Zootaxa 3956 (2): 258 270 www.mapress.com/zootaxa/ Copyright 2015 Magnolia Press Article http://dx.doi.org/10.11646/zootaxa.3956.2.6 http://zoobank.org/urn:lsid:zoobank.org:pub:270500f3-c33e-434b-b5f1-1fdb7a856ad9 ISSN 1175-5326 (print edition) ZOOTAXA ISSN 1175-5334 (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. E-mail: 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
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 KP939072 KP939078. 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 0.2 0.3 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 80222 (field tag AA 01384), adult male (Fig. 1), Thailand, Ubon Ratchathani Province, SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 259
Sirindhorn District, Kham Khuen Kaew Subdistrict, 15 17 47.6 N 105 28 22.0 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 01104 09 (seven adult males): same data as holotype. ZMKU AM 01111 (one adult male): same data as holotype except coll. 30 August 2012. ZMKU AM 00552 53, ZMKU AM 00586 92 (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 266148/THNHM 05185, FMNH 266154/THNHM 05189 (two adult males), FMNH 266147/THNHM 05184, FMNH 266150/THNHM 05025 (two adult females): Thailand, Ubon Ratchathani Province, Na Chaluai District, Phu Jong-Na Yoi National Park, Huay Luang Noi Stream, 14º26 15.9 N 105º16 48.2 E, 360 m elev., coll. 15 September 2004 by Yodchaiy Chuaynkern, Bryan L. Stuart, Chatchay Chuechat, and Sunchai Makchai. FMNH 266151/THNHM 05026 (one adult female): same data as FMNH 266147/THNHM 05184 except 14 25 43.9"N 105 16 51.0"E, 350 m elev. FMNH 266152/THNHM 05187 (one adult female): same data as FMNH 266147/ THNHM 05184 except 14 26 18.6"N 105 16 04.5"E, 325 m elev. Referred material. ZMKU AM 00593 (one juvenile male), same data as ZMKU AM 00586. 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 30.5 42.0, females with SVL 32.9 37.9; 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.
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_005055 None China, Yancheng Liu et al. (2005) Q. spinosa NC_013270 None China, Zhejiang Prov., Jinhua Zhou et al. (2009) L. bannaensis NC_012837 None China, Yunnan Prov., Simao Zhang et al. (2009) L. dabanus AF206496 ROM 22081 Vietnam, Yok Don Chen et al. (2005) L. dabanus GU934329 FMNH 261937 Cambodia, Mondolkiri Prov., Pichrada Dist. Inger & Stuart (2010) L. doriae GU934330 CAS 208425 Myanmar, Pegu State, Bago Yoma Inger & Stuart (2010) L. fragilis AY899241 ZNAC 11006 Not available GenBank submission L. fujianensis AY974191 Not available Not available GenBank submission L. gyldenstolpei AF183124 PWRC 002 Thailand, Loei Prov., Phu Luang Emerson et al. (2000) L. gyldenstolpei AY880440 MNHN 1998.4150 Vietnam Delorme et al. (2004) L. gyldenstolpei GU934331 FMNH 266203 Thailand, Sa Kaeo Prov., Mueang Sa Kaeo Dist. Inger & Stuart (2010) L. hascheanus GU934337 FMNH 270118 Thailand, Nakhon Si Thammarat Prov., Khao Luang Inger & Stuart (2010) L. hascheanus GU934349 LSUHC 6777 Malaysia, Penang Inger & Stuart (2010) L. kadarsani AY313693 LSUMZ 81722 Indonesia, Lombok Island Evans et al. (2003) L. kohchangae GU934332 FMNH 263210 Cambodia, Kampong Speu Prov., Phnom Sruoch Dist. Inger & Stuart (2010) L. khasianus AB981414 KUHE 23158 Thailand, Narathiwat Prov., Bala Matsui et al. (2014) L. lauhachindai sp. nov. KP939072 NCSM 80222 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939073 ZMKU AM 01104 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939074 ZMKU AM 01105 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939075 ZMKU AM 01106 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939076 ZMKU AM 01107 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939077 ZMKU AM 01109 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. lauhachindai sp. nov. KP939078 NCSM 81269 Thailand, Ubon Ratchathani Prov., Sirindhorn Dist. This study L. leporinus AY313691 AMNH 167165 Indonesia, Kalimantan Timor Prov., Kutai Evans et al. (2003) L. leytensis JX911319 USNM 222545 Philippines, Leyte Island Oaks et al. (2013) L. limborgi AB981417 KUHE 15614 Malaysia, Janda Baik Matsui et al. (2014) L. limborgi GU934344 FMNH 262817 Cambodia, Mondolkiri Prov., Samling Inger & Stuart (2010) L. macrognathus AB971138 KUHE 23923 Thailand, Ranong Prov. Matsui & Nishikawa (2014) L. macrognathus AB981416 KUHE 23923 Thailand, Ranong Prov. Matsui et al. (2014) L. macrognathus KJ720984 FMNH 270104 Thailand, Nakhon Si Thamarat Prov., Lambertz et al. (2014) L. malesianus AY313692 Not reported Malaysia, Sarawak Prov., Gunung Buda Evans et al. (2003) L. microdiscus AY313688 LSUMZ 81739 Indonesia, Java, Sukabumi Evans et al. (2003) L. plicatellus AB530624 IABHU 21120 Malaysia, Langkawi Island Hasan et al. (2014) L. plicatellus KJ720981 LSUHC 6710 Malaysia, Pulau Pinang, Empangan Air Hitam Lambertz et al. (2014) L. plicatellus KJ720982 LSUHC 6582 Malaysia, Selangor Dist., Gombak Swamp Lambertz et al. (2014) L. plicatellus KJ720983 LSUHC 4001 Malaysia, Selangor Dist., Kepong Lambertz et al. (2014) L. poilani DQ283378 AMNH A163717 Vietnam, Quang Nam Prov., Tre My Dist. Frost et al. (2006) L. woodworthi JX911331 RMB 4092 Philippines, Luzon Island Oaks et al. (2013) SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 261
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.
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 35.9 30.5 42.0; 33.8 ± 2.8 32.9 37.9; 35.4 ± 2.6 HDL 16.8 13.7 20.6; 15.8 ± 1.5 13.8 15.8; 14.8 ± 0.9 HDW 16.6 13.6 20.0; 15.3 ± 1.4 13.1 15.3; 14.2 ± 1.0 SNT 6.3 4.7 6.8; 5.6 ± 0.6 5.2 5.8; 5.5 ± 0.3 EYE 5.0 4.0 5.7; 4.5 ± 0.4 4.7 5.1; 4.9 ± 0.2 IOD 3.6 2.9 4.2; 3.6 ± 0.3 2.1 3.6; 2.9 ± 0.6 TMP 4.2 3.0 5.0; 3.7 ± 0.4 2.9 3.4; 3.1 ± 0.2 IND 3.5 3.0 4.0; 3.4 ± 0.2 2.8 3.7; 3.3 ± 0.4 SHK 17.0 13.9 19.1; 15.8 ± 1.2 17.1 18.6; 17.7 ± 0.8 TGH 17.8 14.2 20.4; 16.7 ± 1.4 15.3 19.0; 17.3 ± 1.8 LAL 7.5 6.3 8.7; 7.0 ± 0.6 6.9 8.2; 7.5 ± 0.6 HND 8.5 6.9 9.8; 7.9 ± 0.7 6.8 9.0; 8.1 ± 1.0 FTL 16.7 14.1 19.2; 15.9 ± 1.1 16.6 19.0; 17.9 ± 1.1 IML 2.0 1.6 2.5; 2.0 ± 0.2 1.7 2.7; 2.2 ± 0.4 IMW 0.9 0.9 1.2; 1.0 ± 0.1 0.8 1.1; 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 00586 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 00552 53, ZMKU AM 00586 87, 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 01104 07, ZMKU AM 01109) are identical in the 16S gene fragment, but have an uncorrected pairwise divergence of 5.98 6.72% from L. dabanus (n=2) and 4.25 8.40% from L. gyldenstolpei (n=3). Calls. Paratype male ZMKU AM 00589 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 27.44 to 4342.91 ms; note duration of 10.29 34.50 ms; interval between notes 182.50 972.41 ms; note repetition rate of 0.98 4.00 notes per second; and dominant frequency of 1.12 2.33 khz. Type 2 (n = 3) was a single, multi-pulsed note lasting 1749.06 2148.03 ms with 31 37 pulses; pulse duration of 21.24 25.00 ms; interval between pulses 33.74 35.36 ms; pulse rate 16.98 17.49 pulses per second; and dominant frequency of 2.15 2.24 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 30.26 s. These calls showed frequency modulation with or without weak harmonics. SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 263
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 131 360 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 1400 2140 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 52.9 65.5 (mean ± SD 59.8 ± 4.0, n = 15) in L. dabanus; SVL 51.1 68.4 (mean ± SD 62.0 ± 5.7, n = 14) in L. gyldenstolpei; SVL 43.2 48.8 (mean ± SD 45.5 ± 3.0, n = 3, this study), types SVL 47 57 (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.
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
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) 1807.15±1216.01; (27.44 4342.91) 1981.08±207.29; (1749.06 2148.03) Intercall interval (ms) 7063.43±6180.57; (361.37 30262.46) Call rate (calls/s) 0.12 0.12 Note 4.63±2.59; (1 10) 1 Note duration (ms) 18.37±6.80; (10.29 34.50) 1981.08±207.29; (1749.06 2148.03) Internote interval (ms) 464.13±189.90; (182.50 972.41) - Note rate (notes/s) 2.16±0.78; (0.98 4.00) - Pulse - 34.67±3.21; (31 37) Pulse duration (ms) - 22.78±1.97; (21.24 25.00) Interpulse interval (ms) - 34.47±0.82; (33.74 35.36) Pulse rate (pulses/s) - 17.25±0.26; (16.98 17.49) Dominant frequency (khz) 2.1±0.29; (1.12 2.33) 2.2±0.05; (2.15 2.24) 266 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.
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 262745). (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 18.8 25.4, mean ± SD 22.1 ± 1.9, n = 9, females with SMALL FANGED FROG FROM THAILAND Zootaxa 3956 (2) 2015 Magnolia Press 267
SVL 20.5 25.0, 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- 1145922), and the Partnerships for Enhanced Engagement in Research (PEER) Science program (grant PGA- 2000003545), 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 1911 1912 and 1914. 3. Batrachians. Kungliga Svenska Vetenskapsakademiens Handlingar, 55, 13 17. Boulenger, G.A. (1916) Description of a new frog from Siam. The Journal of the Natural History Society of Siam, 2, 103 104. Boulenger, G.A. (1917) Descriptions of new frogs of the genus Rana. Annals and Magazine of Natural History, Series 8, 20, 413 418. 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, 1 223. http://dx.doi.org/10.5962/bhl.title.12471 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, 231 243. Cocroft, R.B. & Ryan, M.J. (1995) Patterns of advertisement call evolution in toads and chorus frogs. Animal Behaviour, 49, 283 303. http://dx.doi.org/10.1006/anbe.1995.0043 Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jmodeltest 2: more models, new heuristics and parallel computing. Nature Methods, 9, 772. http://dx.doi.org/10.1038/nmeth.2109 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, 53 64. 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, 131 142. http://dx.doi.org/10.1006/mpev.2000.0778 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, 794 819. http://dx.doi.org/10.1080/10635150390251063 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, 1 370. http://dx.doi.org/10.1206/0003-0090(2006)297[0001:tatol]2.0.co;2 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, 389 411. http://dx.doi.org/10.3906/zoo-1308-36 Inger, R.F. & Stuart, B.L. (2010) Systematics of Limnonectes (Taylorana) Dubois. Current Herpetology, 29, 51 68. http://dx.doi.org/10.3105/018.029.0201 268 Zootaxa 3956 (2) 2015 Magnolia Press AOWPHOL ET AL.
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