Pristimantis guianensis, Mônico & Ferrão & Chaparro & Fouquet & Lima, 2022

Monico, Alexander Tamanini, Ferrao, Miqueias, Chaparro, Juan Carlos, Fouquet, Antoine & Lima, Albertina Pimentel, 2022, A new species of rain frog (Anura: Strabomantidae: Pristimantis) from the Guiana Shield and amended diagnosis of P. ockendeni (Boulenger, 1912), Vertebrate Zoology 72, pp. 1035-1065 : 1035

Pristimantis guianensis sp. nov.

Figs 7 , 8 , 9 , 10 , 11 , 15 and 16

Eleutherodactylus ockendeni - Zimmerman and Rodrigues (1990), Heyer and Hardy (1991), Lima et al. (2006), Menin et al. (2007)

Pristimantis ockendeni - Rojas-Ahumada and Menin (2010); Lima et al. (2012), Fouquet et al. (2013)

Pristimantis cf. ockendeni - Vacher et al. (2020), Azevedo et al. (2021), Fouquet et al. (2022b)

Holotype.

INPA-H 43918 (field number APL22783), adult male, collected at Reserva Florestal Adolpho Ducke, municipality of Manaus, state of Amazonas, Brazil (2°55′52.0″S; 59°58′25.8″W, 74 m elevation), on 19 November 2019 by A.T. Mônico and I.Y. Fernandes.

Paratopotypes.

Seventeen adult specimens (fourteen males and three females), same locality as the holotype: one male INPA-H 43917 (field number APL22782) collected in November 2019; nine males INPA-H 43919, - INPA-H 439120, INPA-H 439121, INPA-H 43923, - INPA-H 43924, INPA-H 43925, INPA-H 44248, INPA-H 44249, and INPA-H 44250 (field numbers APL22784-86, 22794-99, respectively) and one female INPA-H 43922 (field number APL22793) collected in December 2019 by A.T. Mônico, I.Y. Fernandes, E.D. Koch and A. P. Lima; two males MPEG 44181 and MPEG 44182 (field numbers APL22804 and 22805, respectively) collected January 2020 by A.T. Mônico and I.Y. Fernandes; two males INPA-H 43942 and INPA-H 43944 (field numbers APL23187 and 23189, respectively) and two females INPA-H 43941 and INPA-H 43943 (field numbers APL23187 and 23189, respectively) collected in 28-29 December 2020 by A.T. Mônico, U.F. Souza and I.Y. Fernandes.

Paratypes.

Thirty-eight adult specimens (28 males and 10 females). BRAZIL: AMAZONAS: municipality of Presidente Figueiredo: Km 144 of the BR-174 Highway [six males MPEG 44183, MPEG 44184, INPA-H 43926, INPA-H 44251, INPA-H 43927 and INPA-H 43928 (field numbers APL22806-11, respectively) collected in January 2020 by A.T. Mônico and E.D. Koch], Suçuarana’s Fall at Vila de Balbina [seven males INPA-H 43929, INPA-H 43930, INPA-H 43931, INPA-H 43932, INPA-H 43933, MPEG 44185, MPEG 44186 (field numbers APL22812-18, respectively) and one female MPEG 44187 (field number APL22819) collected in January 2020 by A.T. Mônico and E.D. Koch]; RORAIMA: municipality of São João da Baliza [seven males INPA-H 43935, MPEG 44188, MPEG 44190, INPA-H 43937, INPA-H 43938, INPA-H 43939 and INPA-H 439340 (field numbers APL 22821, 22823, 22825 to 22829, respectively) and three females INPA-H 43934, INPA-H 43936 and MPEG 44189 (field numbers APL22820, 22822, and 22824, respectively) collected in June 2020 by A.T. Mônico, I.Y. Fernandes, M. Ferrão and A. P. Lima]. SURINAME: Sipaliwini [three males MNHN-RA-2020.0115-0117 (field numbers AF2255, 2256 and 2289, respectively) and two females MNHN-RA-2020.0113 and 2020.0114 (field numbers AF2155 and 2258, respectively) collected in April 2014 by A. Fouquet and J. P. Vacher]. GUYANA: Mabura Hill Forest Reserve [four males SMNS11987, 11989, 11993, 11994 and three females SMNS 11990, 11991 and 11995 collected between April 2003 and June 2004; and one male MTD 47769 and one female MTD 47770 collected in August 2010 by. R. Ernst].

