Redescription and Range Extension of Microhyla Sholigari Dutta & Ray

Zootaxa 4208 (6): 547–560 ISSN 1175-5326 (print edition) http://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2016 Magnolia Press ISSN 1175-5334 (online edition) http://doi.org/10.11646/zootaxa.4208.6.3 http://zoobank.org/urn:lsid:zoobank.org:pub:88974C4D-F084-4625-81B3-10CB0F6FC05F Redescription and Range Extension of Microhyla sholigari Dutta & Ray (Amphibia: Anura: Microhylidae) from South West India

KADABA SHAMANNA SESHADRI1,8,*, H. PRITI2,3,*, G. RAVIKANTH3, M. K. VIDISHA4, K. K. VINEETH5, RAMIT SINGAL6, R. R. SARMA3, N. A. ARAVIND3 & KOTAMBYLU VASUDEVA GURURAJA4,7,8,* 1Department of Biological Sciences, National University of Singapore, Singapore 117453. E-mail: [email protected] 2Manipal University, Manipal 576104, India. E-mail: [email protected] 3Ashoka Trust for Research in Ecology and the Environment (ATREE), Royal Enclave, Sriramapura, Jakkur (P.O), Bengaluru 560064, India. E-mail: [email protected]; [email protected]; [email protected] 4Gubbi Labs LLP, # 2-182, II Cross, Extension, Gubbi 572216, India. E-mail: [email protected] 5Department of Applied Zoology, Mangalore University, Mangalagangotri, Mangaluru 574199, India. E-mail: [email protected] 6B-14, Law Apartments, Karkardooma, Delhi 110092, India. E-mail: [email protected] 7Srishti Institute of Art, Design and Technology, N4, Yelahanka New Town, Bengaluru 560064. E-mail: [email protected] 8Corresponding author *Authors contributed equally

Abstract

Microhyla sholigari is an endangered frog described in the year 2000. The original description was based on non-adult types and lacked information on several morphological characters, call structure, genetic material and photographs of the animal in life. The absence of such information posed challenges in field identification of this species. Since the original description, there is one other reported sighting of this species from Kerala in 2001. We encountered specimens that we confer to this species based on morphological similarity to the subadult holotype, from several new localities within and outside the Western Ghats of Karnataka. We here redescribe the species based on additional adult vouchers, provide molecular data, describe the advertisement call and report a range extension. Based on its current distribution, we assess the threat status of the species and suggest listing it as Least Concern according to IUCN Red List criteria. Our paper bridges an important gap in the knowledge of the genus Microhyla in India and highlights the importance of systematic surveys in documenting and understanding amphibian diversity in the region.

Key words: IUCN Red List; range extension; Western Ghats, acoustics; morphology, integrative taxonomy

Introduction

Members of the genus Microhyla Tschudi, 1838, occur across South and Southeast Asia with 40 extant species (Frost, 2016). Despite many species of Microhyla being widespread, they are poorly studied, and information on their distribution is sparse. In India, nine species of Microhyla have been recorded: M. berdmorei (Blyth); M. butleri Boulenger; M. chakrapanii Pillai; M. heymonsi Vogt; M. ornata (Duméril & Bibron); M. pulchra (Hallowell); M. rubra (Jerdon); M. sholigari Dutta & Ray and M. laterite Seshadri, Singal, Priti, Ravikanth, Vidisha, Saurabh, Pratik & Gururaja (Seshadri et al. 2016). Among these, M. ornata, M. rubra M. laterite and M. sholigari are found in South India. Microhyla sholigari was known from two localities in the Western Ghats and is currently listed as Endangered (Biju et al. 2004). Though M. sholigari was described 16 years ago (Dutta & Ray, 2000) and later reported from Kerala in 2001 (Biju, 2001), it lacked morphological measurements of adults, molecular data, photographs in life, and advertisement call description, making field identification a challenge. We encountered a Microhyla species during surveys between 2012 and 2016, the identity of which was difficult to deduce. In this paper we (1) ascertain the identity of the Microhyla species as M. sholigari and provide a redescription using new, adult voucher

Accepted by M. Vences: 28 Oct. 2016; published: 21 Dec. 2016 547 specimens collected from the type locality; (2) provide photographs of live specimens, morphological measurements and genetic divergence of M. sholigari with other congeners using 12S and 16S rRNA gene sequences, (3) describe advertisement calls, and (4) report geographic range extension and re-assess the IUCN Red List status using the Red List criteria.

