Acta Herpetologica 16(2): 63-87, 2021 ISSN 1827-9635 (print) © Firenze University Press ISSN 1827-9643 (online) www.fupress.com/ah DOI: 10.36253/a_h-10742 A new species of the genus Noblella (Amphibia: Strabomantidae) from Ecuador, with new information for Noblella worleyae Carolina Reyes-Puig1,2,3,4,*, Juan M. Guayasamin2,5, Claudia Koch6, David Brito-Zapata1, Matthijs Holland- ers7, Melissa Costales8, Diego F. Cisneros-Heredia1,2,3 1 Universidad San Francisco de Quito USFQ, Colegio de Ciencias Biológicas & Ambientales COCIBA & Instituto de Diversidad Biológi- ca Tropical iBIOTROP, Museo de Zoología/ Laboratorio de Zoología Terrestre, Campus Cumbayá, Quito, Ecuador 2 Universidad San Francisco de Quito USFQ, Colegio de Ciencias Biológicas y Ambientales COCIBA, Instituto BIOSFERA, Campus Cumbayá, Quito, Ecuador 3 Instituto Nacional de Biodiversidad INABIO, Unidad de Investigación, Quito, Ecuador 4 CIBIO/InBIO, Centro de Investigação em Biodiversidade e Recursos Genéticos da Universidade do Porto, Instituto de Ciências Agrári- as de Vairão, R. Padre Armando Quintas, 4485-661 Vairão, Portugal 5 Universidad San Francisco de Quito USFQ, Colegio de Ciencias Biológicas y Ambientales COCIBA, Laboratorio de Biología Evolutiva, Campus Cumbayá, Quito, Ecuador 6 Zoologisches Forschungsmuseum Alexander Koenig ZFMK, Leibniz-Institut zur Analyse des Biodiversitätswandels, Bonn, Germany 7 Southern Cross University, School of Environment, Science and Engineering, Lismore, Australia 8 University of New Brunswick, Department of Biology, Fredericton, Canada *Corresponding autor. E-mail: creyesp@usfq.edu.ec Submitted on: 2021, 31st March; revised on: 2021, 22nd July; accepted on: 2021, 23rd July Guest Editor: Aaron M. Bauer Abstract. We describe a new species of terrestrial-breeding frog of the genus Noblella from the northwestern slopes of the Andes of Ecuador, in the province of Pichincha, Ecuador, and report a new locality for the recently described N. worleyae. We include a detailed description of the osteology of both species and discuss their phylogenetic relation- ships. The new species is differentiated from other species of Noblella by having discs of fingers rounded, without papillae; distal phalanges only slightly T-shaped; toes slightly expanded and rounded distally, without papillae; dorsum uniform brown with irregular suprainguinal dark brown marks; venter yellowish cream, ventral surfaces of legs and thighs reddish to brownish cream; and dark brown throat. The new locality for N. worleyae is located in Los Cedros Reserve, an area highly threatened by mining. We highlight the importance of protecting endemic species of small vertebrates in northwestern Ecuador. Keywords. Frog, Los Cedros Biological Reserve, endemism, Imbabura, Mindo, Pichincha, phylogeny. INTRODUCTION The amphibian diversity in the tropical Andes is outstanding (Duellman, 1988; Myers et al., 2000; Hut- ter et al., 2013, 2017). Each year, several species are described from montane forests of this biodiversity hot- spot (e.g., Rojas-Runjaic et al., 2018; Guayasamin et al., 2019; Paez and Ron, 2019; Reyes-Puig et al., 2019b; San- ta-Cruz et al., 2019; Yanez-Muñoz et al., 2019; Acevedo et al., 2020; Ospina-Sarria et al., 2020; Lehr et al., 2021). Most described species from Ecuador belong to the hyper-diverse genus Pristimantis (Paez and Ron, 2019; Reyes-Puig et al., 2020a), but diversity in other anuran taxa has also increased considerably (e.g., Osornophryne, Hyloscirtus, Noblella, Centrolenidae; Mueses-Cis- neros et al., 2010; Cisneros-Heredia and Gluesenkamp, 64 C. Reyes-Puig et alii 2010; Yánez-Muñoz et al., 2010a; Páez-Moscoso and Guayasamin, 2012; Almendáriz et al., 2014; Guayasamin et al., 2017a, 2019; Reyes-Puig et al., 2019c). Terrestrial-breeding frogs of the genus Noblella Barbour 1930 are minute-size anurans (SVL < 22 mm), morphologically differentiated by having terminal discs on digits not or barely expanded, discs and circum- ferential grooves present distally (except in N. duellm- ani), terminal phalanges narrowly T-shaped, pointed tips of at least Toes III‒IV, and an inner tarsal tubercle (De La Riva et al., 2008; Hedges et al., 2008; Duellman and Lehr, 2009). However, phylogenetic relationships of Noblella are not fully resolved and its monophyly is uncertain (De la Riva et al., 2017; Santa-Cruz et al., 2019). As currently defined, Noblella includes 16 spe- cies, fourteen