Referred material.

Available in Appendix 1.

Diagnosis.

The new species is characterized by the following unique combination of characters: (1) dorsal skin shagreened, frequently with distinctly tubercles, with or without W-shaped on scapular region; (2) tympanum visible, tympanic membrane poorly differentiated, tympanum diameter 24-34% of eye diameter and annulus partially visible externally; supratympanic black band; (3) snout subacuminate to sub-rounded in dorsal view and rounded in in lateral profile, loreal region concave; (4) upper eyelid tubercles present; dark bar between the eyes, and three oblique black streaks below the eye; cranial crests absent; (5) nostril ovoid, slightly protuberant, directed laterally; interorbital distance 29-37% of head width; (6) tongue cordiform to ovoid; (7) absence of dentigerous processes of vomers (Fig. 7 ); (8) males with vocal slits, vocal sac median to subgular; nuptial pads absent; (9) Finger I slightly more shorter than II; finger discs ovoid to expanded (finger disc I less expanded compared to finger disc II, III and IV); (10) fingers without lateral fringes; (11) three to five enlarged ulnar tubercles, barely visible in fixed specimens (easily visible in individuals when in life); (12) tibia length 48-56% of SVL; (13) heel tubercle absent; tarsal tubercles aligned, ovoid to elliptical; tarsal fold short, barely visible; (14) thenar tubercle ovoid to elliptical; small palmar tubercle ill-defined, less than 30% of thenar tubercle; (15) toes without lateral fringes; rudiment of web absent; (16) toes I and II almost of the same size; toes discs ovoid (toe disc I and II, especially) to expanded (III, IV and V); (17) belly skin smooth to areolate, and ventral region of the femur externally areolate; (18) in life, translucent groin with small, scattered dark melanophores and absence of bright colored blotches or marks; (19) in life, iris is dichromatic, brown lower part and cream upper part; (20) SVL in adult males of 16.2-20.7 mm (n = 43) and in females of 21.4-25.7 mm (n = 13); and (21) advertisement call with average call duration of 232 ± 42 ms, inter-note interval of 44 ± 5 ms (68 ± 11 ms), minimum frequency of 2,827-3,695 Hz, maximum frequency of 4,333-6,688 Hz and dominant frequency of 3,466-4,521 Hz.

Comparisons with other species.

The new species is compared to other Pristimantis of the P. unistrigatus species group occurring in the Guiana Shield: P. abakapa Rojas-Runjaic, Salerno, Señaris & Pauly, 2013; P. aureoventris Kok, Means & Bossuyt, 2011; P. crepitaculus Fouquet, Peloso, Jairam, Lima, Mônico, Ernst & Kok, 2022; P. espedeus Fouquet, Martinez, Courtois, Dewynter, Pineau, Gaucher, Blanc, Marty & Kok, 2013; P. grandoculis (van Lidth de Jeude, 1904); P. guaiquinimensis ( Schlüter & Rödder, 2007); P. imthurni Kok, 2013; P. inguinalis (Parker, 1940); P. jamescameroni Kok, 2013; P. jester Means & Savage, 2007; P. marmoratus (Boulenger, 1900), P. memorans (Myers & Donnelly, 1997), P. pulvinatus (Rivero, 1968); P. saltissimus Means & Savage, 2007; and P. sarisarinama Barrio-Amorós & Brewer-Carias, 2008 and to P. ockendeni , which occurs in southwestern Amazonia. Diagnostic characters of compared species are shown in parentheses unless stated otherwise.

Pristimantis guianensis sp. nov. differs from P. ockendeni by the absence of dentigerous processes of vomers (present) and smaller SVL: male 16.2-20.7 mm (n = 43 specimens) in P. guianensis sp. nov. (18.4-22.8 mm, n = 09 in P. ockendeni ) and female 21.4-25.7 mm (n = 13 specimens) in P. guianensis sp. nov. (30.4-30.6 mm, n = 02 in P. ockendeni ). Additionally, the advertisement call of P. guianensis sp. nov. has average call duration of 232 ± 42 ms (540 ± 150 ms), inter-note interval of 44 ± 5 ms (68 ± 11 ms) and is emitted at minimum frequency of 2,827-3,695 Hz (2,047-2,610 Hz), maximum frequency of 4,333-6,688 Hz (2,965-3,504 Hz) and dominant frequency of 3,466-4,521 Hz (2,519-3,144 Hz).

