Amazophrynella javierbustamantei, Rojas, Rommel R., Chaparro, Juan C., Carvalho, Vinicius Tadeu De, Avila, Robson W., Farias, Izeni Pires, Hrbek, Tomas & Gordo, Marcelo, 2016

Rojas, Rommel R., Chaparro, Juan C., Carvalho, Vinicius Tadeu De, Avila, Robson W., Farias, Izeni Pires, Hrbek, Tomas & Gordo, Marcelo, 2016, Uncovering the diversity in the Amazophrynellaminuta complex: integrative taxonomy reveals a new species of Amazophrynella (Anura, Bufonidae) from southern Peru, ZooKeys 563, pp. 43-71 : 49-56

Taxon classification Animalia Anura Bufonidae

Amazophrynella javierbustamantei sp. n.

Holotype

(Fig. 4). MHNC 8331 (Genbank 16S rRNA: KR905184). Adult male, collected at Quebrada Guacamayo (12°54'24.5"S; 69°59'32.7"W, 215 m a.s.l.) km 105 of the highway Puerto Maldonado–Cusco City, District Inambari, Province Tambopata, Department Madre de Dios, Peru, on 27 October 2009 by Juan C. Chaparro and Oscar Quispe.

Paratypes

(Fig. 5). Twenty-two specimens (males= 09, females= 13). MHNC 8363, MHNC 8245, MHNC 8238, adult males, MHNC 8316, MHNC 8484, MHNC 8362, MHNC 8354, adult females, collected with the holotype (12°28'25"S, 69°12'36"W, 205 m a.s.l.). MHNC 11001, adult male, MHNC 11002, MHNC 11003, MHNC 11004, adult females collected by E. Aguilar on 17 May 2009, from La Pampa km 107 highway Puerto Maldonado–Cusco City, Department Madre de Dios (12°40'14.14"S, 72°27'30"W, 250 m a.s.l.). MHNSM 17993, adult male collected by A. Angulo in 1999; from Province Manu, locality of Inambari, Department Madre de Dios (13°02'29.28"S, 70°22'46.65"W, 306 m a.s.l.). MHNSM 25651, adult female, collected by D. Rodriguez on April 2007, from Province La Convención, locality of Camana, Department Cusco (12°05'9.25"S, 73°03'2.61"W, 680 m a.s.l.). MHNC 9939, MHNC 9940, adult females, collected by J. Delgado on 17 January 2010 from Province La Convención, locality of Mapi, Department Cusco (11°31'19.17"S, 73°28'29.83"W, 708 m a.s.l.). MHNC 9387, adult male, collected by G. Estrada on 21 January 2010, from locality of Tambo Poyeni near Quebrada Mayapo, Department Junin (11°19'29.9"S, 73°32'16.7"W, 388 m a.s.l.). MHNC 9754, MHNC 9756, adult males, MHNC 9626, MHNC 9679, MHNC 9680, MHNC 9757, adult females, collected by A. Pari on January 2010, from locality of Tsoroja, Department Junin (11°18'56.06"S, 73°32'32.11"W, 399 m a.s.l. and 11°23'14.50"S, 73°29'43.00"W, 450 m a.s.l.).

Diagnosis.

The new species is part of Amazophrynella based on molecular phylogenetic relationships (Fig. 1) and morphological synapomorphies ( Fouquet et al. 2012a).

Amazophrynella javierbustamantei sp. n. is characterized by: (1) skin on dorsum tuberculate, with many subconical tubercles disperse on arms, legs, head and body; ventral skin coarsely areolate, throat and chest aerolate; (2) tympanic membrane and tympanic annulus not apparent through the skin; (3) snout long, subacuminated, protruding in lateral views; (4) upper eyelid with smaller tubercles, cranial crests absent; (5) dentigerous process of vomers absent; (6) vocal sac, vocal slits and nuptial pads absent; (7) finger I shorter than finger II, tips of digits rounded; (8) fingers lacking lateral fringes; (9) ulnar tubercles present; (10) heel bearing eight or more small low tubercles, tarsus with small tubercles and lack of folds; (11) plantar surfaces of feet bearing one metatarsal tubercle, the inner 2.5x larger than the outer, outer subconical; supernumerary plantar tubercles round and low; (12) toes lacking lateral fringes; webbing basal; toe III equal than toe V, tips of digits rounded; (13) dorsally is dark brown to light brown, and gray to black in some, ventrally, cream with yellow to orange marks, with black to dark brown spots; (14) SVL 16.39-22.25 mm in females, 12.79-16.42mm in males; (15) hand length is the greatest of all species of Amazophrynella : 3.6 mm in males (n= 26) and 4.6 mm in females (n=20), see Fig. 3; (16) thirteen molecular autapomorphies in the 16S rDNA gene.

