HARVARD UNIVERSITY Ernst Mayr Library of the Museum of Comparative Zoology May °3 2® Volume 10 • 2004 Editor Er-Mi Zhao Chengdu Institute of Biology, Academia Sinica, Chengdu, Sichuan, China Associate Editors Raul E. Diaz Museum of Vertebrate Zoology, University of California, Berkeley, California, USA J. Robert Macey Joint Genome Institute, Walnut Creek, California, USA Theodore J. Papenfuss Museum of Vertebrate Zoology, University of California, Berkeley, California, USA James F. Parham Joint Genome Institute, Walnut Creek, California, USA; Museum of Paleontology, University of California, Berkeley, California, USA Editorial Board Robert F. Inger Field Museum, Chicago, Illinois, USA kraig Adler Cornell University, Ithaca, New York, USA Natalia B. Ananjeva Zoological Institute, St. Petersburg, Russia Steven C. Anderson University of the Pacific, Stockton, California, USA Aaron Bauer Villanova University, Villanova, Pennsylvania, USA Christopher Bell University of Texas, Austin, Texas, USA Leo Borkin Zoological Institute, St. Petersburg, Russia Bihui Chen Anhui Normal University, Wuhu, Anhui, China l-Jiunn Cheng Institute of Marine Biology, National Taiwan Ocean University, Keelung, Taiwan, China Ilya Darevsky Zoological Institute, St. Petersburg, Russia Indraneil Das Madras Crocodile Bank, Vadanemmeli Perur, Madras, India William E. Duellman University of Kansas, Lawrence, Kansas, USA Hajime Fukada Sennyuji Sannaicho, Higashiyamaku, Kyoto, Japan Carl Gans University of Michigan, Ann Arbor. Michigan, USA Xiang Ji Hangzhou Normal College, Hangzhou, Zhejiang, China Pi-peng Id Yantai Normal College, Yantai, Shandong, China Robert W. Murphy Royal Ontario Museum, Toronto, Ontario. Canada Goren Nilson University of Goteborg, Goteborg. Sweden Nikolai Orlov Zoological Institute, St. Petersburg, Russia Hidetoshi Ota Department of Biology, University of the Ryukyus, Nishihara, Okinawa, Japan Soheila Shafii University of Shahid Bahonar, Kerman, Iran Hai-tao Shi Hainan Normal University, Haikou. Hainan, China Xiu-ling Wang Xinjiang Normal University, Urumqi. Xinjiang, China Yue-zhao Wang Chengdu Institute of Biology. Academia Sinica, Chengdu. Sichuan. China Yehudah Werner Hebrew University, Jerusalem, Israel ken-tang Zhao Suzhou Railway Teacher's College, Suzhou, Jiangsu. China Asiatic Herpetological Research is published by the Asiatic Herpetological Research Society (AHRS) and the Chinese Society' for the Study of Amphibians and Reptiles (SCSSAR) at the Museum of Vertebrate Zoology, University of California. The editors encourage authors from all countries to submit articles concerning, but not limited to, Asian herpetology. All correspondence with- in China should be sent to Ermi Zhao, Editor, Chengdu Institute of Biology, P.O. Box 416. Chengdu, Sichuan Province, China. Authors should consult Guidelines for Manuscript Preparation and Submission at the end of this issue. Subscriptions and Membership Asiatic Herpetological Research is published by the Asiatic Herpetological Research Society (AHRS) and the Chinese Society for the Study of Amphibians and Reptiles (CSSAR). Volume 1 1 will be published in China and distributed by the Chengdu Institute of Biology, P. O. Box 416, Chengdu 610041, China. Subscriptions and memberships are $30 per year ($5*0 for libraries). For postage outside of China, please add $5 per issue for surface mail or $10 per issue for airmail. For "subscriptions outside of China, checks or money orders payable in US currency should be sent to: Bibliomania, P.O. Box 58355. Salt Lake City'. UT 84158-0355 USA. Phone/Fax 801-562-2660. Payment can be made by Credit Card (Visa. MasterCard. Discover, or American Express) through the Bibliomania web site, which has a secure server for credit cards (http://www.herplit.com). Asiatic Herpetological Research Volume 10 succeeds Volume 9 (2001), Volume 8 (published in 1999), Vol. 7 (1997), Vol.6 (1995), Vol.5 (1993), Vol. 4 (1992), Vol. 3 (1990), and Chinese Herpetological Research Vol. 2, which was published at the Museum of Vertebrate Zoology. 