Copeia 107, No. 4, 2019, 708–723

On the Uncertain Taxonomic Identity of Adenomera hylaedactyla (Cope, 1868) and the Composite Type Series of A. andreae (Muller,¨ 1923) (Anura, Leptodactylidae)

Thiago R. de Carvalho1, Ariovaldo A. Giaretta2, Natan M. Maciel3, Diego A. Barrera4,Cesar´ Aguilar-Puntriano4,5,Celio´ F. B. Haddad1, Marcelo N. C. Kokubum6, Marcelo Menin7, and Ariadne Angulo4,5

Adenomera andreae and A. hylaedactyla are two widespread Amazonian frogs that have been traditionally distinguished from each other by the use of different habitats, toe tip development, and more recently through advertisement calls. Yet, taxonomic identification of these species has always been challenging. Herein we undertake a review of type specimens and include new phenotypic (morphology and vocalization) and mitochondrial DNA information for an updated diagnosis of both species. Our morphological analysis indicates that the single type (holotype) of A. hylaedactyla could either belong to lineages associated with Amazonian forest-dwelling species (A. andreae clade) or to the open-formation morphotype (A. hylaedactyla clade). Given the holotype’s poor preservation, leading to the ambiguous assignment of character states for toe tip development, as well as a vague type locality encompassing a vast area in eastern Ecuador and northern Peru, the identity of this specimen is uncertain. Morphology of toe tip fragments and the original species description suggest that A. hylaedactyla could correspond to at least two described species (A. andreae or A. simonstuarti) or additional unnamed genetic lineages of the A. andreae clade, all bearing toe tips expanded into discs. Analysis of morphometric data, however, clustered the holotype with the Amazonian open-formation morphotype (toe tips unexpanded). While additional data can be obtained from the holotype of A. hylaedactyla, at this time this type cannot be unequivocally assigned to any species of Adenomera distributed across eastern Ecuador and Peru’s northernmost region of Loreto. For the time being, the name A. hylaedactyla still accommodates the only Amazonian open-habitat species. As to the type series of Adenomera andreae, a forest-associated species with toe tips fully expanded (developed into small discs) from eastern Brazilian Amazonia, we found that one of its paratypes shares a morphotype with the open-habitat species and is reassigned to A. hylaedactyla. With the taxonomic identity of A. hylaedactyla unresolved, formal descriptions of cryptic species complexes within the A. andreae clade distributed across the type locality of A. hylaedactyla run the risk of a possible future synonymization with A. hylaedactyla. Yet, not naming more circumscribed and potentially threatened cryptic species puts them at risk, as they would probably not qualify for

conservation funding. Given the current fire crisis in the Amazon Basin, the risk of losing species before they are described far outweighs the risk of synonymization. We recommend that researchers prioritize descriptions based on the potential extinction risk of new species.

Adenomera andreae y A. hylaedactyla son dos especies de ranas amazonicas´ con una amplia distribucion´ que han sido tradicionalmente diferenciadas una de otra por el uso de diferentes ha´bitats, desarrollo de los d´ıgitos posteriores y ma´s recientemente a traves´ de sus cantos de anuncio. Sin embargo, la identificacion´ taxonomica´ de estas especies siempre ha sido dif´ıcil. Realizamos una revision´ del material tipo e incluimos nuevas informaciones fenot´ıpicas (morfolog´ıa y vocalizaciones) y de secuencias de ADN mitocondrial para sus diagnosticos´ actualizados. Nuestro ana´lisis morfologico´ indica que el tipo unico´ (holotipo) de A. hylaedactyla podr´ıa pertenecer a linajes asociados con especies de la selva amazonica´ (A. andreae clade) o al morfotipo de formacion´ abierta (A. hylaedactyla clade). Dada la pobre preservacion´ del holotipo, lo que lleva a la asignacion´ ambigua de estados de caracteres para el desarrollo de la punta del dedo del pie, as´ı como a una localidad tipo vaga que abarca una vasta a´rea en el este de Ecuador y el norte de Peru,´ la identidad de este especimen´ es incierta. La morfolog´ıa de los fragmentos de la punta del dedo del pie y la descripcion´ original de la especie sugieren que A. hylaedactyla podr´ıa corresponder al menos a dos especies descritas (A. andreae, A. simonstuarti)o linajes geneticos´ adicionales sin nombre del clado de A. andreae, todos con puntas de los pies expandidas en discos. Sin embargo, el ana´lisis de los datos morfometricos´ agrupo´ el holotipo con el morfotipo amazonico´ de formacion´ abierta (puntas de los dedos no expandidas). Si bien se pueden obtener datos adicionales del holotipo de A. hylaedactyla, en este momento este tipo no se puede asignar inequ´ıvocamente a ninguna especie de Adenomera distribuida en el este de Ecuador y la region´ ma´s al norte de Loreto en Peru.´ Por el momento, el nombre A. hylaedactyla todav´ıa alberga la unica´ especie de ha´bitat abierto de la Amazon´ıa. En cuanto a la serie tipo de A. andreae, una especie asociada al bosque con

1 Laboratorio´ de Herpetologia, Departamento de Zoologia e Centro de Aquicultura, Universidade Estadual Paulista (UNESP), Rio Claro 13506- 900, Sa˜o Paulo, Brazil; Email: (TRC) [email protected]; and (CFBH) [email protected]. Send reprint requests to TRC. 2 Laboratorio´ de Taxonomia e Sistema´tica de Anuros Neotropicais, Faculdade de Cienciasˆ Integradas do Pontal, Universidade Federal de Uberlaˆndia, Ituiutaba, Minas Gerais, Brazil; Email: [email protected]. 3 Laboratorio´ de Herpetologia e Comportamento Animal, Instituto de Cienciasˆ Biologicas,´ Universidade Federal de Goia´s, Campus Samambaia, Goiaˆnia, Goia´s, Brazil; Email: [email protected]. 4 Departamento de Herpetolog´ıa, Museo de Historia Natural de San Marcos (MUSM), Lima, Peru; Email: (DAB) [email protected]; and (CAP) [email protected]. 5 UCN SSC Amphibian Specialist Group, Toronto, Canada; Email: (AA) aangulo@amphibians.org. 6 Laboratorio´ de Herpetologia, Unidade Academicaˆ de Cienciasˆ Biologicas,´ Centro de Saude´ e Tecnologia Rural, Universidade Federal de Campina Grande, Patos, Para´ıba, Brazil; Email: [email protected]. 7 Laboratorio´ de Taxonomia e Ecologia de Anf´ıbios e Repteis´ and Programa de Pos-Gradua¸´ ca˜o em Zoologia, Instituto de Cienciasˆ Biologicas,´ Universidade Federal do Amazonas, Manaus, Amazonas, Brazil; Email: [email protected]. Submitted: 21 May 2019. Accepted: 1 October 2019. Associate Editor: B. Stuart. Ó 2019 by the American Society of Ichthyologists and Herpetologists DOI: 10.1643/CH-19-237 Published online: 22 November 2019 de Carvalho et al.—On the type specimens of Amazonian Adenomera 709

puntas de los dedos completamente expandidas (desarrolladas en pequenos˜ discos) desde el este de la Amazon´ıa brasilena,˜ descubrimos que uno de sus paratipos comparte un morfotipo con la especie de ha´bitat abierto y es reasignado a A. hylaedactyla. Con la identidad taxonomica´ de A. hylaedactyla sin resolver, las descripciones formales de los complejos de especies cr´ıpticas dentro del clado de A. andreae distribuidos en la localidad tipo de A. hylaedactyla corren el riesgo de una posible sinonimizacion´ futura con A. hylaedactyla. Sin embargo, no nombrar especies cr´ıpticas ma´s circunscritas y potencialmente amenazadas las pone en riesgo, ya que probablemente no calificar´ıan para fondos de conservacion.´ Dada la actual crisis de incendios en la cuenca del Amazonas, el riesgo de perder especies antes de que se describan supera con creces el riesgo de sinonimizacion.´ Recomendamos que los investigadores prioricen las descripciones basadas en el riesgo potencial de extincion´ de nuevas especies.