distributed in the Andes of Ecuador, Peru, and Bolivia, and two (N. losamigos and N. myrmecoides) in the Amazonian lowlands from southeastern Colom- bia, Ecuador, Peru, Bolivia, and western Brazil (Frost, 2021). During the last 15 years, the number of species in the genus has doubled; and four new species have been described since 2019 (Catenazzi and Ttito, 2019; Reyes- Puig et al., 2019c, 2020b; Santa-Cruz et al., 2019). Cur- rently, the total number of species of the genus Noblella is 16, distributed in ten species in Peru, seven in Ecua- dor, three in Bolivia, and one in Colombia and Brazil (Frost, 2021). Andean species of the genus Noblella show a high level of endemicity, with very restricted distributions. While some species of Noblella may apparently be able to survive in environments modified by humans (e.g., N. duellmani, N. losamigos, N. lochites, N. nature- trekii; Duellman and Lehr, 2009; Reyes-Puig et al., 2019c; Santa-Cruz et al., 2019); most species (e.g., N. coloma, N. heyeri, N. personina, N. pygmaea; Lynch, 1986; Guayasamin and Terán-Valdez, 2009; Harvey et al., 2013) seem to depend on undisturbed forest. Three species of Noblella have been described from western Ecuador, all from mature mountain forests: Noblella heyeri (Lynch, 1986) occurs in southwestern Ecua- dor and extreme northwestern Peru; Noblella coloma Guayasamin and Terán-Valdez, 2009 is known from its type locality and surroundings (Rio Guajalito and Chiriboga area; Ron et al., 2019); and Noblella worleyae, a recently described species is known just from seven specimens, all found in mature forest in the Río Man- duriacu Reserve, province of Imbabura, Ecuador (Reyes- Puig et al., 2020b). While the Ecuadorian Andes have suffered serious habitat destruction and fragmentation caused by expan- sion of deforestation, agriculture, mining, among others (Castellanos et al., 2011; Roy et al., 2018; Guayasamin et al., 2019; Lessmann et al., 2019; Ortega et al., 2021), there are still some areas with mature forests that have not been exploited due to their complex topography, difficult access, private protection, or preservation for touristic activities. Unfortunately, all such sites are under strong anthropogenic pressure, including mining concessions and the expansion of agricultural boundaries, among others (Cuesta et al., 2017; Roy et al., 2018; Guayasamin et al., 2019; Ortega et al., 2021). These privileged areas have proven to keep an extremely high cryptic diver- sity of small vertebrates and contain the last remnant populations of numerous threatened species (Cisneros- Heredia and Yanez-Muñoz, 2010; Reyes-Puig et al., 2010, 2019a, 2019b; Yánez-Muñoz et al., 2010b, 2018; Guayasamin et al., 2018, 2019, 2020; Sánchez-Nivicela et al., 2018; Barrio-Amorós et al., 2020). During the last five years, we have carried out sur- veys on the western slopes of the Andes in the provinces of Imbabura and Pichincha, Ecuador. As a result of this continuous effort, we found a new species of leaf-litter frog of the genus Noblella, which we describe herein based on a combination of morphological, molecular, and osteological features. We also document new infor- mation on distribution, external morphology and osteol- ogy for the recently described Noblella worleyae, infor- mation that was not described in detail in the original description. We also include intraspecific variation that will allow complete full with members of the same genus in the future. MATERIALS AND METHODS Taxonomy We followed the family taxonomy proposed by Heinicke et al. (2018) and, also we revised De la Riva et al. (2017) and Bar- rietos et al. (2021). For identifying species, we assumed the uni- fied species concept (De Queiroz, 2005, 2007). Information for species comparisons was extracted from the original descrip- tions and cited once at the beginning of the comparison. Study area and fieldwork Over the last three years (i.e., 2018–2020), we have car- ried out field surveys at several localities in montane forests of northwestern Ecuador, mainly in the provinces of Imbabura and Pichincha. Specimens of two different species of Noblella were found in Mindo (province of Pichincha) and Los Cedros Biological Reserve (province of Imbabura). Mindo is a small town renowned for its adventure and nature-based touris- tic activities; thus the area has numerous reserves that protect cloud forests (Arteaga-Navarro et al., 