1988-1989. as the journal for the Chinese Society for the Study of Amphibians and Reptiles. Volume 2 succceeded Chinese Herpetological Research 1987. published for the Chengdu Institute of Biology by the Chongqing Branch Scientific and Technological Literature Press, Chongqing, Sichuan. China. Acta Herpetologica Sinica ceased publication in June, 1988. Cover: The cover image is of Gonocephalus chamaeleontinus from Pulau Tioman, Pahang, West Malaysia. Photo by L. Lee Grismer. 2004 Asiatic Herpetological Research Vol. 10, pp. 1-7 A New Species of Dibamuc (Squamata: Dibamidae) from West Malaysia Raijl E. Diaz1’2’*, Ming Tzi Leong3, L. Lee Grismer1, and Norsham S. Yaakob4 MCZ i 1 IRRAF Department of Biology, La Sierra University, Riverside, CA 925/5-8247, USA ~ Museum of Vertebrate Zoology’, University of California, Berkeley, CA 94 720, USA * Corresponding author E-mail: rauldiaz1 a herkeley.edu Department of Biological Sciences, National University of Singapore, Kent Ridge, ir. \ / CT w ?C Singapore 119260, Republic of Singapore 4 Forest Research Institute Malaysia, Kepong, 52109 Kuala Lumpur, Malaysia Abstract. - A new lizard of the genus Dibamus is described from Pulau Tioman and Pulau Tulai, Pahang, West Malaysia. This species most closely resembles D. novaeguineae, D. kondaoensis, D. leucurus and D. montanus , but differs from all congeneric species in exhibiting the following combination of characters: postoculars 1, scales bor- dering first infralabial 4, SVL 123 mm, 25-26 midbody scale rows, frontonasal and rostral sutures complete, and the presence of slightly posteriorly notched cycloid body scales as an adult. Key words. - Dibamus, Dibamus tiomanensis , new species, Dibamidae, Pulau Tioman, West Malaysia. Introduction The genus Dibamus presently contains 18 species (see Greer, 1985; Darevsky, 1992; Das, 1996; Honda et al., 1997; Ineich, 1999; Honda et al., 2001; Das and Lim, 2003; Das and Yaakob, 2003), a two-fold difference from the detailed review of the group by Greer (1985). Species of the genus Dibamus collectively range throughout southeast Asia, from southern China and the Philippines through Indonesia. Dibamus alfredi was described by Taylor (1962) from Thailand. D. alfredi were later found on the island of Nias, off the west coast of Sumatra (Greer, 1985) and from Danum Valley in Sabah State, Borneo (Tan, 1993; Das and Yaakob, 2003). A large gap was then left between Thailand and Borneo. Lim and Lim (1999) reported D. cf. alfredi from Pulau Tioman. Upon examination of their specimen, one from Pulau Tioman, and another from P. Tulai, we conclude that these specimens constitute a new species described herein. Pulau Tioman lies between longitudes 104° 7' to 104° 15' E and latitudes 2° 44' to 2° 54' N (Bullock and Medway, 1966). Finding another endemic population on this island provides another reason for its conservation as well as further studying its rich herpetofauna. Material and Methods Single females from both Pulau Tulai and Pulau Tioman were collected, fixed in 10% formalin, and preserved in 70% ethanol. Both specimens were deposited in the Zoological Reference Collection (ZRC) at the Rattles Museum of Biodiversity Research. Sliding calipers were used for all length measurements. Terminology used fol- lows Greer (1985) and Honda et al. (1997). Individuals were sexed externally under a dissecting microscope: males were identified by having two small, flap-like limbs (one on each side of the vent) (Dumeril and Bibron, 1839). Taxonomy Dibamus tiomanensis , new species Figs. 1, 2 Holotype. - ZRC. 2. 