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1 Zootaxa 3956 (2): Copyright 2015 Magnolia Press Article ISSN (print edition) ZOOTAXA ISSN (online edition) A new caruncle-bearing Limnonectes (Anura: Dicroglossidae) from northeastern Thailand ANCHALEE AOWPHOL 1, ATTAPOL RUJIRAWAN 1, WUT TAKSINTUM 1, YODCHAIY CHUAYNKERN 2 & BRYAN L. STUART 3,4 1 Kasetsart University, Faculty of Science, Department of Zoology, Chatuchak, Bangkok, 10900, Thailand 2 Khon Kaen University, Faculty of Science, Department of Biology, Mueang Khon Kaen, Khon Kaen, 40002, Thailand 3 North Carolina Museum of Natural Sciences, 11 West Jones Street, Raleigh NC 27601, USA 4 Corresponding author. bryan.stuart@naturalsciences.org Abstract A new species of the dicroglossid frog genus Limnonectes is described from Ubon Ratchathani Province, northeastern Thailand. Limnonectes lauhachindai sp. nov. differs from its congeners by having males with a low-profiled, U-shaped caruncle with free posterior margin that completely occupies, but does not extend beyond, the interobital region. The new species is most closely related to L. gyldenstolpei and L. dabanus. Its description brings the total number of caruncle-bearing species of Limnonectes to five. Key words: caruncle, Limnonectes gyldenstolpei, Limnonectes dabanus, Ubon Ratchathani Introduction Adult males of four species of Southeast Asian dicroglossid frogs in the genus Limnonectes exhibit extreme secondary sexual characteristics, including enlarged odontoid processes on the lower jaw, hypertrophied heads, and, most unusually, ornamentation consisting of a swollen or cap-like structure (caruncle; Lambertz et al. 2014) on top of their heads (Boulenger 1916; Boulenger 1917; Boulenger 1920; Lambertz et al. 2014; Rowley et al. 2014; Smith 1922; Stuart et al. 2006b). These include L. dabanus (Smith 1922), L. gyldenstolpei (Andersson 1916), L. macrognathus (Boulenger 1917), and L. plicatellus (Stoliczka 1873), four species that represent a monophyletic group (Lambertz et al. 2014). The caruncles in these four species are homologous structures consisting of a dense pad of connective tissue on top of the frontoparietal bones (Lambertz et al. 2014). Caruncles are species-specific in their shape, ranging from a low-profile, domed structure without a free posterior edge (in L. macrognathus), to a large, flap-like, U-shaped structure with a free posterior edge (in L. gyldenstolpei), to a highprofile, horn- or knob-like process (in L. plicatellus) or dome-like structure (in L. dabanus; Lambertz et al. 2014). The function of the caruncles remains unknown, but they may serve a function in male-male combat (Lambertz et al. 2014; Rowley et al. 2014). Our fieldwork in 2004 (reviewed in Stuart et al. 2006a), 2011 and 2012 at two localities in Ubon Ratchathani, northeastern Thailand, revealed an additional species of Limnonectes having males with distinct, cap-like caruncles, but that differed morphologically and genetically from all other known species. Herein, we describe this species as new. Material and methods Sampling. Specimens were collected by hand and fixed in 10% buffered formalin after preserving liver in 20% dimethyl sulfoxide salt-saturated storage buffer or 95% ethanol. Specimens were later transferred to 70% ethanol. 258 Accepted by J. Rowley: 25 Mar. 2015; published: 8 May 2015

2 Specimens and tissue samples were deposited at the Field Museum of Natural History (FMNH), Zoological Museum Kasetsart University, Bangkok, Thailand (ZMKU), and North Carolina Museum of Natural Sciences (NCSM). FMNH specimens were cross-cataloged in the Thailand Natural History Museum (THNHM). Comparative material (Appendix) was examined in the holdings of these institutions and the Natural History Museum, London (BMNH), Muséum national d Histoire naturelle, Paris (MNHN), Museum of Vertebrate Zoology, University of California, Berkeley (MVZ), and La Sierra University Herpetological Collection (LSUHC). Morphology. Measurements were taken to the nearest 0.1 mm with dial calipers: snout-vent length (SVL); head length from tip of snout to rear of jaws (HDL); maximum head width (HDW); snout length from tip of snout to anterior corner of eye (SNT); eye diameter (EYE); interorbital distance (IOD); internasal distance (IND); shank length (SHK); thigh length (TGH); forearm length, from elbow to base of palmar tubercle (LAL); manus length from tip of third digit to base of palmar tubercle (HND); pes length from tip of fourth toe to base of inner metatarsal tubercle (FTL); inner metatarsal tubercle length (IML); and inner metatarsal tubercle width (IMW). Terminology for the cap-like structure, caruncle, follows Lambertz et al. (2014). Molecules. Total genomic DNA was extracted from liver using the PureGene Animal Tissue DNA Isolation Protocol (Gentra Systems, Inc.). A 551 basepair fragment of mitochondrial (mt) DNA that encodes part of the 16S rrna gene (16S) was amplified, sequenced, and edited following Inger & Stuart (2010). Sequences were deposited in GenBank under accession numbers KP KP Additional 16S sequences were downloaded from GenBank of all available caruncle-bearing Limnonectes, representatives of all major clades of