Bioacoustics of Dendropsophus ozzyi

Etymology . The specific epithet is used as a noun in the genitive case and honors John Michael "Ozzy" Osbourne, a famous British rock singer, former vocalist of the rock band Black Sabbath, for its contribution to modern music and culture. Ozzy is commonly associated with bats because of the famous episode in which, on stage while he was performing a gig, he bit off a bat’s head thrown by a fan. He later stated that he thought it was a plastic toy. When calling, this new Dendropsophus species can be vaguely associated with the high pitch sounds emitted by some bat species that are hearable to the human ear. When we heard this species in the field we immediately pictured a calling bat. Because of this “bat association” we take the opportunity to name this species after one of the biggest music legends of all time.

Diagnosis. Dendropsophus ozzyi is diagnosable by the following combination of character states: (1) a medium sized member of the Dendropsophus microcephalus Group, (SVL 18.5–21.5 mm in adult males, N = 37); (2) head wider than body; (3) snout truncate in dorsal and lateral views; (4) nostrils protuberant; (5) large prominent eyes (EL/HW 0.3–0.5, N = 37); (6) nictitating membrane bearing brownish pigmentation on its border; (7) small tympanum (TD/ED 0.3–0.5, N = 37); (8) axillary membrane present; (9) glandular nuptial pads on finger II; (10) hands webbing formula II 2 - – 2- III 2 - – 2- IV 2 + – 2+ V; (11) feet webbing formula I 2 – 2 + II 1 + – 3- III 1 +– 2+ IV 2 + – 1+ V; (12) fingers and toes bearing pointed discs; (13) no inner tarsal fold; (14) calcar tubercle absent, heel with small, inconspicuous tubercles; (15) a dorsal sheath covering 1/3 of the cloacal opening; (16) in life, dorsal surfaces plain light brown with randomly distributed dark smudges; palmar, plantar and ventral surfaces of the axillary membrane, thigh, and shank vivid orange, belly creamy white with clear white spots and a rectangular bright white area, gular region orangish cream with brown flecks, vocal sac transparent; (17) advertisement call composed of a single, not pulsed, tonal, and highly pitched (dominant frequency around 9 kHz) note. FIGURE 3. Dendropsophus ozzyi sp. nov. holotype (MPEG 27811). (A) Dorsal, and (B) lateral views of head; (C) palmar view of right hand, and (D) plantar view right foot. Scale bar = 5 mm . Comparison with other species . Dendropsophus ozzyi differs from all other species of Dendropsophus , except D. shiwiarum —also in the D. microcephalus Group—by having pointed discs on fingers and toes. In the following paragraphs we first diagnose our new species from D. shiwiarum and then provided additional diagnostic characters that differentiate D. ozzyi from all other species of the genus arranged by species groups.

Description of holotype : Adult male, SVL 19.4 mm ; head wider than long (HW/HL 1.18), widest below eyes; snout truncate in both dorsal and lateral views, eye-nostril distance longer than eye diameter ( END /ED 1.17); canthus rostralis indistinct, straight; loreal region slightly concave; lips thin; internarial area slightly depressed; nostrils protuberant, directed dorsolaterally; interorbital area flat; IOD/ED 1.48, IOD/HW 0.49; eyes large and protuberant (ED/HL 0.40, ED/HW 0.33); ELW 1.38 mm . Pupil horizontally elliptical; nictitating membrane transparent, its border slightly pigmented in the same pattern of the eyelid. Supratympanic fold barely visible, semi-circular