2013). Los Cedros Bio- logical Reserve is a protected area that contains 6,879 hectares 65A new species of the genus Noblella from Ecuador of premontane humid tropical forest and cloud mountain forest. This reserve is located south of the Cotocachi-Cayapas Ecologi- cal Reserve (state protected area), and is also recognized by its endemic microfauna (Hutter and Guayasamin, 2015). Collected specimens were euthanized with benzocaine, fixed in 8% forma- lin, and preserved in 75% ethanol. Liver and leg muscle tissue samples were collected from all individuals prior to preserva- tion. Tissues were preserved in 95% ethanol and stored at -20°C at the Laboratorio de Biología Evolutiva USFQ. Specimens were deposited in the Museo de Zoología, Universidad San Francisco de Quito, Ecuador (ZSFQ). DNA extraction, amplification, and sequencing We obtained new DNA sequences of Noblella sp. nov. (ZSFQ 050–051). DNA was extracted from muscle or liver tis- sue following the protocol by Peñafiel et al. (2019). Standard polymerase chain reaction (PCR) was performed to amplify a fragment of the mitochondrial gene 16S rRNA, using a combi- nation of the following primers: 16L10, 16H36E, 16L34, 16H47 (Heinicke et al., 2007). Amplicons were sequenced in both directions by the Macrogen Sequencing Team (Macrogen Inc., Seoul, Korea). The new sequences were assembled and edited with Geneious 7.1.7 (GeneMatters Corp). After assemblage, the sequences were combined with sequences from GenBank for all species of Noblella and representatives of the genera within the Terrarana clade (sensu Hedges et al., 2008), including Bary- cholos Heyer 1969, Bryophryne Hedges, Duellman & Heinicke 2008, Craugastor Cope 1862, Haddadus Hedges, Duellman & Heinicke 2008, Holoaden Miranda-Ribeiro 1920, Ischnocnema Reinhardt & Lutken 1862, Lynchius Hedges, Duellman & Hein- icke 2008, Niceforonia Goin & Cochran 1963, Oreobates Jimé- nez de la Espada 1872, Qosqophryne Catenazzi, Mamani, Lehr, von May 2020, Phrynopus Peters 2873, Pristimantis Jiménez de la Espada 1870, Psychrophrynella Hedges, Duellman & Heinicke 2008, and Microkayla De la Riva, Chaparro, Castroviejo-Fisher, Padial 2017. GenBank codes are shown in our inferred phyloge- netic tree (Fig. 1). Phylogenetic analyses Phylogenetic relationships were inferred using maximum likelihood as the optimality criterion. The final matrix, with 52 terminals, was aligned with MAFFT v.7 (Multiple Align- ment Program for Amino Acid or Nucleotide Sequences: http:// mafft.cbrc.jp/alignment/software/), with the Q-INS-i strat- egy. MacClade 4.07 (Maddison and Maddison, 2005) was used to visualize the alignment, which contained a total of 492 bp. Phylogenetic analyses were performed under the ML criteria in GARLI 2.01 (Genetic Algorithm for Rapid Likelihood Infer- ence; Zwickl, 2006) for the mitochondrial gene 16S. GARLI uses a genetic algorithm that finds the tree topology, branch lengths, and model parameters that maximize lnL simultaneously (Zwickl, 2006). Individual solutions were selected after 10,000 generations with no significant improvement in likelihood, with the significant topological improvement level set at 0.01. Then, the final solution was selected when the total improvement in likelihood score was lower than 0.05, compared to the last solu- tion obtained. Default values were used for other GARLI set- tings, as per recommendations of the developer (Zwickl, 2006). Bootstrap support was assessed via 1,000 pseudoreplicates under the same settings used in tree search. Pairwise genetic distances between species (uncorrected-p) for gene 16S were calculated with PAUP 4a (Swofford et al., 1996). External morphology Diagnosis and description of the new species follow for- mats proposed by Duellman and Lehr (2009) and Lynch and Duellman (1997). For comparisons, we examined specimens of other species of Noblella (see Appendix I). We followed the sequence of characters proposed by Guayasamin and Terán-Val- dez (2009). We measured preserved specimens using digital cal- ipers to the nearest 0.01 mm. These measurements are: snout to vent length (SVL), from the tip of the snout to the cloaca; head length (HL), measured from tip of snout to anterior edge of tympanum; head width (HW), measured at midorbital region; horizontal diameter of the eye (ED); eye–nostril distance (EN), from anterior ocular angle to posterior edge of nostril; horizon- tal diameter of tympanum (TD); minimum interorbital distance (MIOD); minimum eyelid width (MWE); hand length (LH), from posterior edge of palmar tubercle to tip of third digit; shank length (LS), from the tip of the ankle to the knee; and foot length (LF), from posterior edge of external metatarsal tubercle to tip of Toe IV. We determined sexual maturity by the presence of vocal slits or extended vocal sacs in males and by the presence of eggs or convoluted oviducts in females. Detailed illustrations of the head, hands and feet were done with Adobe InDesign ©. Osteology Osteological descriptions were based on one specimen of the new species (ZSFQ 050) and one of Noblella worleyae (MZUTI 1709). Both specimens were scanned using a high- resolution micro-computed tomography (micro-CT) desktop device (Bruker SkyScan 1173, Kontich, Belgium) at the Zoolo- gisches Forschungsmuseum Alexander Koenig (ZFMK, Bonn, Germany). To avoid movements during scanning, specimens were placed in a small plastic container and mounted with sty- rofoam. Acquisition parameters comprised: An X-ray beam (source voltage 43 kV and current 114 µA) without the use of a filter; 800 projections of 500 ms exposure time each with a frame averaging of 5 recorded over a 180° continuous rotation (rotation steps of 0.3 degrees), resulting in a scan duration of 49 min; a magnification setup generating data with an isotrop- ic voxel size of 19.16 µm (MZUTI 1709) and 14.55 µm (ZSFQ 050), respectively. The CT-dataset was reconstructed with N-Recon software (Bruker MicroCT, Kontich, Belgium) and rendered in three dimensions using CTVox for Windows 64 bits version 2.6 (Bruker MicroCT, Kontich, Belgium). Osteological 66 C. Reyes-Puig et alii terminology follows Trueb (1973), Duellman and Trueb (1994), Fabrezi and Alberch (1996), Guayasamin and Terán-Valdez (2009), Scherz et al. (2017), and Suwannapoom et al. (2018). Cartilage structures were omitted from the osteological descrip- tions because micro-CT does not render cartilage. RESULTS Phylogenetic relationships and genetic distances (Fig. 1) The inferred phylogeny shows that the new spe- cies described herein is part of a clade composed of taxa distributed along the western slopes of the Ecua- dorian Andes. This clade is composed by the new spe- cies Noblella sp. nov., Noblella coloma Guayasamin and Terán-Valdez, 2009, and N. worleyae Reyes-Puig, Maynard, Trageser, Vieira, Hamilton, Lynch, Culebras, Kohn, Brito and Guayasamin, 2020. Uncorrected p genetic distances are as follow: N. coloma (QCAZ 40579) and the new species (ZSFQ 050) = 5.1%; N. coloma (QCAZ 40579) and N. worleyae (ZSFQ 550–551) = 8.3%; N. worleyae (ZSFQ 550–551) and the new species (ZSFQ 050) = 1.2%. Generic placement We place the new species in the genus Noblella based on morphological and molecular evidence (Fig. 1). Morphologically, we assign the new species to the genus Noblella, as defined by Hedges et al. (2008), based on possession of the following traits: head not wider than body; cranial crests absent; tympanic membrane differentiated (undifferentiated in N. duellmani, N. naturetrekii and N. madreselva); dentigerous processes of vomers absent; terminal discs on digits not or barely expanded; discs and circumferential grooves present dis- tally (absent in N. duellmani); terminal phalanges nar- rowly T-shaped; Finger I shorter than, or equal in length to, Finger II; Finger IV containing two phalanges in Noblella carrascoicola (De la Riva and Köhler, 1998), N. lochites (Lynch, 1976b), N. losamigos (Santa-Cruz et al., 2019), N. myrmecoides (Lynch, 1976b), N. naturetrekii (Reyes-Puig et al., 2019c), and N. ritarasquinae (Köhler, 2000) and three phalanges in N. coloma (Guayasamin and Terán-Valdez, 2009), N. duellmani (Lehr, Aguilar, and Lundberg, 2004), N. heyeri (Lynch, 1986), N. sp. nov., N. lynchi (Duellman, 1991), N. madreselva (Cat- enazzi, Uscapi, and von May, 2015), N. personina (Har- vey, Almendáriz, Brito-M., and Batallas-R., 2013), N. peruviana (Noble, 1921), N. pygmaea (Lehr and Cat- enazzi, 2009), and N. thiuni (Catenazzi and Ttito, 2019); Toe III shorter than Toe V (except in N. naturetrekii and N. worleyae); tips of at least toes III–IV acuminate; sub- articular tubercles not protruding; dorsum pustulate or shagreen; venter smooth; SVL less than 22 mm. Systematic accounts Noblella mindo new species. Noblella coloma Arteaga et al. (2013). Figs. 2–8 LSID urn:lsid: lsid:zoobank.org:act:3B7741EF-BF26- 