DENOMERA andreae (Muller,¨ 1923) and A. hylaedac- recorded and collected A. andreae from two other localities tyla (Cope, 1868) are two leptodactylid frog species in eastern Brazilian Amazonia, state of Para´ (Belem:´ A with widespread distributions in Amazonia; the latter 1.4497608S, 48.4443518W; and Santa Ba´rbara do Para´: is also distributed throughout the Cerrado savanna region 1.1930568S, 48.2694448W), as well as from other Amazonian and northern Atlantic Forest in Brazil (Heyer, 1973; Fouquet regions in Brazil (Manaus, state of Amazonas: 3.0919648S, et al., 2014). These species are usually distinguished by their 59.9712348W; and Serra do Navio, state of Amapa´: morphology and the use of different habitats: forests (A. 0.8958708N, 52.0009208W) and Peru (Tambopata National andreae) and open formations (A. hylaedactyla; Heyer, 1973, Reserve, Region of Madre de Dios: 12.8359308S, 1977, 1984). Since its description, the vocalization of A. 69.2728708W). Localities are mapped in Figure 1. andreae has been described from different Amazonian regions We recorded and collected A. hylaedactyla from many (e.g., Zimmerman and Bogart, 1984; Boistel et al., 2006). On regions throughout South America, but only specimens and the other hand, acoustic data and DNA sequences remain associated sound recordings from the following localities in unknown for the type locality, originally designated as the Brazilian and Peruvian Amazonia were included in this ‘‘Peixeboi (a. d. Bragan¸cabahn), Staat Para`, Brasilien’’ by study (Fig. 1): Brazil (state of Roraima, Canta´: 2.7665318N, Muller¨ (1923). The exact type locality was a railway station, 60.6089288W; state of Amazonas: Sa˜o Gabriel da Cachoeira: the ‘‘Esta¸ca˜o de Bragan¸ca,’’ which was located in the small 0.1362068S, 67.0875178W, and Manaus: 3.0919648S, town of Peixe-Boi, in the Brazilian state of Para´ (eastern 59.9712348W; state of Acre: Cruzeiro do Sul: 7.6149778S, Amazonia). In the case of A. hylaedactyla, the type locality is 72.7013968W; and Feijo:´ 8.1612498S, 70.3601648W); Peru briefly described by Cope (1868) as being ‘‘from the Napo or (Tambopata National Reserve, Region of Madre de Dios: Upper Maranon’’, comprising a vast Amazonian region 12.8367508S, 69.2936908W). entailing eastern Ecuador and northeastern Peru. This Specimens collected by us were deposited in the following vagueness likely comes from the fact that the only type herpetological collections: AAG-UFU (Universidade Federal specimen for A. hylaedactyla was collected in the course of a de Uberlaˆndia, in Uberlaˆndia, Brazil), CFBH (Universidade general scientific expedition led by James Orton in 1867– Estadual Paulista, in Rio Claro, Brazil), LHUFCG (Laboratorio´ 1868 into the equatorial Andes and Amazon rainforest. Calls de Herpetologia da Universidade Federal de Campina and DNA sequences of specimens assigned to A. hylaedactyla Grande, in Patos, Brazil), and MUSM (Museo de Historia from Peruvian Amazonia have been published elsewhere Natural de la Universidad Nacional Mayor de San Marcos, in (Schluter,¨ 1980; Angulo et al., 2003; Fouquet et al., 2014). Lima, Peru). Other specimens examined, including genetic In 2013, one of us (TRC) conducted a field expedition to and call vouchers, are listed in the Material Examined the type locality region of A. andreae (eastern Brazilian section. Institutional abbreviations followed Sabaj (2016). Amazonia) and succeeded in collecting a new series of specimens and in recording vocalizations. In 2005, one of Morphology.—Morphometric measurements were taken with us (MNCK) visited the same locality and obtained calls and an ocular micrometer (0.1 mm scale) fitted to a stereomicro- specimens of a sympatric form assigned to A. hylaedactyla scope (except SVL, THL, TL, and FL, taken with digital (but see Discussion). Here we undertake a review of the calipers with precision of 0.1 mm) as follows: snout–vent phenotypic traits traditionally employed in the diagnosis of length (SVL), thigh length (THL), tibia length (TL), foot the sympatric A. andreae and A. hylaedactyla, as well as a length (FL), hand length (HAL), eye diameter (ED), tympa- detailed characterization of calls and mtDNA sequences for num diameter (TD), and eye–nostril distance (EN). Other both species. Additionally, we address the uncertain taxo- traits (e.g., head length and width) were not measured from nomic identity of the single type specimen of A. hylaedactyla some specimens or assessed through different methods; thus, and the composite type series of A. andreae. they are not included in descriptive statistics (Table 1) and morphometric analysis (see below). Morphometric measure- ments followed the definitions and terminology of Watters et MATERIALS AND METHODS al. (2016), except HAL, measured as the distance between the Study sites.—We visited the original type locality of A. andreae base of the inner metacarpal tubercle to the tip of finger III. in eastern Brazilian Amazonia (Bragan¸ca railroad station, Snout shape was assessed according to Heyer et al. (1990). located in the town of Peixe-Boi, state of Para´: 1.1933088S, We followed and modified the definitions of toe tip 47.3162088W) and found that there is a small commercial development (character states) of Heyer (1973). The defini- center where the railroad station used to be. Old (tall-tree) tion of character state D refers to toe tips developed into forest fragments still remain on the outskirts of the type flattened discs. However, the dorsolateral flattening of toe locality, but we were not able to record any A. andreae in tips is ambiguous and sometimes hardly discernible in those remnants. Despite this, we found calling males of A. preserved specimens of Adenomera, given that toe tips can andreae in a forest fragment approximately 18 km southwest be easily desiccated because of inappropriate fixation and/or of the type locality, in the limits of a neighboring town (Nova poor preservation conditions. Therefore, we assign character Timboteua: 1.2737538S, 47.4608718W; Fig. 1). We also state D to fully expanded toe tips (a condition that is present 710 Copeia 107, No. 4, 2019

Fig. 1. Political map of northern South America with some of the major river drainages in the Amazon River Basin. Symbols indicate localities with associated call vouchers. Circles ¼ Adenomera andreae: 1, Peixe-Boi (type locality); 2, Nova Timboteua; 3, Santa Ba´rbara do Para´; 4, Belem;´ 5, Serra do Navio. Squares ¼ A. hylaedactyla: 1, Tambopata; 2, Iquitos; 3, Cruzeiro do Sul; 4, Feijo;´ 5, Tabatinga; 6, Sa˜o Gabriel da Cachoeira; 7, Manaus (sympatric call vouchers of A. andreae from this locality); 8, Canta´. State limits of Brazil and region limits of Peru are shown. Abbreviations: Brazil (AC, Acre; AM, Amazonas; AP, Amapa´; PA, Para´; RR, Roraima); Peru (LOR, Loreto; MDD, Madre de Dios). among all members of the A. andreae clade) even in cases Moderately expanded toe tips (character C of Heyer, 1973) where dorsolateral flattening is not clearly observed (previ- refers to swollen, knob-shaped toe tips, and can be observed ously referred to as C–D). Fully expanded, not flattened toe in A. engelsi (Kwet et al., 2009: 96, fig. 1E). Character states A– tips can be seen in A. lutzi (Kok et al., 2007: 49, fig. 2), and B (Heyer, 1973) refer to slightly tapered (state A) or fully expanded toe tips develop into small discs in A. unexpanded to slightly swollen (state B) toe tips. Such simonstuarti (Angulo and Icochea, 2010: 362, fig. 7). character states can be observed, for example, in A. martinezi

Table 1. Morphometric traits (adult males only) of two species of Adenomera (adult males) from Brazilian and Peruvian Amazonia, and the Guianan savanna ecotone in the Brazilian state of Roraima (A. andreae and A. hylaedactyla), corresponding to collection points of Figure 1; and the two male type specimens of A. simonstuarti from the type locality (see Angulo and Icochea, 2010). Values are presented as X6SD (min–max). Morphometric traits are defined in Materials and Methods. A series of seven specimens of A. andreae from the type locality region was included in the descriptive statistics (see Materials and Methods for detailed information). Adenomera hylaedactyla Adenomera andreae Adenomera simonstuarti Trait (mm) n ¼ 25 Holotype (ANSP 2240) Other specimens (n ¼ 25) n ¼ 2

SVL 20.760.8 (19.3–23.0) 24.2 24.061.1 (22.2–26.5) 26.160.2 (25.9–26.2) EN 1.860.2 (1.4–2.0) 1.8 2.060.2 (1.6–2.3) 2.060.1 (1.9–2.1) ED 2.060.2 (1.6–2.3) 2.2 2.260.3 (1.7–2.8) 2.860.3 (2.6–3.0) TD 1.260.1 (1.0–1.4) 1.4 1.360.1 (1.2–1.5) 1.7 HAL 4.460.3 (4.0–5.1) 4.7 5.360.3 (4.7–6.1) 5.4 THL 9.660.4 (8.6–10.7) 9.8 10.560.8 (9.0–12.1) 10.260.8 (9.6–10.7) TL 9.860.4 (9.3–10.7) 11.4 10.960.7 (9.8–12.5) 11.260.6 (10.8–11.6) FL 10.360.4 (9.4–11.2) 11.4 12.360.7 (11.3–13.6) 11.760.8 (11.1–12.2) de Carvalho et al.—On the type specimens of Amazonian Adenomera 711

