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A New Species of Rain Frog from Namaqualand, South Africa (Anura: Brevicipitidae: Breviceps)

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A New Species of Rain Frog from Namaqualand, South Africa (Anura: Brevicipitidae: Breviceps) Zootaxa 3381: 62–68 (2012) ISSN 1175-5326 (print edition) www.mapress.com/zootaxa/ Article ZOOTAXA Copyright © 2012 · Magnolia Press ISSN 1175-5334 (online edition) A new species of Rain Frog from Namaqualand, South Africa (Anura: Brevicipitidae: Breviceps) ALAN CHANNING Biodiversity and Conservation Biology Department, University of the Western Cape, Private Bag X17, Bellville, 7525, South Africa. E-mail: [email protected] Abstract Breviceps branchi sp. nov. is described from coastal Namaqualand, South Africa. It is most similar to Breviceps na- maquensis in colour pattern and overall form, from which it differs by hand and foot morphology and 16S rRNA sequence. Key words: Breviceps, new species, Namaqualand, 16S rRNA, South Africa Introduction The genus Breviceps is known from South Africa northwards to Kenya, and as far west as Angola, with the closely related Balebreviceps found in Ethiopia (IUCN 2011). There are presently 15 species recognised (Frost 2011). The early taxonomy of the genus Breviceps was reviewed by Power (1926), by which time seven species were already known, including the Namaqualand endemics, B. macrops and B. namaquensis. Power (1926) discussed a number of characters that might be useful in separating species of rain frogs. On the basis of differences in 16S rRNA and morphology, I describe a new species of Breviceps from Namaqualand. Material and methods Sampling. A single specimen was collected in Namaqualand, South Africa. A small tissue sample was removed from thigh muscle, and the specimen was fixed in formalin for 24 h, then transferred to 70% ethanol for deposition in the herpetological collection of the Museum für Naturkunde, Leibniz Institute for Research on Evolution and Biodiversity at the Humboldt University, Berlin (ZMB). The specimen was compared to B. macrops, B. gibbosus and the types and additional material of B. namaquensis. Measurements. The following measurements were taken and descriptors recorded: Snout-urostyle length SUL, head width at angle of mouth (HW), interorbital distance measured across the top of the head (IO), eyelid length (EL), distance between the anterior corners of the eyes (EE), internarial distance (NN), length of inner meta- tarsal tubercle (IMT), length of median flange on inner metatarsal tubercle (FGL), length of first toe from inner metatarsal tubercle (T1), length of foot from tip of fourth toe including the outer metatarsal tubercle (F), length of hand from tip of third finger including the palmar tubercle (H), number of tubercles under the third finger (F3T). DNA extraction, amplification and sequencing. Tissues were digested using standard Proteinase-K protocol, and DNA was extracted using phenol-chloroform (Hillis et al. 1996). A 550 bp fragment of the mt 16S gene was amplified using the primers 16SaR-F and 16SbR-R of Kocher et al. (1989), as modified by Bossuyt & Milinko- vitch (2000). Forward and reverse strands were sequenced. Purification and sequencing of both strands was carried out by the Central Analytical Facility of Stellenbosch University. Both sequences were checked against the chro- matograms, trimmed, and combined into a single contig for each fragment using Sequencher 4.9 (GeneCodes Cor- poration). Sequences were checked using BLAST to confirm their placement in the genus (http:// blast.ncbi.nlm.nih.gov/). The sequence is deposited in GenBank, accession number JQ965934. A comparative sequence from B. namaquensis was determined and deposited in GenBank, accession number JQ965933. 62 Accepted by M. Vences: 24 May 2012; published: 6 Jul. 2012 A comparison of the 16S rRNA sequence was made with those of B. macrops (Channing & Wahlberg 2011), B. namaquensis (this study), B. fuscus (Van der Meijden et al. 2005) and B. mossambicus (Van der Meijden et al. 2005, Loader et al. 2004, Frost et al. 2006, Greenbaum et al. 2011) which were available in GenBank. Results Breviceps branchi sp. nov. Branch's rain frog Figs. 1 & 3 Holotype. A male, ZMB 77781, field number AC 3053 (Fig 1), collected at the gate to the diamond security area 600 m west of the Holgat River bridge that carries traffic from Port Nolloth to Alexander Bay in Namaqualand, South Africa, 16 October 2008, 28°55'58.4" S; 16°46'08.9" E. For comparison a specimen of Breviceps namaquen- sis is included in Figure 1. The type locality is shown in Figure 2. Diagnosis. A brevicipitid frog, placed in the genus Breviceps as it does not have squared terminal discs on the phalanges as Callulina; nor a red V on the head and strong transverse ridges posteriorly on the palate as Splaeophryne; nor strongly granular skin and a downturned vent