Materials and methods

Abbreviations. amsl: above mean sea level; ATREE: Ashoka Trust for Research in Ecology and the Environment; BNHS: Bombay Natural History Society, Mumbai; BRTTR: Biligiri Rangaswamy Temple Tiger Reserve; HP: H Priti; IUCN: International Union for Conservation of Nature; KSS: K S Seshadri; KVG: K V Gururaja; NAA: N A Aravind; RRS: R R Sarma; VKK: Vineeth K K. Study areas. Between 2012 and 2016, we conducted 16 expeditions within and outside the Western Ghats of Karnataka. The specimens used in this study were collected from inside the type locality Biligiri Rangaswamy Temple Tiger Reserve (BRTTR) from around human settlements (11.00°–12.15° N and 77.00°–77.27° E; 600– 1800 m amsl). The habitat in BRTTR consists of a forest matrix ranging from dry deciduous to evergreen forests. The region receives an annual rainfall of 600–3000 mm. Subsequently, we also collected individuals M. sholigari from the courtyard of a community hall in Bisle Village, Sakleshpura Taluk, Hasana District, Karnataka State, India (12.72°–12.73° N and 75.68°–75.69° E, 835–885 m amsl). This area has paddy fields surrounded by wet evergreen forests, and the mean annual rainfall is about 5000 mm. Voucher collection. Specimen collection and tissue sampling protocol followed guidelines for the use of live amphibians and reptiles in field research by the American Society for Ichthyologists and Herpetologists (Beaupre et al. 2004) and was approved by the Gubbi Labs Internal Committee on Animal Welfare and Ethics. Animals were caught by hand and transported to field station within 30 minutes inside a moist cloth bag. Animals were photographed first and euthanized by applying a small amount of 20% Benzocaine gel on the ventral side. A small portion of thigh muscle tissue was extracted and stored in molecular grade ethanol. Specimens were later fixed in 4% formalin for 24h and then transferred to 70% alcohol for storage as museum specimens. In total, we collected 12 individuals for this study. Five adult females of M. sholigari were collected from ATREE field station within BRTTR on 14th October 2012 between 19:00–20:00 h by KVG, HP, RRS, and NAA. Also, seven individuals (four males and three females) were collected by VKK on 13th July 2015 from the region surrounding Bisle Community Hall. Specimens used for morphological comparisons are deposited in BNHS Museum, Mumbai. The morphological comparison was based on two adult females from BRTTR (BNHS 5968– 5969) and three adult males (BNHS 5970–5972) and two adult females from Bisle (BNHS 5973–5974). Specimens used for molecular comparisons are deposited in the museum collection of ATREE Conservation Genetics lab, Bengaluru. For molecular analysis we used three adult females from BRTTR (ATREE_MISH_1–3) and one male and one female from Bisle (ATREE_MISH_4 and ATREE_MISH_5 respectively). Molecular analysis. DNA extraction protocol followed Vences et al. (2012). The 12S and 16S rRNA genes were amplified following Gururaja et al. (2014). Amplified products were sequenced at Chromous Biotech, Bangalore, India and Chromas Lite 2.01 (http://www.technelysium.com.au/chromas_lite.html) was used to manually check the sequences. MAFFT algorithm (Katoh et al. 2002) was used for sequence alignment and sequences were trimmed using MEGA 5.10 (Tamura et al. 2011). Sequences are deposited in GenBank (KT600665–KT600669; KT600672–KT600676). The final dataset consisted of 1357 base pairs, and sequences of 21 species of genus Microhyla were retrieved from GenBank on the basis of Howlader et al. (2015; Table 2) for calculating genetic distances. Genetic divergence was computed using the uncorrected pairwise genetic distance between the species using MEGA 5.10. Morphological Measurements. Measurements were made to the nearest 0.1 mm using a Mitutoyo® digital slide caliper. Measurements and terminology are abbreviated as: (SVL); head depth, height of the head measured at post-orbital region (HD); head width, at the angle of the jaws (HW); head length, from the rear of the mandible to the tip of the snout (HL); inter upper eyelid width, i.e. the shortest distance between the upper eyelids (IUE); maximum upper eyelid width (UEW); snout length, measured from the tip of the snout to the anterior orbital border of the eye (SL); eye length, i.e. the horizontal distance between the bony orbital borders of the eye (EL); distance from the rear of the mandible to the nostril (MN); distance from the rear of the mandible to the anterior orbital border of the eye (MFE); distance from the rear of the mandible to the posterior orbital border of the eye (MBE);

548 · Zootaxa 4208 (6) © 2016 Magnolia Press SESHADRI ET AL. distance between anterior corner of eyes, i.e. the shortest distance between the anterior orbital borders of the eyes (IFE); distance between posterior corner of eyes, i.e. the shortest distance between the posterior orbital borders of the eyes (IBE); internarial distance, i.e. least distance between the inner margins of nares (IN); nostril–snout distance, i.e. distance between middle of nostril and tip of snout (NS); eye to nostril distance, i.e. distance between anterior-most point of eye and middle of nostril (EN); forelimb length, measured from the elbow to the base of the outer palmar tubercle (FLL); hand length, measured from the base of the outer palmar tubercle to the tip of the third finger (HAL); finger disc width (FD1–FD4); finger width measured at the base of the disc (FW1–FW4); finger length measured from tip of finger disc to proximal palmar tubercle (FIL–FIVL); thigh length (TL); shank length (ShL); tibia width, i.e. width of tibia at its widest region (TW); foot length, measured from the base of the inner metatarsal tubercle to the tip of the fourth toe (FOL); distance from the heel to the tip of the fourth toe (TFOL); toe disc width (TD1–TD5); toe width measured at the base of the disc (ToW1–ToW5); toe length measured from tip of toe disc to proximal sub-articular tubercle (TIL–TVL); length of inner metatarsal tubercle (IMT); length of outer metatarsal tubercle (OMT); distance from distal edge of metatarsal tubercle to maximum incurvature of web between fourth and fifth toe (MTFF); distance from distal edge of metatarsal tubercle to maximum incurvature of web between third and fourth toe (MTTF); distance from maximum incurvature of web between fourth and fifth toe to tip of fourth toe (FFTF); distance from maximum incurvature of web between third and fourth toe to tip of fourth toe (TFTF). Fingers and toes were photographed using a Nikon® D90 Digital camera with a Nikkor® 105 mm macro lens. ImageJ® was then used to measure fingers and toes using hand length and foot length as a reference. QGIS® Pisa Ver. 2.10 was used for illustrations of hand and foot. A high quality image of hand and feet were opened in QGIS® and outlines were digitized and saved as a vector image. Advertisement calls recording and description. Calls were recorded using an Sennheiser® ME66 unidirectional microphone fitted to a Zoom H4n handy sound recorder at 44.1kHz and 16 bit and saved as .wav format. Thirteen calls from three individuals of M. sholigari having low signal to noise ratio were used for call description, and these individuals were not collected. Call terminology follows Kok & Kalamandeen (2008). Call duration, dominant frequency and number of pulses in each call were analyzed using Audacity Ver.1.3 (Beta) and Raven Pro Ver.1.5. Air temperature and relative humidity were recorded using a Brunton® ADC Pro weather meter. Comparisons. Morphological comparisons were based on publications by Jerdon (1853); Boulenger (1882); Parker (1934); Dutta & Ray (2000); Howlader et al. (2015); Seshadri et al. (2016) and Wijayathilaka et al. (2016). We made bioacoustic comparisons based on Heyer (1971); Kanamadi et al. (1994); Grosselet et al. (2004); Kuramoto & Joshy (2006); Seshadri et al. (2016) and Wijayathilaka & Meegasakumbura (2016). Maps and geographic range estimation. Geographic range and distribution maps were generated using QGIS® Pisa Ver. 2.10. Open source data from Global Administrative areas (www.gadm.org) was used for an administrative boundary, and SRTM 90 m database (http://srtm.csi.cgiar.org) was used for elevation. The area under minimum convex hull was computed by connecting the outermost occurrence points to estimate the extent of occurrence as defined by the IUCN (2001).