The absence of dentigerous processes of vomers easily distinguishes Pristimantis guianensis sp. nov. from P. aureoventris , P. crepitaculus , P. espedeus , P. grandoculis , P. imthurni , P. jamescameroni , P. jester , P. marmoratus and P. saltissimus (present in all species); and the presence of vocal slits in males (absent in P. abakapa , P. aureoventris , P. grandoculis , P. imthurni , P. guaiquinimensis , P. jamescameroni , P. jester and P. saltissimus ).

Pristimantis guianensis sp. nov. differs from P. espedeus , P. guaiquinimensis , P. imthurni , P. jamescameroni and P. pulvinatus by having smaller male SVL of 16.2--20.7 mm (SVL 20.7-24.8 mm in P. espedeus ; 33.4--34.7 mm in P. guaiquinimensis ; 22.9 mm in P. imthurni ; 22.9 mm in P. jamescameroni ; 23.0-26.1 mm in P. pulvinatus ; 22.6-25.8 mm in P. sarisarinama ); from P. espedeus , P. jamescameroni , P. guaiquinimensis and P. memorans by smaller female SVL of 21.4-25.7 mm (SVL 29.4 mm in P. espedeus ; 26.3-27.5 mm in P. jamescameroni ; 32.4-33.6 mm in P. guaiquinimensis ; 31-32 mm in P. memorans ); by having a translucent groin with small, scattered dark melanophores and absence of bright colored blotches or marks in life, P. guianensis sp. nov. can be distinguished from P. abakapa (chocolate brown groin with small white dots), P. aureoventris (black or brown, sometimes with some small golden spots), P. crepitaculus (dark grey groin), P. espedeus (reddish orange groin), P. grandoculis (dark grey groin), P. imthurni (brown groin), P. inguinalis (bright yellow spots on groin), P. jamescameroni (bright orange groin) and P. marmoratus (yellow or pale green wash on groin).

Description of holotype.

INPA-H 43918 (field number APL 22783), an adult male (Fig. 8 ), SVL 17.7 mm; head slightly longer than wide (HL 105% of HW); head width 36.0% of SVL; head length 37.9% of SVL; cranial crest absent; snout subacuminated in dorsal (Fig. 8A ) and rounded in lateral (Fig. 8C ) views; snout short, END 77.3% of ED; nostril ovoid, slightly protuberant, directed laterally; internarial region slightly concave; canthus rostralis almost straight in dorsal view, rounded in profile; loreal region concave; lips rounded; small tubercles on upper eyelid; interorbital region straight, IOD 32.2% of HW; large eye (ED/TD = 3.4); supratympanic fold distinct, extending from posterior margin of eyelid angling posteroventrally behind tympanic annulus; tympanum visible and rounded, 28.5% of ED; tympanic membrane poorly prominent, directed laterally; tympanic annulus distinct, obscured anteriorly, dorsally, and posteriorly by the supratympanic fold; two small postrictal tubercles, poorly visible; choanae of moderate sized, round, not concealed by palatal sheath of maxilla; dentigerous processes of vomers absent; tongue cordiform, longer than wide; vocal slits present; vocal sac median, simple and subgular.

Forearm slightly longer than hand (FAL 102.6% of HAND); four ulnar tubercles enlarged and aligned, poorly defined; size Finger I<II<IV<III (Fig. 8D ); finger discs rounded (Finger I and II) to expanded (Finger III and IV); thenar tubercle ovoid; palmar tubercle bifid and poorly defined, almost twice the width of elongate thenar tubercle; subarticular tubercles well defined, most prominent on fingers I and IV, rounded in dorsal and lateral view; all fingers with ventral pads well defined by circumferential grooves.

