Research Article |
Corresponding author: Juan C. Cusi ( jcarloscusim@gmail.com ) Academic editor: Anthony Herrel
© 2017 Juan C. Cusi, Jiří Moravec, Edgar Lehr, Václav Gvoždík.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Cusi JC, Moravec J, Lehr E, Gvoždík V (2017) A new species of semiarboreal toad of the Rhinella festae group (Anura, Bufonidae) from the Cordillera Azul National Park, Peru. ZooKeys 673: 21-47. https://doi.org/10.3897/zookeys.673.13050
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A new semiarboreal species of the Rhinella festae group is described from montane forests of the Cordillera Azul National Park between 1245 and 1280 m a.s.l. in the Cordillera Oriental, San Martín region, northern Peru. The new species is morphologically and genetically compared with members of the Rhinella acrolopha group (former genus Rhamphophryne) and members of the R. festae group. The new species is characterized by its large size (female SVL 47.1–58.3 mm, n = 4), eight presacral vertebrae, fusion of the sacrum and coccyx, long protuberant snout, snout directed slightly anteroventral in lateral view, cranial crests moderately developed, absence of occipital crest, presence of tympanic membrane, dorsolateral rows of small conical tubercles extending from parotoid gland to groin, hands and feet with long digits, fingers basally webbed and toes moderately webbed. Phylogenetically it is a member of the R. festae group which is most closely related to R. chavin and R. yanachaga from Peru. Morphologically the new species shares similarities with R. tenrec and R. truebae, members of the R. acrolopha group from Colombia.
Amphibia , phylogeny, Rhinella acrolopha group, Rhinella lilyrodriguezae new species
The family Bufonidae Gray, 1825 comprises a clade of neobatrachian anurans commonly known as “true toads” of approximately 600 species and more than 50 genera (
The former genus Rhamphophryne Trueb, 1971, now a part of Rhinella, comprised several species of bufonid toads distributed in South American tropical forests. Most of the diversity of this genus occurs in montane forests, with nine species in northern Colombia and eastern Panama (R. acrolopha Trueb, 1971; R. lindae Rivero & Castaño, 1990; R. macrorhina Trueb, 1971; R. nicefori Cochran & Goin, 1970; R. paraguas Grant & Bolívar-G., 2014; R. rostrata Noble, 1920; R. ruizi Grant, 2000“1999”; R. tenrec Lynch & Renjifo, 1990; and R. truebae Lynch & Renjifo, 1990) and one species extending also to the upper Amazonian Basin of Ecuador (R. festae Peracca, 1904). From early on, the monophyly of the genus was not supported by
Recently, based on morphological and molecular data, the generic name Rhamphophryne was synonymized with Rhinella Fitzinger, 1826 by
The Cordillera Azul National Park (herein CAZNP) located in the Cordillera Oriental of the Andes, Loreto, San Martín, Huánuco and Ucayali regions, is one of the most diverse natural protected areas in northern Peru. CAZNP has an extension of more than 13,000 km2 with an altitudinal range from 200 to 2400 m a.s.l. between the Huallaga and Ucayali rivers (
Here, a thorough comparison of this new bufonid from the Cordillera Azul National Park with other related Rhinella species is provided, its phylogenetic position based on DNA barcoding data elucidated, and the taxon formally described as a new species of the Rhinella festae group.
Fieldwork and deposition of specimens. Six specimens of the new species were collected inside the CAZNP (Fig.
Map showing the type localities of Rhinella lilyrodriguezae sp. n. (○, yellow), selected species of the former genus Rhamphophryne (⊡, blue; with numbers 1 R. acrolopha 2 R. tenrec 3 R. lindae 4 R. nicefori 5 R. rostrata 6 R. ruizi 7 R. paraguas), species of the Rhinella festae group (⊙, red; with letters a R. macrorhina b R. festae c R. chavin d R. yanachaga e R. manu, f: Rhinella cf. nesiotes) and type localities of Rhinella nesiotes (★, green star) and Rhinella tacana (✙, cross, green). Abbreviation PRC: Park Rangers Center.