Limnonectes (based on Pyron & Wiens 2011), and the outgroups Fejervarya limnocharis and Quasipaa spinosa (based on Pyron & Wiens 2011; Table 1). Sequences were aligned using MAFFT v. 7 (Katoh & Standley 2013). The model of sequence evolution that best described the data (GTR+I+G) was inferred using the Akaike Information Criterion as implemented in jmodeltest 2 (Darriba et al. 2012). Four independent Bayesian analyses were performed using MrBayes 3.2 (Ronquist et al. 2012). In each analysis, four chains were run for 20 million generations using the default priors, trees were sampled every 4,000 generations, and the first 25% of trees were discarded as burn-in. A 50% majority-rule consensus of the sampled trees was constructed to calculate the posterior probabilities of tree nodes. Uncorrected pairwise distances were calculated using PAUP* 4.0b10 (Swofford 2002). Calls. Advertisement calls were recorded at 2230 h at an ambient air temperature of 24.0 C on 19 August 2011 from one male (ZMKU AM 00589) that was calling on the ground in deciduous dipterocarp forest with wet grassy understory. Calls were recorded under natural conditions at a distance of approximately m from the animal using an Edirol R-09 WAVE/MP3 Recorder (44.1 khz sampling rate and 24-bit encoding) and a Røde NTG-2 condenser shotgun microphone. Ambient air temperature was measured using an Extech Instruments digital thermometer (accuracy ±1 C). Calls were analyzed using Raven Pro 1.4 for Mac OS X (Cornell Lab of Ornithology). Calls were edited with a sampling rate of 44.1 khz and 24 bits per sample. The audio spectrograms were achieved using Hanning windows function (512-point fast Fourier transform, 50% overlap, 86.1 Hz resolution) in the spectrum section plot. Temporal and spectral parameters were modified from Cocroft & Ryan (1995) and Thomas et al. (2014) as follows: call duration (the time from beginning to end of one call); intercall interval (duration of the interval between consecutive calls); call rate (the total number of calls-1 divided by the time from beginning of first call to beginning of last call); note (the number of notes in a call); note duration (time between the onset and the offset of the middle note in a call); internote interval (the time interval of the middle notes in a call); note rate (the number of note per second); pulse (the number of pulses in a call); pulse duration (the time between the onset and the offset of the middle pulse in a call); interpulse interval (the time interval of the middle pulse in a call); pulse rate (the number of pulses per second); and dominant call frequency (the frequency of maximum amplitude measured from a power spectrum generated using Raven s selection spectrum function over the duration of the entire call). Results Limnonectes lauhachindai sp. nov. Holotype: NCSM (field tag AA 01384), adult male (Fig. 1), Thailand, Ubon Ratchathani Province, SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 259

3 Sirindhorn District, Kham Khuen Kaew Subdistrict, N E, 131 m elev., coll. 29 August 2012 by Anchalee Aowphol, Siriporn Yodthong, Natee Ampai, and Attapol Rujirawan. FIGURE 1. Holotype (NCSM 80222) of Limnonectes lauhachindai sp. nov. in preservative. (A) Dorsal view. (B) Ventral view. (C) Plantar view of right foot. (D) Profile view of head. Paratypes: NCSM 81269, ZMKU AM (seven adult males): same data as holotype. ZMKU AM (one adult male): same data as holotype except coll. 30 August ZMKU AM , ZMKU AM (nine adult males; Fig. 2): same data as holotype except coll. 19 August 2011 by Attapol Rujirawan, Wut Taksintum, Virayuth Lauhachinda, Anchalee Aowphol, and Siriporn Yodthong. FMNH /THNHM 05185, FMNH /THNHM (two adult males), FMNH /THNHM 05184, FMNH /THNHM (two adult females): Thailand, Ubon Ratchathani Province, Na Chaluai District, Phu Jong-Na Yoi National Park, Huay Luang Noi Stream, 14º N 105º E, 360 m elev., coll. 15 September 2004 by Yodchaiy Chuaynkern, Bryan L. Stuart, Chatchay Chuechat, and Sunchai Makchai. FMNH /THNHM (one adult female): same data as FMNH /THNHM except "N "E, 350 m elev. FMNH /THNHM (one adult female): same data as FMNH / THNHM except "N "E, 325 m elev. Referred material. ZMKU AM (one juvenile male), same data as ZMKU AM Etymology. The specific epithet is a patronym for Associate Professor Dr. Virayuth Lauhachinda, Kasetsart University, and co-collector of the new species, in recognition of his contributions to herpetology in Thailand. Suggested common names. Lauhachinda's Fanged Frog (English), Kob Ngon Arjarn Virayuth (Thai). Diagnosis. Assigned to the genus Limnonectes on the basis of molecular evidence (Fig. 3), the presence of fanglike odontoid processes on the lower jaw (Emerson et al. 2000; Lambertz et al. 2014), and males with hypertrophied heads (Lambertz et al. 2014). A small-sized Limnonectes having males with SVL , females with SVL ; males with low-profiled, U-shaped caruncle with free posterior margin that completely occupies, but does not extend beyond, interobital region; males with hypertrophied heads; both sexes with enlarged odontoid processes on anterior margin of lower jaw; and webbing on toes. 260 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.