Results

We encountered populations of Microhyla from 14 localities within and outside the Western Ghats of Karnataka (Table 1; Fig. 1), which appeared to match the description of M. sholigari. We compared specimens collected in this study from the type locality (BRTTR) with the original description of M. sholigari in Dutta & Ray (2000), and found similarity in the following suite of characters: very small sized frogs (both males and females in the range of 15.9 to 19.2 mm); pointed snout protruding beyond mouth in ventral view; head wider than long; tongue oval with complete margin without lingual papilla; indistinct tympanum; presence of both inner and outer metatarsal tubercle and reduced webbing in feet and disc on toes with circum-marginal groove, cover bifurcate distally. We found following differences from original description: presence of disc on fingers with circum-marginal groove, cover notched distally, venter buff white, dorsum smooth, interspersed with tubercles increasing in intensity towards vent; anterior part of the thigh with a distinct black band starting from knee and terminating short of the groin. Specimens from Bisle were also compared with the individuals from the type locality to confirm the identity. In several localities, M. sholigari co-occurred with M. ornata and M. rubra, but M. sholigari emerges as a distinct species based on morphology, genetic divergence and calling pattern. Overall, the genetic divergence values across

REDESCRIPTION OF MICROHYLA SHOLIGARI Zootaxa 4208 (6) © 2016 Magnolia Press · 549 the 22 species of Microhyla based on a combined dataset of 16S and 12S rRNA genes varied between 9.61–14.61% (Table 3). The genetic divergence values within M. sholigari individuals ranged from 0.00–0.42%.

TABLE 1. Locality records for Microhyla sholigari. Locality Longitude (°E) Latitude (°N) Altitude (m amsl) Kundaadri Betta 75.1684 13.5493 650 Jaipura 75.3725 13.4019 700 Balehonnur 75.4650 13.3489 752 Mudigere 75.6406 13.1343 963 Hassan 75.8116 12.9478 929 Bisle* 75.6915 12.7189 837 Somwarpet 75.8504 12.5943 1106 Napoklu 75.6885 12.3095 913 Siddapura 75.8751 12.2974 900 Virajpet 75.8039 12.1950 910 Gonikoppa 75.9260 12.1845 850 Kutta 76.0478 11.9677 950 BRTTR* 77.1430 11.9881 1177 Doddasampige# 77.1836 11.9473 1190 Shivanalli 77.5582 12.7188 906 Waynad Wildlife Sanctuary## 76.3640 11.6460 NA

*specimen collected for morphometric and molecular work; All records were from this study except #Dutta & Ray 2000 and ##Biju et al. 2004.

FIGURE 1. Map showing current distribution records of M. sholigari. Orange circles are from the present study. Blue rhombuses are records from the literature. Grey line indicates Western Ghats boundary. Light green areas with gray boundaries represent protected areas. Maps were generated using QGIS® Pisa Ver. 2.10. Data was sourced from www.gadm.org for the administrative boundary of India and Shuttle Radar Topography Mission (SRTM) 90 m database (http://srtm.csi.cgiar.org) for elevation.

550 · Zootaxa 4208 (6) © 2016 Magnolia Press SESHADRI ET AL. TABLE 2. Species used for phylogenetic analysis, their voucher numbers and GenBank accession numbers. Sl. No Species Voucher 16S 12S 1 M. achatina MZBamp16401 AB598335 AB598311 2 M. annectens KUHE52438 AB634659 AB634601 3 M. berdmorei KUHE AB598338 AB598314 4 M. butleri KUHE33557 AB201189 AB201178 5 M. fissipes KUHE35165 AB201186 AB201175 6 M. fowleri KUHE21992 AB634667 AB634609 7 M. heymonsi KUHEK1845 AB201190 AB201179 8 M. malang KUHE42597 AB598322 AB598298 9 M. mantheyi KUHE52556 AB598334 AB598310 10 M. marmorata KUHE32455 AB611955 AB634610 11 M. mixtura CIB AB634669 AB634611 12 M. nilphamariensis MZH-2360-66 KP072787 NA 13 M. mukhlesuri IABHU3880 AB543609 NA 14 M. okinavensis KUHE12840 AB201184 AB201173 15 M. ornata ZSI-A9119 AB201188 AB201177 16 M. palmipes MZBAAmp16323 AB634671 AB634613 17 M. perparva KUHE53675 AB634673 AB634615 18 M. petrigena KUHE53743 AB634675 AB634617 19 M. pulchra KUHE35119 AB201191 AB201180 20 M. rubra NA AB201192 AB201181 21 M. superciliaris KUHE52558 AB634682 AB634624 22 M. sholigari_1* ATREE_MISH_1 KT600674 KT600667 23 M. sholigari_2* ATREE_MISH_2 KT600675 KT600668 24 M. sholigari_3* ATREE_MISH_3 KT600676 KT600669 25 M. sholigari_4# ATREE_MISH_4 KT600672 KT600665 26 M. sholigari_5# ATREE_MISH_5 KT600673 KT600666

* specimens from BRTTR; # specimens from Bisle.

Redescription of Microhyla sholigari Dutta & Ray, 2000 Figure 2a–j; 3 and Table 4.

Suggested common name: Sholiga narrow-mouthed frog Voucher specimens: BNHS 5968–5969, 2 ex., adult females, collected near the kitchen of ATREE field station (11.9879° N, 77.1433° E, elevation 1177 m) within BRTTR, Chamarajanagara District, Karnataka State, collected by KVG, HP, RRS and NAA, 14th October 2012; BNHS 5970–5972, 3 ex., adult males, BNHS 5973– 5974, 2 ex., adult females, courtyard of Bisle Community Hall (12.7192° N, 75.6915° E, elevation 837 m), Bisle Village, Sakleshpura Taluk, Hasana District, Karnataka State, collected by VKK, 13th July 2015. Diagnosis. Dutta & Ray (2000) provided a diagnosis on a subadult female individual (ZSI A9061). A comprehensive diagnosis is provided here based on an adult male individual of Microhyla sholigari and it can be distinguished from all other congeners in the Indian subcontinent by the following suite of characters: (i) A very small sized adult frog (male: 15.9–16.2 mm, N = 3 and female: 16.5–19.2 mm, N = 4); (ii) snout pointed in dorsal and ventral view with indistinct canthus rostralis, snout protrudes beyond mouth in ventral view; (iii) tongue oval with complete margin lacking a lingual papilla; (iv) tympanum indistinct; (v) head wider than long; (vi) skin smooth on dorsum and venter; (vii) throat buff colored with brown pigmentation; (viii) reduced webbing in feet,

REDESCRIPTION OF MICROHYLA SHOLIGARI Zootaxa 4208 (6) © 2016 Magnolia Press · 551 reaching proximal tubercle on the fourth toe on the inside; (ix) disc on fingers with circum-marginal groove, cover notched distally and disc on toes with circum-marginal groove, cover bifurcate distally.