34 10, adult male collected at Kampung Paya, Pulau Tioman, Pahang, West Malaysia (Fig. 2) on 16 September 1995. Paratypes. - Adult female (ZRC 2.5092) collected along the trail to Bukit Bakau, Pulau Tulai, West Malaysia (Fig. 2) collected 14 July 2001. Juvenile female (ZRC. 2. 5260) collected along the Tekek-Juara Cross Island Trail, Pulau Tioman, West Malaysia collected 1 1 July 2001. Diagnosis. - Dibamus tiomanensis differs from all other species of Dibamus in having cycloid scales which are slightly notched posteriorly as an adult and flat cycloid light brown dorsal scales with cream borders as a juve- nile. It also differs from other Dibamus in having the fol- lowing combination of characters: rostral sutures incom- plete; nasal and labial sutures complete; scales bordering posterior edge of first infralabial 4; postocular 1; trans- verse scale rows just posterior to head 29, at midbody 25, proximally anterior to vent 21; subcaudals 45; snout blunt in lateral profile (Fig. 1; Table 1); presacral verte- brae 124; postsacral vertebrae 23 (Table 3). © 2004 by Asiatic Herpetological Research Vol. 10, p. 2 Asiatic Herpetological Research 2004 Figure 1 . Photograph of Dibamus tiomanensis, new species, on forest leaf litter. Description of holotype. - Snout-vent length 92.5 mm; tail length 13.1 mm; midbody diameter 2.5 mm. Snout bluntly rounded; nostril lateral; rostral pad with large number of evenly distributed sensory papillae; rostral sutures incomplete; nasal sutures complete from nostril Figure 2. Lateral (A), dorsal (B), and ventral (C) view of head of Dibamus tiomanensis , new species, (f: frontal, fn: frontonasal, ip: interparietal, if: first infralabial, I: labial suture, m: mental, n: nasal suture, o: ocular, po: postoc- ular, si: supralabial) to ocular; labial sutures complete from anterior part of nasal suture to mouth; frontonasal six times wider than long; frontal approximately 1.05 times wider than fron- tonasal; interparietal bordered posteriorly by four slight- ly smaller nuchal scales; postocular one; supralabial one; scales bordering posteromedial edge of first infral- abaial four; ear opening absent; eyes dimly visible through ocular; body scales notched posteriorly; trans- verse scale rows just posterior to head 23, at midbody 25, at just anterior to vent 23; subcaudals 50; tip of tail blunt, not terminating in a spine; hind limb length 2.6 mm. Description of paratypes. - The paratypes (both females) are similar to the holotype in all aspects except the following: transverse scale rows posterior to head 29, transverse scale rows anterior to vent 2 1 and 22, and subcaudals 45 and 48. Variation. - Paratype ZRC.2.5260 is the only juvenile. It shows a possible ontogenetic change in scale mor- phology. Juveniles have cycloid, flat, and light brown- cream bordered scales. Adults have posteriorly notched brown scales. Color in life. - Adults have a brown ground color both dorsally and ventrally, except on the snout and jaws which are a lighter shade of brown. Juveniles have a cream-colored snout and jaws which contrast well with the darker spotted sensory papillae and body scales which are light-brown bordered with cream. Manthey and Grossman (1997:205) present a color photograph of the holotype. 