4 TABLE 1. Fejervarya, Quasipaa, and Limnonectes 16S sequences used in the molecular analyses. Institutional abbreviations of vouchers follow Sabaj-Pérez (2014), with the additions of PWRC = Phu Luang Wildlife Research Centre Museum; RMB = Rafe M. Brown field series; and ZNAC = Anhui Normal University. Species GenBank No. Voucher Locality Source F. limnocharis NC_ None China, Yancheng Liu et al. (2005) Q. spinosa NC_ None China, Zhejiang Prov., Jinhua Zhou et al. (2009) L. bannaensis NC_ None China, Yunnan Prov., Simao Zhang et al. (2009) L. dabanus AF ROM Vietnam, Yok Don Chen et al. (2005) L. dabanus GU FMNH Cambodia, Mondolkiri Prov., Pichrada Dist. Inger & Stuart (2010) L. doriae GU CAS Myanmar, Pegu State, Bago Yoma Inger & Stuart (2010) L. fragilis AY ZNAC Not available GenBank submission L. fujianensis AY Not available Not available GenBank submission L. gyldenstolpei AF PWRC 002 Thailand, Loei Prov., Phu Luang Emerson et al. (2000) L. gyldenstolpei AY MNHN Vietnam Delorme et al. (2004) L. gyldenstolpei GU FMNH Thailand, Sa Kaeo Prov., Mueang Sa Kaeo Dist. Inger & Stuart (2010) L. hascheanus GU FMNH Thailand, Nakhon Si Thammarat Prov., Khao Luang Inger & Stuart (2010) L. hascheanus GU LSUHC 6777 Malaysia, Penang Inger & Stuart (2010) L. kadarsani AY LSUMZ Indonesia, Lombok Island Evans et al. (2003) L. kohchangae GU FMNH Cambodia, Kampong Speu Prov., Phnom Sruoch Dist. Inger & Stuart (2010) L. khasianus AB KUHE Thailand, Narathiwat Prov., Bala Matsui et al. (2014) L. lauhachindai sp. nov. KP NCSM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP ZMKU AM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP ZMKU AM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP ZMKU AM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP ZMKU AM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP ZMKU AM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP NCSM Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. leporinus AY AMNH Indonesia, Kalimantan Timor Prov., Kutai Evans et al. (2003) L. leytensis JX USNM Philippines, Leyte Island Oaks et al. (2013) L. limborgi AB KUHE Malaysia, Janda Baik Matsui et al. (2014) L. limborgi GU FMNH Cambodia, Mondolkiri Prov., Samling Inger & Stuart (2010) L. macrognathus AB KUHE Thailand, Ranong Prov. Matsui & Nishikawa (2014) L. macrognathus AB KUHE Thailand, Ranong Prov. Matsui et al. (2014) L. macrognathus KJ FMNH Thailand, Nakhon Si Thamarat Prov., Lambertz et al. (2014) L. malesianus AY Not reported Malaysia, Sarawak Prov., Gunung Buda Evans et al. (2003) L. microdiscus AY LSUMZ Indonesia, Java, Sukabumi Evans et al. (2003) L. plicatellus AB IABHU Malaysia, Langkawi Island Hasan et al. (2014) L. plicatellus KJ LSUHC 6710 Malaysia, Pulau Pinang, Empangan Air Hitam Lambertz et al. (2014) L. plicatellus KJ LSUHC 6582 Malaysia, Selangor Dist., Gombak Swamp Lambertz et al. (2014) L. plicatellus KJ LSUHC 4001 Malaysia, Selangor Dist., Kepong Lambertz et al. (2014) L. poilani DQ AMNH A Vietnam, Quang Nam Prov., Tre My Dist. Frost et al. (2006) L. woodworthi JX RMB 4092 Philippines, Luzon Island Oaks et al. (2013) SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 261