Hindlimbs slender, with transversal bars complementary, being four on thigh, three on tibia and two on tarsus; tibia length 54% of SVL; heel without tubercles; tarsus with row of small, low tubercles, poorly visible; tarsal folds absent; foot length 41% of SVL; subarticular tubercles well defined (except in Toe IV), round in dorsal and lateral view; toes lacking lateral fringes and webbed; toes lengths I <II <III <V <IV (Fig. 8E ); Toe I only slightly shorter than II; Toe V slightly longer than Toe III (Toe III extending to border of toe discs V); discs well defined by circumferential grooves; toe discs ovoid (toes I, II and III) to expanded (toes IV and V); thenar tubercle ovoid and palmar tubercle small and ill-defined; all toes with ventral pads well defined, most prominent in toes IV and V.

Dorsal skin shagreened, with a W-shaped mark on scapular region, a dark blotch between the eyes and other ill-defined near the groin (Fig. 8A ); upper eyelid shagreened, with several distinctly enlarged tubercles on each eyelid; upper and posterior surfaces of hindlimbs smooth; skin on flanks and chest smooth; slightly areolate on belly; and ventral region of the femur externally areolate; dorsolateral folds absent; pectoral and discoidal folds not visible; cloaca not protuberant, cloacal region without tubercles.

Morphometric measurements are presented in Table 4 .

Coloration of holotype in preservative: After 30 months in 70% ethanol, colours of the holotype faded, notably glandular supracarpal pad (Fig. 8 ). The snout is dorsally dark brown and well-delimited by a transversal light gray interorbital band extending on half of the eyelids; dark bands below the eyes; supratympanic black band; black blotches in the tympanic region (Fig. 8C ); a dark brown mark is well-defined on the scapular region posteriorly followed by two oblique traversal dark brown bands. Similar, transversal dark brown bands are present on the arms, hands and legs and feet. Arms, belly and ventral surfaces of thighs, tibia and tarsus are cream with brown melanophores, arm and belly skin are the lightest, with melanophores sparsest, while ventral surfaces of hindlimbs have a greater amount of melanophores (Fig. 8B ); upper arm without bands (only few melanophores) and lower arm with one dark transversal band; black blotch in the cloaca region; more colors details are shown in Fig. 8 .

Coloration of holotype in life: In life, yellow dorsal surface with orange blotches and tubercles all over the dorsal surface; three horizontal dark bands on dorsum, the anterior one on interorbital region, the medial one bordering the W-shaped mark on the scapular region, and posterior one ill-defined near the groin (Fig. 10A ). Black blotches located in the tympanic region and dark bands below the eyes. Upper surface of legs and arms with light brown transversal band. Ventral surface is cream covered with minute gray melanophores, while the throat region is slightly yellow. The arm and belly skin are the lightest and the melanophores are the sparsest. Ventral surfaces of thighs, tibia and tarsus are gray, due to the high amount of melanophores. During the day the coloration turns darker. The iris is dichromatic: lower part brown and upper part golden color with dark brown reticulation. More colors details are shown in Fig. 8A .

I ntraspecific variation.

Pristimantis guianensis sp. nov. is small, SVL in adult males of 16.2-20.7 mm (n = 43; Table 4 ) and in females of 21.4-25.7 mm (n = 13; Table 4 ). Dorsal pattern of males is variable: most specimens of the type series (73% of type specimens) show irregular dark markings present on dorsum, similar to holotype (Fig. 9A ), slightly dark middorsal and other ill-defined near the groin (12% of types; Fig. 9B ), with snout-vent stripe (only one specimen; Fig. 9C ), dorsal view coloration strongly delimited and different from sideview (13% of types; Fig. 9D , 9E and 9F ), lighter (Fig. 9D ) or darker (Fig. 9F ), with or without dark transverse stripes on dorsum (Fig. 8E ). Specimens of dorsal pattern with irregular dark markings (Fig. 9A and 9C ) and slightly dark middorsal (Fig. 9B ) has interorbital region is covered by a dark band (85.7% of specimens). In addition, these two patterns are closely linked to the presence of a W-shaped on scapular region (66% of types), even if not very evident (16% of types). Dorsal skin texture is fairly variable too (probably depending on activity, as observed in many species of the genus (e.g., Guayasamin et al. 2015; Kok et al. 2018), from shagreened to granular, with or without tubercles.