2004 Asiatic Herpetological Research Vol. 10, p. 3 Table 1. Comparison of several scale characters and measurements within Dibamus. The size of the frontal is meas- uied relative to the frontonasal and the interparietal relative to the surrounding anterior body scales. Sample sizes for postoculars and scales on posterior edge of infralabials are given in parentheses. Entries for midbody scale rows and subcaudal scales are as follows from top to bottom: range, mean, and sample size (modified from Greer, 1985). (*= 1 & 3 refer to 1 scale present on the left infralabial and 3 on right; **=tail regenerated; *** = text in Das and Yaakob (2003) mentions 3 scales bordering the infralabials in diagnosis, whereas 4 scales are mentioned in description of holotype). Dibamus Post- oculars Scales on posterior edge of infralabial Mid-body scale rows Subcaudal Scales Males Females Relative size of: Frontal Interparietal Max. SVL Tail Length (% of SVL) alfredi 2(4) 3(3) 20-21 46-47 41-46 1. 4-2.0 1. 7-2.2 135 17-18 4(1) 20.3 46.5 43.5 3 2 2 bogadeki 1(1) 2(1) 23 51 _ . . 177** 22.5 23 51 1 1 booliati 1(2) 3(2)*** 20 _ 24-39 _ _ 102.7 9.4-13.0 20 31.5 1 2 bourreti 1(1) 2(1) 24 _ 52+ 2.3 4.5 151 23+ 24 52+ 1 1 celebensis 2(10) 3(6) 26-30 38-40 35-40 1.2-2. 3 1. 0-2.9 188 10-13 3(3) 4(7) 27.4 39.3 38.0 13 3 4 deharvengi 1(1) 2(1) 16 57 _ 1.3 1.4 92 22.4 16 57 1 1 greeri 1(3) 1&3(2)* 20 53 54 _ _ 86 23-28 20 53 54 1 1 1 ingeri 2(1) 3(1) 20 36 _ 1.5 1.0 96 14.8 20 36 1 1 kondaoensis 2(1) 3(1) 23 59 _ 1.03 1.0 112.4 19.4 23 59 1 1 leucurus 1(23) 3(21) 20-23 48-52 41-47 1.2-4. 2 1. 0-3.1 136 16-20 4(2) 21.0 49.5 43.5 23 4 4 montanus 1(2) 2(2) 22 49 43 2.0 2.2 130 15-18 22 49 43 2 1 1 nicobaricum 1(6) 4(6) 23-25 34-38 31-36 _ _ 134.7 8.7-18.3 24.6 35.6 34.3 6 3 3 novaeguineae 2(92) 3(53) 22-26 42-45 37-42 1. 0-3.0 0.7-2. 4 158 9-19 3(2) 4(41) 24.5 43.0 39.6 5(1) 107 6 9 seramensis 4(1) 4(1) 33 - 40 0.7 1.2 203 11 33 40 1 1 smithi 1(1) 2(5) 18-19 59 59-61 1.5-2. 3 1. 3-2.0 108 21-24 2(4) 18.8 59 60.0 5 1 3 Vol. 10, p. 4 Asiatic Herpetological Research 2004 Table 1. Continued. Dibamus Post- oculars Scales on posterior edge of infralabial Mid-body scale rows Subcaudal Scales Males Females Relative size of: Frontal Interparietal Max. SVL Tail Length (% of SVL) somsaki 1(4) 2(4) 18-19 44**-58 27**-57 1.1-1.27 1.0-2.16 106 18-24 18.5 51 42 4 2 2 taylori 3(13) 2(2) 22-28 41-55 41-52 0.2-1. 3 1.0-1. 2 169 14-19 4(6) 3(14) 23.4 48.4 48 4(4) 22 5 7 tiomanensis 1(3) 4(3) 25-26 50 45-48 1.2 1.8 123 15-16 25.3 50 46.5 3 1 2 vorisi 2(2) 3(2) 20 33 11 1.2 1.0 89.2-90.1 6.1-16.8 20 33 11 2 1 1 Figure 3. Map of Southeast Asia showing the distribution of known Dibamus tiomanensis specimens. (1) Trail to Bukit, Pulau Tulai, (2) Tekek-Juara Cross island trail on Pulau Tioman (from Kg. Tekek to Kg. Juara) Etymology. - This species is named after the type local- ity for the holotype (Pulau Tioman = Tioman Island) Distribution. - Endemic to Pulau Tioman and adjacent Pulau Tulai (Fig. 2). Comparisons. - Dibamus tiomanensis was listed as D. cf. alfredi owing to its geographic proximity to D. alfre- di , which occurs in Peninsular Malaysia and Thailand (Manthey and Grossman, 1997; Taylor 1963). The pres- ence of four scales bordering the first infralabial posteri- orly differentiates the new species, Dibamus tiomanen- sis, from D. bogadeki, D. booliati , D. bourreti , D. dehar- vengi, D. greeri , D, ingeri, D. kondaoensis, D. mon- tanus, D. smithi, D. somsaki, and D. vorisi. In having one post ocular present, D. tiomanensis differs from D. alfredi, D. celebensis, D. novaeguineae, D. seramensis, and D. taylori. From the remaining two congeners, D. i 1 i i 1 mm 1 mm Figure 4. Lateral (A), dorsal (B), and ventral (C) view of heads of Dibamus alfredi (left) and Dibamus novaeguineae (right). Figures from Greer (1985). [f: frontal, fn: frontonasal, ip: interparietal, if: first infralabial, I: labial suture, m: mental, n: nasal suture, o: ocular, po: postocular, si: supralabial] tiomanensis differs from D. montanus Smith, 1921 (Langbian Plateau, Vietnam) in having more pre-sacral vertebrae (124 vs. 112-114) and D. leucurus (Bleeker, 1860) (Sumatra, Borneo) in the presence of slightly pos- teriorly notched cycloid scales as an adult. 