(Carvalho and Giaretta, 2013a: 217, fig. 4C) and A. juikitam Call frequencies in Adenomera are distributed over harmonic (Carvalho and Giaretta, 2013b: 546, fig. 7C), respectively. A series. In order to facilitate interspecific comparisons, the further consideration is that different toes have different fundamental frequency (f0) is referred to herein as the character states. Toes II–IV are usually more defined in species fundamental harmonic (H1), and the dominant frequency, with moderately or fully expanded toe tips (character states which coincides with second harmonic (2f0), is referred to C–D), whereas toes I and V could be slightly swollen or even herein as the dominant harmonic (H2). We produced sound unexpanded. figures using Soundruler with the following parameters: spectrogram (window type ¼ Hanning, FFT size ¼ 256 points, Multivariate statistics.—We conducted a Principal Compo- and FFT overlap ¼ 90%), the strength of frequency nent Analysis (PCA) on a morphometric (log10-transformed components were indicated by their darkness; amplitude values) dataset of eight traits from adult male specimens of spectrum (FFT size ¼ 1024 points). three Amazonian species of Adenomera (A. andreae; A. hylaedactyla, including the holotype; A. simonstuarti) that DNA sequencing and phylogenetic analysis.—We extracted are known to occur in the type locality region of A. total DNA from muscle tissue preserved in ethanol using a hylaedactyla. The analysis was performed using vegan 2.5-2 tissue-storage buffer from the DNeasy Tissue Kit (Qiagen). It (Oksanen et al., 2018), implemented in R 3.5.0 (R Core Team, is important to mention that we had access to tissue 2018). Scores, eigenvalues, and explained variation of the fragments from the four type specimens of A. andreae, but first three principal components are provided in Appendix 1. were unable to extract viable DNA using the standard extraction protocol described earlier. We do not know if Acoustics.—We recorded calls in the field (see localities in the those specimens were fixed and kept in formalin or ethanol. Study Sites section) using digital recorders (Marantz PMD Either way, genetic material is likely degraded, so specific 660, 661, 670, 671; M-audio Microtrack 2; sampling rates: extraction protocols would likely be required to assess DNA 44.1 or 48.0 kHz; bit depths: 16 or 24) and Sennheiser information. ME66/K6 or ME67/K6 unidirectional microphones. We We used the following primers for amplification of the stored recordings as uncompressed WAV files. Furthermore, cytochrome c oxidase subunit I (COI) gene: dgLCO1490 (50– we had access to two additional recordings (same parame- GGTCAACAAATCATAAAGAYATYGG–30; Meyer, 2003), ters as above) of A. hylaedactyla from the following localities dgHCO2198 (50–TAAACTTCAGGGTGACCAAARAAYCA–30; in southwestern Amazonia: (1) Estacion´ Biologica´ Jose´ Meyer, 2003), T3-AnF1 (50–ATTAACCCTCACTAAAGA A´ lvarez Alonso (approximate coordinates: 3.9659568S, CHAAYCAYAAAGAYATYGG–30; Lyra et al., 2017), and T7- 73.4172148W), region of Loreto, northern Peru, by Giussepe AnR1 (50–AATACGACTCACTATAGCCRAARAATCARAA Gagliardi-Urrutia; (2) Tabatinga (approximate coordinates: DARRTGTTG–30; Lyra et al., 2017). DNA amplification and 4.2320178S, 69.9369088W), state of Amazonas, northern

purification followed Lyra et al. (2017). We sent PCR Brazil, by Rommel R. Rojas. Sound recordings are deposited products to Macrogen Inc. (Seoul, South Korea), where they in the acoustic repositories of AAG-UFU and CFBH collec- were sequenced with a BigDye Terminator Cycle Sequencing tions; information on the files is provided in Appendix 2. Kit (version 3.0, Applied Biosystems) in an ABI 3730 We analyzed calls using an interface built between Soundr- automated DNA sequencer (Applied Biosystems). Taxon uler 0.9.6.1 (Gridi-Papp, 2007) and Matlab 6.5.2 (Matlab, sampling included all named and confirmed candidate 2004, The language of technical computing, version 6.5.2., species of Fouquet et al. (2014) in the A. andreae clade (A. The MathWorks, Inc., Natick, MA). We applied a high-pass andreae, A. chicomendesi, A. simonstuarti, and lineages referred filter up to 300 Hz to sound files in Soundruler prior to to as sp. C, sp. D, and sp. T) plus nominal species from each conducting the acoustic analysis to reduce background of the remaining Adenomera clades. The A. hylaedactyla clade noise. We quantified acoustic traits through automated is comprised of A. coca, A. diptyx, and A. hylaedactyla; all three analysis in the time domain (notes, pulses, intervals, and were included in the dataset. We also added new sequences of pulse rate) and in the frequency domain (fundamental and the following specimens (respective GenBank accession dominant frequencies, and frequency modulation), except numbers) to the COI dataset: A. andreae—AAG-UFU 2791, note rate per minute (quantified manually in Audacity 2797 (MK959183–84); A. cf. andreae—MUSM 39464, 39469– 2.1.1; Audacity Team, 2017). Acoustic traits are given as 71 (MK959185–88); A. hylaedactyla—AAG-UFU 3859–61, averaged means for the males analyzed and their corre- 5540–41, 5895, 5897, 5907–09 (MK959189–98), MUSM sponding standard deviation as standard deviations of the 39459–60 (MK959199–200). We used Lithodytes lineatus averaged mean values. Ranges include the span of values (KU494587), Hydrolaetare caparu (KC603988), Leptodactylus from the raw dataset. In the case of ‘‘pulse duration,’’ given fuscus (KU494530), and Scythrophrys sawayae (KU494787) as that acoustic signals analyzed had more than one pulse, we outgroup taxa, and rooted on Physalaemus nattereri first averaged the duration of each pulse of a given note and (KU494649). then obtained the averaged mean for each male analyzed We used CodonCode Aligner (CodonCode Corporation) to from the mean duration of call pulses. Spectrogram edit sequences, MAFFT 7 (Katoh et al., 2017) under default parameters are as follows: FFT size ¼ 1024 points, FFT parameters for sequence alignment, and jModelTest 2 overlap ¼ 90%, window type ¼ Hanning, contrast ¼ 70%. (Darriba et al., 2012) for testing evolutionary models using Specific parameters for automated recognition in Soundru- Akaike Information Criterion (AIC). The best-fitting nucleo- ler are: Adenomera andreae—detection (smoothing ¼ 500, tide substitution model selected was the Tamura-Nei (TrN; resolution ¼ 1), delineation (smooth factor ¼ 1, smoothing ¼ Tamura and Nei, 1993) with gamma distribution of rates 13, and resolution ¼ 1), critical amplitude ratio ¼ 0.9; across sites. We reconstructed phylogenetic trees using Adenomera hylaedactyla—detection (smoothing ¼ 300, reso- Bayesian inference optimality criterion with two indepen- lution ¼ 1), delineation (smooth factor ¼ 1, smoothing ¼ 13, dent runs of 2.0 3 106 generations, starting with random and resolution ¼ 1), critical amplitude ratio ¼ 1.3. Acoustic trees and four Markov chains (one cold), sampled every 1,000 definitions and terminology follow Carvalho et al. (2019a). generations in MrBayes 3.2.6 (Ronquist et al., 2012) and 712 Copeia 107, No. 4, 2019

Fig. 2. Photographs of the holotype of Adenomera hylaedactyla, ANSP 2240. (A, B) Body in dorsal and ventral view, respectively; (C–D) right and left foot, respectively. discarding 25% of generations and trees as burn-in. We used have observed incorrect character coding among examined the standard deviation of split frequencies (,0.01) and specimens with varying degrees of desiccation and for which Estimated Sample Size (.200) to assess run convergence. we had other reliable data for correct species assignment

We computed uncorrected pairwise distances using ape 3.4 (calls and/or DNA sequences), such as the presence of lateral (Paradis et al., 2004) and spider 1.3-0 (Brown et al., 2012), expansions in A. hylaedactyla (which could be interpreted to implemented in R 3.5.0 (R Core Team, 2018), treating gaps be small discs) and poorly preserved specimens of A. andreae using the pairwise deletion option. with weak or no indication of toe tip expansions. Lastly, it is important to highlight that Heyer (1973) sexed the holotype RESULTS as a male. Its snout tip is a little bit misshaped, which makes it difficult to confirm gender—reproductive adult males of Holotype of Adenomera hylaedactyla.—The original descrip- Adenomera usually have a shovel-shaped snout ending with a tion by Cope (1868) was based on a single specimen, the fleshy ridge, giving their snout an acuminate shape in profile holotype (ANSP 2240; see Malnate, 1971). Cope stated that (females tend to have a more rounded snout in profile). ‘‘the toes have distinct dilatations at the end’’. However, given poor preservation conditions (Fig. 2A–B), the exami- Nevertheless, there appears to be a very weak ridge on the nation of character state(s) for toe tips in this specimen is snout tip. Thus, we regard the holotype of A. hylaedactyla as challenging, as most of the toes are broken off (Fig. 2C–D). an adult male (see Table 1 for morphometric traits). One toe tip fragment appears to be laterally expanded Considering all available evidence, the identity of A. (character states C–D), and in line with Cope’s description. hylaedactyla is a complicated matter. Based on toe tip This means that the holotype of A. hylaedactyla would have expansions, the holotype could correspond to nominal fully expanded toe tips, as in A. andreae (Heyer, 1973). Other forms of two Amazonian forest-dwelling species (A. andreae toe tips could be considered to be unexpanded or slightly or A. simonstuarti) or additional undescribed species in the A. expanded (character states B–C). On the other hand, the PCA andreae clade distributed in Amazonian northern Peru clustered the holotype of A. hylaedactyla with the Amazonian (potentially unnamed genetic lineages highlighted by Fou- open-formation species of Adenomera (Fig. 3), which has toe quet et al., 2014; Carvalho, unpubl. data). However, since tips unexpanded compared with forest-associated morpho- Heyer (1973) associated the name A. hylaedactyla with an types with toe tips expanded (Fig. 4). open-formation species, the name has been linked to a When Heyer (1973) examined this holotype, there were morphotype with snout usually sub-elliptical from above in still some intact toe tips, and he recognized that terminal toe males (Figs. 4A–B, 5A) and toe tips unexpanded or slightly digits were somewhat desiccated but that they were dilated swollen (character state B; Fig. 4D), in contrast to forest- (character states B–C) and not flattened (i.e., developed into associated species (e.g., A. andreae), which share a morpho- small discs; character state D). Subsequently, one of us (AA) type with snout usually sub-ovoid from above in males (Figs. also examined the holotype and took photographs that 4E–F, 5B) and toe tips, except toe I, fully expanded (Fig. 4H). confirm that the still extant toe tip fragment and other toe On the other hand, our exploratory PCA applied to a tips are slightly and/or moderately expanded. Of note, morphometric dataset (Fig. 3) supports Heyer’s (1973) different degrees of desiccation might mislead to ambigu- designation of the holotype of A. hylaedactyla to the ous/equivocal character states of toe tips. For example, we Amazonian open-habitat species (although it is important de Carvalho et al.—On the type specimens of Amazonian Adenomera 713

Fig. 3. Scatterplot of the first two principal component axes from a PCA applied to a morphometric dataset of three Amazonian species of Adeno- mera and the holotype of A. hylae- dactyla (all adult males). Individual symbols represent specimens of a given species. Adenomera andreae and A. hylaedactyla are encircled by solid-line and dashed-line convex polygons, respectively. Additional in- formation is presented in Appendix 1. to note that the PCA may change with the inclusion of a emitted at a repetition rate of 84–215 per second larger sample size and data of undescribed candidate species (140.3610.7). The fundamental frequency (H1) ranges from of the A. andreae clade). Given all available evidence, this 1852–2211 (2021.0697.0) Hz; the dominant frequency (H2) holotype cannot be unambiguously assigned to any of the ranges from 3680–4457 (4071.16213.8) Hz. Notes can have a forest-associated lineages (A. andreae clade) or the open- modest or pronounced frequency modulation, either positive habitat species (A. hylaedactyla; sensu Heyer, 1973) distributed or negative, varying from –328 to 656 (209.76151.9) Hz. over the vast type locality region of the name-bearing type.