The specimens were compared with species previously assigned either to the Rhinella festae group or the R. acrolopha group (see Appendix) and original species descriptions. The coordinates of the type and reference localities of species of the R. acrolopha group were obtained using Global Gazetteer version 2.3 (http://www.fallingrain.com/world/index.html). The geographic coordinates were based on the datum WGS84 and maps were designed using ArcGIS version 10.0.
Morphological characters. Morphometric measurements in millimeters (mm) were taken with a digital caliper Mitutoyo (nearest to 0.1 mm). Measurement abbreviations used throughout the text are:
SVL snout–vent length;
HW head width (at level of angle of jaw);
HL head length (from angle of jaw to tip of snout);
ED horizontal eye diameter;
IOD interorbital distance;
EW upper eyelid width;
EL eyelid length (upper eyelid length);
IND internarial distance;
E–N eye-nostril distance (straight line distance between anterior corner of orbit and posterior margin of external nares);
NSD nostril-snout distance;
SL snout length (between anterior corner of eye and tip of snout);
FL forearm length (between flexed elbow and proximal edge of palmar tubercle);
HNDL hand length (between proximal edge of palmar tubercle and tip of Finger III);
FEML femur length;
TL tibia length;
FOOTL foot length (distance from proximal margin of inner metatarsal tubercle to tip of Toe IV).
Fingers and toes are numbered preaxially to postaxially from I–IV and I–V, respectively. We determined comparative lengths of Toes III and V by adpressing both toes against Toe IV; lengths of fingers I and II were determined by adpressing the fingers against each other. Condition of the tympanum was assessed by visual examination under stereoscope. Specimens were sexed by examination of gonads and secondary sex characters. Ovarian eggs number and coloration were observed by dissection of a gravid female. The measurements of each egg were obtained of the maximum diameter, calculating the mean and standard deviation. Format of the description and diagnosis follows the standards of
Webbing formula follows
Taxon sampling. Three specimens of the putative new species from the Rhinella festae group from the Cordillera Azul National Park were compared with the taxa from the dataset from
Species, sample localities, museum numbers and GenBank accession numbers for DNA sequences used in the phylogenetic analysis of all samples of the Rhinella festae group, and new material of the R. margaritifera, R. veraguensis and R. marina groups. For the species group affiliation see Fig.
Species | Locality | Museum No. | GenBank Accession No. | Reference |
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16S Cytb | ||||
Rhinella lilyrodriguezae sp. n. | Peru: San Martín, Bellavista, Alto Biavo, Cordillera Azul National Park |
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KY912598 | This study |
Rhinella lilyrodriguezae sp. n. | Peru: San Martín, Bellavista, Alto Biavo, Cordillera Azul National Park |
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KY912599 | This study |
Rhinella lilyrodriguezae sp. n. | Peru: San Martín, Bellavista, Alto Biavo, Cordillera Azul National Park |
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KY912600 | This study |
Rhinella yanachaga | Peru: Pasco, Oxapampa, Cordillera Yanachaga: Quebrada Yanachada, 2900 m | FMNH 282819 | KF992148 |
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Rhinella yanachaga | Peru: Pasco, Oxapampa, Cordillera Yanachaga: Quebrada Yanachada, 2900 m |
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KF992149 |
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Rhinella chavin | Peru: Palma Pampa | MTD 43789 | DQ158441 |
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Rhinella sp. C (= Rhinella sp. acrolopha group sensu |