5 FIGURE 2. Paratype male (ZMKU AM 00586) of Limnonectes lauhachindai sp. nov. in life. Description of holotype. Habitus moderately stocky; body tapering to groin. Head broad and depressed; head length and width subequal. Snout obtusely pointed in dorsal view, round in profile, projecting well beyond lower jaw in profile; nostril dorsolateral, much closer to tip of snout than to eye, below canthus, internarial distance subequal to interorbital distance; canthus rostralis indistinct, rounded, slightly constricted behind nostrils; lores concave, oblique; eye diameter 80% snout length, interorbital distance greater than upper eyelid width; pineal ocellus visible; tympanum round, not elevated from side of head, annulus weakly visible, tympanum diameter about 82% eye diameter and greater than distance between tympanum and eye; small, slit-like vocal sac openings on floor of mouth near lateral margin of tongue; vomerine teeth on two oblique ridges, equal in distance to each other as to choanae; two large odontoid processes at front of mandible; median triangular protuberance at mandibular symphisis. Forelimb moderately robust. Fingers moderately slender, without webbing; tip of fingers rounded, weakly expanded into discs; relative finger lengths II < IV < I < III; moveable flap of skin on preaxial sides of Fingers II IV; distinct subarticular tubercles, one on Fingers I II, two on Fingers III IV; distinct palmar tubercles, one at base of Finger I, two in contact as base of Fingers II IV; nuptial pad absent. Hindlimb moderately robust. Toes moderately slender; tips of toes rounded, expanded into small discs; relative toe lengths I<II<V<III<IV; webbing on Toe I to base of disc, on preaxial side of Toe II to midway between subarticular tubercle and tip and continuing as a fringe to base of tip, on postaxial side of Toe II to base of tip, on preaxial side of Toe III to level of distal subarticular tubercle continuing as a fringe to base of tip, on postaxial side of Toe III to base of tip, on preaxial and postaxial sides of Toe IV to level of distal subarticular tubercle and continuing as a fringe to base of tip, and on Toe V to base of tip; moveable flap of skin on outer margins of Toes I and V; distinct fold on distal half of tarsus; distinct, elongate, oval, inner metatarsal tubercle, length about 44% distance between tip of toe I and tubercle; no outer metatarsal tubercle. Skin above shagreened with irregular rows of large oval warts, most concentrated near flank and lower back; low-profiled, U-shaped caruncle with free posterior margin that fully occupies, but does not extend beyond, the 262 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.

6 interorbital region; distinct supratympanic ridge from posterior corner of eye to axilla; large rictal gland; no dorsolateral fold; skin on venter smooth. Measurements of holotype given in Table 2. TABLE 2. Measurements (mm) of types of Limnonectes lauhachindai sp. nov. Abbreviations defined in the text. Measurement Holotype male n=1 All males n=20 Range; Mean ± SD Paratype females n=4 Range; Mean ± SD SVL ; 33.8 ± ; 35.4 ± 2.6 HDL ; 15.8 ± ; 14.8 ± 0.9 HDW ; 15.3 ± ; 14.2 ± 1.0 SNT ; 5.6 ± ; 5.5 ± 0.3 EYE ; 4.5 ± ; 4.9 ± 0.2 IOD ; 3.6 ± ; 2.9 ± 0.6 TMP ; 3.7 ± ; 3.1 ± 0.2 IND ; 3.4 ± ; 3.3 ± 0.4 SHK ; 15.8 ± ; 17.7 ± 0.8 TGH ; 16.7 ± ; 17.3 ± 1.8 LAL ; 7.0 ± ; 7.5 ± 0.6 HND ; 7.9 ± ; 8.1 ± 1.0 FTL ; 15.9 ± ; 17.9 ± 1.1 IML ; 2.0 ± ; 2.2 ± 0.4 IMW ; 1.0 ± ; 1.0 ± 0.1 Color of holotype in preservative. Dorsum dark brown; tips of dorsal warts white, warts encircled with black. Venter cream, chin dark brown, throat with large brown spots, belly and ventral surfaces of limbs with very small dark spots. Color of paratype ZMKU AM in life. Dorsum brown, with irregular black spots, becoming brassy on dorsal surfaces of limbs and upper flank; continuous black streak under canthus and supratympanic fold, extending from nostril to upper half of tympanum; lips brown with broad black bars; iris bronze; broad cream-yellow vertebral stripe from anterior margin of upper jaw to vent; upper surfaces of hindlimb with broad black bands; lower flank beige and gray (Fig. 2). Variation. Females lack