FIGURE 2. Plate depicting morphology of M. sholigari, an adult male, BNHS 5970. (a) Dorsal view, (b) ventral view, (c) ventral view of throat, (d) lateral profile showing tympanic region, (e) ventral view of hand, (f) line drawing of hand depicting palmar and subarticular tubercles, (g) circum-marginal groove on third fingertip, (h) fourth toe tip showing bifurcated circum- marginal groove, (i) ventral view of feet, (j) line drawing of feet depicting webbing, tarsal and subarticular tubercles.

Differential diagnosis. Microhyla sholigari is compared with congeners from India, namely M. berdmorei, M. butleri, M. chakrapanii, M. heymonsi, M. laterite, M. ornata, M. pulchra and M. rubra. Microhyla sholigari differs from M. berdmorei in size (SVL, male: 15.9–16.2 mm, female: 16.5–19.2 mm vs. male: 25–28 mm, female: 27–45 mm); webbing in feet (reaching proximal tubercle on fourth toe on the inside vs. full webbing); number of pulse per call (64–72 vs. 3–9) and dominant frequency (3375–3704 Hz vs. 1500–1800 Hz). Microhyla sholigari differs from M. butleri in size (SVL, male: 15.9–16.2 mm, female: 16.5–19.2 mm vs. male: 21–23 mm, female: 23–26 mm); snout (pointed vs. rounded); webbing in feet (reaching proximal tubercle on fourth toe on the inside vs. nearly half); number of pulse per call (64–72 vs. 6–7) and dominant frequency (3375–3704 Hz vs. 1200–4500 Hz). Microhyla sholigari differs from M. chakrapanii in size (SVL, male: 15.9–16.2 mm, female: 16.5–19.2 mm vs. male: 22 mm); snout (pointed vs. rounded) and discs on finger (present vs. absent). Microhyla sholigari differs from M. heymonsi in size (SVL, male, 15.9–16.2 mm vs. 21.5–24.4 mm); no lateral band from snouth to groin vs. a continuous dark lateral band from the end of the snout to the groin and number of pulse per call (64–72 vs. 11–17). Microhyla sholigari differs M. laterite in vocal sac (cream with sparse brown pigmentation vs. purplish black); webbing in feet (reaching proximal tubercle on fourth toe on the inside vs. reaching distal tubercle on fourth toe on the inside); dorsal pattern (strongly constricted vs. less constricted) and number of pulse per call (64–72 vs. 90–

552 · Zootaxa 4208 (6) © 2016 Magnolia Press SESHADRI ET AL. 126). Microhyla sholigari forms a sister relationship with M. laterite and a detailed comparison is provided in Seshadri et al. (2016). Microhyla sholigari differs from M. ornata in size (SVL, male, 15.9–16.2 mm vs. 18–21 mm); discs on fingers and toes (present vs. absent); vocal sac (cream with sparse brown pigmentation vs. greyish black) and number of pulse per call (64–72 vs. 8–14). Microhyla sholigari differs from M. pulchra in size (SVL, male: 15.9–16.2 mm, female: 16.5–19.2 mm vs. male: 35 mm); discs on fingers and toes (present vs. absent) and webbing in feet (reaching proximal tubercle on fourth toe on the inside vs. nearly half). Microhyla sholigari differs from M. rubra in size (SVL, male, 15.9–16.2 vs. 25.8–29.6 mm); discs on fingers and toes (present vs. absent); vocal sac (cream with sparse brown pigmentation vs. black) and number of pulse per call (64–72 vs. 15–21). Description of an adult male (Voucher No. BNHS 5970, Fig. 2, all measurements in mm). A small sized adult (SVL = 16.2 mm), head wider than long (HW = 4.9; HL = 3.9). Snout acute in both dorsal and ventral views, upper jaw protrudes slightly in ventral view. Snout acuminate in lateral profile, 1.5 times longer the eye length (SL = 2.4; EL = 1.6). Canthus rostralis rounded. Loreal region concave. Interorbital space sloping towards snout, 2.28 times larger than upper eyelid width, and wider than internarial distance (IUE = 2.0; UEW = 0.9; IN = 1.4). The distance between posterior margins of eyes 1.84 times that of anterior margins (IBE = 4.2; IFE = 2.3). Nostrils rounded, without flap, closer to the tip of snout than to eye (NS = 1.1; EN = 1.3). Symphysial knob present, weak. Tongue relatively large, oval, free at base, margin entire. Lingual papillae absent. Vomerine teeth absent. Tympanum indistinct, moderate supratympanic fold. Single subgular vocal sac with a pair of openings at the base of lower jaw without skin folds. Eyes small (EL = 1.6), pupil-rounded. Forelimb shorter than hand (FLL = 3.2; HAL = 4.3). Dermal fringe present on fingers. Webbing between fingers absent. Relative lengths of fingers I