While additional data can potentially be acquired from the Phylogenetic placement.—We assign A. hylaedactyla to the holotype of A. hylaedactyla, such as DNA sequence data or only open-formation morphotype distributed throughout other techniques that might help with its identification, we Amazonian formations. Our call voucher specimens from suggest that (1) the name A. hylaedactyla is kept in use to southwestern Peruvian and Brazilian Amazonia, eastern accommodate the open-habitat species, bearing in mind the Brazilian Amazonia, and intermediate Amazonian regions uncertainty associated with species identity; and (2) the fall within the A. hylaedactyla clade of Fouquet et al. (2014; decision to formally describe new forest-associated species (A. Fig. 7). andreae clade) from the type locality region of A. hylaedactyla (eastern Ecuador and Peru’s northernmost region of Loreto) Updated diagnosis and comparisons with Amazonian conge- be assessed based on the potential extinction risk of the new ners.—Specimens assigned to A. hylaedactyla in the present species, given that conservation funding is tied to described study agree with the previous morphological and color species. characterization provided by Heyer (1973). Adenomera hylae- dactyla is characterized by the following combination of Accounts of Adenomera hylaedactyla (Cope, 1868).—Heyer features: (1) medium to large male size for the genus (SVL ¼ (1973) characterized and described morphological variation. 22.2–26.5 mm); (2) toe tips unexpanded or barely expanded The advertisement call associated with this morphotype has (character state B); (3) mid-dorsal pinstripe incomplete, when been described a few times from different regions in South present, extending from cloacal region to scapular region or America (e.g., Straughan and Heyer, 1976; Schluter,¨ 1980; shorter; (4) absence of nearly solid dark-colored stripe on Schneider et al., 1988; Menin et al., 2009). In the next underside of forearm; (5) absence of antebrachial tubercle; (6) sections, we provide a new characterization of the advertise- use of open habitats; (7) advertisement call composed of ment call, phylogenetic position of call vouchers, and an single notes; (8) call notes formed by partly fused pulses; (9) updated diagnosis of the species name applied to accommo- call dominant frequency coinciding with the second har- date the Amazonian open-formation morphotype of Adeno- monic; (10) high note rate (.100 notes/min; on average, mera. around 150 notes/min). Adenomera hylaedactyla is distinguished from species with Advertisement call of A. hylaedactyla.—Based on calls of 24 moderately swollen or fully expanded toe tips (character males recorded from different Amazonian regions in Brazil states C–D: A. andreae [Heyer, 1973], A. chicomendesi and Peru (see Appendix 2 for locality data; Table 2). [Carvalho et al., 2019b], A. heyeri [Boistel et al., 2006], A. Descriptive statistics are given as min–max (X6SD). The lutzi [Kok et al., 2007], and A. simonstuarti [Angulo and advertisement call (Fig. 6A–D) consists of single, pulsed notes Icochea, 2010]) by having toe tips unexpanded or barely (n ¼ 1074 notes; n ¼ 7196 pulses) given at a repetition rate of expanded (character state B; Fig. 4D). Adenomera hylaedactyla 107–242 per minute (153.5631.7). Note duration is 41–89 can be further distinguished from A. simonstuarti (Angulo (59.468.7) ms, and the rise time is 6–64% (31.5610.3). Notes and Icochea, 2010) by the absence of nearly solid dark- are formed by 4–10 (6.761.0) partly fused pulses that are colored stripe on underside of forearm, and from A. lutzi and 714 Copeia 107, No. 4, 2019

Fig. 4. Dorsum and venter of entire specimens of Adenomera (scale Fig. 5. Live adult males of (A) Adenomera hylaedactyla and (C) A.cf. bars ¼ 5 mm), and detail of ventral view of hand and foot (scale bars ¼ 1 andreae (Forest Call III of Angulo et al., 2003) from southwestern Peruvian mm): (A–D) A. hylaedactyla (MUSM 39460; SVL ¼ 23.4 mm) from Amazonia, and (B) A. andreae from eastern Brazilian Amazonia. (A) Tambopata, in Madre de Dios, southeastern Peru; and (E–H) A. andreae MUSM 39461, SVL ¼ 23.9 mm and (C) MUSM 39470, SVL ¼ 20.2 mm, (AAG-UFU 2789; SVL ¼ 20.3 mm) from Nova Timboteua, in the Brazilian from Tambopata National Reserve/Explorer’s Inn Ecolodge, region of state of Para´ (ca. 18 km from the type locality in Peixe-Boi). Tambopata, in Madre de Dios. (B) AAG-UFU 2789, SVL ¼ 20.3 mm, from Nova Timboteua, in Para´ (ca. 18 km from the type locality in Peixe-Boi).

A. phonotriccus by the absence of antebrachial tubercle (Kok et dwellers (Boistel et al., 2006; Angulo and Icochea, 2010; al., 2007; Carvalho et al., 2019a). Adenomera hylaedactyla Carvalho et al., 2019a, 2019b). occupies open habitats, whereas A. andreae, A chicomendesi, A. The advertisement call of A. hylaedactyla (Fig. 6A–D; Table heyeri, A. lutzi, A. phonotriccus, and A. simonstuarti are forest 2) is composed of single notes, whereas A. simonstuarti de Carvalho et al.—On the type specimens of Amazonian Adenomera 715

Table 2. Geographic variation in advertisement call traits of Adenomera hylaedactyla from Brazilian and Peruvian Amazonia, and the Guianan savanna ecotone in the Brazilian state of Roraima. See Appendix 2 for additional information. Values are presented as X6SD (min–max); n ¼ males recorded. ND ¼ note duration; NR ¼ note rate; RT ¼ note rise time; PN ¼ pulses per note; PR ¼ pulse rate; H1 ¼ fundamental harmonic; H2 ¼ second harmonic (¼ dominant frequency); FM ¼ linear frequency modulation. SW/NW Amazonia comprises localities in northern (Loreto) and southeastern Peru (Madre de Dios), and northwestern Brazil (Tabatinga, Feijo,´ and Cruzeiro do Sul); SGC ¼ Sa˜o Gabriel da Cachoeira. Adenomera hylaedactyla

SW/NW Amazonia SGC (Amazonas, Brazil) Manaus (Amazonas, Brazil) Canta´ (Roraima, Brazil) Traits (Peru and Brazil) n ¼ 15 n ¼ 2 n ¼ 6 n ¼ 7

ND (ms) 57.869.8 (41–77) 52.762.4 (47–60) 69.063.1 (45–89) 55.364.6 (44–66) NR/min 157.9638.8 (125–242) 170.661.7 (169–172) 127.9612.9 (107–142) 164.9627.6 (136–204) RT (%) 29.1612.2 (7–64) 37.8613.4 (13–58) 29.767.4 (6–59) 34.369.9 (15–62) PN 6.561.2 (4–10) 7.460.4 (6–10) 7.061.1 (4–10) 6.560.7 (4–8) PR/s 136.468.0 (84–167) 163.861.3 (128–215) 134.864.7 (96–164) 143.369.1 (91–163) H1 (Hz) 2054.16103.8 (1852–2211) 1933.9651.9 (1863–2024) 1929.0629.8 (1855–1986) 2082.1658.9 (1981–2202) H2 (Hz) 4133.76268.7 (3682–4457) 3885.36223.0 (3680–4055) 3917.0632.4 (3758–4037) 4175.96126.1 (3941–4457) FM (Hz) 264.86134.4 (–86–603) 456.0684.4 (47–656) 83.76153.0 (–328–388) 176.4641.6 (–129–388)

produce multi-note calls (Carvalho et al., unpubl. data). The from A. andreae (H1: 2221–2588 Hz; H2: 4336–5233 Hz; Table call of A. hylaedactyla is pulsed, in contrast to the nonpulsed 3). Adenomera hylaedactyla (41–89 ms; Table 2) differs from A. call of A. lutzi (Kok et al., 2007). From the pulsed-call species, coca (110–145 ms; Angulo and Reichle, 2008), A. chicomendesi A. hylaedactyla can be distinguished from A. phonotriccus (154–247 ms; Carvalho et al., 2019b), and A. heyeri (137–185 (complete pulses; Carvalho et al., 2019a) by having notes ms; Boistel et al., 2006) by its shorter note duration. From the with partly fused pulses. Adenomera hylaedactyla is distin- broadly sympatric Amazonian A. andreae (24–68 notes/min; guished by lower fundamental (H1) and dominant (H2) Table 3), A. hylaedactyla can be further distinguished by a frequencies (H1: 1852–2211 Hz; H2: 3680–4457 Hz; Table 2) higher note rate (107–242 notes/min; Table 2).