Dark bands and blotches located below the eyes are present in the most specimens of type series, not very evident or absent (91%, 4%, and 5% of types, respectively). Tr ansversal dark brown bands are present on the arms and hands, intermediary, or absent (89%, 4%, and 7% of types, respectively). In legs and feet, transversal dark bands are present in most specimens of the type series, intermediary, or absent (80%, 9%, and 11% of types, respectively). The three to five ulnar tu bercles are small in 9% of types. Supratympanic black band in all species.

Ventral surface can have different shades, varying according to the concentration of melanophores: light (Fig. 9G ), intermediary (Fig. 9H ) or dark (Fig. 9I ). Most type specimens (57%) had the intermediate concentration, while light and dark were less frequent (29% and 14% of types).

Dorsal background coloration in life is widely variable, from gr eenish yellow (Fig. 10A , 10B and 10F ) and light orange (Fig. 10C , 10F and 10G ) to brown (Fig. 10E , 10H , 10I ), while females do not vary as much. Snout-vent stripe can be present (Fig. 9G ). The iris is dichromatic: brown to dark copper metallic lower part and cream to silver upper part with dark brown reticulation (Fig. 10 ). When active, the ventral surface is with cream background, cover by white granules and brown melanophores (nocturnal, Fig. 11A ), and gray at the day, which is the result of melanophores expansion (diurnal, Fig. 11B ).

Advertisement call.

We recorded 111 calls of 34 males from 2 m above the ground underwood vegetation at temperatures between 23-25°C and 82-99% relative air humidity. The list of call recordings is disponible in Appendix 4, and descriptive statistics of call parameters are in Table 5 . The advertisement call of Pristimantis guianensis sp. nov. is short (mean 232 ms ± standard deviation 42, range 158-371 ms) and composed of 4-6 notes (4.6 ± 0.7). Calls with four notes were the most common arrangement, corresponding to 51% of all analysed calls (n = 57 calls), followed by calls with five notes (38%; n = 42 calls) and six notes (11%; n = 12 calls). Note duration averaged 16 ± 5 ms (5-30 ms; n = 510 notes). Inter-note intervals are homogeneous within calls (average = 44 ± 5 ms; range 14-56 ms; n = 399 inter-note intervals), i.e., inter-note the same temporal structure. Notes lack true harmonics. The average minimum frequency is 3,210 ± 152 Hz (2,827-3,695 Hz), while average maximum frequency is 5,264 ± 639 Hz (4,333-6,688 Hz), and the dominant frequency is 3,970 ± 179 Hz (3,466-4,521 Hz) (n = 111 calls) (Fig. 12A and 12B , Table 5 ).

We provide additional acoustic variables in Appendix 5 .

Courtship call repertoire.

We observed that when a female is present, males calls consist of 7-8 notes (7.75 ± 0.5; n = 4 calls of two males), with call duration of 359 ± 31 ms (321-391 ms), note duration of 13 ± 2 ms (10-14 ms) with inter-note intervals of 39 ± 1 ms (38-40 ms). The average minimum frequency is 3,097 ± 55 Hz (3,025-3,158 Hz), while average maximum frequency is 5,080 ± 678 Hz (4,134-5,861 Hz), and the dominant frequency is 3,914 ± 191 Hz (3,790-4,199 Hz).

We recorded the vocal repertoire of a male (INPA-H 43944, FNJV 58762) until the amplexus with a female (INPA-H 43943) for ~80 s (Fig. 13 ). The male jumped and positioned close to the female (approximately 20 cm) and then emitted one call with 8 notes (Fig. 13A ), two-harmonic structure and more high-pitched than advertisements calls (maximum frequency reaching 8,182 Hz). The call duration was 386 s, where notes lasted 15 ± 1 ms (13-17 ms) with inter-note intervals of 38 ± 2 ms (35-41 ms). The call was emitted with a dominant frequency of 3,790 ± 172 Hz, minimum frequency of 3,231 ± 168 Hz and maximum frequency of 5,489 ± 602 Hz. The male came even closer to female and, after 47 s emitting the previous call, emitted a second call (Fig. 13B ), less high-pitched and longer than the previous one (Fig. 9A ): 11 notes, call duration 757 ms, note duration 20 ± 5 ms (17-33 ms), inter-note intervals 54 ± 19 ms (28-85 ms), minimum, maximum and dominant frequency of 3,144 ± 111 Hz, 4,780 ± 158 Hz and 3,704 ± 151 Hz, respectively. Finally, the male emitted two calls with inter-call interval of 860 ms between them (Fig. 13C ): the former call has call duration of 761 ms and consisted of 13 notes with note duration of 19 ± 3 ms (14-22 ms), inter-note interval of 45 ± 16 ms (31-87 ms) s, and frequencies were similar to previous call (minimum 3,032 ± 104 Hz, maximum 4,845 ± 245 Hz and dominant of 3,876 ± 213 Hz); while the latter call lasted 547 ms and was composed of 9 notes, with note duration of 18 ± 3 ms (15-25 ms), inter-note interval of 39 ± 4 ms (32-45), except for last note (107 ms), dominant frequency was 3,962 ± 219 Hz, minimum frequency 2,995 ± 76 Hz and maximum frequency 4,705 ± 214 Hz. The female then bent down and formed the amplexus with the male.