REDESCRIPTION OF MICROHYLA SHOLIGARI Zootaxa 4208 (6) © 2016 Magnolia Press · 553 21 10.13 – 10.38 20 8.44 9.61 – 9.85 19 8.92 10.58 11.83 – 12.08 18 11.82 10.83 9.85 12.84 – 13.09 17 5.46 12.06 12.53 11.06 14.35 – 14.61 16 9.63 9.63 9.87 9.90 8.45 11.35 – 11.60 15 8.47 11.56 11.10 8.91 8.23 9.65 10.84 – 11.09 14 10.34 10.36 12.30 12.59 8.44 9.66 11.37 13.59 – 13.85 1. M. achatina; 2. M. annectens; 3. M. berdmorei; 4. M. butleri; 5. M. M. 5. butleri; M. 4. berdmorei; M. 3. annectens; M. 2. achatina; 1. M. 13 6.40 8.67 8.67 10.58 10.13 8.23 8.46 8.43 11.56 – 11.81 ; 13. M. mukhlesuri; 14. M. okinavensis; 15. M. ornata; 16. M. palmipes; 17. M. palmipes; ornata; M. 16. 15. okinavensis; 13.mukhlesuri;M. 14. M. M. ; 12 8.69 9.88 4.99 8.71 12.30 11.35 8.45 6.14 9.15 10.34 – 10.58 11 8.21 4.32 3.23 9.85 9.15 11.06 10.82 7.74 9.40 9.39 11.32 – 11.57 M. sholigari_5. sholigari_5. M. 10 9.13 10.10 9.63 11.07 9.88 9.40 6.57 7.27 10.58 9.61 10.60 11.86 – 12.12 – 9 11.07 6.14 9.86 7.28 7.29 12.04 11.81 13.04 12.56 10.35 9.62 10.59 11.81 – 12.06 8 6.37 9.85 6.80 10.10 6.82 7.97 11.30 11.07 12.04 12.06 9.88 9.62 11.06 12.55 – 12.81 26. M. sholigari_1 M. 26. – 7 5.44 7.73 10.09 5.66 10.10 5.67 7.06 10.12 9.14 10.58 10.35 7.97 9.64 8.90 11.81 – 12.06 species species for analysis.used Genetic distances are in percentage. 6 9.15 9.87 9.14 11.05 9.14 9.13 10.11 9.14 10.57 11.55 12.55 11.32 7.49 9.85 10.57 10.34 – 10.59 ntheyi; 10. M. marmorata; 11. M. mixtura; nilphamariensis M. M. 12. 11. marmorata;ntheyi; M. 10. Microhyla 5 9.62 5.89 7.04 7.28 9.62 4.10 9.17 1.92 5.69 9.62 7.96 11.31 10.62 8.23 8.69 9.14 11.31 – 11.56 4 10.33 10.82 10.81 10.33 11.07 9.37 9.37 9.14 9.85 11.81 11.07 9.89 10.32 11.30 9.85 9.39 7.26 11.38 – 11.63 3 10.58 8.91 2.13 9.38 10.35 9.62 10.81 8.90 8.90 9.40 9.38 9.84 11.56 12.55 10.85 7.03 10.10 10.33 11.07 – 11.32 2 9.13 10.32 11.30 10.09 10.81 12.29 12.03 6.12 11.29 11.30 11.55 13.03 11.56 10.36 8.01 7.73 11.55 12.03 11.30 12.58 – 13.09 1 11.78 7.49 11.31 7.28 7.50 7.76 7.09 5.92 10.82 6.34 9.14 6.82 7.04 10.57 11.57 13.29 12.30 7.99 9.63 11.31 12.58 – 13.09 2 – 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 6 fissipes; 6. M. fowleri; 7. M. heymonsi; 8. M. malang; 9. M. 8. malang; M. ma heymonsi; fissipes; M. 7. 6. fowleri; M. 22 superciliaris; M. 21. rubra; M. 20. pulchra; M. 19. petrigena; M. 18. perparva; TABLE 3. distancesTABLE pairwise Un-corrected genetic of

19.2 19.2 – 5.5 4.6 5.2 2.6 1.3 2.5 2.0 3.9 3.0 1.5 1.8 3.2 5.0 1.2 1.6 3.9 5.5 0.3 0.4 0.5 0.5 0.3 0.3 0.4 0.4 1.3 1.7 3.1 2.2 10.7 – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – Range 16.5 5.1 3.7 3.5 2.0 0.9 1.9 1.4 3.2 2.4 1.3 1.5 2.8 4.4 1.0 1.1 3.2 4.2 0.2 0.3 0.4 0.2 0.2 0.3 0.3 0.2 0.9 1.1 2.2 1.5 8.0 ……continued on the next page next on the ……continued Average ± SD 1.32 ± 18.06 0.19 ± 5.37 0.44 ± 4.25 0.85 ± 4.33 0.29 ± 2.27 0.24 ± 1.10 0.29 ± 2.20 0.25 ± 1.70 0.29 ± 3.53 0.26 ± 2.63 0.10 ± 1.47 0.13 ± 1.61 0.20 ± 2.93 0.29 ± 4.70 0.08 ± 1.08 0.25 ± 1.33 0.40 ± 3.57 0.60 ± 4.81 0.06 ± 0.29 0.06 ± 0.34 0.05 ± 0.40 0.09 ± 0.36 0.04 ± 0.27 0.03 ± 0.30 0.05 ± 0.32 0.08 ± 0.31 0.20 ± 1.05 0.29 ± 1.34 0.36 ± 2.65 0.37 ± 1.81 1.20 ± 9.31

5969 5969 Female 16.5 5.1 3.7 3.5 2.1 0.9 1.9 1.6 3.2 2.6 1.3 1.5 2.8 4.4 1.0 1.1 3.2 4.5 0.2 0.3 0.5 0.2 0.2 0.3 0.4 0.2 1.0 1.1 2.6 1.5 8.0 October 2012 October th 5968 5968 14 BRTTR Female 17.5 5.3 4.1 3.7 2.0 0.9 2.0 1.7 3.6 2.6 1.5 1.8 3.2 4.5 1.1 1.1 3.3 4.2 0.3 0.3 0.4 0.4 0.3 0.3 0.3 0.3 0.9 1.2 2.2 1.5 8.7 5974 5974 Female 19.1 5.5 4.6 4.9 2.5 1.3 2.5 2.0 3.9 3.0 1.5 1.6 2.8 4.9 1.2 1.6 3.9 5.1 0.3 0.4 0.4 0.5 0.3 0.3 0.3 0.4 1.3 1.4 2.6 2.2 9.8 July 2015 2015 July th 5973 5973 13 Bisle Female 19.2 5.5 4.6 5.2 2.6 1.3 2.4 1.4 3.5 2.4 1.5 1.6 2.9 5.0 1.1 1.5 3.9 5.5 0.2 0.4 0.4 0.4 0.2 0.3 0.3 0.3 1.1 1.7 3.1 2.1 10.7 16.2 16.2 – 5.1 4.1 4.2 2.0 1.4 2.4 2.0 3.5 2.7 1.4 1.5 2.6 4.5 1.1 1.3 3.4 4.5 0.4 0.5 0.5 0.5 0.4 0.5 0.4 0.4 1.2 2.1 2.9 2.4 8.9 For abbreviations, see methods. Prefix abbreviations,For BNHS to methods. numbers. see voucher – – – – – – – – – – – – – – – – – – – – – – – – – – – – – – Range 15.9 4.9 3.9 3.5 1.9 0.9 2.2 1.6 3.4 2.2 1.1 1.4 2.5 4.2 1.0 1.2 3.2 3.9 0.3 0.4 0.4 0.3 0.2 0.3 0.3 0.3 0.9 1.4 1.9 1.6 8.5 Microhyla sholigari. Average ± SD 0.16 ± 15.98 0.10 ± 4.99 0.12 ± 4.01 0.35 ± 3.77 0.07 ± 1.96 0.25 ± 1.17 0.12 ± 2.27 0.25 ± 1.72 0.07 ± 3.45 0.24 ± 2.47 0.20 ± 1.28 0.04 ± 1.45 0.10 ± 2.58 0.15 ± 4.33 0.03 ± 1.03 0.04 ± 1.24 0.11 ± 3.25 0.29 ± 4.22 0.06 ± 0.32 0.08 ± 0.40 0.08 ± 0.48 0.11 ± 0.40 0.09 ± 0.30 0.07 ± 0.38 0.05 ± 0.37 0.08 ± 0.33 0.16 ± 1.01 0.35 ± 1.67 0.57 ± 2.55 0.44 ± 1.85 0.20 ± 8.65 5972 5972 Male 15.9 5.1 4.0 4.2 2.0 1.4 2.2 2.0 3.4 2.5 1.4 1.5 2.6 4.3 1.0 1.2 3.4 4.5 0.3 0.4 0.5 0.3 0.3 0.4 0.4 0.3 1.0 1.5 2.8 1.7 8.9 5971 5971 Male 15.9 5.0 4.1 3.5 1.9 1.3 2.2 1.6 3.5 2.2 1.1 1.4 2.6 4.5 1.0 1.2 3.2 3.9 0.3 0.4 0.4 0.3 0.2 0.3 0.3 0.3 0.9 1.4 1.9 1.6 8.5 July 2015 2015 July th 5970 5970 13 Bisle Male 16.2 4.9 3.9 3.6 2.0 0.9 2.4 1.6 3.5 2.7 1.4 1.4 2.5 4.2 1.1 1.3 3.2 4.3 0.4 0.5 0.5 0.5 0.4 0.5 0.4 0.4 1.2 2.1 2.9 2.4 8.6 Morphometric measurements of Morphometric (mm) TABLE 4. Voucher Date Locality Sex SVL HW HL HD IUE UEW SL EL MN MFE MBE IN IFE IBE NS EN FLL HAL FD1 FD2 FD3 FD4 FW1 FW2 FW3 FW4 FIL FIIL FIIIL FIVL TL