in outline, less visible on the posterior portion; tympanum small (TD/HL 0.17), distinct, nearly round, separated from eye by a distance of 1.3 tympanum diameter; choanae small, oval, not concealed by palatal shelf of maxillary arch; vomerine odontophores very small, the right bearing two small, unaligned teeth while the left bears three; tongue cordiform, notched behind, posterior 1/3 not adhered to floor of mouth; vocal slits long, extending from midlateral base of tongue to almost the angle of jaws; vocal sac developed, single, and subgular, extending over the forearm (Figs. 3–4). Arm slender, not hypertrophied; axillary membrane reaching half arm, delimited by the m. pectoralis portio axillaris (seen by transparency). Lateral border of arm with fimbria composed of small tubercles, not all clearly individualized. Fingers short, bearing discs; finger discs are slightly pointed, albeit disc of Finger II is slightly less, close to round; relative length of fingers II <III <V <IV; subarticular tubercles less wide than the digits, round on all fingers, bifid on penultimate articulation of Finger IV, most prominent on fingers II, III and IV; supernumerary tubercles present; inner metacarpal tubercle flat, elliptic; outer metacarpal tubercle bifid, nearly not visible; glandular nuptial pad covering the area corresponding to the prepollex in mesial view; webbing basal between fingers II and III; webbing formula II 2 -–2- III 2 -–2- IV 2 +–2+ V (Fig. 3C). Hind limb long and slender (TL/SVL 0.53); no tarsal fold, but outer margins of tarsus and feet with fimbriae similar to the one found in arm; calcar tubercle absent, heel with small, inconspicuous tubercles; toes bearing pointed discs, smaller than those on fingers; feet slightly dehydrated, right one more than the left; relative lengths of toes I <II <III <V <IV; subarticular tubercles, round, bifid on penultimate articulation of Toe V, shallow; small supernumerary tubercles present on toes III and IV; inner metatarsal tubercle flat, elliptical; outer metatarsal tubercle indistinct, nearly not visible; webbing formula I 2–2 + II 1 +–3- III 1 +–2+ IV 2 +–1+ V. Toe IV bears a gland, visible at both dorsal and ventral views at the level of subarticular tubercles II and III; dorsally, a glandular patch is visible on Toe III ( Fig. 4 ). Skin on dorsum, head, dorsal surfaces of forearms and thighs, flanks and groin smooth, finely shagreen on head, shank and tarsus; skin on belly and ventral surfaces of thighs granular. Cloacal opening directed posteriorly, above the thighs, covered by a small cloacal sheath dorsally; cloacal tubercles absent. Pectoral region, belly, ventral surfaces of forearm and thigh with many minute acini filled with dark yellow secretion. Pectoral acini not arranged as a glandular patch; loosely distributed over the area ( Fig. 4 ) Coloration in preservative : All dorsal surfaces yellowish brown, except thighs, fingers II, III, IV and toes I, II, III, IV that are cream. Dorsal ground color is darker at the head, fading posteriorly. Dorsum bears 13 brown smudges of irregular shape. Canthus rostralis, eyelid and the anterior third of the flanks have brown and white markings. Iris black with many minute silver spots of variable diameters. Ventral areas creamy white, except for the submandibular area that bears various brown flecks, palmar and plantar surfaces have a pattern resembling the dorsal pattern. The belly also presents a bright white rectangular area, roughly