Boophis entingae, Glaw & Köhler & Riva & Vieites & Vences, 2010

Glaw, Frank, Köhler, Jörn, Riva, Ignacio De La, Vieites, David R. & Vences, Miguel, 2010, Integrative taxonomy of Malagasy treefrogs: combination of molecular genetics, bioacoustics and comparative morphology reveals twelve additional species of Boophis 2383, Zootaxa 2383 (1), pp. 1-82 : 22-26

Boophis entingae sp. nov.

( Fig. 10 , Appendix 9)

Remark. This species has been referred to as Boophis brachychir by Glaw & Vences (1992, 1994, 2007:162– 163) and as Boophis sp. 11 by Vieites et al. (2009). The pictures 3, 4a and 4b in Glaw & Vences (2007:163) all show B. entingae (see account above for B. brachychir ).

Holotype. ZSM 2083 /2007 ( FGZC 1057 ), adult male (call voucher), from Montagne d'Ambre National Park , northern Madagascar, ca. 12°29'38''S, 49°10'55'' E, app. 750 m a.s.l., collected on 25 February 2007 by P. Bora, H. Enting, F. Glaw, A. Knoll and J. Köhler.

Paratypes. ZSM 897 /2003 ( FGMV 2002–908 ), juvenile, Montagne d'Ambre , collected on 18 February 2003 by F. Glaw , R. Randrianiaina and A. Razafimanantsoa ; ZSM 624 /2001 ( FGMV 2001.158 ), adult male (call voucher), from Tsaratanana massif, Manarikoba forest , Andampy (Camp 0), 14°02'32''S 48°45'42''E, ca. 730 m a.s.l., collected on 10–11 February 2001 by F. Andreone, F. Mattioli, J. Randrianirina and M. Vences ; ZSM 830 /2003 ( FG / MV 2002–0775 ) and ZSM 831 /2003 ( FGMV 2002–0776 ), two adult males from Manongarivo , Camp 0, collected on 1 February 2003, by F. Glaw , R. D. Randrianiaina and M. Vences ; ZSM 852 / 2003 ( FG / MV 2002–0822 ), adult male, and ZSM 858 /2003 ( FG / MV 2002–0829 ), juvenile, both from Manongarivo , Camp 1, collected on 3 February 2003 by F. Glaw , R. D. Randrianiaina and M. Vences ; ZSM 899 /2003 ( FG / MV 2002–0911 ), female, Montagne d'Ambre , collected on 17–18 February 2003 by F. Glaw , R. Randrianiaina and A. Razafimanantsoa ; ZSM 214 /2004 ( FGZC 418 ), Montagne d'Ambre , collected on 21 February 2004 by F. Glaw, M. Puente , R. Randrianiaina and A. Razafimanantsoa ; ZSM 2081 /2007 ( FGZC 1055 ), from Montagne d'Ambre , 12°29'38''S, 49°10'55'' E, ca. 750 m a.s.l., collected on 25 February 2007 by P. Bora, H. Enting, F. Glaw, A. Knoll and J. Köhler ; ZFMK 53637–53639 , three adult males from near Benavony , collected on 16 February 1992 by F. Glaw and J. Müller-Jung .

Etymology. We dedicate this species to Hildegard Enting (Frankfurt/Main). Her help with logistics, field work and preparation of specimens during two Madagascar expeditions contributed significantly to their success.

