litonedis, G. pseustes, and G. turnerorum, but it is absent in G. cuencana and G. plumbea. Gastrotheca pseustes differs further from G. litonedis by having a complex call structure (long and short pulsed-notes), which is simple in G. litonedis (only short pulsed-notes). The sister species of G. litonedis is the parapatric G. cuencana. Distinctive morphological and coloration features (compare Figs. 26–27, vs 2–4) are (character states for G. cuencana in parentheses): Gastrotheca litonedis is larger than G. cuencana (SVL student’s t-test: t = -6.725, df = 99, p G. litonedis has a higher note rate, longer note duration, shorter inter-note interval, higher dominant frequency, and higher 90% bandwidth frequency compared with G. cuencana. Finally, these two species also differ by having a genetic distance of 1.2 % (in a DNA dataset of 438 bp, 16S gene). Gastrotheca litonedis is syntopic with G. lojana and G. pseustes 2, from which it differs notably by the texture of skin on dorsum, ventral coloration, and call structure. The skin on the dorsum is smooth in G. litonedis, whereas it is finely granular in G. lojana and coarsely granular in G. pseustes 2; furthermore, both species have cream venters with dark flecks, spots, or mottling, and a complex call structure with long and short pulsed-notes (G. lojana call in Fig. 17 H–N). Variation. Morphometric variation of 22 females and 13 males (from Duellman 2015) is summarized in Table 3. Females are larger than males (58.0± 2.4 mm; 53.8± 2.8 mm). The skin on the dorsum is mostly smooth in most specimens; however, in some specimens it is weakly areolate. All adults have a moderate supratympanic fold, which usually extends from the upper part of the tympanum to a point above the insertion of the forelimb. Some individuals have several small rounded tubercles anterolateral to the tympanum. Each dentigerous vomerine process has 2–10 teeth (5.4±1.6, n = 34). Color variation in preservative. In most specimens the dorsum is bluish gray without marks; in a few specimens faint paravertebral marks are present. The narrow canthal stripe, flanks, and posterior surfaces of thighs are dark brown. A creamy white supralabial stripe from the posterior margin of the lip to the insertion of the forelimb is prominent in all specimens. Creamy white dorsolateral, supracloacal and heel stripes are present. On the posterior part of the flanks the coloration varies from small white flecks to mottling on a dark brown ground color. The anterior surfaces of the thighs are usually gray with black flecks or spots. The ventral surfaces are uniform dull gray in most specimens; some have a pale gray venter with evenly distributed dark flecks and spots, and in CJ 1412 the venter is pale gray with black markings. Some specimens have a faint tinge of pale blue on the ventral surfaces of the shanks. Males have a cream nuptial pad on the medial surface of the thumb. Color variation in life. (Figs. 26, 27). The dorsum varies from uniform green (QCAZ 42858), to brown (CJ 404); in some individuals dark paravertebral marks and dark brown flecks are present (QCAZ 49977). The ventral surfaces are pale gray in most specimens; some have evenly distributed dark flecks and spots, and some have a faint bluish tinge on the ventral surfaces (QCAZ 49977). A narrow dark brown canthal stripe is present; a cream supralabial stripe extends from the posterior margin of the lip to the insertion of the forelimb. The tympanum is brown, tan, or olive green. The iris is reddish bronze with dense black reticulations. The posterior surfaces of the thighs are dark brown or cream with orange flecks. A cream dorsolateral stripe is evident in most individuals. The flanks, groin, and anterior surface of the thighs usually are bronze-brown or tan with cream or dark flecks and spots. In some individuals a pink-orange tinge is present in the groin (Fig. 27). The supracloacal and heel stripes are cream or tan, and the ventral surfaces of the shanks have a faint pale blue tinge. Tadpoles. Tadpoles belong to Type IV tadpole of Orton (1953), and the exotroph, benthic ecomorphological guild. All measurements are expressed in millimeters. The following description is based on a specimen in Stage 37 (CJ 6558), from a series of 79 tadpoles (CJ 6557) born and reared in laboratory conditions, from a mother collected when gravid in the field (CJ 6018) at Laguna de Busa, 2780 m, Azuay Province, Ecuador, by Manuel A. Morales-Mite on 0 6 January 2016 (Fig. 5A). Total length 52.8; body length 19.4 (37% of total length). Body ovoid in dorsal and lateral views, slightly depressed; throat slightly concave in lateral profile, sloping from tip of snout to belly; body width at the level of spiracle 12.5, and height at same position 10.3, head width at level of eyes 10.0. Lateral line system present but barely visible, supraorbital and infraorbital