Fig. 6. Advertisement calls of Adenomera hylaedactyla (A–D) from Iquitos (in Loreto, northern Peru), A. andreae (E–H) from Nova Timboteua (in Para´, northern Brazil), and A. cf. andreae (I–L) from Tambopata (in Madre de Dios, southeastern Peru). (A, E, I) Time-domain sections (ca. 20 s). (B– D) Oscillogram, spectrogram, and amplitude spectrum of the note delimited in A. (F–H) Oscillogram, spectrogram, and amplitude spectrum of the note delimited in E. (J–L) Oscillogram, spectrogram, and amplitude spectrum of the note delimited in I. Spectrograms/oscillograms are equally scaled (ca. 0.15 s). Amplitude spectra are given in logarithmic dB scale. Additional information is presented in Appendix 2. 716 Copeia 107, No. 4, 2019

Fig. 7. The 50% majority-rule consensus tree from Bayesian inference of partial COI gene showing the phylogenetic positions of A. andreae, A. cf. andreae (¼Forest Call III of Angulo et al., 2003), and A. hylaedactyla within Adenomera. GenBank accession numbers are noted on the terminals. Posterior probabilities are given near the nodes, and branch scale is indicated in number of substitutions per site. Asterisks (*) indicate maximum support. Within-species support values are not shown. For clarity, some clades were collapsed. See Data Accessibility for tree file.

Table 3. Geographic variation in advertisement call traits of Adenomera andreae from eastern and central Brazilian Amazonia, and from eastern Guiana Shield in the Brazilian state of Amapa´; and A. cf. andreae (Forest Call III of Angulo et al., 2003) from southeastern Peru. See Appendix 2 for additional information. Values are presented as X6SD (min–max); n ¼ males recorded. ND ¼ note duration; NR ¼ note rate; RT ¼ note rise time; PN ¼ pulses per note; PR ¼ pulse rate; H1 ¼ fundamental harmonic; H2 ¼ second harmonic (¼ dominant frequency); FM ¼ frequency modulation; NA ¼ not applicable. Adenomera andreae Adenomera cf. andreae

Eastern Amazonia (Para´) Central Amazonia Eastern Guiana Shield Tambopata Traits n ¼ 15 (Amazonas) n ¼ 5 (Amapa´) n ¼ 5 (Madre de Dios) n ¼ 8

ND (ms) 57.264.5 (45–69) 62.365.4 (48–75) 53.765.5 (41–76) 30.362.3 (21–41) NR/min 46.3611.5 (24–66) 57.867.7 (48–66) 56.769.8 (41–68) 50.366.5 (40–59) RT (%) 48.866.2 (14–62) 46.1613.4 (7–77) 36.4611.0 (12–60) 37.6614.7 (20–70) PN 6.061.4 (3–10) 5.561.2 (3–9) 5.860.7 (4–9) NA PR/s 129.2624.8 (46–208) 102.4622.4 (39–167) 132.4618.1 (75–181) NA H1 (Hz) 2387.7671.7 (2221–2588) 2371.0671.4 (2260–2527) 2380.7675.9 (2222–2534) 2560.7695.1 (2422–2745) H2 (Hz) 4838.26165.5 (4336–5233) 4732.86148.4 (4490–5180) 4731.16261.6 (4242–5233) 5157.56184.4 (4845–5534) FM (Hz) 583.76138.7 (234–1077) 390.96132.0 (86–703) 618.96138.1 (215–904) 744.6628.1 (188–1034) de Carvalho et al.—On the type specimens of Amazonian Adenomera 717

Fig. 8. Photographs (dorsal view of body and foot) of type specimens of Adenomera andreae. (A–B) Neotype, ZSM 1911/145-4; (C–D) paratopo- type, ZSM 1911/145-2, reassigned to A. hylaedactyla.

Type series of A. andreae.—The holotype (Munich 136/1911), hylaedactyla, which has toe tips unexpanded or barely a 20 mm SVL adult male originally designated by Muller¨ expanded (Heyer, 1973, 1984; Fig. 4). (1923), was lost during World War II (U. Gruber, pers. comm. We had access to high-quality photographs of four type to W. R. Heyer; Heyer, 1973). Heyer (1973) thus designated a specimens of A. andreae, the neotype (ZSM 145/1911-4; 21 mm SVL female type as the species’ neotype (ZSM 145/ adult female; Fig. 8A–B) and three paratopotypes (ZSM 1911-4; Fig. 8A–B); the other three paratopotypes are ZSM 145/1911-1 to 145/1911-3; adult female and male, and 145/1911-1 to 145/1911-3. In addition to these, Muller¨ juvenile, respectively). Based on a more sub-elliptical head (1923) also designated two females from Utinga (today, a in dorsal view, relatively slender and rocket-shaped body, state park within the city of Belem,´ ca. 130 km from the type mid-dorsal pinstripe, and unexpanded/slightly swollen toe locality; 1.4259138S, 48.4443118W) and two other females tips (character state B) of the paratopotype ZSM 145/1911- from the vicinity of Belem´ (coordinates of our collection 2 (Fig. 8C–D), we reassign this specimen to A. hylaedactyla, point in Belem:´ 1.4497608S, 48.4443518W) as additional type which occurs sympatrically with A. andreae in eastern specimens (originally referred to as co-types). However, we Brazilian Amazonia. ZSM 145/1911-3 does not have well- could not track down these latter four female specimens. The developed toe tip expansions, but it clearly corresponds to most useful feature described from both the lost holotype a juvenile. It is a real challenge to identify juvenile (Muller,¨ 1923) and subsequently designated neotype (Heyer, Adenomera, because morphological features might be 1973) is the shape of the toe tips, characterized as clearly barely observed or even absent (e.g., antebrachial tubercle, widened (Muller,¨ 1923) and expanded into toe discs (Heyer, toe tip development, snout shape). Based on overall body/ 1973). This morphological feature alone has been used to snout shape, dorsal coloration, and toe tips, we judge that distinguish A. andreae from the only known sympatric this specimen should probably be conspecific with A. species of Adenomera in eastern Brazilian Amazonia, A. andreae. 718 Copeia 107, No. 4, 2019