Diagnosis. Assigned to the genus Boophis based on the presence of an intercalary element between ultimate and penultimate phalanges of fingers and toes (verified by external examination), presence of nuptial pads and absence of femoral glands in males, absence of gular glands in males, enlarged terminal discs of fingers and toes, lateral metatarsalia separated by webbing, absence of outer metatarsal tubercle, molecular phylogenetic relationships (see Vieites et al. 2009 for a complete molecular analysis of Boophis ), and overall similarity to other Boophis species. Assigned to the Boophis goudoti group based on the following combination of characters: relatively large size (male SVL 47–58 mm); brownish dorsal ground colour; non-transparent ventral skin, absence of red ventral colour; presence of dermal flaps or tubercles on heels and elbows; presence of webbing between fingers; single subgular vocal sac; presence of vomerine teeth; presence of white tubercles ventrally under the cloacal opening; presence of a sharp canthus rostralis, and molecular phylogenetic relationships (see Vieites et al. 2009). Boophis entingae differs from all described species in the B. goudoti group by strong genetic differentiation (see below) and from B. boehmei , B. rufioculis , B. axelmeyeri , B. reticulatus , B. madagascariensis , B. goudoti , B. obscurus , and B. periegetes by advertisement calls: in brief, calls of B. goudoti , B. madagascariensis and B. obscurus are irregular croaks and moans rather than the regular series of 2–4 moans of B. entingae ; the calls of B. brachychir and B. periegetes are melodious whistles rather than moans of noisy structure; calls of B. boehmei are series composed of two different note types; calls of B. axelmeyeri and B. rufioculis are composed of series of much shorter notes than those of B. entingae . The strongest similarities are with calls of B. reticulatus which however emits regularly a second call type (a rapid series of short notes) unknown from B. entingae . Boophis entingae furthermore differs from B. boehmei , B. rufioculis , B. burgeri , B. reticulatus and B. axelmeyeri by larger size (SVL of males 46.5–58.5 mm vs. 27–43 mm) and iris colouration. It differs from B. goudoti and B. periegetes by presence of distinct dermal flaps on heels (vs. absence). It differs from B. madagascariensis by presence of a usually distinct, sharply outlined brown marking that extends from upper eyelids and the interorbital region to the posterior dorsum (vs. absence). It differs from B. brachychir by its larger size (SVL of males 46.5–58.5 mm vs. 38.8–46.9 mm) and a reticulated pattern on posterior surfaces of thighs which is absent in B. brachychir . From B. obscurus , resurrected herein, B. entingae is distinguished by the absence of dorsal spicules in breeding males (vs. presence in B. obscurus ), presence of a heel tubercle (vs. absence), and apparently smaller size of males (46.5–58.5 mm vs. 42–82 mm). From B. roseipalmatus it differs by the smaller size of males (46.5–58.5 mm vs. 59–68 mm, see Appendix 2) and absence of pinkish colour on webbing and iris periphery.

Description of holotype. Adult male, SVL 53.3 mm. Body moderately slender; head longer than wide, wider than body; snout long, sub-elliptical in dorsal view, obtuse to acuminate in lateral view; nostrils directed laterally, nearer to eye than to tip of snout; canthus rostralis sharp in cross section, straight in dorsal view, reaching the tip of snout; loreal region concave posteriorly, almost straight anteriorly; eye large; tympanum distinct, rounded, TD 61% of ED; supratympanic fold narrow, prominent; vomerine odontophores distinct, well separated in two elongated patches, positioned median between choanae; choanae medium-sized, rounded. Tongue removed for tissue sample. Arms moderately slender; a small pointed dermal appendage on elbow; thin dermal fold on outer edge of lower arm; subarticular tubercles single, round; inner palmar tubercle narrow, elongated, partly fused with nuptial pad; fingers moderately webbed and with lateral dermal fringes; webbing formula 1(1.5), 2i(1.5), 2e(1), 3i(2), 3e(1.25), 4(1); relative length of fingers 1<2<4<3 (finger 2 distinctly shorter than finger 4); finger discs enlarged. Hind limbs slender; a pointed dermal appendage on heel; tibiotarsal articulation reaching between nostril and tip of snout when hind limb is adpressed along body; lateral metatarsalia separated by webbing; inner metatarsal tubercle medium-sized, distinct, elongated; no outer metatarsal tubercle; toes moderately webbed; webbing formula 1(0), 2i(0.75), 2e(0), 3i(0.75), 3e(0), 4i(1), 4e(1), 5(0.25); relative length of toes 1<2<3=5<4; toe discs enlarged. Skin smooth on dorsal surfaces, smooth on throat and chest, coarsely granular on belly, granular on ventral surface of thighs, prominent scattered tubercles around cloaca.