4589-B231-73F198AA1218 Proposed standard English name. Mindo Leaf Frog Proposed standard Spanish name. Rana Noble de Mindo Holotype. ZSFQ 050 (Fig. 2–6), adult female, col- lected in El Cinto, 11 Km E from Mindo town, Mindo (0.09022°S, 78.818581°W; 1,673 m; Fig. 2), province of Pichincha, República del Ecuador, by Melissa Costales, Matthijs Hollanders and Emilia Peñaherrera on 08 July 2017. Paratypes (2 females, 2 males). Adult males (ZSFQ 049, 051) and adult females (ZSFQ 304–305) collected at the type locality (same data as holotype), by Melissa Cos- tales on 04 October 2015. Etymology The specific name “mindo” is a word of unknown meaning in Panzaleo, an extinct pre-Columbian lan- guage of northern Ecuador (Jijón y Caamaño 1940). It is used as a noun in apposition, and alludes to the valley of Mindo, where the type locality of the new species is located. The remnant forests of this emblematic valley protect several species of endemic amphibians and rep- tiles such as Pristimantis mindo, Noblella mindo, and Anolis proboscis. Diagnosis Noblella mindo sp. nov. (Figs. 3–8) presents the fol- lowing characteristics: (1) skin of dorsum finely sha- green, skin on venter smooth, discoidal fold slightly defined, discoidal and thoracic folds absent; (2) tympan- ic annulus and membrane visible externally, supratym- panic fold inconspicuous (Figs. 3, 4); (3) snout short (eye-to-nostril distance 57% of eye diameter), rounded in dorsal and lateral views (Fig. 3); (4) eyelids without tubercles; (5) dentigerous processes of vomers absent; (6) vocal slits and sac present, nuptial pads not visible; (7) fingers not expanded distally, finger tips rounded, with- out papillae (Fig. 3); Finger I shorter than Finger II (Fig. 3); (8) distal phalanges slightly T-shaped, phalangeal for- mula of hands: 2-2-3-3 (Fig. 7); (9) supernumerary pal- mar tubercles present (slightly visible) mostly at the base 67A new species of the genus Noblella from Ecuador of the digits, ulnar tubercles diminutive and rounded, subarticular tubercles rounded; circumferential grooves absent; (10) one tarsal tubercle elongated and subconi- cal (Fig. 3); two prominent metatarsal tubercles (inner tubercle 3–4 times size of external); toes slightly expand- ed and rounded distally, without papillae; (11) Toe V shorter than Toe III, supernumerary plantar tubercles absent, distal portions of circumferential grooves not visible; (12) phalangeal formula of feet: 2-2-3-4-3 (Fig. 7); (13) in life, uniform brown dorsum, cream middor- sal, longitudinal line distinct and present in all individu- als, dark brown suprainguinal marks, white rictal gland, Noblella thiuni (MK072732, CORBIDI 18723) Noblella madreselva (MN056356, CORBIDI 15770) Noblella madreselva (MN064565, CORBIDI 15769) Psychrophrynella usurpator (KY652662, AC186-09) Psychrophrynella usurpator (KU884559, CORBIDI 16495) Psychrophrynella glauca (MG837565, CORBIDI 18729) Noblella losamigos (MN100040, —) Noblella losamigos (MN366392, MVZ 292687) Noblella losamigos (KY652644, RvM3-12) Noblella pygmaea (KY652645, MUSM 24536) Noblella sp (KY652646, MUSM 27582) Bryophryne bustamantei (KT276293, MHNC6019) Bryophryne cophites (EF493537, KU173497) Bryophryne phuyuhampatu (MF419259, —) Bryophryne tocra (MF186398, MUBI5419) Barycholos pulcher (EU186709, KU 217781) Barycholos ternetzi (KU495150, MALCX17P10) Noblella coloma (MT710932, QCAZ 40579) Noblella coloma (MT710931, QCAZ 32702) Noblella mindo sp. nov. (MT710934, ZSFQ-050) Noblella mindo sp. nov. (MT710935, ZSFQ-051) Noblella worleyae (MT710935, ZSFQ 551) Noblella worleyae (MT710934, ZSFQ 550) Noblella worleyae (MT710933, ZSFQ 2502) Noblella lochites (EU186699, KU177356) Noblella personina (MK838465, QCAZ 58818) Noblella heyeri (JX267463, QCAZ 31471) Noblella myermecoides (JX267464, QCAZ 40180) Noblella naturetrekii (MK838462, DHMECN 13307) Ischnocnema bilineata (JX267324, MNRJ 46476) Holoaden bradei (EF493366, USNM 207945) Holoaden luederwaldti (EU186710, MZUSP 131872) Ischnocnema colibri (MH538418, CFBH_41810) Ischnocnema parnaso (MH538421, CFBH 41812) Niceforonia brunnea (EF493357, KU178258) Niceforonia peraccai (EF493710, KU178266) Niceforonia dolops (EF493394, —) Lynchius parkeri (MK423937, QCAZ 61015) Lynchius simmonsi (JF810005, QCAZ 41640) Oreobates barituensis (JF810000, MCN1360) Oreobates quixensis (QCAZ 31186, JF810003) Phrynopus barthlenae (AM039653, —) Phrynopus inti (MF651906, UMMZ 