Etymology.

The specific epithet " guianensis " refers to the region of occurrence of the new species, widely distributed in the western Guiana region (the lowlands of the eastern part of the Guiana Shield).

Distribution, Natural history and Conservation.

Pristimantis guianensis sp. nov. is likely endemic to the western portion of the Guiana region (Fig. 14 ) which is bounded by the Amazon and the Negro Rivers on the South, the Branco River on the southwest. Similarly distributed species are Allobates sumtuosus (Morales, 2002) ( Simões et al. 2013), Synapturanus mirandaribeiroi Nelson & Lescure, 1975 ( Fouquet et al. 2021a), Amazophrynella manaos Rojas, Carvalho, Ávila, Farias & Hrbek, 2014, and Anomaloglossus stepheni (Martins, 1989) ( Vacher et al. 2017). In Brazil, it is known from Manaus and Presidente Figueiredo (Amazonas), São João da Baliza (Roraima) and Oriximiná ( Pará); in Guyana from the Mabura Hill Reserve, and in Suriname from Sipaliwini.

Pristimantis guianensis sp. nov. is crepuscular/nocturnal and inhabits the understory of unflooded (terra firme) forests. Its breeding activity takes place in the rainy season, between November and February in the state of Amazonas, and between May and August in the state of Roraima, both in Brazil. Calling males of P. guianensis sp. nov. aggregate in groups of about eight, separated by 2-3 m distance. At Reserva Ducke (Manaus, Brazil), understory areas rich in Marantaceae are preferentially used (Fig. 15A ). In other localities (e.g., São João da Baliza, Brazil), the species is found mainly on the edge of forest fragments. The new species occurs in sympatry with congeneric P. koehleri Padial & De la Riva, 2009 and P. zimmermanae (Heyer & Hardy, 1991) in Manaus and Presidente Figueredo, P. zeuctotylus (Lynch & Hoogmoed, 1977) in São João da Baliza, P. chiastonotus (Lynch & Hoogmoed, 1977), P. zeuctotylus and P. inguinalis in Sipaliwini ( Fouquet et al. 2015).

Males call perched on vegetation (Fig. 15B ) a bout 2 m above the ground. They start calling at dusk (~18:00h) and remain active until ~19:00h, their activity then remains sporadical until around midnight. When the weather is rainy, the activity is sustained throughout the night. The amplexus (n = 2) is axillary (Fig. 15C ). Juveniles and females are rarely found. We registered three clutches in situ in São João da Baliza (Roraima, Brazil), all within dead leaves hung in the vegetation above the ground. Two clutches were laid in the same nest (Fig. 14D ) about 1 m above the ground, one was recent and the other in a more advanced developmental stage; the third clutch (about 40 cm above the ground) contained dead eggs, apparently due to fungal infestation. The first two clutches contained 11 and 13 large eggs. In the surroundings of the nests (up to 5 m), we observed adult males (n= 6 individuals) and females (n = 3).

On 29 December 2020 we found an amplecting pair (Fig. 14C ) from Reserva Florestal Adolpho Ducke (Tinga basin), Manaus municipality. Male (INPA-H 43944) and female (INPA-H 43943) remained amplected overnight in the same collection container. Oviposition of 13 eggs took place at ~9:30 on the next day. In laboratory, we observed egg development until hatching (Fig. 16 ) in a sterile and moist container (at room temperature, between 24.8 and 28.3°C). Egg diameter increased from ~4.1 mm at oviposition to 5.8 mm before froglet hatching. The first froglet has hatched on the 25th day, and it lasted seven days (33rd day) to all the remnant froglets hatch. The SVL of froglets at hatching was ~4.3 mm.

As previously mentioned, Pristimantis guianensis sp. nov. has a wide geographic distribution, from Manaus, Brazil in the South to Mabura Hill Reserve, Guyana in the North (i.e., 800 km linear distance), and potentially 256 thousand km². The species inhabits primary and secondary forests, as well as edge areas. Furthermore, their populations are locally abundant. Therefore, we believe that the species fits into the category "Least Concern" of the International Union for Conservation of Nature (IUCN).

Figures

Figure 7. Pristimantis guianensis sp. nov. volumetric renderings of μ-CT scans of the skull in ventral views. A Male (Paratype, MNHN-RA- 2020.0115) and B Female (Paratype, MNHN-RA- 2020.0114). Absence of dentigerous processes of vomers can be noted in the doted red circle.

Figure 8. Preserved holotype of Pristimantis guianensis sp. nov. (INPA-H 43918) from Reserva Florestal Adolpho Ducke, Manaus municipality, Amazonas state, Brazil. A dorsal and B views of body; C lateral view of head; and ventral view of D hand and E foot. Photographs: A. T. Monico.

Figure 9. Preserved Pristimantis guianensis sp. nov. dorsal and ventral surface pattern variation. A MPEG 44181; B INPA-H 43929; C INPA-H 43928; D INPA-H 43939; E MPEG 44185; F INPA-H 43940; G MPEG 44181; H INPA-H 43929; and I INPA-H 43940. Photographs: A. T. Monico.

Figure 10. Males of Pristimantis guianensis sp. nov. in life. A Holotype INPA-H 43918 (SVL 17.7 mm); B Paratype MPEG 44188 (SVL 18.4 mm); C Paratopotype INPA-H 44250 (SVL 17.6 mm); D Paratype INPA-H 43939 (SVL 17.6 mm); E Paratype MPEG 44185 (SVL 17.9 mm); F Paratopotype MPEG 44181 (SVL 17.8 mm); G Paratype INPA-H 43928 (SVL 17.9 mm); H Paratype INPA-H 43931 (SVL 17.7 mm); and I Paratype INPA-H 43926 (SVL 16.9 mm). Photographs: A. T. Monico, except (C) E. D. Koch.

Figure 11. Ventral coloration of Pristimantis guianensis sp. nov. in life. A Paratopotype INPA-H 43944 (nocturnal); and B Paratype MNHN-RA- 2020.0115 (diurnal). Photographs: (A) A. T. Monico and (B) A. Fouquet.

Figure 12. Pristimantis guianensis sp. nov. advertisement calls spectrograms and oscillograms with five and six notes, respectively. A Holotype INPA-H 43918 (FNJV 58730); B Paratopotype INPA-H 43919 (FNJV 58731), air temperature 23.2 ° C.

Figure 13. Pristimantis guianensis sp. nov. courtship call repertoire spectrograms for 80 seconds before the amplexus. Male INPA-H 43944, FNJV 58762; air temperature 24.8 ° C and humidity 97 %. A First call after approaching the female; B second call, issued after 47 s of previous call; C last two calls before forming the amplexus.

Figure 14. Geographic distribution of individuals of Pristimantis guianensis sp. nov. (red circles) and Pristimantis ockendeni sensu stricto (yellow triangles) used in our analyses. Stars indicate type localities of each species.

Figure 15. Pristimantis guianensis sp. nov. natural history and breeding aspects. A Example of habitat used by the new species; B calling male (INPA-H 43942); C amplectant pair (female INPA-H 43943, male INPA-H 43944), from RFAD, Manaus, Amazonas; D suspended litter spawning about 1 m above the ground, the two clutches are at different stages (left, more developed; right, recent oviposition), from Sao Joao da Baliza municipality, Roraima, Brazil. Photographs: A. T. Monico, except (B) I. Y. Fernandes.

Figure 16. Pristimantis guianensis sp. nov. eggs and embryos from Reserva Florestal Adolpho Ducke (Manaus, Brazil). Photographs: A. T. Monico.