Measurements (in mm): SVL 53.3, HW 18.2, HL 19.7, ED 7.0, END 3.9, NSD 4.6, NND 5.7, TD 4.3, TL 28.5, HAL 17.3, FOL 24.3, FOTL 39.4.

After six months in preservative, ground colour of upper surface of head, dorsum and limbs greyish brown, with few irregularly scattered black spots; irregular, indistinct dark blotch posterior to eyelids; supratympanic fold bordered with dark brown; upper lip creamy white; outer edges of lower arm with thin white line, bordered by thin brown line ventrally; dorsal surfaces of limbs indistinctly barred; flanks pale beige, with some brown spots in groin; two whitish dots of enlarged tubercles in cloacal region; indistinct beige supracloacal line; posterior surfaces of thighs greyish pale with dark brown reticulation; ventral surfaces creamy beige, with some pale greyish mottling on belly and ventral surfaces of thighs.

In life, ground colour of upper surface of head, dorsum and legs pale grey, with few irregularly scattered black spots; sharply outlined creamy pale brown blotch on middorsum, its corners extending to interorbital region and upper eyelids anteriorly, posteriorly fading at level of urostyl; supratympanic fold bordered with black; loreal region with greyish brown shade; outer edges of lower arm and tarsus with thin white line, bordered by thin blackish line ventrally; dorsal surfaces of limbs with creamy pale brown transversal bars; flanks whitish, with some brown spots in groin; two white dots corresponding to enlarged tubercles in cloacal region; indistinct white supracloacal line; posterior surfaces of thighs light blue with blackish reticulation; throat, chest and venter creamy white, with some indistinct pale greyish mottling on belly; ventral surfaces of thighs, palmar and plantar surfaces with weak bluish to pinkish tint and indistinct pale grey mottling. Iris copper, with some fine black spotting, outer iris area black, iris periphery turquoise green.

Variation. Morphological variation is given in Appendix 2. In preservative, the ground colouration of dorsal surfaces varies from grey to brown, with a variable pattern from mostly uniform to a pattern consisting of diffuse dark brown blotches and minute black flecks. Specimen ZFMK 53639 (male) lacks the dark stripe under the supratympanic fold, and has well-marked minute spicules on dorsal surface. Some specimens (ZSM 624/2001, 830–831/2003, 852/2003, males) have a pale triangle between eyes and snout and two faint, broad dorsolateral bands of the same colour delimiting a darker inverted triangle on middorsum. There is no remarkable variation in ventral colouration. Likewise, there is no sexual dimorphism in colour pattern, but females are larger than males (maximum SVL in females 86.5 vs. 58.5 mm in males). Calling specimens during call emission had a single subgular vocal sac that was only weakly inflated.

Natural history. Males call at night along rainforest streams, from perch heights about 1– 2 m. Streams where this species was found were often relatively slow-moving and with a sandy bottom.