caruncles and have narrower heads in dorsal view than males. The largest male paratype (ZMKU AM 01111; SVL 42.0 mm) is noticeably larger than the second largest (NCSM 81269; SVL 37.7 mm). Seven (NCSM 81269, ZMKU AM 01106, ZMKU AM , ZMKU AM , ZMKU AM 00593) of 25 (28%) specimens have a pale vertebral stripe, a polymorphic character seen in other Limnonectes, including L. dabanus and L. kohchangae (Stuart & Emmett 2006; Stuart et al. 2006b). Measurements are summarized in Table 2. Molecules. The aligned dataset contained 1,624 characters. The new species was recovered as sister to a clade containing L. dabanus and L. gyldenstolpei (Fig. 3). The holotype and six paratypes (NCSM 81269, ZMKU AM , ZMKU AM 01109) are identical in the 16S gene fragment, but have an uncorrected pairwise divergence of % from L. dabanus (n=2) and % from L. gyldenstolpei (n=3). Calls. Paratype male ZMKU AM had two different advertisement call types, referred to here as Type 1 and Type 2 (Fig. 4). Type 1 (n = 30) had 1 10, one-pulsed notes lasting to ms; note duration of ms; interval between notes ms; note repetition rate of notes per second; and dominant frequency of khz. Type 2 (n = 3) was a single, multi-pulsed note lasting ms with pulses; pulse duration of ms; interval between pulses ms; pulse rate pulses per second; and dominant frequency of khz (Table 3). The advertisement calls were repeated at a rate of approximately 0.12 calls per second and intercall interval varied from 0.36 to s. These calls showed frequency modulation with or without weak harmonics. SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 263

7 FIGURE 3. Fifty percent majority-rule consensus phylogram resulting from Bayesian analysis of the mitochondrial 16S gene from dicroglossid frogs. Numbers at nodes are Bayesian posterior probabilities. Numbers in parentheses are GenBank accession numbers. The source of sequences are given in Table 1. Distribution and natural history. Limnonectes lauhachindai sp. nov. is only known from Na Chaluai and Sirindhorn Districts, Ubon Ratachathani Province, Thailand (Fig. 5), from m elevation. Those from Na Chaluai District were taken on the bank or in the water of a shallow stream flowing over bedrock in semi-evergreen forest on a single day between h. Those from Sirindhorn District were taken on the ground in deciduous dipterocarp forest with wet grassy understory (Fig. 6). Eggs and larvae are unknown. Comparisons. Limnonectes lauhachindai sp. nov. differs from all other species of Limnonectes except L. dabanus, L. gyldenstolpei, L. macrognathus and L. plicatellus by having mature males with a caruncle on top of the head. Limnonectes lauhachindai sp. nov. further differs from these four species by having males with a lowprofiled, U-shaped caruncle with free posterior margin that fully occupies, but does not extend beyond, the interorbital region (caruncle high-profiled and domed in L. dabanus; caruncle extending beyond interorbital region in L. gyldenstolpei; caruncle lacking U-shape and free posterior margin in L. macrognathus; caruncle high-profiled and horned in L. plicatellus; Fig. 7). Limnonectes lauhachindai sp. nov. further differs from L. dabanus, L. gyldenstolpei, and L. macrognathus by having much smaller males [SVL (mean ± SD 59.8 ± 4.0, n = 15) in L. dabanus; SVL (mean ± SD 62.0 ± 5.7, n = 14) in L. gyldenstolpei; SVL (mean ± SD 45.5 ± 3.0, n = 3, this study), types SVL (n = 2; Boulenger 1920) in L. macrognathus], and from L. plicatellus by lacking dorsal rugosities arranged in distinct, longitudinal rows parallel to the body axis (present in L. plicatellus). 264 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.