2004 Asiatic Herpetological Research Vol. 10, p. 5 Table 2. Matrix of diagnostic characters and their states for species of Dibamus (modified from Greer, 1985). Dibamus Rostral suture complete and separate (+), complete and meeting (-), incomplete (0) or absent (1) Nasal suture complete (+), reduced (-), or absent Labial suture complete (+), incomplete dor- sally to varying degrees (-), or absent (0) First infralabial surrounded by 1 ("*■)» 2 (-). 3(0), 4(1), or 5(2) scales No. of post ocular scales 1(+),2(-), 3(0), or 4(1) Body scales cycloid (+) or cycloid and slightly knotched posteriorly (-) in adults alfredi 0 - - 0,1 - + bogadeki 0 + + - + + booliati 1 - + - + + bourreti + + 0 - + + celebensis 0 + + 0,1 -,o + deharvengi - + + - + + greeri 0 - + 0* + + ingeri 0 + + 0 - + kondaoensis 0 + + 0 - + leucurus 0 - + 0,1 + + montanus - + + 0 + + nicobaricum + + + 1 + + novaeguineae 0 + + 0,1,2 -,0 + seramensis 0 + + 1 1 + smithi 0 - 0 - + - » + somsaki - + + - + + taylori 0 + + -,0,1 0,1 + tiomanensis 0 + + 1 + - vorisi 0 + 0 0 - + * = See Table 1 for information on this character state. D. nicobaricum is included in this study following Das' ( 1 996) redescription and reevaluation of the species (in which it is inaccurately referred to as D. nicobaricus through parts of the paper) despite Honda et al. (2001) avoidance of it's recognition as a nominate species. Great difficulty arises in finding specimens of Dibamus for study due to their fossorial lifestyle. As a result, many descriptions are based on 2-5 individuals. An unusually large collection of D. novaeguineae from Mt. Canlaon, Negros Island, Philippines (Greer, 1985:150) has given a unique insight to how variable morphological characters can be within a single popula- tion (See Table 1). Further studies are needed in study- ing variation within this family as slight character state variances have warranted the recognition of new species [See Das and Lim (2003), Das and Yaakob (2003), and this paper] which may prove to be a variant of an already described taxon. Vol. 10, p. 6 Asiatic Herpetological Research 2004 Table 3. Sacral vertebrae count of described species of Dibamus. Dibamus Sacral vertebrae pre-sacral post-sacral alfredi 116-126 22-26 bogadeki 134 25 booliati 113-120 11-25 bourreti 115-129 12-40+ celebensis 117-132 17-22 deharvengi 120 36 greeri 96-111 28-31 ingeri 97 21 kondaoensis 140 33 leucurus 106-135 21-28 montanus 112-114 24-27 nicobaricum 124 23 novaeguineae 109-125 18-24 seramensis 130 18 smithi 130-137 30-34 somsaki 119 31 taylori 112-135 21-29 tiomanensis 124 23 vorisi 97 20 Natural History The holotype was found under a large stone in Kampung Paya, Pulau Tioman. ZRC 2.5092 was found beneath leaf litter in loose dirt adjacent to a large rock and bam- boo stands in secondary forest along the trail to Bukit Bakau on Pulau Tulai at 20 m elevation. ZRC.2.5260 was found beneath a decaying log one meter from the cross-island trail in primary forest on Pulau Tioman. Adult Dibamus tiomanensis displayed a behavior most likely intended to ward off a predator. When picked up or startled, the body scales flare up at an angle almost perpendicular to the body. When viewed, the smooth surface appears rugose, resembling the bristle- covered epidermis of an earthworm. It is possible that a non-palatable species