-- | -- | KT221613 | Machado et al. 2016 |
Rhinella festae | Ecuador: Pastaza, Petrolera Garza | KU 217501 | DQ158423 |
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Rhinella festae | Ecuador: Napo, Estacion Biologica Jatun Sacha | QCAZ 18203 | KR012624 | dos Santos et al. 2015 |
Rhinella macrorhina (before “R. rostrata” in GenBank) | Colombia, Antioquia, 0.5 km W (by road) Medellin† | MVZ:Herp:231697 (FC-13112) | AF375533 | Gluesenkamp, unpublished |
Rhinella macrorhina‡ | Colombia, Antioquia, 0.5 km W (by road) Medellin† | MVZ:Herp:150267 (FC-13113) | AF375532 | Gluesenkamp, unpublished |
Rhinella cf. nesiotes | Bolivia: La Paz, Caranavi, Serranía de Bella Vista | UTA 53310 | DQ158478 |
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Rhinella yunga | Peru: Junín, area of Rio Huatziroki, buffer zone of the Pui Pui Protected Forest, 1950 m |
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KY912601 | This study |
Rhinella yunga | Peru: Junín, area of Rio Huatziroki, buffer zone of the Pui Pui Protected Forest, 2230 m |
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KY912602 | This study |
Rhinella yunga | Peru: Junín, area of Rio Huatziroki, buffer zone of the Pui Pui Protected Forest, 2075 m | NMP6V 75552 | KY912603 | This study |
Rhinella cf. margaritifera | Peru: Junín, Ayte, buffer zone of the Pui Pui Protected Forest, 2007 m |
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KY912604 | This study |
Rhinella cf. margaritifera | Peru: Junín, Ayte, buffer zone of the Pui Pui Protected Forest, 2007 m |
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KY912605 | This study |
Rhinella cf. margaritifera | Peru: Junín, Ayte, buffer zone of the Pui Pui Protected Forest, 2007 m | IWU 334 | KY912606 | This study |
Rhinella cf. margaritifera | Peru: San Martín, Rioja, Pardo Miguel, Alto Mayo Protected Forest |
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KY912607 | This study |
Rhinella cf. margaritifera | Peru: San Martín, Rioja, Pardo Miguel, Alto Mayo Protected Forest |
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KY912608 | This study |
Rhinella cf. margaritifera | Bolivia: Polpebra | CBF 5800 | KY912609 | This study |
Rhinella cf. leptoscelis | Peru: Junín, area of Rio Bravo, buffer zone of the Pui Pui Protected Forest, 1721 m |
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KY912610 | This study |
Rhinella cf. leptoscelis | Peru: Junín, area of Rio Bravo, buffer zone of the Pui Pui Protected Forest, 1721 m | PE 008A | KY912611 | This study |
Rhinella cf. leptoscelis | Peru: Junín, area of Rio Bravo, buffer zone of the Pui Pui Protected Forest, 1721 m | PE 008B | KY912612 | This study |
Rhinella cf. leptoscelis | Peru: Junín, area of Rio Bravo, buffer zone of the Pui Pui Protected Forest, 1721 m | PE 008C | KY912613 | This study |
Rhinella poeppigii | Perú: Junin, La Merced, Pampa del Carmen, old swimming pool |
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KY912614 | This study |
Laboratory protocol and bioinformatics. A fragment of the mitochondrial 16S rRNA gene (~ 550 bp) was targeted. DNA extraction, PCR amplification and sequencing followed the methods described in
Phylogenetic analysis and systematics. The maximum likelihood (ML) analysis and Bayesian phylogenetic inference produced trees with the same topologies and strong support for the main clades. Our phylogenetic tree supported the R. festae group (Bayesian posterior probabilities 1.00/ML bootstrap 97) and the distinctiveness of the morphologically identified new species of Rhinella from the Cordillera Azul National Park. The new species was most closely related to a sister group composed of R. yanachaga and R. chavin (Fig.
The Bayesian consensus tree resulting from analysis of the mitochondrial 16S rRNA gene dataset for South American Rhinella species based on the data from
Rhinella
cf.