8 FIGURE 4. Advertisement call of paratype male (ZMKU AM 00589) of Limnonectes lauhachindai sp. nov. recorded at an ambient air temperature of 24.0 C. (A) 26 s waveform showing five representative calls with two different call types: a) Type 1 and b) Type 2, (B) waveform of Type 1 call, (C) spectrogram of Type 1 call, (D) waveform of Type 2 call, and (E) spectrogram of Type 2 call. SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 265

9 FIGURE 5. Map illustrating the type (star) and paratype (circle) localities of Limnonectes lauhachindai sp. nov. in Ubon Ratchathani Province, Thailand. TABLE 3. Measurements of advertisement call parameters of paratype male (ZMKU AM 00589) of Limnonectes lauhachindai sp. nov. Parameter values are given as mean ± SD (and ranges). Call parameter Call type (number of calls) Type 1 (n = 30) Type 2 (n = 3) Note description 1 note = 1 pulse 1 note = multi-pulses Call duration (ms) ± ; ( ) ±207.29; ( ) Intercall interval (ms) ± ; ( ) Call rate (calls/s) Note 4.63±2.59; (1 10) 1 Note duration (ms) 18.37±6.80; ( ) ±207.29; ( ) Internote interval (ms) ±189.90; ( ) - Note rate (notes/s) 2.16±0.78; ( ) - Pulse ±3.21; (31 37) Pulse duration (ms) ±1.97; ( ) Interpulse interval (ms) ±0.82; ( ) Pulse rate (pulses/s) ±0.26; ( ) Dominant frequency (khz) 2.1±0.29; ( ) 2.2±0.05; ( ) 266 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.

10 FIGURE 6. Type locality of Limnonectes lauhachindai sp. nov. in Kham Khuen Kaew Subdistrict, Sirindhorn District, Ubon Ratchathani Province, Thailand. FIGURE 7. Dorsal view of caruncle on top of head (above), with approximate margin of caruncle indicated in red (below). (A) Limnonectes lauhachindai sp. nov. (holotype). (B) L. dabanus (FMNH ). (C) L. gyldenstolpei (NCSM 79550). Limnonectes lauhachindai sp. nov. superficially resembles L. hascheanus (Stoliczka 1870), L. limborgi (Sclater 1892), and L. kohchangae (Smith 1922), but differs from all of them by having cephalic caruncles in males (absent in L. hascheanus, L. limborgi, and L. kohchangae). Limnonectes lauhachindai sp. nov. further differs from L. hascheanus by having larger body size [males with SVL , mean ± SD 22.1 ± 1.9, n = 9, females with SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 267

11 SVL , mean ± SD 23.0 ± 1.6, n = 9, in L. hascheanus (Inger & Stuart 2010)], and from L. hascheanus and L. limborgi by having fully webbed toes (greatly reduced toe webbing in L. hascheanus and L. limborgi). Male secondary sexual characters are unknown in L. khammonensis (Smith 1929), which is known only from the female holotype, but females of L. lauhachindai sp. nov. differ by having a distinct tympanum (indistinct in L. khammonensis) and less toe webbing (Toe IV webbed to distal subarticular tubercle, continuing as fringe to base of disc, and all remaining toes webbed to base of disc in L. khammonensis). Acknowledgements Virayuth Lauhachinda, Natee Ampai, Chatchay Chuechat, Sunchai Makchai, and Siriporn Yodthong provided invaluable assistance with fieldwork. Alan Resetar (FMNH), Barry Clarke (BMNH), Annemarie Ohler (MNHN), Jim McGuire and Carol Spencer (MVZ), and Lee Grismer (LSUHC) provided access to specimens in their care. David McLeod, Jim McGuire, and Jodi Rowley improved the manuscript. Somphouthone Phimmachak produced the map. The John D. and Catherine T. MacArthur Foundation, the U.S. National Science Foundation (grant DEB ), and the Partnerships for Enhanced Engagement in Research (PEER) Science program (grant PGA ), which is a partnership between the U.S. Agency for International Development (USAID) and the National Science Foundation, supported this research. References Andersson, L.G. (1916) Zoological results of the Swedish zoological expeditions to Siam and Batrachians. Kungliga Svenska Vetenskapsakademiens Handlingar, 55, Boulenger, G.A. (1916) Description of a new frog from Siam. The Journal of the Natural History Society of Siam, 2, Boulenger, G.A. (1917) Descriptions of new frogs of the genus Rana. Annals and Magazine of Natural History, Series 8, 20, Boulenger, G.A. (1920) A monograph of the South Asian, Papuan, Melanesian