as Probreviceps (Channing & Howell 2006). The latter three genera are restricted to montane forests on the eastern side of the continent, while Breviceps is wide- spread and common in arid areas in the western parts of the continent. The new species most closely resembles Breviceps namaquensis in form and colour pattern, from which it can- not be distinguished on differences in body proportions, such as the ratios of HW/SUL, F/SUL, and EE/HW. Based on the description of the type series of Breviceps namaquensis (Power 1926), the following differences are appar- ent (B. namaquensis values in parentheses): Interorbital space 40% of upper eyelid (50%), fourth finger 50% length of second (60–80%), double subarticular tubercles under third and fourth toes (single conical tubercles), soles of feet granulated (smooth), inner metatarsal tubercle well developed, 20 degrees to axis of fourth toe (not well devel- oped, 30–50 degrees), body with even granulations (smooth at least anteriorly), four spots mid-dorsally (two spots), elongated patch on either side of vertebral line in sacral region (spot on either side). B. branchi sp. nov. is further distinguished from B. namaquensis by the many small granules under the fourth toe (23) compared to the 4–9 of B. namaquensis. The third finger is also more granulated, with 24 small tubercles under the third finger, but no more than 10 in B. namaquensis (Fig. 3). The dark band under the eye extends back to the arm insertion, but does not reach the arm in most B. namaquensis. I examined all the specimens of B. namaquensis in South African institutions (see material listed below), and found that this species never has such granular digits. The granular fingers and toes separate this species from B. acutirostris, B. adspersus, B. bagginsi, B. fichus, B. fuscus, B. gibbosus, B. macrops, B. montanus, B. mossambicus, B. rosei, B. sopranus, B. sylvestris and B. verrucosus when compared to the illustrations in Du Preez & Carruthers (2009) and Channing & Minter (2004). The 16S mt rRNA fragment differs by 4.5% from B. namaquensis, 5.2% from B. macrops, 8.0% from B. fus- cus, and 13% from B. mossambicus. This is within the range of differences between other congeneric species of amphibians (Fouquet et al. 2007). Description of the Holotype. A male, snout-urostyle length 41.3 mm. Top of head smooth, upper eyelid slightly granular, with six pale tubercles along margin of eyelid. Lower eyelid white with transparent patches and fine dark speckling. Snout protruding and angular in outline, nostrils thin slits with pale patch on the outer rim. Internostril distance 2 mm. Measurements (mm) of the holotype and a summary of the measurement of 14 B. namaquensis (in parentheses) follow: SUL 41.3 (19.5–51.9, mean 37.1); HW 12.1 (8.2–14.6, mean 12.5); NN 2.0 (1.7–2.5, mean 2.1); IO 2.4 (2.1–4.5, mean 3.6); EL 6.0 (4.8–7.6, mean 6.4); EE 6.6 (4.4–8.5, mean 7.1); IMT 3.8 (1.3–4.7, mean 3.3); FGL 1.9 (0.9–2.8, mean 1.9); T1 5.6 (2.7–7.2, mean 5.2); F 13.8 (7.2–17.5, mean 13.4); H 9.9 (5.5–12.9, mean 10.0); F3T 24 (2–10, mean 6.6). Fingers well developed, smooth above. Relative finger lengths 3>2>1>4. Fingers with double subarticular and supernumerary tubercles, with 24 tubercles under the third finger (Fig. 3). Palm very granulated. Inner metacarpal tubercle double, rounded, only slightly protruding. Outer metacarpal tubercle flat, divided. Toes well developed, smooth above, but very granulated below. Double subarticular tubercles present on toes 2, 3, and 4. Only the tip of toe 1 extends beyond the fleshy webbing and sole. Fleshy webbing between toes 3 and 4 extends beyond the level NEW SPECIES OF BREVICEPS FROM NAMAQUALAND Zootaxa 3381 © 2012 Magnolia Press · 63 of toe 2. Inner metatarsal tubercle elongated, protruding, aligned at 20° to the fourth toe axis. Outer metatarsal tubercle well developed, almost in line with the inner metatarsal tubercle. Throat and ventrum with minute white asperities, and a transparent belly patch. Sides of body with numerous small, white-tipped glands, each with three or more openings. There are 18 of these glands counted along a straight line between arm and leg. FIGURE 1. A—Holotype of Breviceps branchi sp. nov (ZMB 77781), Photo courtesy K. Wahlberg, B—Breviceps namaquen- sis from Farm Brasil, Namaqualand, South Africa (ZMB 77780). 64 · Zootaxa 3381 © 2012 Magnolia Press CHANNING FIGURE 2. Map showing the collecting locality (square symbol) of Breviceps branchi sp. nov. AB–Alexander Bay, PN–Port Nolloth, South Africa. Map courtesy Google Earth. Colour in life. The back is a pale yellow-brown, with darker brown markings. There are paired pale paradorsal patches, with a pale bar from eye to eye over the head. A dark band runs from below the eye to the arm insertion. Numerous small white glandular warts are present on the skin, most conspicuous along the sides (Fig.
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