covering the area of the sternum. Dissections of paratype CFBH 35764 demonstrated that it is a layer of iridophores at the perimysium of the abdominal muscles. Coloration in life (based on paratype MPEG 27810) : Dorsal surfaces of head and body brown. Flanks, arms, hands (excluding fingers II to IV and finger disc of Finger V), tibia, and the external region of feet (including toes IV and V) light brown. Thighs, fingers II to IV and finger disc of Finger V, and internal region of feet (including toes I to III and toe discs of toes IV and V) vivid orange. Palmar, plantar and ventral surfaces of the axillary membrane, thigh and shank vivid orange. Ventrolateral areas of tarsus and feet dark brown. Belly creamy white with clear white spots and a rectangular bright white area (as described above). Gular region orangish cream with brown flecks. Iris copper with dark reticulations. Lateral areas of the head and flanks light brown, nearly white, composing a diffuse stripe starting at the loreal region and extending posteriorly to the groin through the trunk. Variation . Measurements are shown in Table 1 . External morphology of the type series is generally concordant with the holotype ; including color ( Fig. 5–6 ). The number and conspicuousness of ventral acini varies from nearly none (MPEG 27809) to many (MPEG 27810, 27812 and MZUSP 154087 as also do the number of clear white flecks (while MPEG 27809 has none, MPEG 27810 has many) and the iridophore area of the abdominal perimysium. Dorsal coloration varies slightly, from lighter (MPEG 27814) to darker (MPEG 27809; CFBH 35764) shades of brown. The number, shape, and position of dorsal smudges also vary from absent (MPEG 27814; MZUSP 154086) to more then 20 (MPEG 27812). Left thigh muscles of MPEG 27809, MZUSP 154084–154088, and the left foot of MPEG 27810 were removed in the field as tissue samples for molecular analyses. Advertisement call . Most of the recordings have more than one individual calling, and in many cases it is impossible to distinguish which one is emitting the note. Nonetheless, we have measured the note duration and spectral parameters for all the notes in the best recording (PLDR 035; N = 11 notes from up to three distinct calling males), and only the first three notes and the intervals between them in the second recording (PLDR 036; certainly emitted by the same individual). One of the individuals in recording PLDR 0 36 was collected (MPEG 27814, field number FPR 205) but the calls emitted by this male are unsuitable for bioacoustical analyses (saturated recording). Nonetheless, the call can be unambiguously identified as belonging to the same species as the ones for which analyses were performed. Analyses of recordings reveal that the advertisement call of Dendropsophus ozzyi consists of an unusually high-pitched single note, emitted at relatively long intervals (Fig. 7). Amplitude modulation within the note shows higher energy at the beginning of the note, with energy decreasing abruptly and then continuing with small energy for the remaining of the note duration. Note duration is 165–386 ms (mean 248 ms; N = 14), interval between notes is 1965–5183 ms ( N = 2), and the dominant frequency is 9130.1–10136.7 Hz (mean 9560.7 Hz; N = 14). Notes are not clearly multi-pulsed, except for a single note with four pulses. FIGURE 7. Waveform (A) and audiospectrogram (B) of advertisement calls (recording PLDR 036) of two individuals of Dendropsophus ozzyi sp. nov. from the type-locality. The first call is from paratype MPEG 27814, the second from an unvouchered specimen. Natural history . Dendropsophus ozzyi was found mainly on bushes close to water bodies, such as ponds and swamps, inside Terra Firme forest. The most commonly used type of vegetation were shrubs and small trees < 5cm DBH (diameter at breast height), where individuals were observed on the leaves and branches in heights that varied from ground level (0 m) to 3.5 m , with prevalence between 1.0 and 1.5 m . No difference was observed in height of perch between sexes. Most individuals were observed 0.5 m away from water bodies. In addition, no individuals were observed farther than 5.0 m away from water bodies. Calling males were observed in the wettest months, from November to June. Time of observation varied overnight, from 18:00 to 23:00 and was more frequent Catalog Number Field Number Status SVL HL HW ED ELW END IOD TD THL TL FL MPEG 27809 FPR 105 Paratopotype 19.80 5.80 6.20 2.50 1.40 2.90 3.60 0.90 9.70 10.40 13.80 MPEG 27810 FPR 140 Paratopotype 20.00 6.50 7.20 2.60 1.40 2.50 3.60 1.40 10.80 11.10 14.10 MPEG 27811 FPR 141 Holotype 19.50 5.80 6.90 2.30 1.50 2.70 3.40 1.00 9.10 10.30 13.40 MPEG 27812 FPR 142 Paratopotype 18.50 5.70 6.50 2.30 1.50 2.90 3.60 1.10 9.60 10.10 13.30 MPEG 27813 FPR 143 Paratopotype 21.50 5.70 7.10 2.90 1.50 2.90 3.60 1.40 9.70 9.70 13.50 MPEG 27814 FPR 205 Paratopotype 19.80 6.10 6.80 2.30 1.50 2.30 3.30 0.80 10.20 9.80 13.50 INPA-H 32742 J123 Paratype 18.30 6.60 6.90 3.00 1.90 2.05 2.50 1.10 10.20 10.50 13.20 MZUSP 154088 J117 Paratype 20.10 7.40 8.00 2.90 1.90 2.00 2.60 1.00 10.20 10.80 13.00 MNRJ 86925 J264 Paratype 18.10 6.80 6.50 2.90 1.50 1.90 2.50 0.95 10.20 10.20 13.20 INPA-H 32744 J121 Paratype 19.00 6.60 7.00 2.80 2.10 2.10 2.50 0.75 10.00 10.40 13.40 CFBH 35764 J205 Paratype 19.40 7.10 7.30 2.90 1.70 1.90 2.50 0.85 10.25 10.50 13.50 CFBH 35762 J129 Paratype 18.60 7.20 7.30 2.85 2.40 2.00 2.10 0.90 10.40 10.90 13.90 MNRJ 86921 J206 Paratype 20.40 7.40 7.40 3.10 1.90 1.85 2.20 1.00 10.50 10.45 13.90 CFBH 35760 J125 Paratype 18.30 7.40 7.30 3.00 1.85 1.80 2.50 0.80 10.30 10.25 13.50 MNRJ 86923 J208 Paratype 20.50 7.10 7.30 3.00 1.90 1.90 2.60 1.00 10.85 10.90 14.10 MNRJ 86922 J207 Paratype 19.20 7.00 7.10 2.65 1.60 1.90 2.20 1.20 9.70 10.40 14.10 MZUSP 154087 J116 Paratype 20.65 7.10 7.60 3.00 2.30 1.85 2.80 1.05 11.00 10.80 14.15 CFBH 35761 J126 Paratype 20.90 7.40 7.20 2.90 2.30 1.90 2.55 0.95 11.70 11.50 13.70 INPA-H 32746 J18 Paratype 19.90 6.90 7.60 2.90 2.20 2.00 2.70 0.90 11.30 11.30 14.65 MNRJ 86924 J263 Paratype 19.50 7.50 7.10 3.00 1.60 1.90 2.30 1.05 10.00 10.45 13.80 MZUSP 154086 J115 Paratype 19.80 6.80 7.70 2.85 1.90 1.80 2.40 0.90 9.90 10.40 13.35 MZUSP 154085 J114 Paratype 19.40 7.10 7.40 2.70 1.70 2.10 2.20 0.95 10.65 10.60 12.70 INPA-H 32747 J19 Paratype 19.20 6.70 6.95 3.05 2.30 1.90 2.50 0.90 10.00 10.20 13.70 INPA-H 32748 J124 Paratype 18.00 6.45 6.85 2.65 1.80 1.75 2.30 0.90 10.15 10.00 12.90 INPA-H 32743 J20 Paratype 19.90 7.20 7.50 2.90 2.10 1.90 2.85 0.90 10.40 10.40 14.35 MZUSP 154084 J113 Paratype 19.30 7.00 7.25 2.90 1.85 2.05 2.60 0.95 10.85 11.05 13.00 INPA-H 32745 J122 Paratype 17.90 6.80 6.85 2.50 1.75 1.85 2.25 1.00 10.40 10.15 13.60 MPEG 22352 JUR246 Paratype 19.70 5.80 6.20 2.50 1.39 2.90 3.50 0.80 9.80 10.50 13.60 MPEG 22353 JUR247 Paratype 19.60 5.80 6.10 2.60 1.41 2.50 3.50 0.90 9.60 10.30 13.40 MPEG 22354 JUR248 Paratype 21.30 6.60 7.10 2.80 1.50 2.90 3.60 