and Australian frogs of the genus Rana. Records of the Indian Museum, 20, Chen, L., Murphy, R.W., Lathrop, A., Ngo, A., Orlov, N.L., Ho, C.T. & Somorjai, I.L.M. (2005) Taxonomic chaos in Asian ranid frogs: an initial phylogenetic resolution. Herpetological Journal, 15, Cocroft, R.B. & Ryan, M.J. (1995) Patterns of advertisement call evolution in toads and chorus frogs. Animal Behaviour, 49, Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jmodeltest 2: more models, new heuristics and parallel computing. Nature Methods, 9, Delorme, M., Dubois, A., Kosuch, J. & Vences, M. (2004) Molecular phylogenetic relationships of Lankanectes corrugatus from Sri Lanka: endemism of South Asian frogs and the concept of monophyly in phylogenetic studies. Alytes, 22, Emerson, S.B., Inger, R.F. & Iskandar, D. (2000) Molecular systematics and biogeography of the fanged frogs of Southeast Asia. Molecular Phylogenetics and Evolution, 16, Evans, B.J., Brown, R.M., McGuire, J.A., Supriatna, J., Andayani, N., Diesmos, A., Iskandar, D., Melnick, D.J. & Cannatella, D.C. (2003) Phylogenetics of fanged frogs: testing biogeographical hypotheses at the interface of the Asian and Australian faunal zones. Systematic Biology, 52, Frost, D.R., Grant, T., Faivovich, J., Bain, R.H., Haas, A., Haddad, C.F.B., de Sá, R.O., Channing, A., Wilkinson, M., Donnellan, S.C., Raxworthy, C.J., Campbell, J.A., Blotto, B.L., Moler, P., Drewes, R.C., Nussbaum, R.A., Lynch, J.D., Green, D.M. & Wheeler, W.C. (2006) The amphibian tree of life. Bulletin of the American Museum of Natural History, 297, Hasan, M., Islam, M.M., Khan, M.M. R., Igawa, T., Alam, M.S., Djong, H.T., Kurniawan, N., Joshy, H., Sen, Y.H., Belabut, D.M., Kurabayashi, A., Kuramoto, M. & Sumida, M. (2014) Genetic divergences of South and Southeast Asian frogs: a case study of several taxa based on 16S ribosomal RNA gene data with notes on the generic name Fejervarya. Turkish Journal of Zoology, 38, Inger, R.F. & Stuart, B.L. (2010) Systematics of Limnonectes (Taylorana) Dubois. Current Herpetology, 29, Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.

12 Katoh, K. & Standley, D.M. (2013) MAFFT Multiple Sequence Alignment software version 7: improvements in performance and stability. Molecular Biology and Evolution, 30, Lambertz, M., Hartmann, T., Walsh, S., Geissler, P. & McLeod, D.S. (2014) Anatomy, histology, and systematic implications of the head ornamentation in the males of four species of Limnonectes (Anura: Dicroglossidae). Zoological Journal of the Linnean Society, 172, Liu, Z.-Q., Wang, Y.-Q. & Su, B. (2005) The mitochondrial genome organization of the rice frog, Fejervarya limnocharis (Amphibia: Anura): a new gene order in the vertebrate mtdna. Gene, 346, Matsui, M. & Nishikawa, K. (2014) Description of a new species of Limnonectes from Sarawak, Malaysian Borneo (Dicroglossidae, Anura). Current Herpetology, 33, Matsui, M., Nishikawa, K. & Eto, K. (2014) A new burrow-utilising fanged frog from Sarawak, East Malaysia (Anura: Dicroglossidae). Raffles Bulletin of Zoology, 62, Oaks, J.R., Sukumaran, J., Esselstyn, J.A., Linkem, C.W., Siler, C.D., Holder, M.T. & Brown, R.M. (2013) Evidence for climate-driven diversification? A caution for interpreting ABC inferences of simultaneous historical events. Evolution, 67, Ohler, A. & Dubois, A. (1999) The identity of Elachyglossa gyldenstolpei Andersson, 1916 (Amphibia, Ranidae), with comments on some aspects of statistical support to taxonomy. Zoologica Scripta, 28, Pyron, R.A. & Wiens, J.J. (2011) A large-scale phylogeny of Amphibia including over 2800 species, and a revised classification of extant frogs, salamanders, and caecilians. Molecular Phylogenetics and Evolution, 61, Ronquist, F., Teslenko, M., van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model spaces. Systematic Biology, 61, Rowley, J.J.L., Le, D.T.T., Hoang, H.D. & Altig, R. (2014) The breeding behaviour, advertisement call and tadpole of Limnonectes dabanus (Anura: Dicroglossidae). Zootaxa, 3881 (2), Sabaj-Pérez, M.H. (Ed.) (2014) Standard symbolic codes for institutional resource collections in herpetology and ichthyology: an online reference. Version 5.0 (22 September 2014). Available from: (accessed 15 January 2015) Sclater, W.L. (1892) On some specimens of frogs in the Indian Museum, Calcutta, with descriptions of several new species. Proceedings of the Zoological Society of London, 1892, Smith, M.A. (1922) The frogs allied to Rana doriae. The Journal of the Natural History Society of Siam, 4, Smith, M.A. (1929) Descriptions of a new skink from Christmas Island and a new frog from Annam. Annals and Magazine of Natural History, Series 10, 3, Stoliczka, F. (1870) Observations on some Indian and Malayan Amphibia and Reptilia. Journal of the Asiatic Society of Bengal, 39, Stoliczka, F. (1873) Notes on some species of Malayan Amphibia and Reptilia. Journal of the Asiatic Society of Bengal, 42, Stuart, B.L. & Emmett, D.A. (2006) A collection of amphibians and reptiles from the Cardamom Mountains, southwestern Cambodia. Fieldiana, Zoology New Series, 109, Stuart, B.L., Chuaynkern, Y., Chan-ard, T. & Inger, R.F. (2006a) Three new species of frogs and a new tadpole from eastern Thailand. Fieldiana, Zoology New Series, 111, Stuart, B.L., Sok, K. & Neang, T. (2006b) A collection of amphibians and reptiles from hilly eastern Cambodia. The Raffles Bulletin of Zoology, 54, Swofford, D.L. (2002) PAUP*: Phylogenetic Analysis Using Parsimony *(and other methods). Version 4.0b10. Sinauer Associates, Sunderland, Massachusetts. Thomas, A., Suyesh, R., Biju, S.D. & Bee, M.A. (2014) Vocal behavior of the elusive purple frog of India (Nasikabatrachus sahyadrensis), a fossorial species endemic to the Western Ghats. PLoS One 9, e Zhang, J.-F., Nie, L.-W., Wang, Y. & Hu, L.-L. (2009) The complete mitochondrial genome of the large-headed frog, Limnonectes bannaensis (Amphibia: Anura), and a novel gene organization in the vertebrate mtdna. Gene, 442, SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 269

13 Zhou, Y., Zhang, J.-Y., Zheng, R.-Q., Yu, B.-G. & Yang, G. (2009) Complete nucleotide sequence and gene organization of the mitochondrial genome of Paa spinosa (Anura: Ranoidae (sic)). Gene, 447, APPENDIX. Comparative specimens examined. Limnonectes dabanus: VIETNAM, Langbian Plateau, Daban, BMNH , BMNH (syntypes); Binh Thuan Province, Bac Binh District, NCSM CAMBODIA, Mondolkiri Province, Keo Seima District, FMNH ; Mondolkiri Province, O Rang District, FMNH , FMNH , FMNH ; Mondolkiri Province, Pichrada District, FMNH , FMNH , FMNH ; Ratanakiri Province, Ta Veng District, FMNH , FMNH ; Ratanakiri Province, Veunsai District, MVZ , MVZ ; Stung Treng Province, Siem Pang District, FMNH , ; Tbong Khmum Province, Memot District ( Mimot ), MNHN (holotype of Rana toumanoffi). Limnonectes doriae: MYANMAR ( Burma ), Tenasserim, Molleyil, FMNH Limnonectes gyldenstolpei: CAMBODIA, Kampong Speu Province, Aural District, NCSM LAOS, Xaignabouli Province, Xaignabouli District, NCSM ; Vientiane Province, Xaysomboun District, NCSM Limnonectes hascheanus: MALAYSIA, Penang, LSUHC 6777, LSUHC 6783, LSUHC , LSUHC THAILAND, Yala Province, FMNH , FMNH ; Trang Province, FMNH Limnonectes khammonensis: LAOS, Napé, BMNH (holotype). Limnonectes kohchangae: THAILAND ( Siam ), Trat Province, Ko Chang District, FMNH , BMNH , BMNH , BMNH (all syntypes). CAMBODIA, Kampong Speu Province, Aural District, NCSM , NCSM ; Kampong Speu Province, Phnom Sruoch District, FMNH , FMNH ; Kampot Province, Kampot District, FMNH ; Pursat Province, Veal Veang District, NCSM 80259; Pursat Province, Kravanh District, FMNH ; Koh Kong Province, Sre Ambel District, FMNH ; Koh Kong Province, Thmar Baing District, FMNH , Limnonectes limborgi: MALAYSIA, Selangor, Bukit Lanjan, FMNH , FMNH , FMNH , FMNH , FMNH , FMNH , FMNH , FMNH THAILAND, Chiang Mai Province, Doi Suthep, FMNH , FMNH Limnonectes macrognathus: THAILAND, Nakhon Si Thammarat Province, Khao Luang National Park, FMNH ; Nakhon Si Thammarat Province, Na Bon, FMNH ; Surat Thani Province, Khao Sok National Park, FMNH , FMNH Limnonectes plicatellus: MALAYSIA, Selangor State, Petaling District, Bukit Lanjan, FMNH Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.