Comparison with other species.

Amazophrynella javierbustamantei sp. n. (Figs 4, 5, 6) differs in the following character states (states of other species in parentheses). From Amazophrynella minuta (Fig. 6A) by having body skin texture tuberculate (roughly granular); relative abundance of spiny granules on the forelimbs (prickly warty skin on axillary region of the forelimbs); absence of large warts on dorsum (presence of large warts); throat and chest cream-grayish (light brown); posterior side of belly color pale orange yellowish with tiny rounded black or dark brown spots (throat and the whole belly intensely orange yellowish); tiny rounded black spots covering the belly (irregular black ocelli or blotches); metatarsal tubercle rounded (oval). From Amazophrynella bokermanni (Fig. 6B) relative size of fingers, with finger I shorter than II (I>II); snout vent length smaller in males (15.8 mm) and females (22.25 mm) ( Amazophrynella bokermanni with maximum 22 mm SVL in males and 28 mm SVL in females, see Izecksohn 1993); smaller snout in males, with 2.2 mm SL, n = 26 (2.7 mm SL, n = 5; see Table 1); posterior side of belly color pale orange yellowish with tiny rounded black or dark brown spots (white coloration with small black dots). From to Amazophrynella vote (Fig. 6C) snout subacuminated in dorsal view (rounded); posterior side of belly color pale orange yellowish with tiny rounded black or dark brown spots (ventral color pattern reddish brown, with presence of small white dots). From Amazophrynella manaos (Fig. 6D) snout subacuminated (snout truncate); dorsal skin finely granular (dorsal surfaces granular); throat and chest grayish (dark coloration); posterior side of belly color pale orange yellowish with tiny rounded black spots (venter cream with black spots or stripes). From to Amazophrynella matses (Fig. 6E) snout subacuminated (snout slightly truncate), edges of nasal protrusion not dilated (dilated in ventral view); shape of palmar tubercle rounded (palmar tubercles elliptical); finger tips unexpanded (expanded), rounded tiny black spots covering the belly (medium-sized black ocelli or streaks); coloration of the belly pale yellow (belly completely yellow). From Amazophrynella amazonicola (Fig. 6F) by the absence of small triangular protrusion on the tip of the snout in both dorsal and ventral views (presence); body surface granular (finely granular), dorsum uncovered with medium-sized granules scattered irregularly (covered with medium-sized granules scattered irregularly); posterior side of belly color pale orange yellowish with tiny rounded black or dark brown spots (orange yellowish with dark red and brown blotches).

Description of the holotype.

Body slender, head triangular, slightly longer than wide; head length 35.5% of SVL, head width 30.9% of SVL. Snout long, subacuminate in dorsal view, protruding in lateral view; canthus rostralis straight and loreal region vertical; without papilla; snout length 39.0% of head length; tympanic membrane and tympanic annulus not apparent through the skin, skin of the tympanic area covered by round sub-conical warts; vocal sac externally not visible, vocal slits absent; eyes prominent 23.8% of head length; upper eyelid covered with small tubercles; those close to the external margin aligned in a more or less distinct row; nostril closer to snout than to eyes; internarial distance smaller than eye diameter; presence of a line of small spiny granules from the outer edge of the mouth to upper arm, choanas small and circular.

Dorsal skin finely tuberculate with several large tubercles scattered sub-conical tubercles on upper arm; texture of ventral skin granular, covered by rounded granules. Dorsolateral surfaces, granular, with presence of large rounded tubercles. Forelimbs slender, upper arm length 29.6% of SVL; edges of lower arm and upper arm finely tuberculate with several large sub-conical and spiny granules; hand length 76.5% of upper arm length; fingers slender, tips not expanded; relative length of fingers I<II<IV<III; supernumerary tubercles and accessory palmar tubercles present, palmar large and rounded, supernumerary tubercles low, small rounded; subarticular tubercles rounded and small, one tubercle on fingers I, II and IV and two on finger III; fingers I and II basally webbed; indistinct nuptial pad. Hind limbs slender; ventral skin from thigh to tarsus covered by spiny tubercles, foot length 66% of thigh length; relative length of toes I<II<V<III<IV: inner metatarsal tubercle oval, 2.5 × larger than outer; outer metatarsal tubercles small, rounded; subarticular tubercles present, rounded, present one on fingers I, II, and two on fingers III, V and three on finger IV; and tip of toes not expanded.

Measurements of the holotype

(in millimeters). SVL 15.1; HW 4.6; HL 5.3; SL 2.1; ED 1.2; IND 1.0; UAL 4.4; HAL 3.4; THL 8.1; TAL 8.1; TL 4.5; FL 5.3.