1.40 10.40 11.10 14.10 MPEG 22355 JUR249 Paratype 18.60 5.70 6.50 2.30 1.42 2.30 3.30 0.90 9.60 10.10 13.30 MPEG 22400 JUR221 Paratype 19.80 5.70 6.20 2.40 1.43 2.30 3.70 1.10 9.70 9.70 13.50 MPEG 22401 JUR222 Paratype 21.40 6.70 7.20 2.90 1.51 2.90 3.40 1.40 10.50 11.20 14.30 MPEG 22402 JUR223 Paratype 19.90 5.70 6.20 2.50 1.42 2.40 3.30 1.10 9.60 9.90 13.10 MPEG 22403 JUR224 Paratype 18.70 5.60 6.10 2.30 1.47 2.30 3.50 0.80 9.50 9.70 13.20 MPEG 22404 JUR225 Paratype 18.60 5.60 6.10 2.40 1.45 2.50 3.30 1.20 9.50 9.80 13.20 CFBH 35762 J130 Paratype 18.75 7.20 7.45 2.95 2.20 2.00 2.45 0.95 10.70 10.90 13.80 29 Species clade Calltype Minimum Maximum Average Reference D. berthalutzae D. bipunctatus * decipiens monophasic 3.7 monophasic 4.8 4.7 5.6 3.8? Forti et al. (2012); Moura et al. (2012) Abrunhosa et al. (2001) D. branneri monophasic 5.9 6.6 6.3 Nunes et al. (2007) D. coffeus diphasic 5.7 6 5.8 Köhler et al. (2005) D. cruzi monophasic 5.2 7 6.3 Bastos et al. (2003) D. decipiens decipiens monophasic 4.7 5.2 5 Abrunhosa et al. (2001) D. elianeae rubicundulus monophasic 2.9 4 3.3 Martins (2004) D. gryllatus diphasic 3.6 4.9 3.8 Duellman (1973) D. haddadi decipiens monophasic 4.3 4.9 4.5 Ruas et al. (2012) D. jimi rubicundulus diphasic 3.6 4.6 4 Martins & Jim (2004) D. joannae diphasic 6.6 7 6.9 Köhler & Lötters (2001a) D. juliani monophasic 3.4 4.1 3.7 Moravec et al. (2006) D. leali ** monophasic 6.1 6.7 6.4 Köhler & Lötters (2001a) and references therein D. meridianus *** monophasic? ? 8 Pombal Jr. & Bastos (1998) D. microcephalus diphasic 5.1 6.2 5.7 Duellman (1968) D. minusculus **** monophasic 9 9.3 9.2 Duellman & Pyles (1983) D. nanus diphasic 2.6 5.5 3.9 Martins & Jim (2003) D. oliveirai decipiens monophasic 5.7 6.7 6.1 Santana et al. (2011) D. ozzyi sp. nov. monophasic 9.1 10.1 9.6 Present study D. phlebodes diphasic 3.2 4 3.6 Duellman (1968) D. reicheli monophasic 6.2 6.6 6.3 Moravec et al. (2008) D. rhodopeplus ** diphasic 3.12 3.9 3.51 Duellman (1972, 1978); Marquez (1993) D. riveroi monophasic 4.3 5.3 4.8 Marquez et al. (1993) D. robertmertensi diphasic 5.1 5.8 5.4 Duellman (1968) D. rubicundulus rubicundulus monophasic? ? 4.4 Cardoso & Vielliard (1985) D. sanborni diphasic 3.8 5.9 5.1 Martins & Jim (2003) D. sartori diphasic 2.9 3.6 3.2 Duellman (1968) D. shiwiarum monophasic 4 5.2 4.4 Ortega-Andrade & Ron (2013) D. tritaeniatus rubicundulus monophasic 3.9 4.5 4.2 Teixeira et al. (2013) D. werneri diphasic 6.3 7.4 6.7 Lingnau & Bastos (2003); Lingnau et al. (2004) = Abrunhosa et al. (2001) report different values in their text and table. We reproduce the largest range reported. **= Marquez (1993) reports dominant frequencies around 120 Hz, a curiously low value. Authors also report additional emphasized frequencies of higher values that are a more robust test to our hypothesis that D. ozzyi sp. nov. has an unusually high dominant frequency. See Orrico et al. (2009) for a discussion on dominant frequency artifacts. ***= Bastos & Pombal (2003) do not report which frequency is dominant, just that there are emphasized harmonics around 3 and 8 kHz. Their figure 11 depicts a call with dominant frequency around 7 kHz. **** Lescure & Marty (2000) and Tárano (2010) report different values (~5 kHz and 3.8 kHz respectively). between 19:00 and 21:00. Amplectant pairs and tadpoles were not observed. All collection sites are between the Tapajós and Madeira Rivers, and we expect more populations of D. ozzyi to be found within this area.