Communicator whistles: A Trek through the taxonomy of the Boophis marojezensis complex reveals seven new, morphologically cryptic treefrogs from Madagascar (Amphibia: Anura: Mantellidae) Author Vences, Miguel 0000-0003-0747-0817 Zoologisches Institut, Technische Universität Braunschweig, Mendelssohnstr. 4, 38106 Braunschweig, Germany Author Köhler, Jörn 0000-0002-5250-2542 Hessisches Landesmuseum Darmstadt, Friedensplatz 1, 64283 Darmstadt, Germany Author Hutter, Carl R. 0000-0001-6381-6339 Museum of Natural Sciences and Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA Author Preick, Michaela 0000-0002-8014-1975 Institut für Biochemie und Biologie, Universität Potsdam, Karl-Liebknecht-Str. 24 - 25, 14476 Potsdam, Germany Author Petzold, Alice 0000-0002-0743-2004 Institut für Biochemie und Biologie, Universität Potsdam, Karl-Liebknecht-Str. 24 - 25, 14476 Potsdam, Germany Author Rakotoarison, Andolalao 0000-0003-2620-440X Mention Environnement, Université de l’Itasy, Faliarivo Ambohidanerana, 118 Soavinandriana Itasy, Madagascar & School for International Training, VN 41 A Bis Ankazolava Ambohitsoa, Antananarivo, 101 Madagascar Author Ratsoavina, Fanomezana M. 0000-0003-1661-1669 Mention Zoologie et Biodiversité Animale, Université d’Antananarivo, BP 906, Antananarivo, 101 Madagascar Author Glaw, Frank 0000-0003-4072-8111 Zoologische Staatssammlung München (ZSM-SNSB), Münchhausenstr. 21, 81247 München, Germany Author Scherz, Mark D. 0000-0002-4613-7761 Natural History Museum of Denmark, University of Copenhagen, Universitetsparken 15, 2100 Copenhagen Ø, Denmark text Vertebrate Zoology 2024 2024-10-14 74 643 681 journal article 10.3897/vz.74.e121110 0228B083-CB4C-4DE3-8332-58DD834E7AC2 Boophis picardi sp. nov. Lineage D Figures 6 , 9 Identity. This species has previously been referred to as B. sp. Ca 68 in Hutter et al. (2018) . It was included in B. marojezensis sensu lato by Glaw et al. (2001) , Glaw and Vences (2007) , Vieites et al. (2009) , and Rosa et al. (2012) , and not explicitly included or mentioned in the studies of Randrianiaina et al. (2012) , and Perl et al. (2014) . Adult specimens from Mandraka considered to represent B. majori by Blommers-Schlösser (1979 b ) are probably to be referred to this species. Holotype . ZSM 264 / 2006 ( ZCMV 1447), adult male , collected by D. R. Vieites , M. Vences , F. Rabemananjara , P. Bora , C. Weldon , and J. Patton on 7–8 February 2006 at An’Ala ( 18.9193 ° S , 48.4880 ° E , 889 m a. s. l. ), Northern Central East of Madagascar . Paratypes . ZSM 262 / 2006 ( ZCMV 1436), ZSM 263 / 2006 ( ZCMV 1437), ZSM 265 / 2006 ( ZCMV 2364), three adult males with same collection data as holotype . ZSM 266 / 2006 ( ZCMV 2403), ZSM 1969 / 2006 ( ZCMV 1494), ZSM 1970 / 2006 ( ZCMV 2417), three adult males with same collecting locality and collectors as holotype, but collected between 7–10 February 2006 . ZSM 326 / 2000 , adult male , collected by F. Glaw on 10 April 2000 at Vohidrazana ( 18.9658 ° S , 48.5103 ° E , 731 m a. s. l.) . ZSM 189 / 2002 ( FGMV 2001.1160), adult male , collected by M. Vences on 26–27 November 2001 at Vohidrazana ( 18.9658 ° S , 48.5103 ° E , 731 m a. s. l.) . ZSM 1021 / 2003 ( FGMV 2002.2359), adult male , collectors unknown, collected in 2003 at the type locality An’Ala (ca. 18.9193 ° S , 48.4880 ° E , ca. 880 m a. s. l.) . KU 340631 ( CRH 261), adult female , and KU 340641 ( CRH 275), adult male , collected by C. R. Hutter , S. M. Lambert , Z. F. Andriampenomanana , and S. Justin on 10 December 2014 at Vohimana ( 18.9209 ° S , 48.5122 ° E , 787 m a. s. l.) . Definition. A small treefrog assigned to the genus Boophis , subgenus Boophis , in the family Mantellidae based on occurrence on Madagascar , presence of intercalary element between ultimate and penultimate phalanx of fingers and toes (verified by external examination), presence of webbing between fingers, presence of nuptial pads in males, and absence of femoral glands in males. Assigned to the Boophis blommersae group based on small body size (male SVL 21.3–23.2 mm ), predominantly brownish dorsal coloration, absence of red color on webbing or ventral side of limbs of many specimens, calling males along streams, and molecular phylogenetic relationships. Within the B. blommersae group, defined by absence of dorsolateral bands, presence of distinct red color in outer iris area, especially its dorsal and ventral edges, in most specimens, and advertisement calls with high dominant frequencies of 4903–5819 Hz consisting of 17–25 whistling notes comprising multiple short (19‒78 ms) and a few long notes (90–225 ms). Also characterized by numerous diagnostic nucleotide positions in the mitochondrial 16 S rRNA gene: MolD identified the following robust diagnostic nucleotide combination compared to all other species in the B. marojezensis complex (sites given relative to the full-length 16 S sequence of Mantella baroni ): “ G ” in the site 177, “ G ” in the site 233, “ C ” in the site 314. Diagnosis. Within the B. blommersae group, distinguished from B. blommersae by calls mainly consisting of frequency-modulated whistles (vs. pulsed trills); and from B. vittatus by calls mainly consisting of frequency-modulated whistles (vs. series of short clicks), and absence of dorsolateral stripes (vs. presence). Furthermore, distinguished from B. marojezensis by presence of red color in outer iris area in most specimens (vs. absence), and advertisement calls consisting of 17–25 notes (vs. 7–8 notes), and from B. kirki sp. nov. by advertisement calls consisting of two types of (short and long) notes (vs. notes of successively increasing duration). For a distinction from other species of the B. marojezensis complex described herein, see accounts of these new species below. Description of the holotype . Adult male, in good state of preservation, SVL 23.0 mm, tissue from right thigh removed as tissue samples for molecular analysis and posterior venter cut open for parasitological examination. Body slender; head wider than long, wider than body; snout rounded in dorsal view, truncate in lateral view; nostrils directed laterally, about equidistant to tip of snout and eye; canthus rostralis indistinct, slightly concave in dorsal view, loreal region slightly concave; tympanum indistinct but recognizable, round, TD about 44 % of ED ; supratympanic fold very indistinct, largely straight; vomerine odontophores weakly developed, well-separated in two very small rounded aggregations, positioned posteromedial to choanae; choanae medium-sized, rounded; maxillary teeth present. Tongue ovoid, posteriorly bifid, free. Arms slender, forearms of slightly larger diameter, subarticular tubercles single, round; metacarpal tubercles not recognizable; fingers weakly webbed and with lateral dermal fringes; webbing formula 1 (traces), 2 i (traces), 2 e (traces), 3 i (traces), 3 e (2), 4 (1.5); relative length of fingers 1 <2 <4 <3 (finger 2 distinctly shorter than finger 4); finger discs enlarged, rounded; nuptial pads indistinct, recognizable as unpigmented weak swelling on first finger. Hindlimbs slender; tibiotarsal articulation reaching nostril when hindlimb is adpressed along body; lateral metatarsalia separated by webbing; inner metatarsal tubercle small, distinct, elongated; no outer