lines not evident at level of snout, infraorbital line present at level of the eye, touching the inferior portion of the orbit, and making contact with supraorbital line immediately behind the eye. Postorbital line represented by three stitches arranged in line and forming a 45-degree angle with the body plane. Angular line, anterior oral line and loreal lines not visible; dorsal body and middle body lines not visible. Nostril medium sized (in proportion to body length), ovoid, protruding, having a fleshy annulus, its opening directed anterolaterally. Snout–nostril distance 4.2; internarial distance 2.8. Eyes dorsally positioned; eye length 1.9, eye width 1.8; interorbital distance 5.2. Spiracle sinistral, located at midbody level, spiracular opening oriented posteriorly; distance from tip of snout to spiracular opening 12.9; end of spiracular tube rounded, attached to body wall, inner wall of spiracular tube not evident; spiracle length 2.7, spiracle transverse width 3.5. Vent tube dextral, opening oriented posteriorly, tube length 2.5, tube transverse width 2.2. Tail length 33.3; caudal musculature slender, narrowing gradually until tail terminus; caudal muscle height 4.1, width 3.6; caudal fins well developed and proportional, dorsal fin arising abruptly near tail-body junction; dorsal fin height 4.1, ventral fin height 3.7; maximum tail height 9.2; tail terminus rounded, caudal musculature not reaching fin terminus. Oral disc small, ventral, located near tip of snout, not protruding laterally beyond body; transverse width 5.8. It is surrounded by a uniserial row of marginal papillae, interrupted medially on upper lip. Lower lip papillae alternate in orientation, giving appearance of two rows; upper lip with 23 papillae on right side and 23 papillae on left side; lower lip bearing 57 marginal papillae. Upper jaw sheath medium-sized, forming a smooth arch and finely serrated, transverse width 2.9 (50% of width of oral width) and height 0.3. Lower jaw sheath V- shaped, open and finely serrated, width 2.6, and height 0.7. Labial tooth row formula 2/3(1); tooth rows lengths: A1: 4.5, A2: 4.4, P1 right row 1.65, P1 left row 2.0, P1 gap 0.2, P2: 4.1, P3: 3.95. (See also Fig. 6A of CJ 5292, Stage 35). Color in life. Based on a specimen in Stage 34 from a series (CJ 1947) obtained from San Fernando, Laguna de Busa, Azuay Province, 2834 m, by Elicio E. Tapia, Sofía Carvajal-Endara, and Henry Grefa on 9 June 2011 (Fig. 6A). In dorsal view, body olive-gray, head and tail suffused with pink and cream. Flanks cream anteriorly to olivegray posteriorly. In ventral view, cream anteriorly in gular region; followed posteriorly by red gills visible troughout the throat, and olive-gray guts. Caudal musculature pink proximally, gradually becoming gray posteriorly; dorsal and ventral fins translucent, with olive stippling distributed regularly along length of tail except distally, where stippling diminishes. Spiracle and oral apparatus nearly translucent. Iris gold. Variation. Variation of 28 meristic characters of tadpoles in Stages 33–40 (CJ 5447, 5449, 5464–67, 6557, 6558) are shown in Table 8. Total length varies between 31.4 (Stage 33) and 61.9 (Stage 40) and tail length proportion varies from 58% to 69% until Stage 40. Number of marginal papillae varies among specimens and Gosner stages; variation in number of ventral papillae at lower lip is high (39–66). We documented changes in coloration during ontogenetic development of CJ 1947 (Fig. 28A–C). At Stage 39, the dorsum and flanks are olive-gray with a diffuse pattern of gray paravertebral marks. By Stage 41, dorsum of body and limbs is green with well-defined dark green paravertebral marks; a cream dorsolateral stripe is bordered below by a fine black line; a creamy white stripe is present on face where upper lip will be formed. The flanks are greenish gray. At Stage 46, the dorsum is nearly uniform green; the paraverterbal marks are barely evident. The flanks are brown, and the iris is red. CJ 6813 is similar to the above described but has scattered cream flecks on a green dorsum. Comparisons. Tadpoles of Gastrotheca litonedis may occur in sympatry with those of G. pseustes 2 and G. lojana. Gastrotheca litonedis differs from them by having less pigmentation on dorsum and venter (compare in Fig. 5). It further differs from G. lojana by having a lowest tail dorsal fin.and from G. pseustes by lacking a reticulated pattern on the flanks. Vocalization. A total of nine individuals of Gastrotheca litonedis were recorded from two locations (6 individuals from San Gerardo and 3 from San Fernando, Laguna de Busa) in Azuay Province (Appendix III). Descriptive statistics of the acoustic variables are provided in Table 5. The advertisement call of G. litonedis is a simple call, composed of a series of short, single-pulsed notes, emitted at regular intervals (Fig. 8 E–H). In