festae
:
Lily Rodriguez’s Beaked Toad
Sapo picudo de Lily Rodríguez
Five individuals (Figs
A large species of the Rhinella festae group confirmed by 16S DNA barcoding. The new species can be diagnosed by the following combination of characters: (1) large size, SVL 47.1–58.3 mm in females (n = 4), males are unknown; (2) eight presacral vertebrae; (3) sacral vertebrae fused with coccyx; (4) snout long, acuminate, pointed to rounded terminally in dorsal view; snout protuberant, directed slightly anteroventral in profile as a “shark snout”; (5) cranial crests moderately developed; (6) canthal, supraorbital, postorbital and supratympanic crests continuous, distinctly elevated in female, slightly elevated in juveniles; pretympanic crest present, occipital crest absent; (7) tympanic membrane present, tympanic annulus weakly defined; (8) mandibular angle not protruding; (9) parotoid glands moderately large, roughly triangular to rounded in outline, slightly swollen laterally, incorporated into lateral row of tubercles; (10) dorsolateral rows of small, conical tubercles extending from parotoid gland to groin; (11) hands and feet with long digits, fingers basally webbed and toes moderately webbed; (12) skin on dorsum smooth with scattered conical tubercles in females; (13) subarticular tubercles diffuse, round to ovoid; (14) supernumerary tubercles present, round but poorly developed; (15) cloacal sheath absent; (16) in life, dorsum light brown to greenish brown with irregular brown, dark brown or black markings; with or without grey white middorsal stripe; venter cream yellow to brownish grey with minute light cream spots; iris silvery greenish with irregular black mottling.
The new species shares similarities with members of the Rhinella festae and R. acrolopha groups. Three species of the R. acrolopha group from Colombia and Panama (R. truebae, R. lindae and R. tenrec) are large-sized toads similarly to R. lilyrodriguezae sp. n. (maximum SVL 65.9 mm in R. truebae, 62.2 mm in R. lindae, 60.8 mm in R. tenrec and 58.3 mm in R. lilyrodriguezae). The new species is most similar morphologically to R. truebae and R. tenrec, but is distinguished by lacking the occipital crest (which is low in R. truebae and R. tenrec). Rhinella lilyrodriguezae sp. n. can also be distinguished from R. tenrec by having a sacrum fused with the coccyx, cranial crests moderately developed and tympanic membrane present (sacrum not fused with the coccyx, cranial crests poorly developed, tympanic membrane absent in R. tenrec); from R. truebae and R. lindae by having a dorsolateral row of small conical tubercles extending from parotoid gland to groin, hands basally webbed, and feet moderately webbed (dorsolateral fold formed by the tubercles fusion, hands and feet extensively webbed in R. truebae and R. lindae). Rhinella lindae possesses a snout slightly directed upwards (snout directed slightly anteroventral as “shark snout” in R. lilyrodriguezae).
Other species of the Rhinella acrolopha group are R. acrolopha, R. nicefori, R. paraguas, and R. ruizi. These species are differentiated from R. lilyrodriguezae sp. n. (characters in parentheses) by the absence of the tympanic membrane (present), hands and feet extensively webbed in R. paraguas and R. ruizi, and reduced webbing in R. acrolopha and R. nicefori (hands basally webbed and feet moderately webbed), sacrum not fused with the coccyx except in R. paraguas (fused), seven presacral vertebrae except in R. paraguas (eight) and the presence of occipital crest except in R. paraguas (absent). Rhinella nicefori and R. ruizi have hands and feet with short digits (long digits). Rhinella paraguas and R. ruizi have cranial crests very low (moderately developed). Rhinella nicefori has a dorsolateral row of enlarged tubercles extending from the posterior margin of the parotoid gland to a point about three-fourths the distance between the axilla and groin (conical tubercles extending to groin). Rhinella acrolopha possesses a snout directed markedly anteroventrally (directed slightly anteroventral as “shark snout”) and a dorsolateral row of depressed tubercles extending from the posterior margin of the parotoid gland posteriorly to a point about two-thirds distance between axilla and groin (conical tubercles extending from parotoid gland to groin).