Advertisement call of A. andreae.—Acoustic characterization which distinguishes it from the open-formation A. hylaedac- is based on calls of 15 males from eastern Brazilian Amazonia tyla (Heyer, 1977; Rodr´ıguez and Duellman, 1994). (see Appendix 2 for locality data; Table 3 for geographic The advertisement call of A. andreae (Fig. 6E–H; Table 3) is variation of the call). Descriptive statistics are given as min– composed of single notes, whereas that of A. simonstuarti max (X6SD). The advertisement call (Fig. 6E–H) is comprised comprises calling bouts (multi-note calls; Carvalho et al., of single, pulsed notes (n ¼ 431 notes; n ¼ 2781 pulses) given unpubl. data). The advertisement call notes of A. andreae are at a repetition rate of 24–66 per minute (46.3611.5). Note pulsed, even though pulsing may be hardly detected some- duration is 45–69 (57.264.6) ms, and the rise time is 14–62% times in the call envelope. To the human ear, the call of A. (48.866.2). Notes are formed by 3–10 (6.061.4) partly fused andreae sounds raspy, which appears to reflect its weak pulsing pulses that are emitted at a repetition rate of 46–208 per throughout the acoustic signal, in contrast to the nonpulsed second (129.2624.8). The fundamental frequency (H1) call of A. lutzi (Kok et al., 2007). From the pulsed-call species, ranges from 2221–2588 (2387.7671.7) Hz; the dominant A. andreae can be distinguished from A. phonotriccus (complete frequency (H2) from 4336–5233 (4838.26165.5) Hz. Notes pulses; Carvalho et al., 2019a) by having notes with partly have a modest or pronounced frequency upsweep varying fused pulses. Adenomera andreae is distinguished from A. from 234–1077 (583.76138.7) Hz. hylaedactyla (H1: 1852–2211 Hz; H2: 3680–4457 Hz; Table 2) by higher frequencies (H1: 2221–2588 Hz; H2: 4242–5233 Hz; Phylogenetic placement.—All specimens of A. andreae from Table 3), which could also reflect its relatively smaller SVL. the region of the type locality were recovered deeply nested Adenomera andreae (41–76 ms; Table 3) is distinguished from A. in the lineage of Fouquet et al. (2014) associated with chicomendesi (154–247 ms; Carvalho et al., 2019b), A. coca nominal A. andreae (Fig. 7). The only sympatric species of (110–145 ms; Angulo and Reichle, 2008), and A. heyeri (137– Adenomera collected in eastern Brazilian Amazonia (voucher 185 ms; Boistel et al., 2006) by its shorter note duration. From ZUFG 101) fell within the open-formation A. hylaedactyla the partly sympatric A. hylaedactyla (107–242 notes/min; Table clade (Fig. 7). The four call vouchers of Forest Call III from 2), A. andreae can be further distinguished by a lower note rate Tambopata, Peru (MUSM 39464, 39469–71; see Angulo et al. (24–68 notes/min; Table 3). [2003] for terminology and description) fell within A. andreae. Genetic distances in COI within A. andreae are Acoustic comparisons between A. andreae and A. hylaedacty- substantial (0.0–11.4%, mean distance: 7%; n ¼ 77), albeit la.—In addition to occupying different habitats across the phenotypic divergence is pronounced only in the Tambopata Amazon rainforest, these species can also be distinguished by population (Forest Call III) based on our acoustic dataset (see their calls (Tables 2, 3). One striking difference is the note Discussion, Table 3, Figs. 5C, 6I–L). repetition rate: A. hylaedactyla gives call notes at a notably higher rate (107–242 notes/min) than does A. andreae (24–68 Updated diagnosis and comparisons with Amazonian conge- notes/min), even though these species have similar note ners.—We collected a new series of A. andreae from a site durations (A. andreae: 41–76 ms; A. hylaedactyla: 41–89 ms). located approximately 18 km from the type locality (Fig. 1), Moreover, both fundamental (H1) and dominant (second which largely agrees with the previous morphological and harmonic; H2) frequencies differ noticeably in their calls: A. Adenomera color characterization provided by Heyer (1973). hylaedactyla has lower frequencies (H1: 1852–2211 Hz; H2: andreae is characterized by the following combination of 3680–4457 Hz) compared to A. andreae (H1: 2221–2588 Hz; features: (1) small to medium size for the genus (adult male H2: 4242–5233 Hz). These differences may reflect different SVL ¼ 19.3–23.0 mm); (2) toe tips fully expanded (character SVLs between A. andreae and A. hylaedactyla (Table 1), state D), especially in toes II–IV; (3) two possible chromotypes comparatively small- and mid-sized species, respectively, (presence/absence) of dorsolateral stripe; (4) absence of nearly based on the assumption of inverse relationship between solid dark-colored stripe on underside of forearm; (5) absence body size and call frequency (see review by Gerhardt and of antebrachial tubercle; (6) forest-dweller; (7) advertisement Huber, 2002). call composed of single notes; (8) call notes formed by partly fused notes—even though notes may be weakly pulsed or Notes on natural history.—Adenomera andreae and A. hylae- even indistinct sometimes; (9) call dominant frequency dactyla are found in sympatry throughout the Amazon coinciding with the second harmonic; (10) low note rate rainforest, but habitat use and calling activity are different, (,100 notes/min; on average, around 55 notes/min). as previously reported by Heyer (1977) and Menin et al. Adenomera andreae is distinguished from medium- to large- (2009). Adenomera andreae occurs within lowland rainforest sized congeners, i.e., A. lutzi and A. simonstuarti (combined and forest edges with well-drained soils (terra firme forests) SVL 25.7–33.5 mm; Table 1; Kok et al., 2007) by having small and often calls during the middle to the end of the day with a to medium-sized adult males (SVL 19.3–23.0 mm). By having crepuscular peak. In contrast, A. hylaedactyla can be found in toe tips fully expanded (character state D; Fig. 4H), A. andreae every kind of open formation, including forest clearings, is distinguished from species with unexpanded and/or alongside river banks, and in completely altered environ- slightly swollen toe tips (character states A–B: A. coca [Angulo ments, such as pastures and gardens, and is observed to call and Reichle, 2008], A. hylaedactyla [Heyer, 1973], and A. during daytime and throughout the night, especially on phonotriccus [Carvalho et al., 2019a]). From A. hylaedactyla drizzly days, with a peak in the first half of the night. (dorsolateral stripe absent), A. andreae can be further distinguished by having two possible chromotypes (pres- DISCUSSION ence/absence) of dorsolateral stripe. Adenomera andreae can be distinguished from A. simonstuarti (Angulo and Icochea, Advertisement call.—Calls previously assigned to A. andreae 2010) by the absence of a nearly solid dark-colored stripe on have been described by Zimmerman and Bogart (1984) and underside of forearm, and from A. lutzi and A. phonotriccus by Boistel et al. (2006). Ma´rquez et al. (1995) described the call the absence of antebrachial tubercle (Kok et al., 2007; of a population assigned at that time to A. andreae; it was, Carvalho et al., 2019a). Adenomera andreae is a forest-dweller, however, later reassigned to A. diptyx (De la Riva et al., 2000; de Carvalho et al.—On the type specimens of Amazonian Adenomera 719 see further discussion on this topic in the next paragraphs). the call characterization for A. hylaedactyla based on a Angulo and Icochea (2003) described the call of a population detailed quantitative analysis and a substantially larger that they referred to as A. cf. andreae, although they made it sample size of 24 males recorded (n ¼ 1074 calls; n ¼ 7196 clear that this was a distinct species based on acoustic pulses). Ma´rquez et al. (1995) described calls from Bolivian features, and Angulo et al. (2003) identified three forest- populations of Adenomera assigned to A. andreae (subsequent- associated calls from Tambopata, in the upper Amazon Basin ly reassigned to A. diptyx; De la Riva et al., 2000) and A. of Peru. hylaedactyla. We had access to the original recordings housed Calls reported by Angulo and Icochea (2003) for a forest- at Fonoteca Zoologica´ in Madrid (FonoZoo) and found that, associated population near R´ıo Camisea (Peru) and assigned based mainly on the dominant frequency and pulse number, to A. cf. andreae differ strikingly from nominal A. andreae in calls assigned to A. andreae by Ma´rquez et al. (1995) should their call envelopes and much lower dominant frequency, actually be assigned to A. hylaedactyla (FonoZoo recording which is the same as the fundamental frequency. Differences #10434). On the other hand, calls assigned to A. hylaedactyla of this order of magnitude are considered indicative of by these authors might correspond to calls of A. diptyx interspecific variation. We believe that the population from (FonoZoo recordings #10574, 10576–77, 10586, and 10629). R´ıo Camisea could correspond to one of the candidate species It is worth adding that calls of the latter species deserve a in the A. andreae clade (see Fouquet et al., 2014). Two detailed description from the type locality, since there is different calls from Cusco Amazonico´ (Peru) assigned to cryptic species richness within A. diptyx (V. Zaracho, pers. Adenomera sp. (Duellman, 2005) were recorded in forest and comm. to T. R. de Carvalho). Therefore, the recordings listed clearing habitats. It may be that a species of the A. andreae above are provisionally assigned to A. cf. diptyx. clade other than nominal A. andreae (possibly Forest Call II of Angulo et al., 2003) and A. hylaedactyla were recorded, Taxonomy.—The holotype of A. hylaedactyla does not have respectively, in that area based on note duration and key features for an unambiguous species assignment. While repetition rate. A figure detailing fine-temporal characteris- we are not able to assess DNA sequence from this old and tics and corresponding frequency structure would help with poorly preserved type specimen, it would appear that the identification. At this time, these suggestions need to be most reasonable and practical taxonomic solutions would be evaluated with additional recordings. Calls from French to (1) keep the name A. hylaedactyla to accommodate the Guiana assigned to A. andreae (Lescure and Marty, 2000) Amazonian open-formation morphotype of Adenomera (sensu correspond to A. heyeri. Heyer, 1973), and to (2) avoid describing new forest-dwelling Angulo et al. (2003) reported three types of forest- species of Adenomera (A. andreae clade) from the vast region associated calls for Adenomera from Tambopata, in south- of Amazonia entailing the type locality of A. hylaedactyla so eastern Peru. Adenomera chicomendesi (¼Forest Call I) and as to reduce the risk of synonymization. There are at least two