REDESCRIPTION OF MICROHYLA SHOLIGARI Zootaxa 4208 (6) © 2016 Magnolia Press · 555 17.1 17.1 11.6 3.2 12.0 0.6 0.7 0.6 0.6 0.5 0.6 0.8 0.8 0.9 0.6 1.1 0.5 – 2.2 3.4 5.5 8.3 3.8 6.1 6.1 5.6 5.9 – – – – – – – – – – – – – – – – – – – – – – – – Range 9.4 2.5 9.5 0.3 0.4 0.4 0.5 0.4 0.3 0.5 0.5 0.5 0.4 0.7 0.5 13.5 1.2 1.7 3.7 6.0 3.3 4.2 4.1 4.5 5.1 Average ± SD 1.06 ± 10.35 0.33 ± 2.78 1.06 ± 10.55 0.12 ± 0.42 0.15 ± 0.50 0.05 ± 0.48 0.03 ± 0.52 0.05 ± 0.43 0.15 ± 0.43 0.18 ± 0.57 0.16 ± 0.63 0.16 ± 0.62 0.11 ± 0.48 0.17 ± 0.83 0.05 ± 0.50 1.66 ± 14.96 0.43 ± 1.51 0.73 ± 2.41 0.84 ± 4.22 1.11 ± 6.63 0.28 ± 3.60 0.96 ± 5.13 0.93 ± 5.13 0.45 ± 5.06 0.34 ± 5.48

5969 Female 9.4 2.5 9.5 0.4 0.5 0.6 0.5 0.4 0.4 0.5 0.6 0.5 0.5 0.8 0.5 13.5 1.4 2.6 3.8 6.1 3.4 4.5 4.1 4.9 5.1 October 2012 October th 5968 14 BRTTR Female 9.6 2.6 10.0 0.3 0.4 0.4 0.5 0.4 0.3 0.5 0.5 0.6 0.4 0.8 0.5 13.8 1.2 1.7 3.7 6.0 3.3 4.2 4.6 4.5 5.3 5974 Female 10.9 2.9 10.7 0.4 0.4 0.5 0.5 0.5 0.3 0.5 0.6 0.5 0.5 0.7 0.5 15.4 1.3 2.0 4.0 6.2 3.8 5.8 5.7 5.2 5.9 July 2015 2015 July th 5973 13 Bisle Female 11.6 3.2 12.0 0.6 0.7 0.5 0.6 0.4 0.6 0.8 0.8 0.9 0.6 1.1 0.5 17.1 2.2 3.4 5.5 8.3 3.8 6.1 6.1 5.6 5.6 14.2 14.2 9.6 9.6 2.5 9.9 0.3 0.4 0.4 0.5 0.4 0.3 0.4 0.5 0.6 0.5 1.5 0.4 – 1.2 1.8 3.4 5.8 3.3 5.2 4.9 5.0 5.1 – – – – – – – – – – – – – – – – – – – – – – – – Range 9.4 2.5 9.6 0.2 0.4 0.3 0.4 0.3 0.2 0.4 0.4 0.5 0.3 0.7 0.3 13.6 1.0 1.5 3.0 5.6 3.1 4.9 4.5 4.7 4.9 Average ± SD 0.07 ± 9.52 0.03 ± 2.50 0.16 ± 9.78 0.02 ± 0.26 0.03 ± 0.37 0.07 ± 0.37 0.05 ± 0.42 0.02 ± 0.35 0.05 ± 0.28 0.01 ± 0.42 0.05 ± 0.48 0.09 ± 0.55 0.07 ± 0.42 0.43 ± 0.98 0.04 ± 0.37 0.30 ± 13.86 0.09 ± 1.05 0.14 ± 1.61 0.21 ± 3.21 0.09 ± 5.68 0.14 ± 3.21 0.18 ± 5.05 0.25 ± 4.62 0.17 ± 4.85 0.09 ± 4.98 5972 5972 Male 9.6 2.5 9.9 0.2 0.4 0.3 0.4 0.4 0.2 0.4 0.5 0.6 0.3 0.8 0.4 14.2 1.2 1.5 3.0 5.6 3.2 5.2 4.9 4.7 4.9 5971 5971 Male 9.4 2.5 9.6 0.3 0.4 0.4 0.4 0.4 0.3 0.4 0.5 0.6 0.5 0.7 0.3 13.6 1.0 1.8 3.4 5.8 3.3 4.9 4.5 5.0 5.1 July 2015 2015 July th 5970 5970 13 Bisle Male 9.6 2.5 9.8 0.3 0.4 0.4 0.5 0.3 0.3 0.4 0.4 0.5 0.4 1.5 0.4 13.9 1.0 1.6 3.3 5.6 3.1 5.0 4.5 4.8 4.9 (Continued) (Continued) TABLE 4. Voucher Date Locality Sex ShL TW FOL ToW1 ToW2 ToW3 ToW4 ToW5 TD1 TD2 TD3 TD4 TD5 IMT OMT TFOL TIL TIIL TIIIL TIVL TVL MTFF MTTF TFTF FFTF