Vocalization. The call of B. entingae from Benavony and Montagne d'Ambre was already briefly described by Glaw & Vences (1992, 1994) as that of B. brachychir . Call type 1 consists of pulsatile notes which might be arranged in series of 2–3. The general harmonic structure becomes unharmonious at the end of the note. Frequency is distributed in a relatively broad frequency bands. Sometimes a second call type is emitted, consisting of short click notes of about 60 ms duration ( Glaw & Vences 1994). In addition to these already published data, we here describe call type 1 recorded at Manongarivo Special Reserve and Andampy (Tsaratanana), as well as the distress call recorded at Montagne d'Ambre National Park. Call type 1 from Manongarivo ( Fig. 11A ): note duration, 232–310 ms (280 ± 30; n = 6); inter-note interval, 226–245 ms (n = 3); note repetition rate (within series), approximately 2 notes/second; dominant frequency range 1700–3200 Hz, maximum call energy at 2200–2360 Hz ( Vences et al. 2006, CD 1, track 61, as B. brachychir ). Call type 1 from Andampy, Tsaratanana, recorded in 2001 at 25–26 °C air temperature is almost identical to call type 1 from Manongarivo in spectral and temporal characters: note duration, 227–331 ms (276 ± 39; n = 10); internote interval (within series), 168–214 ms (190 ± 20; n = 5); note repetition rate (within series), 2.0–2.3 notes/ second; dominant frequency range 1900–3300 Hz, maximum call energy at 2530–2600 Hz. The distress call ( Fig. 11B ) consists of a long pulsed note. Note duration is 589–712 ms (641 ± 43; n = 8). Frequency is distributed in a very broad band of approximately 500–15000 Hz. Several distinct frequency bands are recognizable. Between 500–10000 Hz, 15 of such bands are present, separated from each other by a frequency gap of approximately 500–700 Hz; maximum call energy is at 4740–6790 Hz ( Vences et al. 2006, CD 1, track 61, as B. brachychir ).

Comparative call data. The call of B. entingae differs significantly from that of B. brachychir longer note duration, longer inter-note intervals and much slower note repetition rate. For a detailed comparison, see the numerical parameters of B. brachychir calls from Manongarivo Special Reserve provided above.

Molecular relationships. The molecular phylogenetic analysis is unable to resolve relationships among species in the B. goudoti group ( Fig. 7 ), probably due to a limited number of informative sites in the short 16S rRNA fragment studied. However, each species is resolved as a clearly and usually strongly supported monophyletic unit (except for B. obscurus , see below). The analysis also indicates that several species as currently understood comprise highly divergent mitochondrial lineages, such as B. reticulatus , B. rufioculis , and B. boehmei (each with differences between specimens from the Andasibe and Ranomafana regions). Boophis brachychir and B. entingae are each monophyletic clades and show high genetic divergences to each other (4.5–5.7%) and to other species in the B. goudoti group (4.5–11.5%), while within each of these species the divergences are low: 1.8% between B. brachychir from Montagne d'Ambre and Manongarivo, and 1.1% between B. entingae from Montagne d'Ambre and Manongarivo. Both species show comparatively low divergences of 4.5–5.3% to B. goudoti and B. obscurus (resurrected below), whereas the divergences to B. madagascariensis and B. roseipalmatus (described below) are higher (8.9–11.5%).

Distribution. Boophis entingae is known from its type locality, Montagne d'Ambre National Park, and from Manongarivo Special Reserve (campsites 0 and 1), Tsaratanana Strict Nature Reserve (Manarikoba forest, Andampy campsite), and Benavony (Appendix 10).

Figures

FIGURE 7. Maximum parsimony 16S rRNA phylogram of species of the Boophis goudoti group. From 510 total characters, 340 were constant and 130 parsimony informative. MP searches retained 3681 trees of which a strict consensus is shown. Consensus support values higher than 50, from 2000 bootstrap replicates, are shown; an asterisk indicates Bayesian posterior probabilities equal or higher than 95%. Species newly described or resurrected herein are in bold.

FIGURE 10. Male holotype of Boophis entingae sp. nov. (ZSM 2083/2007): (A) dorsolateral view; (B) ventral view; (C) detail showing colouration of posterior surfaces of thigh.

FIGURE 11. Spectrograms and waveforms of calls of Boophis entingae sp. nov.: (A) advertisement call from Manongarivo Special Reserve (recorded on 2 February 2003, air temperature 22°C); (B) distress call from Montagne d'Ambre National Park (recorded on 14 March 1994, air temperature 21.2°C).