The remaining species of the R. acrolopha and R. festae groups from Colombia and Ecuador (R. festae, R. macrorhina and R. rostrata) are distinguished from R. lilyrodriguezae sp. n. by lacking the tympanic membrane (present), by having hands and feet extensively webbed (hands basally webbed and feet moderately webbed), seven presacral vertebrae (except for R. macrorhina which has eight), snout directed markedly anteroventrally in R. festae and R. macrorhina and straight in R. rostrata (directed slightly anteroventral). Rhinella festae and R. macrorhina have occipital crest well developed (absent). Rhinella macrorhina is distinct in having a sacrum not fused with the coccyx (fused), and a dorsolateral row of small tubercles extending from posterior margin of parotoid gland posteriorly to a point about one-half distance between axilla and groin (tubercles extending from parotoid gland to groin). Rhinella festae possesses dorsolateral row of slightly enlarged, conical tubercles extending from posterior margin of skull to a point about three-fourths distance between axilla and groin (tubercles extending from parotoid gland to groin) and hands with short digits (long digits).
Peruvian species of the R. festae group (R. chavin, R. manu, R. nesiotes and R. yanachaga) are distinguished from R. lilyrodriguezae sp. n. by having smaller females (maximum SVL 21.4 mm in R. manu, 23.6 mm in R. nesiotes, 45.7 mm in R. yanachaga, except in R. chavin with 54.8 mm; vs. 58.3 mm in R. lilyrodriguezae) and by having webbing of hands and feet fleshy (membranous in R. lilyrodriguezae). Rhinella chavin possesses a snout rounded in lateral view (snout protuberant, directed slightly anteroventral as “shark snout”), large [about twice ED], ovoid parotoid glands (moderately large [about same size as ED] and triangular), dorsolateral row of large, nearly round elevated tubercles beginning above insertion of forelimb extending to inguinal region (small, conical tubercles extending from parotoid gland to groin), elevated, elongate glands on forearm, tibia and outer dorsal margin of the foot and hand (glands absent) and hands and feet with relatively short digits (long digits). Rhinella nesiotes has a snout rounded in lateral view (snout protuberant, directed slightly anteroventral as “shark snout”), lacks cranial crests (moderately developed), low ovoid parotoid glands (moderately large and triangular), lacks dorsolateral row of tubercles (present) and hands and feet with relatively short digits (long digits). Rhinella manu has a snout pointed in lateral view (snout protuberant, directed slightly anteroventral as “shark snout”), large [about twice ED], oblong parotoid glands to the point of being nearly spherical (moderately large [about same size as ED] and triangular) and inconspicuous cranial crests (moderately developed). Rhinella yanachaga is most similar to R. lilyrodriguezae sp. n. from cloud forests of central Peru, both have cranial crests, tympanic membrane distinct, moderately large [about same size as ED] parotoid glands, dorsolateral rows of small, conical tubercles extending from parotoid gland to groin, fingers and toes relatively long and long, slender extremities. Nevertheless, R. yanachaga differs from R. lilyrodriguezae sp. n. by having a snout slightly protruding in lateral view (snout protuberant, directed slightly anteroventral as “shark snout”), well developed webbing on hands and feet (hands basally webbed, and feet moderately webbed), dorsal skin smooth without keratin-tipped tubercles in females (dorsum smooth with scattered conical tubercles in females) and dorsal coloration in life is dark brown with small, irregular, green spots and markings (dorsum light brown to greenish brown with irregular brown, dark brown or black markings).
Gravid female; body robust; SVL 58.3 mm; head triangular in dorsal view; head wider than long (HW 1.12 times HL), head width 30% of SVL; head length 27% of SVL; head narrower than body; snout acuminate, rounded terminally in dorsal view; not bulbous at tip; distance from the nostril to the tip of the snout (3.2 mm) is noticeably less than the distance from the nostril to the eye (5.8 mm), constituting 20% of head length; snout long, protuberant, directed slightly anteroventral as “shark snout” in profile (Fig.
SVL: 58.3; HW: 17.8; HL: 15.9; ED: 4.4; IOD: 9.3; EW: 3.9; EL: 6.4; IND: 4.3; E–N: 5.8; NSD: 3.2; SL: 9.1; FL: 15.4; HNDL: 16.3; FEML: 23.7; TL: 23.4; FOOTL: 22.6.
(Fig.
(Fig.