245218) Phrynopus kauneorum (AM039655, —) Phrynopus sp (AM039660, —) Phrynopus pesantesi (AM039656, —) Phrynopus mariellaleo ( MH538299, CORBIDI 11658) Pristimantis cedros (KT210170, MZUTI 1710) Pristimantis ornatissimus (KU574613, MZUTI 4329) Pristimantis subsigillatus (KU999217, MZUTI 1999) Haddadus aramunha (KF740845, UFBA 283) Haddadus binotatus (KF740846, USP/TCX51ST09) Craugastor aenigmaticus (MK211617, UCR 22737) Craugastor tarahumaraensis (EU186702, —) Craugastor_longirostris (EF493395, KU 177803) 0.05 length units M i c r o k a y l a + Q o s q o p h r y n e C l a d e “N o b le lla ” S o u th e rn C la d e N o b le lla N o rth e rn C la d e 5 8 9 5 9 8 5 8 7 4 7 6 1 0 0 5 9 9 3 9 9 1 0 0 9 5 7 4 8 0 9 6 9 8 8 4 5 3 7 0 6 8 5 0 7 8 1 0 0 6 8 7 6 5 6 9 5 7 8 6 7 6 8 1 0 0 Fig. 1. Phylogeny of Noblella (light gray boxes) showing the relationships of N. mindo sp. nov. The phylogeny was inferred based on mito- chondrial (16S) DNA sequences (16S; 52 terminals, 492 bp) and under the Maximum likelihood criterion. For each individual, museum catalog number or, if unavailable, GenBank accession number is shown. 68 C. Reyes-Puig et alii flanks with dark brown band narrowing towards groin, dotted with white, light groin, with low concentration of melanophores, dark brown throat, chest and ventral surfaces of arms with a white cross formed by a longi- tudinal, fine line running from chin to chest crossing a similar line departing from midventral surface of each forelimb, yellowish-cream venter, reddish-copper iris with minute turquoise scattered dots (Fig. 4); (14) SVL in adult males 16.5–17.0 mm (mean 16.7 mm, n = 2), SVL in adult females 18.3–19.5 mm (mean 19.0 mm, n = 3). Comparisons (Fig. 6, Table 1) Noblella mindo sp. nov. differs from its congeners by the presence of rounded fingertips, without papillae; dis- tal phalanges slightly T-shaped; toes slightly expanded and rounded distally, without papillae; dorsum uniform brown with middorsal cream line, suprainguinal marks dark brown, rictal gland white, light groin, throat and chest dark brown with white cross, and venter yellow- ish cream. Noblella mindo sp. nov. is most similar and closely related to N. coloma and N. worleyae (Fig. 1), but they differ as follows (characters of N. mindo sp. nov. in parentheses): Noblella coloma has all finger tips acumi- nate (all rounded), dark middorsal line (light), dark ric- tal gland (white), orange to reddish-venter (yellowish- cream), dark groin (light), uniform dark brown throat, chest and ventral surfaces of arms (dark brown with white cross), ulnar tubercles absent (diminutive and rounded), and smaller body size of 16.0 mm SVL in adult female (18.3–19.5 mm SVL in adult females); the new species (characters of N. mindo sp. nov. in parentheses) is distinguished from Noblella worleyae has finger tips slightly acuminate on Fingers I and IV and acuminate on Fingers II and III (rounded), T-shaped distal phalanges (slightly T-shaped), prootic and exoccipital fused to form otoccipital (separated), sphenethmoid well-ossified and ventrally fused at midline (moderately ossified, ventrally fused at midline in posterior half and separated in anteri- or half). For more comparison’s information see Table 1. Noblella mindo sp. nov. has three phalanges on Fin- ger IV like N. duellmani (Lehr, Aguilar and Ludenberg, 2004), N. heyeri (Lynch, 1986), N. lynchi (Duellman, 1991), N. madreselva (Catenazzi, Uscapi and von May, 2015), N. personina (Harvey, Almendáriz, Brito, and Batallas, 2013), N. peruviana (Noble, 1921), N. pygmaea (Lehr and Catenazzi, 2009), and N. thiuni (Catenazzi and Ttito, 2019), but they differ as follows (characters of N. mindo sp. nov. in parentheses): Noblella duellmani has dorsal skin pustular (finely shagreen), tympanum membrane and annulus absent (present), upper eyelid bearing small tubercles (absent), ulnar tubercles coa- lesced into low folds (diminutive and round, not form- ing a fold), outer edge of tarsus bearing row of low and elongate tubercles (absent), tips of Fingers I-II slightly expanded and tips of Fingers III–IV slightly acuminate (all finger tips rounded), venter brown with tan mottling (yellowish-cream), and larger body size of 20.0 mm SVL in adult female (18.3–19.5 mm SVL in adult females); Noblella heyeri has dorsal skin weakly pustulate (finely shagreen), snout subacuminate in dorsal view (round), ulnar tubercles distinct