Advertisement calls description. Calls of M. sholigari were recorded on 11th June 2016, between 21:00 h and 23:45 h at Bisle, 12.7192° N, 75.6915° E, elevation 837 m. The air temperature was 23.4 ± 0.06° C and relative humidity of 94.33 ± 0.58%. Advertisement calls sound like ‘Zeeeeee…..Zeeeee…..Zeeeee…’ and are heard as a chorus. Calls of M. sholigari had 52–67 pulses (Mean ± SE, 62.92 ± 3.86; N = 13) in each call. Average dominant frequency was 3596.08 ± 98.46 Hz (range: 3375–3704) and call duration was 0.76 ± 0.04 s (range: 0.65–0.81 s). A single call with a duration of 0.76 s is given in Figure 4. A video clip of advertisement call of M. sholigari is at https://youtu.be/Zx7qQ8T75U8. Geographic range extension and IUCN status. We encountered this species in localities in Western Ghats including one locality in the outskirts of the city of Bengaluru (Shivanalli), which is outside the Western Ghats boundary (Fig. 1, Table 1). From the type locality, the new range of M. sholigari extends to about 284 km to the north-west. The localities vary from forests inside protected areas to open fallow land outside of the Western Ghats. The geographic extent of occurrence as determined from the minimum convex hull was 28,304.6 km2. This species is locally abundant (> 50 calling male individuals/100 m2 were observed at Bisle, BRTTR and Shivanalli). There is however, limited information on population size, fluctuation and trends, the number of individuals and generation length. M. sholigari being widespread, does not occur in any one geographically and distinct ecological location. Additionally, there appear to be no sub-population effects as indicated by the low genetic divergence between the two locations sampled in this study (Table 3). In light of the extended geographic range, this species no longer meets the requirements to be classified as Endangered under the criteria of B1ab(iii) or any other category, and thus, we suggest down-listing M. sholigari to Least Concern (LC) instead.

Discussion

Microhyla sholigari was described by Dutta & Ray (2000) based on a subadult type series without information on the sexes of any specimen except for the subadult female holotype. The original description also lacks a photograph of live individuals making field identification challenging. Although Biju (2001) reported a sighting from Kerala, no subsequent sightings were reported for about fifteen years until its re-discovery from the type locality (Gururaja, 2015). The absence of molecular data for several extant species is a common phenomenon across taxa, leading to a ‘Darwinian shortfall’ where phylogenetic information is absent for most organisms, thus inhibiting a robust understanding of phylogenetic relationships within a particular group (Diniz-Filho et al. 2013). Microhyla sholigari is one such extant species that lacked molecular data. Using 12S and 16S rRNA we provided the genetic divergence of M. sholigari to other congeners, thus filling an important knowledge gap. Another challenge is to

558 · Zootaxa 4208 (6) © 2016 Magnolia Press SESHADRI ET AL. understand systematics of a group of taxa, known as the ‘Linnean shortfall’ (Bini et al. 2006) where several cryptic and undescribed lineages exist. Studies from the Indian subcontinent have reported the existence of cryptic and undescribed lineages in the genus Microhyla (Howlader et al. 2015). Subsequently, a new species of Microhyla was described as M. laterite by Seshadri et al. (2016) which forms a sister relationship with M. sholigari in our analysis. Yet another direct implication of our work is the extension of geographic range and re-assessment of its threat status as per IUCN Red List criteria. Microhyla sholigari is currently listed as Endangered on the IUCN Red List, pending re-assessment (Biju et al. 2004). It was known from two locations within the Western Ghats and was listed as endangered under the criterion B1ab(iii) where the extent of occurrence is < 5000 km2, the habitat is severely fragmented, and there is an inferred or projected decline in area, extent, and/or quality of habitat (Biju et al. 2004; IUCN 2001). From our extensive surveys along the Western Ghats and other parts of Karnataka, we were able to detect this species from over 14 localities including forested areas near the city of Bengaluru and the current geographic extent of occurrence is about five times higher (28,304.6 km2). Our observations suggest that the extent and quality of habitat in the areas where these frogs occur is not declining. The up-listing or down-listing of species from one threat category to another of the IUCN Red List requires an assessment against all the five criteria (A–E, with 11 sub-criteria) but only one criterion needs to be fulfilled for designation of threat categories (IUCN 2001). In the case of M. sholigari, the information on geographic distribution was the most accurate and reliable among all other criteria and was hence used for a conservative estimate of the extent of occurrence (Criteria B, B1). This also happens to be an important criterion used by Biju et al. (2004) for the earlier assessment. With all the updated information presented here, down-listing the threat status from Endangered to Least Concern is justified as no other criteria are met. Finally, our work emphasizes the importance of systematic surveys within and outside the Western Ghats for furthering our knowledge and understanding of amphibians. We stress that such initiatives are necessary to overcome limitations we currently face in understanding amphibian ecology and evolution in the region.

Acknowledgements

We are deeply indebted to the members of the ‘Bisle Kappe Team’, the field staff of ATREE and the Karnataka Forest Department provided logistical support in BRTTR. Godwin Rodny D’Souza helped VKK collect and transport voucher specimens. Rahul Khot, curator BNHS assisted in deposition of vouchers. We thank Madhukara S Putty, Catharina Karlsson, David Bickford, Mark D. Scherz and Miguel Vences for their constructive criticisms that improved the manuscript quality. KSS was supported by Mohamed Bin Zayed Species Conservation Fund and Chicago Board of Trade (CBOT) endangered species fund. RS was supported by Rufford Small Grants. KVG was supported by Earthwatch Institute, India.