(Figs
The remaining paratypes show some variation in nocturnal color pattern. The overall dorsal coloration of the juveniles (
Morphometric measurements in mm of the holotype and paratypes of Rhinella lilyrodriguezae sp. n. See text for abbreviations.
Measurements | Holotype | Paratypes | ||||
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Sex | Female | Female | Female | Female | Juvenile | Juvenile |
SVL | 58.3 | 57.5 | 55.5 | 47.1 | 34.9 | 27.8 |
HW | 17.8 | 18.0 | 17.2 | 11.4 | 11.6 | 9.4 |
HL | 15.9 | 15.1 | 14.1 | 8.9 | 9.0 | 9.2 |
ED | 4.4 | 4.3 | 4.3 | 4.2 | 3.7 | 3.2 |
IOD | 9.3 | 8.8 | 8.8 | 8.4 | 5.6 | 4.9 |
EW | 3.9 | 3.9 | 3.9 | 3.7 | 3.4 | 2.8 |
EL | 6.4 | 6.8 | 6.0 | 6.2 | 3.9 | 3.2 |
IND | 4.3 | 4.4 | 4.3 | 4.1 | 2.5 | 2.1 |
E-N | 5.8 | 5.7 | 5.6 | 5.5 | 3.4 | 3.9 |
NSD | 3.2 | 3.2 | 3.0 | 2.9 | 1.7 | 1.3 |
SL | 9.1 | 9.0 | 8.7 | 8.1 | 5.8 | 4.9 |
FL | 15.4 | 15.2 | 14.1 | 13.5 | 9.1 | 7.2 |
HNDL | 16.3 | 16.4 | 15.1 | 13.7 | 8.7 | 6.5 |
FEML | 23.7 | 23.5 | 22.0 | 20.3 | 13.2 | 11.7 |
TL | 23.4 | 23.2 | 21.4 | 19.9 | 13.1 | 11.3 |
FOOTL | 22.6 | 23.3 | 22.0 | 20.9 | 13.5 | 10.8 |
The specific epithet lilyrodriguezae is a noun in the genitive case and a patronym for Dr. Lily Rodriguez, for her contributions to the knowledge of the Peruvian amphibians and her initiatives that have promoted the creation of numerous natural protected areas in Peru, such as the Cordillera Azul National Park.
Rhinella lilyrodriguezae sp. n. is only known from the Shapaja sector within the CAZNP in northern Peru, at elevations between 1245 and 1280 m a.s.l. (Fig.
One gravid female (
An early comprehensive revision of the former genus Rhamphophryne (= Rhinella acrolopha group) based on external morphology and osteology hypothesized multiple evolutionary histories and complex morphological variation within the genus (
Our 16S rRNA analysis supports the monophyly of the Rhinella festae clade of
Finally, the monophyly of the Rhinella festae clade as proposed by
We are grateful to A. Catenazzi and anonymous reviewers for their comments that improved our manuscript J. C. Cusi is sincerely grateful to K. Tasker for funding the fieldwork and for the opportunity to participate in the research within the natural protected area. We thank Tatiana Pequeño and Jorge Martinez at CIMA (Investigación y Manejo de Areas Naturales: Cordillera Azul) for providing access, permits and information on localities in the Cordillera Azul National Park. We are grateful to J. Córdova for granting access at the Museo de Historia Natural, Universidad Nacional Mayor de San Marcos, Lima, Peru (
Specimens examined
Rhinella chavin (9 specimens). PERU: HUÁNUCO, Pachitea, Chaglla, Palma Pampa, 20 km southeast of Chaglla, 3010 m:
Rhinella manu (8 specimens). PERU: CUSCO: Paucartambo, Kosñipata:
Rhinella nesiotes (4 specimens). PERU: HUANUCO, Puerto Inca, Yuyapichis, Comunidad El Sira:
Rhinella yanachaga (4 specimens). PERU: PASCO: Oxapampa, Oxapampa, Yanachaga-Chemillén National Park (PNYCH), W side of the Cordillera Yanachaga near Río San Alberto, 2600 m: