Pseudopaludicola serrana, Toledo, 2010

Toledo, Luís Felipe, 2010, Description of a new species of Pseudopaludicola Miranda-Ribeiro, 1926 from the state of São Paulo, Southeastern Brazil (Anura, Leiuperidae), Zootaxa 2681, pp. 47-56 : 48-55

Pseudopaludicola serrana sp. nov.

Holotype. ZUEC 16442 , an adult male collected in a temporary pond in the Condomínio Retiro das Pedras (private area), Serra da Moeda , municipality of Brumadinho, state of Minas Gerais, southeastern Brazil (20º06'06.7" S and 43º59'20.8" W; approximately 1.450 m above sea level), by J. Tolledo, N. R. Silva, P. G. Taucce, and P. Pinheiro on 01 December 2009 ( Figure 1A ).

Paratopotypes. Four adult males ( ZUEC 16396-98 ; 16443) , one female ( ZUEC 16399 ) , and two juveniles ( ZUEC 16394-5 ) all collected with the type .

Diagnosis. Pseudopaludicola serrana is assigned to the genus Pseudopaludicola due to the presence of hypertrophied antebrachial tubercle (see Lynch, 1989). Pseudopaludicola serrana is an intermediate size species for the genus and is characterized by the following combination of characters: (1) long hindlimbs and absence of T-shaped terminal phalanges, (2) single, dark, and subgular vocal sac with dark longitudinal folds in males; (3) immaculate to light brown, nuptial pads in males that cover the external part of finger I; (4) SVL/ HL greater than 2.7; and (5) advertisement call with dominant frequency above 5 kHz, pulse duration range between 13 and 23 ms, and mean interval between notes of 177 ms.

Comparison with other species. Pseudopaludicola serrana is distinguished from the species of the P. pusilla group ( P. canga , P. ceratophryes , P. llanera , and P. pusilla ) by the absence of T-shaped terminal phalanges (the paratype ZUEC 16396 had its fingers dissected). From the species of the P. falcipes group, except from P. murundu and P. saltica , it distinguishes by having very long hindlimbs, with tibio-tarsal articulation reaching beyond the end of the snout. From P. saltica it is distinguished by having larger and clearer nuptial pads (smaller and darker in P. saltica ), longitudinal folds in the vocal sac (absent in P. saltica ), and a darker vocal sac (immaculate in P. saltica ). From P. saltica and P. murundu it distinguishes by having the head not as long (SVL/HL = 2.99 ± 0.30; 2.71–3.34 / HL/HW = 0.97 ± 0.06; 0.86–1.02) as in P. saltica (SVL/HL = 2.28 ± 0.11; 2.00–2.44 / HL/HW = 1.15 ± 0.08; 1.01–1.33) and P. murundu (SVL/HL = 2.42 ± 0.12; 2.25–2.62 / HL/HW = 1.11 ± 0.07; 1.01–1.24) ( Table 1 ). The physical characteristics of the advertisement calls also distinguish P. serrana From P. saltica and P. murundu . Significant differences can be observed in pulse duration (between the three species), interval between pulses (between the three species, although the ranges of variation are overlapped), interval between notes (significant difference between P. serrana and the two other species), and dominant frequency (significant difference between P. saltica and the two other species) (see Tables 2 , 3 , and 4 ).

Description of the holotype. Body elliptic and broad. Head triangular, slightly longer than wider. Snout sub-elliptical in dorsal view and rounded in profile ( Figure 2A, 2B ). Nostrils slightly protuberant, directed anterolaterally. Mouth opening ventral. Canthus rostralis rounded. Loreal region slightly concave. Choanae rounded. Eye protuberant, its diameter almost the same size of the interorbital distance. Interorbital area flat. Tympanum indistinct. Vocal sac single, externally expanded, large, and with longitudinal folds; vocal slits present. Vomerine teeth absent. Tongue elliptical, longer than wide. Finger length I<IV<II<III. Toe length I<II<III=V<IV. Finger and toe tips without disks ( Figure 2C, 2D ). Thumb with keratinized pale beige nuptial pad. Finger webbing absent and toe webbing reduced I-II 2-3 III 3-4 IV-V. Finger and toe subarticular tubercles conical and single. A large subconical ulnar tubercle. Few rounded supernumerary tubercles in the hand in the area delimited by the first subarticular tubercles, the elliptical internal metacarpal tubercle, and the ovoid external metacarpal tubercle. Hindlimbs robust and long. Thigh shorter than tibia; foot longer than thigh and slightly shorter than tibia. Supernumerary tubercles absent in the foot. Metatarsal tubercles present, elliptical; internal larger than the external; external more protuberant than the internal. A well developed fold from the internal metatarsal tubercle to the mid-ventral tarsus. Skin of belly smooth; ventral surface of thigh granular. Dorsum of head, body, and limbs smooth with scattered tubercles; the skin on the scapula region has two arc shaped granular folds. Flanks without tubercles. Cloacal region not granular. Measurements of the holotype are presented in table 1.

Color of the holotype. In life, dorsum and limbs are brown with green and brown spots or stripes; vertebral line from the snout to vent absent; belly white; ventral view of legs yellowish white and vocal sac beige with dark dots; iris reddish-brown superiorly and grayish-brown inferiorly; superior labium with vertical wide green stripes (see Figure 1A ). In preservative, dorsum and dorsal part of limbs grayish brown with dark brown spots or stripes; belly and ventral surface of legs light beige; vocal sac grayish.

Variation. Females larger than males, lacking vocal sac and nuptial pads (present in males). Males present arms proportionally thicker than females. Nuptial pad can be brownish to light cream color. In 76.5 % of the individuals (13 out of 17 examined, including material not included in type series) the vertebral line is absent. In life this line can be yellow or red (see Figure 1B and C ). Dorsal pigmentation varies considerably among individuals in the number and form of the dorsal spots, and in the general pattern, from brown to grayish, but always with some green blotches ( Figure 1A, B, and C ).

Natural history notes. Males were observed calling in open fields in the domain of Cerrado (Brazilian savannah). The species was observed calling in riparian fields (Campo Rupestre). Males called from the sandy, muddy or rocky ground, near shallow (from 2 to 5 cm of depth) slow-flow or still water bodies. Males were observed calling in the sundown and during the first hours of the night. Two defensive behaviors were observed: ceasing the calling activity and flee against human approach. Gravid females present ovules with black animal pole and yellowish-beige vegetative pole. A couple entered in axillary amplexus inside a plastic bag where they laid 32 eggs, which developed into tadpoles in laboratory until Gosner´s stage 25.

Geographic distribution. The new species is known from three localities at the Espinhaço mountain range: Serra do Cipó, municipality of Santana do Riacho (approximately 19°08’- 43°40’; 1071 m; ZUEC 2323), Serra da Moeda, municipality of Brumadinho (type locality), and Serra do Lenheiro, municipality of São João del Rei (approximately 21°08’ – 44°17’; 1081 m; ZUEC 16445–53; 16455–56), all higher than 1,000 m above sea leavel and in the state of Minas Gerais, Southeastern Brazil ( Figure 3 ).

Etymology. The specific epithet is the feminine form from the Portuguese adjective “serrano” that means “the one who lives in the mountains”.

Description of the advertisement call. The advertisement call of Pseudopaludicola serrana is composed of groups of pulsed notes (with one to four pulses) that are emitted in mean intervals of 0.18 s. Calls are generally long, varying from about 14 to 45 seconds in the analyzed series. Calls are high pitched, with frequencies varying between about 3.0 and 8.5 kHz. The mean rate of notes emitted per minute is 227. The pulses within a note generally present an ascending modulation, i.e., the first pulse presents lower frequencies than the second, which, in turn, presents lower frequencies than the third, and so on ( Figure 4 ). Further information of the physical characteristics of the advertisement call is in Table 2 . Comparisons with other species are presented in table 3.

Tadpoles. Larvae were obtained from eggs laid in plastic bags after collection of adult specimens in type locality. The following description is based on 16 tadpoles (ZUEC 16457) in developmental stages 25 ( Gosner, 1960). Body elliptic in dorsal and ventral views ( Figure 5 ), depressed/globular with flattened venter; body wider than high; snout rounded; eyes medium-sized, dorsolateral; nostrils dorsal, small, and oval; nostrils closer to the eyes than to the tip of snout; spiracle sinistral, its opening in the middle of body; cloacal tube large, medial; caudal musculature robust; dorsal fin originating on the anterior part of the tail; dorsal fin wider than ventral fin ( Figure 5B ). Oral disc directed ventrally, emarginated, and bordered by one row of marginal papillae, interrupted along a large area on the anterior labium; tooth row formula 2(2)/2(1); upper and lower jaw sheathes strongly developed and serrate. Some of the tadpoles present unkeratinized mouthparts, probably due to Batrachochytrium dendrobatidis infection, acquired in the laboratory, as individuals collected in the same locality are apparently uninfected. In preservative and in life, dorsum pale brown; throat and belly transparent; caudal musculature with very scattered pale brown pigmentation; fins transparent with few scattered pale brown pigmentation ( Figure 5 ). Lateral lines were not observed.

Three tadpoles at Gosner’s stage 25 measured [mean ± SD (range); size in mm] total length 9.34 ± 1.12 (8.05–10.05); body length 4.38 ± 0.56 (3.73–4.71); tail length 4.96 ± 0.56 (4.32–5.35); maximum body height 1.83 ± 0.31 (1.50–2.10); maximum body width 2.53 ± 0.39 (2.11–2.89); tail maximum width 1.61 ± 0.67 (1.00–2.32); internarial distance 0.48 ± 0.13 (0.34–0.60); interorbital distance 0.57 ± 0.07 (0.49–0.63); eyenostril distance 0.31 ± 0.08 (0.26–0.40); eye diameter 0.46 ± 0.06 (0.40–0.56).

Figures

FIGURE 1. Live specimens of Pseudopaludicola serrana sp. nov.: the holotype (adult male ZUEC 16442) (A), an adult red-striped female (ZUEC 16455) (B), and an adult white-striped male (ZUEC 16456) (C).

FIGURE 2. Holotype of Pseudopaludicola serrana sp. nov. Dorsal (A) and lateral (B) views of the head; ventral views of hand (C) and foot (D).

FIGURE 3. Geographic distribution of Pseudopaludicola serrana sp. nov. The triangle indicates type locality (Serra da Moeda) and the circles represent the other known localities of occurrence: Serra do Cipó (to the North) and Serra do Lenheiro (to the South). In the up left, map of Brazil indicating the locality of the state of Minas Gerais (in grey). SP: São Paulo; MG: Minas Gerais; RJ: Rio de Janeiro; ES: Espírito Santo.

FIGURE 4. Waveform (below) and spectrogram (above) of the first two seconds of the advertisement call of Pseudopaludicola serrana sp. nov. recorded at type locality: Serra da Moeda, Minas Gerais.

FIGURE 5. Tadpole of Pseudopaludicola serrana sp. nov. at Gosner’s stage 25 (ZUEC 16457) in dorsal (A), lateral (B), and ventral (C) views.

Tables

TABLE 1. Measurements of the type series (holotype, males including the holotype, and females) of Pseudopaludicola serrana sp. nov. Values presented in millimeters as mean ± standard deviation (range).

	Holotype	Paratypes
Measurement	Male	Males (N = 5)	Female
Snout-vent length	15.2	15.22 ± 0.31 (15.0–15.8)	18.9
Axial-groin length	4.8	5.07 ± 0.36 (4.7–5.6)	6.7
Head length	5.5	5.13 ± 0.57 (–)	6.7
Head width	5.4	5.30 ± 0.54 (4.5–6.0)	7.3
Eye diameter	1.3	1.34 ± 0.18 (1.2–1.6)	1.9
Upper eyelid width	1.2	1.13 ± 0.11 (1.0–1.2)	1.5
Interorbital distance	1.2	1.43 ± 0.19 (1.2–1.7)	1.7
Internarial distance	1.5	1.55 ± 0.12 (1.4–1.7)	2.1
Eye-nostril distance	1.3	1.42 ± 0.11 (1.3–1.6)	1.7
Snout-eye distance	3.0	2.88 ± 0.11 (2.8–3.0)	3.4
Humerus length	2.3	2.47 ± 0.27 (2.1–2.8)	2.3
Radio-ulna length	2.3	2.37 ± 0.22 (2.1–2.6)	3.2
Hand length	4.6	4.44 ± 0.20 (4.2–4.7)	4.6
Thigh length	8.5	8.85 ± 0.42 (8.4–9.4)	10.9
Tibia length	10.3	10.25 ± 0.39 (9.9–10.9)	12.9
Foot length	9.6	9.73 ± 0.23 (9.6–10.1)	12.5

TABLE 2. Physical characteristics of the advertisement calls of Pseudopaludicola saltica (n = 12 notes/ 1 call/ 1 male; air temperature 24°C), P. murundu (n = 14 notes/ 2 calls/ 2 males; air temperature = 25.5°C), and P. serrana sp. nov. (n = 15 notes/ 3 calls/ 3 males; air temperature = 25°C); all recorded at their type localities and analyzed for the present study. Values presented as mean ± standard deviation (range).

Species	Frequency (kHz)			Duration (ms)
	Dominant	Minimum	Maximum	Note	Pulse
P. saltica	4.60 ± 0.18 (4.39–4.91)	3.05 ± 0.24 (2.52–3.34)	6.35 ± 0.35 (5.92–6.93)	76.33 ± 18.59 (31.0–97.0)	33.0 ± 4.43 (24.0–38.0)
P. murundu	5.72 ± 0.32 (5.17–6.37)	5.03 ± 0.20 (4.71–5.33)	6.82 ± 0.26 (6.44–7.25)	98.86 ± 8.57 (87.0–112.0)	11.21 ± 2.29 (6.0–14.0)
P. serrana	5.75 ± 0.11 (5.51–6.03)	3.56 ± 0.44 (3.12–4.50)	7.98 ± 0.41 (7.01–8.49)	85.33 ± 22.00 (25.0–115.0)	18.87 ± 3.04 (13.0–23.0)

continued.

TABLE 3. Statistical outputs of the comparisons between physical characteristics of advertisement call of the species of Pseudopaludicola of the group of P. saltica, made by means of Kruskall-Wallis tests, followed by Mann-Whitney comparisons (Bonferroni corrected \ uncorrected). Bold values indicate significant differences (P <0.05).

Pulse duration
	P. murundu		P. saltica		P. serrana
P. murundu				0.00002		0.00008
P. saltica		0.00005				0.00005
P. serrana		0.00023		0.00015
Interval between notes
	P. murundu		P. saltica		P. serrana
P. murundu				0.66200		0.00023
P. saltica		1.00000				0.00040
P. serrana		0.00068		0.00121
Dominant frequency
	P. murundu		P. saltica		P. serrana
P. murundu				0.00009		0.21200
P. saltica		0.00026				0.00001
P. serrana		0.63600		0.00004
Interval between pulses
	P. murundu		P. saltica		P. serrana
P. murundu				0.00109		0.00209
P. saltica		0.00327				0.03379
P. serrana		0.00628		0.10140