and round (diminutive, round), toe tips slightly acuminate (round), venter brown with cream fleck (yellowish-cream), and smaller body size of 13.1–15.9 mm SVL in adult females (18.3–19.5 mm SVL in adult females); Noblella lynchi has dorsal skin pus- tular (finely shagreen), snout subacuminate in dorsal view (round), ulnar tubercles coalesced into low folds (diminutive and rounded, not forming a fold), outer edge of tarsus bearing row of low and elongate tubercles (absent), toe tips weakly acuminate (round), and venter brown with fine cream flecks (yellowish cream); Noblella madreselva has dorsal skin with small tubercles (finely shagreen), tympanic membrane not differentiated and tympanic annulus barely visible below skin (well-differ- entiated), upper eyelid with minute tubercles (absent), toe tips weakly acuminate (rounded), venter black with large and irregularly shaped white mark (yellowish- cream), and smaller body size of 17.6 mm SVL in adult female (18.3–19.5 mm SVL in adult females); Noblella Fig. 2. Distribution of Noblella mindo sp. nov and N. worleyae in Ecuador. 69A new species of the genus Noblella from Ecuador personina has dorsal skin smooth with pustules (finely shagreen), snout subtruncate in profile (round), finger and toe tips acuminate with papillae (round, lacking papillae), venter white (yellowish-cream), and smaller body size of 15.6–17.9 mm SVL in adult females (18.3– 19.5 mm SVL in adult females); Noblella peruviana has tympanic membrane not differentiated (differentiated), toe tips slightly acuminate (round), and venter tan (yel- lowish cream); Noblella pygmaea has dorsal skin tuber- cular (finely shagreen), thoracic fold present (absent), dorsolateral fold on anterior half of body present (absent), upper eyelid bearing small tubercles (absent), minute tubercle on heel present (absent), toe tips pointed (round), venter pale grayish brown (yellowish-cream), and smaller body size of 11.3–12.4 mm SVL in adult females (18.3–19.5 mm SVL in adult females); Noblella thiuni has thin dorsolateral folds visible on anterior half of body (absent), tympanic membrane not differentiated (differentiated), fingertips bulbous (round), ulnar tuber- cles absent (present, diminutive and round), venter cop- per reddish with a profusion of silvery spots (yellowish cream), and smaller body size of 11.0 mm SVL in male (16.5–17 mm in adult females). Noblella carrascoicola (De la Riva and Köhler, 1998), N. lochites (Lynch, 1976b), N. losamigos (Santa-Cruz et al., 2019), N. myrmecoides (Lynch, 1976b), N. naturetrekii (Reyes-Puig et al., 2019c), and N. ritarasquinae (Köhler, 2000) are easily differenti- ated from N. mindo sp. nov. by having two phalanges on Finger IV instead of three. Description of the holotype Adult female (ZSFQ 050); head narrower than body, its length 40.8% of SVL; head longer than wide; head width 31.1% of SVL; snout round in dorsal and lateral Fig. 3. Noblella mindo sp. preserved holotype, ZSFQ 050, adult female, SVL = 18.3 mm. (A) Foot in ventral view. (B) Hand in ven- tral view. (C) Head in dorsal view. (D) Head in lateral view. Illus- trations by Carolina Reyes-Puig. Fig. 4. Color patterns of Noblella mindo sp. nov. in life. (A, C) Dorso-lateral and ventral patterns of holotype, ZSFQ 050, adult female, SVL = 18.3 mm. (B, D) Dorsolateral and ventral patterns of paratype, ZSFQ 051, adult male, SVL = 16.9 mm. Photos by Matthijs Hollanders. 70 C. Reyes-Puig et alii views; canthus rostralis straight, slightly concave in pro- file; loreal region slightly concave; upper eyelid 45.6% of interorbital distance; eye-nostril distance 54.8% of eye diameter; tympanum visible externally, tympanic mem- brane differentiated from surrounding skin; supratym- panic fold indistinct. Dentigerous processes of vom- ers absent and vomerine teeth absent; choanae laterally oriented; tongue longer than wide, elongated, partially notched posteriorly. Skin of dorsum finely shagreen, evident tubercles absent; skin on flanks smooth; venter smooth; discoidal fold slightly visible, dorsolateral folds and thoracic folds absent; diminutive rounded ulnar tubercles; palmar tubercle oval, about 2 times the size of the thenar tuber- cle; supernumerary palmar tubercles present, mainly at the base of the digits; proximal subarticular tubercles prominent, rounded; phalangeal formula 2-2-3-3; fingers not expanded distally, finger tips rounded, circumferen- tial grooves absent; relative lengths of fingers: I < II < IV < III; forearm lacking evident tubercles. Hindlimb lengths moderate, tibia length 49.3% of SVL; foot length 46.1% of SVL; dorsal surfaces of hindlimbs shagreen; tubercles on the heel absent; one prominent elongated tarsal tubercle on ventral surface of tarsus; two metatarsal tubercles, inner elongated con- spicuous, outer subconical; proximal and distal subar- ticular tubercles well-defined; supernumerary tubercles absent. Toes slightly expanded and rounded distally; distal portions of circumferential grooves not visible; phalangeal formula 2-2-3-4-3; relative lengths of toes: I < II < V < III < IV. Measurements of holotype (in mm) SVL= 18.3, HL= 7.5, HW= 5.7, ED= 2.4, EN= 1.3, MWE= 1.5, TD= 0.9, MIOD= 3.4, LH= 3.4, LS= 9.0, LF= 8.4. For measurements of the type series (mm) see Table 2. Color of holotype in life (Fig. 4) Dorsum brown, grayish brown towards the flanks; well-defined cream middorsal stripe, extending from interparietal region to cloaca and continuing along pos- terior surfaces of hindlimb. Loreal region black, extend- ing as homogeneous dark band to upper insertion of arm and into body flanks, narrowing towards groin and limit- ed dorsally with a lighter brown line; flanks strongly light flecked; groin dark. Rictal gland white. Venter and ven- tral surfaces of hindlimbs yellowish cream; throat dark brown with large irregular yellowish cream marks and Fig. 5. Color variation of preserved Noblella mindo sp. nov. in (A–E) dorsal and (F–J) ventral views: (A, F) ZSFQ 305, paratype, adult female, SVL = 19.5 mm; (B, G) ZSFQ 304, paratype, adult female, SVL = 19.2 mm; (C, H) ZSFQ 050, holotype, adult female, SVL = 18.3 mm; (D, I) ZSFQ 051, paratype, adult male, SVL = 17.0 mm; (E, J) ZSFQ 049, paratype, adult male, SVL = 16.5 mm. Photos by David Brito-Zapata and Carolina Reyes-Puig. 71A new species of the genus Noblella from Ecuador medium longitudinal line. Forelimbs ventrally yellowish cream with dark brown marks, dorsally light brown with dark brown marks; iris reddish copper with minute scat- tered turquoise dots. Hindlimbs like dorsum. Color of holotype in ethanol (Fig. 5) Dorsum brown, darker towards middorsum, well- defined middorsal line cream, extending from inter- parietal region to cloaca where stripe continues along posterior surface of thighs and pes. Dorsal surfaces of forelimbs brown with black spots. Labial bars absent; rictal gland light brown. Loreal region black, extend- ing as homogeneous dark band to upper insertion of arm and into body flanks, narrowing towards groin; flanks strongly light flecked; groin dark. Dorsal surfaces of hindlimbs lighter brown than dorsum to cream with dark fleck and spots. Throat dark brown with cream irregular marks and medium longitudinal stripe. Chest, venter and ventral surfaces of thigh and crus cream. Variation of color patterns and external morphology (Figs. 4–5) Adult females ZSFQ 050, 304–305 and the adult male ZSFQ 051 exhibit a cream middorsal stripe extend- ing from interparietal region to cloaca; stripe of ZSFQ 305 is thinner and faintly defined. Dark suprainguinal marks are faint in ZSFQ 049. Throat, chest and ventral surfaces of forelimbs are dark brown with a white cross formed by a longitudinal, fine line running from chin to chest, crossing a similar line departing from midventral Fig. 6. Ventral views of (A–D) hands and (E–H) feet from three species of Noblella: (A, E) Noblella coloma, extracted from Guayasamin and Terán-Valdez (2009); (B, F) N. worleyae (holotype); (C, G) N. worleyae (ZSFQ 3851); (D, H) N. mindo sp. nov. (holotype). Scale bars = 1 mm. Illustrations by Carolina Reyes-Puig. 72 C. Reyes-Puig et alii Ta bl e 1. M ai n di ag no st ic c ha ra ct er s of th re e sp ec ie s of N ob le lla fr om n or th w es te rn E cu ad or . Sp ec ie s C ha ra ct er s So ur ce Fi ng er ti ps To e tip s D is ta l ph al an ge s To es pa pi lla e U ln ar tu be rc le s V en te r an d th ro at co lo ra tio n Pr oo tic an d ex oc ci pi ta lS ph en et hm oi d Le ng th o f tr an sv er se pr oc es se s of Pr es ac ra ls N eu ra l a rc h of Pr es ac ra ls N ob le lla co lo m a A cu m in at e Sl ig ht ly e xp an de d an d ac um in at e di st al ly T- sh ap ed A bs en t A bs en t V en te r or an ge w ith m in ut e w hi te a nd b ro w n sp ot s Se pa ra te d W el l- os si fie d, ve nt ra lly no t f us ed a t m id lin e II , V II I< V – V II