Coloration of the holotype.

In life: dorsum of the holotype mostly light brown with dark brown in the dorsum; dorsolaterally creamish-brown with scattered black blotches; dorsal surfaces of hands and feet creamish-brown, and gray on arms and legs; belly creamish-gray with black dots, and the throat gray; fingers, toes and plantar surfaces reddish-black; groin with orange marks; iris with a bronze ring; cloaca with orange flap, black pupil and bronze iris. In alcohol: dorsum brownish-grey; venter cream with black and brown dots; orange surfaces turned cream, with a white longitudinal stripe on upper jaw extending from nostril to forearm.

Variation.

The new species is phenotypically variable. In some individuals (e.g. MHNC 8245 and MHNC 11002, see Fig. 5) patterning on the dorsum varies, with these specimens presenting brown chevrons extending from the head to the vent. Some individuals showed a white line extending from the tip of the nose to the upper arm. Another specimen (MHNC 9739, see Fig. 5) presented a yellow pale coloration in the axillary region (in ventral view). In some individuals, the coloration of the throat extended onto the chest (e.g. MHNC 11002, MHNC 9739 and MHNC 8245, see Fig. 5). The pale yellow coloration of the belly surface may extend from thighs to the chest or just to the middle of the belly (e.g. MHNC 8362, see Fig. 5 and Fig. 7B). In some individuals, the thighs are abundantly covered by rounded tiny spots extending to the shank (Fig. 7B). In preserved specimens the dorsum becomes light brown and the belly coloration vary from white to yellow pale (e.g. MHNSM 31255 and MHNSM 17993, see Fig. 5). The color of the finger becomes pale red and in other individuals the red coloration of the fingers became brown or orange (Fig. 5).

Bioacoustics.

The following values are presented as: min-max (average ± SD, number of notes). The call is a trill type call issued during continuous and regular intervals (Fig. 8). Each note had a duration of between 0.03 to 0.08 seconds (0.05 ± 0.01 seconds, n = 20). The number of pulses varied between 8 to 18 pulses per note (10.4 ± 2.6 pulses/note, n = 20). The silence between notes varied from 0.4 to 1.6 seconds (0.8 ± 0.3 seconds, n = 20). The dominant frequency varied from 3962.1 to 3789.8 kHz (3927.6 ± 70.7 kHz, n = 20), and coincides with the fundamental frequency. Time to peak amplitude was around 0.014 to 0.04 seconds (0.02 ± 0.01 seconds, n = 20).

Distribution, ecology and conservation.

Amazophrynella javierbustamantei sp. n. is known from the Department of Cusco, in the lower Urubamba river basin and Department of Madre de Dios (Inambari, Candamo and Nueva Arequipa) in Peru (Fig. 9). Its distribution can vary from 215 m a.s.l. to 708 m a.s.l. Additional specimens were recorded at Los Amigos Biological Station, Tapir Lodge, and Explorers Inn, in Tambopata National Reserve. Individuals were active during the day, jumping on leaf litter, at night they were sleeping on leaves around 30 cm above ground. This species breeds close to the edges of permanent oxbow lakes, males call during the day while perched above streams in tangles ( Cocroft et al. 2001). Three of the localities, km 105, 107 and 117 of the highway Puerto Maldonado–Cusco, Department Madre de Dios, show evidence of serious environmental impacts due to illegal gold mining activities, with forest and soil removed, and environmental pollution via organic and inorganic chemicals and heavy metal (specially mercury) poisoning. In addition, the new species is distributed inside of territories where oil companies are operating. On the other hand, the species is present in two protected areas, the Tambopata Natural Reserve and Machiguenga Communal Reserve. The conservation status of this species remains unknow, but was listed in 2008 as Least Concern on the IUCN red list (2015), because it was confused with Amazophrynella minuta , and because Amazophrynella minuta s.l. had wide distribution at that time, apparent tolerance of a certain degree of habitat modification, presumed large population, and because it is unlikely to be declining, and thus did not qualify for listing in a more threatened category. With recent studies the genus, the species complex of Amazophrynella minuta , was split in five species, three of them are now formally described for Peru ( Amazophrynella matses , Amazophrynella amazonicola and Amazophrynella javierbustamantei sp. n.). The recognition of these new species will require the reevaluation of the conservation status of these species. It should also act as an impetus for additional field and laboratory studies of Peruvian amphibians, in order to understand the real conservation status of this fauna.

Etymology.

The species is named after Dr. Javier Bustamante, a Peruvian residing in United States, to whom we dedicate this species in recognition of his friendship and support of herpetological taxonomy and systematics research and amphibian conservation in Peru.