Forest Call II are several orders of magnitude longer, have species of Adenomera occurring sympatrically in the vast and more pulses, and have unique call envelopes; thus, they are vague type locality of A. hylaedactyla (A. andreae and A. considered to be two undescribed species of the A. andreae simonstuarti), plus additional candidate species associated clade (Carvalho et al., 2019b). On the other hand, Forest Call with the A. andreae clade (related to nominal A. andreae and III (referred to as A. cf. andreae in our study; Fig. 5C) shares A. simonstuarti; Carvalho, unpubl. data). However, it is most acoustic features with A. andreae, even though they important to note that not describing species has conse- differ markedly in the duration of their calls (Table 3). Also, quences for their conservation given that conservation the envelope of A. cf. andreae does not have incomplete funding is tied to described species and is often directed pulses and could possibly be regarded as a nonpulsed note, in towards threatened species. An undescribed and potentially contrast with nominal A. andreae from eastern Brazilian threatened cryptic species runs the risk of going extinct Amazonia, whose pulse variation ranges from indistinctly because it is not acknowledged as a valid species. With pulsed to partly pulsed within and between males (see Fig. 6). rampant fires currently burning over a vast swath of It is worth noting that Fouquet et al. (2014) had attributed Amazonia, this is not a farfetched possibility. Thus, the risk Forest Call III to nominal A. andreae, which was classified of losing species far outweighs that of synonymization, and into three clades based on genetic structure. The genetic we recommend that descriptions be prioritized based on the diversity associated with the name A. andreae, combined with potential extinction risk of new species. acoustic variation in the Tambopata population referred to as There are noticeably different acoustic patterns associated Forest Call III, support the hypothesis that A. andreae might with the highly conservative open-formation morphotype of constitute a species complex which diverged acoustically but A. hylaedactyla, as already reported by Heyer (1984). In to a lesser extent when compared with other species addition to the pattern described for nominal A. hylaedactyla complexes in the genus (Carvalho and Giaretta, 2013a; in the present study (Table 2), we have recorded distinct calls Carvalho et al., 2019a). Nonetheless, we preliminarily assign throughout Brazilian Amazonia, as well as across the Cerrado Tambopata Forest Call III to A. cf. andreae while additional savanna in mainland Brazil and in the northern Atlantic phenotypic data are acquired. Forest. We assigned one of these call patterns to nominal A. Calls have been assigned to A. hylaedactyla in previous hylaedactyla based on the correspondence with the species’ studies. We analyzed calls from the same regions as the type locality region in Peruvian and Brazilian Amazonia (Fig. previous descriptions (Brazilian and Peruvian Amazonia), but 1). The A. hylaedactyla clade has an intricate evolutionary did not include calls from central-western Brazil or eastern history and distribution pattern accompanied by pro- Bolivia, given the taxonomic uncertainty surrounding pop- nounced acoustic diversity, which can potentially represent ulations from those regions (Heyer, 1973; Straughan and an additional case of species complex in Adenomera,as Heyer, 1976; Ma´rquez et al., 1995). Acoustic traits provided previously discussed by Fouquet et al. (2014). Therefore, we inthepresentstudy(Table2)largelyagreewiththe decided to set aside all data accumulated over the past decade descriptions by Schluter¨ (1980), Schneider et al. (1988), for this species complex towards a future contribution, so as Angulo et al. (2003), and Menin et al. (2009), but we refined to evaluate in detail the taxonomic status of genetic lineages 720 Copeia 107, No. 4, 2019 under the name A. hylaedactyla, previously indicated by Adenomera lutzi: INPA-H 6247, Brazil, Roraima, Upper Mau´ Fouquet et al. (2014), combined with our acoustic data from River; MZUSP 149404–09, 150799–804, Guyana, Potaro- throughout South America east of the Andes, and the Siparuni. lengthy nomenclatural history under this species. Moreover, the taxonomic status of A. coca should be carefully examined. Adenomera phonotriccus: MPEG 41155 (holotype), CFBH The genetic vouchers associated with A. coca are well nested 43130–31, MPEG 41156 (paratypes), Brazil, Para´, Palestina within A. hylaedactyla (Fouquet et al., 2014; Fig. 7); however, do Para´. there are noticeable differences in acoustic temporal traits between A. coca (Angulo and Reichle, 2008) and A. Adenomera simonstuarti: MUSM 18218 (holotype), MUSM hylaedactyla (Table 2). Additional sound recordings of A. coca 18220–21, 18229 (paratypes), Peru, Cusco, La Convencion,´ are needed for further quantitative characterization and Echarate, R´ıo Camisea. comparisons with the acoustic variation in A. hylaedactyla. A. andreae With regard to , the pan-Amazonian distribution DATA ACCESSIBILITY attributed to this species (Heyer, 1973; Fouquet et al., 2014) will require future evaluation with the inclusion of prelim- Supplemental material is available at https://www. inary acoustic evidence presented in this study for the copeiajournal.org/ch-19-237. Tambopata population (Table 3; Fig. 6). It is of special importance, whenever possible, to use complementary ACKNOWLEDGMENTS sources of information other than morphology for species assignments in the A. andreae clade, i.e., forest dwellers We received financial support through research grants from distributed throughout Amazonia, assuming that there are Sa˜o Paulo Research Foundation (FAPESP #2013/50741-7) and still some undescribed species in the clade (Carvalho et al., from National Council for Scientific and Technological 2019b). To our knowledge, cryptic species richness associated Development (CNPq #302518/2013-4 and #446935/2014- with the morphotype of A. andreae is expected to be 0). MM and NMM also thank CNPq for research grants. The concentrated in Amazonian regions across north-central taxonomic review of species of Adenomera was part of the first Peru, eastern Ecuador, southern Colombia and Venezuela, author’s Ph.D. dissertation, with financial support from a and northwestern Brazil (Fouquet et al., 2014). The combined doctoral and a postdoctoral fellowship from FAPESP (#2012/ use of acoustic and genetic data will help researchers better 15763-7 and #2017/08489-0, respectively). Collecting per- understand species complexes in the A. andreae clade and mits were issued by Instituto Chico Mendes, Brazil (ICMBio/ reduce the knowledge gap between species composition and SISBIO #30059), and Sernanp and Serfor, Peru (#17-2018- richness in Amazonia, the most species-rich terrestrial SERNANP-JEF and #192-2019-MINAGRI-SERFOR). We are grateful to B. F. V. Teixeira, D. L. Bang, C. E. Costa-Campos, ecosystem on the planet. L. B. Martins, P. Azarak, and the Explorer’s Inn team for their MATERIAL EXAMINED assistance during fieldwork; Centro de Estudos de Insetos Sociais (CEIS, UNESP/Rio Claro) and M. L. Lyra for providing Adenomera andreae: AAG-UFU 2788–94, Brazil, Para´, Nova workspace and help with DNA sequencing protocols; Timboteua; AAG-UFU 2797–98, Brazil, Para´, Belem;´ AAG- Fonoteca Zoologica´ (MNCN-CSIC, Madrid), the Macaulay UFU 4282–83, INPA-H 34045, 34048, 34073–74, 34076, Library at the Cornell Lab of Ornithology (ML), G. Gagliardi- 34081–82, 34084–86, 34090, LHUFCG 189, 194, 197–204, Urrutia, I. De la Riva, and R. R. Rojas for enabling access to ZUEC 3937, 3969, 3973–74, 7799, Brazil, Amazonas, Manaus; sound recordings of Adenomera; and D. R. Frost for insightful AAG-UFU 5994, 6006–07, CFBH 43259, 43265, Brazil, comments. We are indebted to Michael Franzen and Frank Amapa´, Serra do Navio; MUSM 27554, Peru, Loreto, Datem Glaw from the Munich museum (ZSM) for providing us with del Maranon;˜ QCAZ 64180–84, 64186–87, 64191–92, 64194– high-quality photographs and sending us tissue samples 99, Ecuador, Orellana, Parque Nacional Yasun´ı. from the type specimens of A. andreae. Ned Gilmore from ANSP allowed AA earlier access to the holotype of A. Adenomera chicomendesi: CFBH 43562 (holotype), AAG-UFU hylaedactyla. The following individuals granted access to 5862–64, CFBH 43563, ZUEC 24528–31 (paratypes), Brazil, specimens under their care: F. P. Werneck and A. Silva (INPA- Acre, Rio Branco, Parque Zoobotaˆnico; MUSM 39462–63, H), N. Pupin and D. B. Delgado (CFBH), L. F. Toledo and K. 39467–68, 39472–74 (paratypes), Peru, Madre de Dios Rabelo (ZUEC), A. L. C. Prudente and F. Sarmento (MPEG), T. Reserva Nacional de Tambopata. Grant and A. S. Benetti (MZUSP), G. F. Pontes (MCP), S. R. Ron (QCAZ). Adenomera heyeri: MPEG 30099–101, Brazil, Para´, Oriximina´, ESEC-Gra˜o-Para´. LITERATURE CITED

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Ronquist, F., M. Teslenko, P. van der Mark, D. L. Ayres, A. Nova Timboteua, Para´.—(1) Unvouchered; sound file: Darling, S. Hohna,¨ B. Larget, L. Liu, M. A. Suchard, and Adenom_andreaeNovaTimboteuaPA1bTRC_AAGm671; 25 J. P. Huelsenbeck. 2012. MrBayes 3.2: efficient Bayesian November 2013, 1808 h, air 27.68C. (2) Voucher AAG-UFU phylogenetic inference and model choice across a large 2788; sound file: Adenom_andreaeNovaTimboteuaPA2TRC_ model space. Systematic Biology 61:539–542. AAGm671; 26 November 2013, 1647 h, air 26.58C. (3) Sabaj, M. H. 2016. Standard symbolic codes for institutional Voucher AAG-UFU 2789; sound file: Adenom_andreaeNova resource collections in herpetology and ichthyology: an TimboteuaPA3TRC_AAGm671; 26 November 2013, 1656 h, Online Reference. Version 6.5 (16 August 2016). Electron- air 26.58C. (4) Voucher AAG-UFU 2790; sound file: Adenom_ ically accessible at http://www.asih.org/, American Society andreaeNovaTimboteuaPA4TRC_AAGm671; 26 November of Ichthyologists and Herpetologists, Washington, D.C. 2013, 1748 h, air 26.58C. (5) Voucher AAG-UFU 2791; sound Schluter,¨ A. 1980. Bio-akustische untersuchungen an lep- file: Adenom_andreaeNovaTimboteuaPA5TRC_AAGm671; todactyliden in einem begrenzten gebiet des tropischen 26 November 2013, 1750 h, air 26.58C. (6) Unvouchered; regenwaldes von Peru. Salamandra 16:227–247. sound file: Adenom_andreaeNovaTimboteuaPA1cBFVT_ Schneider, H., G. Joermann, and W. Hodl.¨ 1988. Calling AAGmt; 25 November 2013, 1827 h, air 27.68C. (7) Voucher and antiphonal calling in four Neotropical anuran species AAG-UFU 2793; sound file: Adenom_andreaeNova of the family Leptodactylidae. Zoologische Jahrbucher¨ TimboteuaPA1bLBM_AAGm671; 25 November 2013, 1915 h, air 28.38C. (8) Unvouchered; sound file: Adenom_ Abteilung fur¨ Allgemeine Zoologie und Physiologie der andreaeNovaTimboteuaPA2LBM_AAGm671; 25 November Tiere 92:77–103. 2013, 1925 h, air 28.38C. Straughan, I. R., and W. R. Heyer. 1976. A functional analysis of the mating calls of the Neotropical frog genera Santa Ba´rbara do Para´, Para´.—(1) Unvouchered; sound file: of the Leptodactylus complex (Amphibia, Leptodactylidae). Adenomera_andreaeStaBarbaraPA2AAGm661MK2; 11 Febru- Papeis´ Avulsos de Zoologia 29:221–245. ary 2019, 1804 h, 28.08C. (2) Unvouchered; sound file: Tamura, K., and M. Nei. 1993. Estimation of the number of Adenomera_andreaeStaBarbaraPA3AAGm661MK2; 11 Febru- nucleotide substitutions in the control region of mito- ary 2019, 1809 h, 28.08C. (3) Unvouchered; sound file: chondrial DNA in humans and chimpanzees. Molecular Adenomera_andreaeStaBarbaraPA4AAGm661MK2; 11 Febru- Biology and Evolution 10:512–526. ary 2019, 1814 h, 28.08C. (4) Unvouchered; sound file: Watters, J. L., S. T. Cummings, R. L. Flanagan, and C. D. Adenomera_andreaeStaBarbaraPA5AAGm661MK2; 11 Febru- Siler. 2016. Review of morphometric measurements used ary 2019, 1818 h, 28.08C. (5) Unvouchered; sound file: in anuran species descriptions and recommendations for a Adenomera_andreaeStaBarbaraPA6AAGm661MK2; 11 Febru- standardized approach. Zootaxa 4072:477–495. ary 2019, 1712 h, 28.08C. (6) Voucher AAG-UFU 6623; sound Zimmerman, B. L., and J. Bogart. 1984. Vocalization of file: Adenomera_andreaeStaBarbaraPA8AAGm661MK2; 11