References

Beaupre, S.J., Jacobson, E.R., Lillywhite, H.B. & Zamudio, K.R. (2004) Guidelines for use of live Amphibians and Reptiles in Field and Laboratory Research. Available from: http://www.asih.org/sites/default/files/documents/resources/ guidelinesherpsresearch2004.pdf (accessed 5 March 2016) Biju, S.D. (2001) A synopsis of frog fauna of the Western Ghats. Indian Society for Conservation Biology, Occasional Publication, Thiruvananthapuram, 24 pp. Biju, S.D., Dutta, S.K., Bhuddhe, G.D., Vasudevan, K. & Srinivasulu, C. (2004) Microhyla sholigari. The IUCN Red List of Threatened Species 2004: e.T57893A11688938. http://dx.doi.org/10.2305/IUCN.UK.2004.RLTS.T57893A11688938.en Bini, L.M., Diniz-Filho, J.A.F., Rangel, T.F.L.V.B., Bastos, R.P. & Pinto, M.P. (2006) Challenging Wallacean and Linnean shortfalls: knowledge gradients and conservation planning in a biodiversity hotspot. Diversity and Distributions, 12, 475– 482. http://dx.doi.org/10.1111/j.1366-9516.2006.00286.x Boulenger, G. (1882) Catalogue of the Batrachia Salientia: Ecaudata. Collection of the British Museum. Second Edition. Taylor and Francis, London, 256 pp. Diniz-Filho, J.A.F., Loyola, R.D., Raia, P., Mooers, A.O. & Bini, L.M. (2013) Darwinian shortfalls in biodiversity conservation. Trends in Ecology and Evolution, 28, 689–695. http://dx.doi.org/10.1016/j.tree.2013.09.003

REDESCRIPTION OF MICROHYLA SHOLIGARI Zootaxa 4208 (6) © 2016 Magnolia Press · 559 Dutta, S.K. & Ray, P. (2000) Microhyla sholigari, A new species of Microhylid frog (Anura: Microhylidae) from Karnataka, India. Hamadryad, 25, 38–44. Frost, D.R. (2016) Amphibian Species of the World: an Online Reference. Version 6.0. American Museum of Natural History, New York, USA. Available from: http://research.amnh.org/herpetology/amphibia/index.html (accessed 28 January 2016) Grosselet, O., Sensgupta, S., Gupta, A., Vauche, M. & Gupta, S. (2004) Microhyla heymonsi Vogt 1911 (Anura: Microhylidae) from mainland India, with bioacoustic analysis of its advertising call. Hamadryad, 29, 131–133. Gururaja, K.V. (2015) In search of an elusive frog. Jungle Lodges and Resorts, Karnataka, India. Available from: http:// jlrexplore.com/explore/from-the-field/in-search-of-an-elusive-frog (accessed 15 December 2015) Gururaja, K.V., Dinesh, K.P., Priti, H. & Ravikanth, G. (2014) Mud-packing frog: a novel breeding behaviour and parental care in a stream dwelling new species of Nyctibatrachus (Amphibia, Anura, Nyctibatrachidae). Zootaxa, 3796 (1), 33–61. http://dx.doi.org/10.11646/zootaxa.3796.1.2 Heyer, W.R. (1971) Mating calls of some frogs from Thailand. Field Museum of Natural History. http://dx.doi.org/10.5962/bhl.title.2934 Howlader, M.S., Nair, A., Gopalan, S.V. & Merila, J. (2015) A new species of Microhyla (Anura: Microhylidae) from Nilphamari, Bangladesh. PLoS ONE, 10 (3), e0119825. http://dx.doi.org/10.1371/journal.pone.0119825 IUCN (2001) The IUCN Red List of Threatened Species Categories and Criteria. Ver. 3.1. Available from: http:// www.iucnredlist.org/static/categories_criteria_3_1 (accessed 15 December 2015) Jerdon, T.C. (1853) Catalogue of reptiles inhabiting the Peninsula of India. Journal of the Asiatic Society of Bengal, 22, 522– 534. Kanamadi, R., Hiremath, C. & Schneider, H. (1994) Courtship, amplexus and advertisement call of the frog, Microhyla rubra. Current Science, 66, 683–684. Katoh, K., Misawa, K., Kuma, K.I. & Miyata, T. (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research, 30, 3059–3066. http://dx.doi.org/10.1093/nar/gkf436. Kok, P.J.R. & Kalamandeen, M. (2008) Introduction to the taxonomy of the amphibians of Kaieteur National park, Guyana. ABC Taxa, 5, 1–278. Kuramoto, M. & Joshy, S.H. (2006) Morphological and acoustic comparisons of Microhyla ornata, M. fissipes and M. okinavensis (Anura: Microhylidae). Current Herpetology, 25, 15–27. http://dx.doi.org/10.3105/1345-5834(2006)25[15:maacom]2.0.co;2 Parker, H. (1934) A Monograph of the Frogs of the Family Microhylidae. Trustees of the British Museum, London, 208 pp. Seshadri, K.S., Singal, R., Priti, H., Ravikanth, G., Vidisha, M.K., Saurabh, S., Pratik, M. & Gururaja, K.V. (2016) Microhyla laterite sp. nov., A New Species of Microhyla Tschudi, 1838 (Amphibia: Anura: Microhylidae) from a Laterite Rock Formation in South West India. PLoS ONE, 11, e0149727. http://dx.doi.org/10.1371/journal.pone.0149727 Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution, 28, 2731–2739. http://dx.doi.org/10.1093/molbev/msr121 Vences, M., Nagy, Z.T., Sonet, G. & Verheyen, E. (2012) DNA barcoding Amphibians and Reptiles. In: Kress, W.J. & Erickson, D.L. (Eds.). DNA Barcodes: Methods and Protocols, Methods in Molecular Biology. Springer Science+Business Media, LLC, pp. 79–108. http://dx.doi.org/10.1007/978-1-61779-591-6_5 Wijayathilaka, N., Garg, S., Senevirathne, G., Karunarathna, N., Biju, S.D. & Meegaskumbura, M. (2016) A new species of Microhyla (Anura: Microhylidae) from Sri Lanka: an integrative taxonomic approach. Zootaxa, 4066 (3), 331–342. http://doi.org/10.11646/zootaxa.4066.3.9 Wijayathilaka, N. & Meegasakumbura, M. (2016) An acoustic analysis of the genus Microhyla (Anura: Microhylidae) of Sri Lanka. PLoS ONE, 11 (7), e0159003. http://doi.org/10.1371/journal.pone.0159003

View publication stats