metatarsal tubercle; toes broadly webbed; webbing formula 1 (0.5), 2 i (0.75), 2 e (0.25), 3 i (1), 3 e (0), 4 i (1.75), 4 e (1.5), 5 (0.5); relative length of toes 1 <2 <3 <5 <4; toe discs enlarged, rounded. Skin smooth on dorsal surfaces, throat, chest, and ventral surface of thighs, finely granular on belly; cloacal region surrounded by an area of distinct, large, white-colored granules. In preservative, 17 years after collection (Fig. 6 ), dorsally reddish brown with a distinct and moderately contrasted dark brown hourglass marking on anterior part of the dorsum, and a dark brown transverse bar on the posterior part of the dorsum. Many dark spots of different sizes are scattered across the dorsum. Limbs light brown with a few rather poorly contrasted darker brown crossbands: 1–2 on forearm, 2–3 on shank, 2–3 on thigh. Ventrally cream, white on belly, with some dark pigment only on ventral side of feet. Color of holotype in life not recorded. Variation. Several paratypes from An’Ala in preservative are characterized by a distinct dorsal hourglass pattern (plus additional patch posterior to it), particularly contrasted in ZSM 1970 / 2006 and 1969 / 2006. ZSM 268 / 2006 has a contrasted pinkish patch above the right eye, ZSM 326 / 2000 has a pinkish marking on the central dorsum and many small white-pinkish spots on the anterior dorsum, whereas ZSM 265 / 2006 has the dorsum covered with numerous larger pink patches (ca. 15 partly fused patches). ZSM 189 / 2002 from Vohidrazana features, in addition to the dorsal hourglass marking, a fine light vertebral line. In life, the dark dorsal pattern is often only weakly recognizable (Fig. 9 ). In one paratype ( KU 340641 ), the hindlimbs have an orange tint in life but no deep red color is recognizable (Fig. 9 B ). The outer iris color can be deep red (Fig. 9 C ; see also Rosa et al. 2012 ) or light orange (Fig. 9 A ). Iris periphery light blue. Figure 9. Individuals of Boophis picardi sp. nov. in life. A, B male paratype KU 340641 ( CRH 275) from Vohimana in dorsolateral and ventral view. C, D male paratypes from An’Ala, not assignable to specific voucher specimens. Etymology. Named after the fictional character Captain Jean-Luc Picard, first portrayed by Sir Patrick Stewart in Gene Roddenberry’s Star Trek: The Next Generation, and later in Akiva Goldsman, Michael Chabon, Kirsten Beyer, and Alex Kurtzman’s Star Trek: Picard. Tadpoles. The tadpole of this species is unknown. Natural history. An arboreal, nocturnal treefrog found in humid rainforests along fast flowing streams. Little is known of the ecology of the species. At Vohidrazana, male specimens were collected calling along a stream, from 1 meter to occasionally several meters above the ground. Many other calling males could be heard calling higher but were unable to be reached. Calls. Advertisement calls of B. picardi sp. nov. recorded at An’Ala on 12 February 1995 (21.5 ° C air temperature) consist of two different note types, namely a series of rather short, quickly repeated notes, followed by 2–3 distinctly longer notes separated by slightly longer intervals. All notes are tonal in character and exhibit a distinct upward frequency modulation, with a frequency shift comprising 400–500 Hz in long notes and about half that in short notes. Amplitude across the entire call is increasing, with the first short notes being relatively soft, reaching maximum call energy at about one third of the call’s duration. Within notes, no distinct amplitude modulation is recognizable. Numerical parameters of three analyzed calls of different individuals are as follows: call duration 1554–1832 ms (1673.3 ± 143.1 ms); notes / call 19–21 (20.3 ± 1.2); short note duration 24–53 ms (34.4 ± 6.6 ms); long note duration 112–200 ms (157.9 ± 26.8 ms); inter-note interval 25–96 ms (41.1 ± 17.5 ms); note repetition rate of short notes within the call vary around 16 calls / second; dominant frequency 4903–5444 Hz (5267 ± 152 Hz); prevalent bandwidth 4200–5600 Hz. Calls of B. picardi sp. nov. recorded at Betampona on 30 October 2007 , 22: 30 h (air temperature 18 ° C) (from Rosa et al. 2011 , 2012 ), are in general agreement with those described from An’Ala above. A slight difference is obvious for overall amplitude modulation across the call, with amplitude of notes slightly decreasing again after having reached maximum call energy at one third of the call’s duration. Also, inter-note intervals are slightly longer compared to calls from An’Ala. Numerical parameters of two analyzed calls are as follows: call duration 1991–2000 ms; notes / call 17; short note duration 28–43 ms (32.8 ± 5.1 ms); long note duration 90–205 ms (140.8 ± 39.4 ms); inter-note interval 42–119 ms (64.6 ± 24.3 ms); note repetition rate of short notes within the call vary around 13 calls / second; dominant frequency 5442–5819 Hz (5667 ± 111 Hz); prevalent bandwidth 4500–6200 Hz. The character of calls of B. picardi sp. nov. from Vohidrazana, recorded on 17 February 2001 (air temperature not recorded), generally agrees with those from An’Ala and Betampona described above, but Vohidrazana calls contain more long notes (6) following the short notes. Numerical parameters of two analyzed calls are as follows: call duration 2362–2388 ms; notes / call 24; short note duration 27–62 ms (37.8 ± 10.9 ms); long note duration 112–162 ms (139.0 ± 17.4 ms); inter-note interval 28–145 ms (57.5 ± 33.7 ms); note repetition rate of short notes within the call vary around 17 calls / second; dominant frequency 5292–5560 Hz (5417 ± 89 Hz); prevalent bandwidth 4900–5800 Hz. Calls recorded at Vohidrazana, in December 2015 and January 2016 (air temperatures 17.9 and 20.3 ° C) and corresponding to call vouchers KU 342939 ( CRH 971) and KU 342967 ( CRH 1044) have the following numerical parameters (six calls analyzed): call duration 2010–2376 ms (2182.3 ± 119.7 ms); notes / call 20–25 (22.2 ± 1.8); short note duration 28–78 ms (38.0 ± 12.6 ms); long note duration 124–209 ms (163.3 ± 29.6 ms); inter-note interval 27–122 ms (55.8 ± 28.0 ms); note repetition rate of short notes within the call vary around 15 calls / second; dominant frequency 4971–5122 Hz (5060 ± 64 Hz); prevalent bandwidth 4200–5500 Hz. Calls recorded at Mantadia, on 14 January 2017 (air temperature 19.1 ° C) and corresponding to the voucher specimen KU 347246 ( CRH 1932) are also in agreement with the calls described above. Numerical parameters of three analyzed calls are as follows: call duration 2012–2330 ms (2165.0 ± 159.3 ms); notes / call 21–24 (23.0 ± 1.7); short note duration 19–75 ms (32.6 ± 14.0 ms); long note duration 123–225 ms (169.3 ± 33.9 ms); inter-note interval 35–89 ms (53.4 ± 17.7 ms); note repetition rate of short notes within the call vary around 15 calls / second; dominant frequency 5033–5388 Hz (5178 ± 119 Hz); prevalent bandwidth 4300–5600 Hz. Distribution. According to the molecular data summarized herein, the species is known from several sites in the wider area around the village of Andasibe, i. e., (1) the type locality, An’Ala, (2) Vohidrazana, (3) Vohimana, and it also has been recorded from (4) Betampona (Sahambendrana and Sahabefoza sites, according to Rosa et al. 2012 ). The elevational range spans from 349 (Betampona, Sahabefoza) to 880 m a. s. l. (An’Ala).