primary forest frog species in the Central Amazon. Acta February 2019, 1743 h, 28.08C. Amazonica 14:473–519. Adenomera andreae (central Amazonia) Manaus, Amazonas.—(1) Unvouchered; sound file: Adenom_ Appendix 1. Scores, eigenvalues, and explained variation of the first andreaeManausAM1TRC_AAGmt; 22 January 2011, 1804 h, three principal components retained from a PCA on a morphometric 26.08C. (2) Voucher LHUFCG 194; sound file: Adenom_ dataset of three Amazonian species of Adenomera: A. andreae; A. andreae_23_1_ReservaDucke_MNCK; 16 December 2005, hylaedactyla, including the species’ holotype as a separate group; and 1834 h, 28.08C. (3) Voucher LHUFCG 195; sound file: A. simonstuarti. Adenom_andreae_24_1_ReservaDucke_MNCK; 16 December Trait PC1 PC2 PC3 2005, 1835 h, 29.08C. (4) LHUFCG 196; sound file: Adenom_andreae_35_1_ReservaDucke_MNCK; 17 December Snout–vent length 1.524 –0.044 –0.006 2005, 1757 h, 24.08C. (5) LHUFCG 202; sound file: Eye–nostril distance 0.959 –1.043 0.576 Adenom_andreae_39_1_ReservaDucke_MNCK; 17 December Eye diameter 0.913 1.186 –0.299 2005, 1813 h, 24.08C. Tympanum diameter 0.874 0.878 0.979 Hand length 1.469 0.034 –0.209 Adenomera andreae (eastern Guiana Shield) Thigh length 1.310 –0.445 –0.157 Tibia length 1.488 –0.007 –0.215 Serra do Navio, Amapa´.—(1) Unvouchered; sound file: Foot length 1.499 –0.161 –0.195 Adenomera_andreaeSNavioAP1bAAGm661MK2; 27 March Eigenvalues 5.167 1.370 0.601 2017, 1750 h, 26.08C. (2) Unvouchered; sound file: Variation (%) 64.6 17.1 7.5 Adenomera_andreaeSNavioAP2AAGm661MK2; 27 March 2017, 1755 h, 26.08C. (3) Unvouchered; sound file: Adenomera_andreaeSNavioAP3AAGm661MK2; 27 March APPENDIX 2 2017, 1759 h, 26.08C. (4) Unvouchered; sound file: TRC114; 31 March 2018, 1834 h, air 25.48C. (5) Voucher Adenomera andreae All the following sound recordings of and CFBH 43265; sound file: TRC116; 31 March 2018, 1848 h, air A. hylaedactyla were obtained in Brazilian Amazonia unless 25.48C. otherwise indicated. Adenomera cf. andreae (southwestern Peruvian Amazo- Adenomera andreae (eastern Amazonia) nia) Belem,´ Para´.—(1) Voucher AAG-UFU 2797; sound file: Tambopata, Madre de Dios.—(1) Voucher USNM 343236; Adenom_andreaeBelemPA1TRC_AAGm671; 28 November sound file: ML198801; 3 November 1990, 1655 h, 26.68C. (2) 2013, 1815 h, 27.58C. Voucher USNM 343232; sound file: ML198802; 3 November de Carvalho et al.—On the type specimens of Amazonian Adenomera 723

1990, 1715 h, 26.28C. (3) Voucher USNM 343237; sound file: 2016, 1823 h, 27.08C. (2) Unvouchered; sound file: ML198804; 3 November 1990, 1820 h, 29.58C. (4) Voucher Adenomera_hylaedactylaCantaRR3aAAGm671; 21 July MUSM 39464; sound file: TRC167; 25 November 2018, 1616 2016, 1831 h, 27.08C. (3) Unvouchered; sound file: h, 26.68C. (5) Unvouchered; sound file: TRC169; 25 Novem- Adenomera_hylaedactylaCantaRR4aAAGm671; 23 July ber 2018, 1626 h, 26.68C. (6) Unvouchered; sound file: 2016, 1813 h, 27.08C. (4) Unvouchered; sound file: TRC178; 27 November 2018, 1619 h, 25.98C. (7) Voucher Adenomera_hylaedactylaCantaRR6aAAGm671; 23 July MUSM 39469; sound file: TRC179; 27 November 2018, 1643 2016, 1819 h, 27.08C. (5) Unvouchered; sound file: h, 25.98C. (8) Unvouchered; sound file: TRC180; 27 Novem- Adenomera_hylaedactylaCantaRR7aAAGm671; 23 July 8 ber 2018, 1648 h, 25.0 C. 2016, 1827 h, 27.08C. (6) Unvouchered; sound file: Adenomera_hylaedactylaCantaRR9aAAGm671; 23 July Adenomera hylaedactyla (central Amazonia) 2016, 1851 h, 27.08C. (7) Unvouchered; sound file: Manaus, Amazonas.—(1) Unvouchered; sound file: Adenom_ Adenomera_hylaedactylaCantaRR15aAAGm671; 23 July hylaedactylaAM1TRC_AAGmt; 22 January 2011, 1828 h, 2016, 1917 h, 27.08C. 26.08C. (2) Voucher AAG-UFU 4740; sound file: Adenom_ hylaedactyla_53_1_ReservaDucke_MNCK; 17 December Adenomera hylaedactyla (southwestern Amazonia) 2005, 1834 h, 28.08C. (3) Voucher AAG-UFU 4736; sound file: Adenom_hylaedactyla_40_2_ReservaDucke_MNCK; 17 Tambopata, Madre de Dios, Peru.—(1) Voucher ROM 40105; December 2005, 1942 h, 22.08C. (4) Voucher AAG-UFU sound file: Peru-hyl-AA9945; 26 January 1999, 1911 h, 4737; sound file: Adenom_hylaedactyla_44_ReservaDucke_ 25.58C. (2) Unvouchered; sound file: TRC160; 24 November MNCK; 17 December 2005, 1942 h, 22.08C. (5) Voucher 2018, 1819 h, 23.58C. (3) Unvouchered; sound file: TRC161; Voucher AAG-UFU 4738; sound file: Adenom_hylaedactyla_ 24 November 2018, 1828 h, 23.58C. (4) MUSM 39460; sound 45_2_ReservaDucke_MNCK; 17 December 2005, 2019 h, file: TRC170; 25 November 2018, 1855 h, 25.58C. 22.08C. (6) Voucher AAG-UFU 4739; sound file: Adenom_ hylaedactyla_52_ReservaDucke_MNCK; 17 December 2005, Iquitos, Loreto, Peru.—(1) Unvouchered; sound file: DR470; 2109 h, 24.08C. 14 June 2014, 2135 h, temperature unspecified.

Cruzeiro do Sul, Acre.—(1) Unvouchered; sound file: Adenomera hylaedactyla (northwestern Amazonia) Adenom_hylaedactylaCruzeiroSulAC1TRC_AAGm671; 6 ˜ Sao Gabriel da Cachoeira, Amazonas.—(1) Unvouchered; February 2017, 1922 h, 25.58C. (2) Unvouchered; sound file: sound file: Adenom_hylaedactylaSGabrCachoe Adenomera_hylaedactylaCruzeiroDoSulAC1aDLB_AAGm671; AM1AAGm671; 26 June 2014, 1815 h, 30.28C. (2) Voucher 6 February 2017, 2212 h, 27.38C. AAG-UFU 3861; sound file: Adenom_hylaedactylaSGabr

CachoeAM2AAGm671; 26 June 2014, 1819 h, 30.28C. Feijo,´ Acre.—(1) Unvouchered; sound file: Adenomera_ hylaedactylaFeijoAC1bDLB_AAGm671; 4 February 2017, Adenomera hylaedactyla (Cerrado-Amazonia transition- 2050 h, 24.08C. al zone) Canta´, Roraima.—(1) Voucher AAG-UFU 5541; sound file: Tabatinga, Amazonas.—(1) Unvouchered; sound file: Adenomera_hylaedactylaCantaRR2aAAGm671; 21 July ZOOM26; date unspecified, 1915 h, 24.08C.