our recordings, the calls have between 1 and 21 notes per call. The notes have a mean duration of 0.075 s (SD = 0.025) and a mean inter-note interval of 1.768 s (SD = 0.286). The mean dominant frequency of the call is 1224.0 Hz (SD = 66.415), with mean 90% bandwidth of 1114.7–1308.1 Hz. The fundamental frequency and harmonics are not clearly recognizable. Comparisons. The advertisement call of Gastrotheca litonedis is similar to that of G. cuencana; all the other species of Gastrotheca species in the southern Ecuadorian Andes have complex calls. However, the call of G. litonedis has a higher note rate, longer note duration, shorter inter-note interval, higher dominant frequency, and higher 90% bandwidth frequency compared with G. cuencana (Table 5). Distribution and ecology. Gastrotheca litonedis is known from three localities (ca. Girón, San Fernando (Laguna de Busa), San Gerardo) on the eastern slope of the Cordillera Occidental in Azuay Province (Fig. 9). Its elevational range is 2750–2854 m in an area of extent of occurrence of about 10.2 km 2. This nocturnal, semiarboreal species inhabits forest and disturbed areas in the Evergreen Montane Forest of Cordillera Occidental of the Andes (Ministerio de Ambiente del Ecuador 2012), where the average annual rainfall is 611–658 mm and the average annual temperature is 12.2–12.5 °C (Fick & Hijmans 2017). Most of the specimens collected at Laguna de Busa were found among totora reeds (Schoenoplectus californicus) at the border of the lagoon. Males were calling from totora reeds at approximately 1–2 m above ground. Females were perched on totora reeds approximately 40 cm above ground and along small streams that were close to the lagoon. Some males also were calling from branches of shrubs around the lagoon approximately 2.5 m above ground. During the day a male and a female were basking on leaves of calla lilies (Zantedeschia aethiopica), which were present along irrigation ditches, bordering pastures. At this locality, Gastrotheca litonedis is syntopic with G. pseustes (sensu lato) and G. lojana; all of them have a biphasic mode of development, in which brooding females release tadpoles in ponds, where they complete their development (Figs. 10–11). In all the visits made to this locality (October 2007 and December 2007, August 2008, June 2011), gravid females were found, as well as tadpoles in small streams, temporary ponds, and at the edge of the lagoon. One of the gravid females collected on 9 June 2011 was transported to the laboratory, where it deposited 96 tadpoles. At San Gerardo, most individuals were found on the border of an irrigation ditch in a pasture. Males were calling from the grass at approximately 50 cm above ground. At this locality, G. pseustes (sensu lato) also was found. A brooding female of G. litonedis, showing heliophilic behavior during most of the incubation time, under captive conditions in an outdoor enclosure, is depicted in Figure 10C. Conservation status. IUCN categorizes Gastrotheca litonedis as Endangered (Coloma et al. 2004). However, the current assessment was based on information combining the two species described in this study (G. litonedis and G. cuencana). Thus, we re-categorize this species as Critically Endangered, according with IUCN criteria and sub-criteria B1ab(i,ii,iii,iv). Currently, the known area of occurrence of G. litonedis is extremely small (10.2 km 2), and its area of occupancy is even smaller; although we suspect that its distribution might increase as further searches are conducted. Its populations are in heavily human populated areas. For example, the surroundings of Laguna de Busa have been deforested and modified for human activities (Fig. 11D). The lake contains two introduced, predatory fish (trout and carp) and its surroundings are greatly altered by the introduction of exotic eucalyptus and conifers. Additional threats at this site are unregulated tourisms activities. Comments. In the original description of Gastrotheca litonedis, Duellman and Hillis (1987) included data on nine males and 15 females. As noted by Duellman (2015), of these specimens, only the holotype, KU 202690, is G. litonedis. The other specimens used by Duellman & Hillis (1987) are identified herein as G. cuencana. In addition, the photograph published as the holotype of G. litonedis by Duellman and Hillis (1987:156: Fig. 9) is not the holotype. The frog in the photograph is actually G. lojana (KU 148794, from 2 km SSW of Saraguro, Loja Province, Ecuador). Also, the frog QCAZ 2692 depicted in Duellman (2015: Fig. 11.12B) is G. lojana (not G. litonedis). Genetic sequences generated in this study from tissues of the holotype differ from the sequences of other specimens generated and published by Wiens et al. (2007) (Genbank numbers: DQ679395, DQ679355, DQ679323, DQ679287), which are more similar to sequences of G. cuencana.