of worm exists in the same area and has served as a model for D. tiomanensis to mimic. Darevsky (1992) mentions that D. greeri has bright blue rings on its body, perhaps mimicking a megascolicid worm inhabiting the same leaf litter. Such mimicking behavior was also recently reported in D. booliati (Das and Yaakob, 2003). Acknowledgments We thank Mr. Sahir bin Othman, Director of Wildlife, Perhilitan, for permission to conduct fieldwork in the Seribuat Archipelago and Peter Ng, Chang Man Tang and Kelvin Kok Peng Lim for the loan of specimens. We also thank Allen Greer and Ted Papenfuss for reviewing the manuscript. We especially thank Indraneil Das for his significant contribution to the manuscript and for providing sacral counts. We would also like to thank all the students of Tropical Field Biology 487e from La Sierra University who provided field assistance, and Karen Klitz for her help with Fig. 2. Literature Cited Bullock, J. A. and L. Medway. 1966. Observations on the Fauna of Pulau Tioman and Pulau Tulai: General Introduction. Bulletin of the National Museum, Republic of Singapore. No. 34, March 1966. Darevsky, I. S. 1992. Two new species of worm-like lizard Dibamus (Sauria: Dibamidae) with remarks on the distribution and ecology of Dibamus in Vietnam. Asiatic Herpetological Research 4:1-12 Das, I. 1996. The validity of Dibamus nicobaricum (Fitzinger in Steindaehner, 1867) (Squamata: Sauria: Dibamidae). Russian Journal of Herpetology3(2): 157-162 Das, I. and K. K. P. Lim. 2003. Two new species of Dibamus (Squamata: Dibamidae) from Borneo. Raffles Bulletin of Zoology 51(1): 137-141 . Das, I. and N. Yaakob. 2003. A new species of Dibamus (Squamata: Dibamidae) from Peninsular Malaysia. Raffles Bulletin of Zoology 51(1): 143- 147 Greer, A. E. 1985. The relationships of the lizard gen- era Anelytropsis and Dibamus. Journal of Herpetology 19:116-156. Honda, M. and J. Nabhitabhata, H. Ota., T. Hikida. 1997. A new species of Dibamus (Squamata: Dibamidae) from Thailand. Raffles Bulletin of Zoology 45(2):275-279 Honda, M., H. Ota, T. Hikida, and I. S. Darevsky 2001. A new species of the worm-like lizard, Dibamus Dumeril & Bibron 1839 (Squamata Dibamidae), from Vietnam. Tropical Zoology 14:119-125. Ineich, I. 1999. Une nouvelle espece de Dibamus (Reptilia: Squamata: Dibamidae) du Vietnam. Bulletin de la Societe zoologique de France 124 (3):279-286 2004 Asiatic Herpetological Research Vol. 10, p. 7 Lim, K. P and L. J. Lim. 1999. The terrestrial herpeto- fauna of Pulau Tioman, Peninsular Malaysia. Raffles Bulletin of Zoology 6:131-155. Manthey, U. and W. Grossmann. 1997. Amphibien & Reptilien Siidostasiens. NTV Verlag, Munster, 512 pp. Taylor, E. H. 1962. New oriental reptiles. University of Kansas Science Bulletin 44( 14):687- 1 077 Taylor, E. H. 1963. The lizards of Thailand. University of Kansas Science Bulletin 44:687-1077. 2004 Asiatic Herpetological Research Vol. 10, pp. 8- A New Species of Leptolalax (Anura: Megophryidae) from Pulau Tioman, West Malaysia L. Lee Grismer, Jesse L. Grismer, and Timothy M. Youmans Department of Biology, La Sierra University, Riverside, CA 92515-8247 Abstract. - A new species of Leptolalax is described from a cave at the top of Gunung Kajang, Pulau Tioman, Pahang, West Malaysia. It differs from all other Malaysian species of Leptolalax in several aspects of coloration and in hav- ing smooth as opposed to rough or pebbled skin on the dorsum. Key words. - Leptolalax , Anura, Tioman, Malaysia. Introduction There are at least eight species of Leptolalax found throughout West Malaysia and Borneo (Berry, 1975; Inger et al., 1995; Inger et al., 1997; Matsui, 1997) but only one of these, L. gracilis, is known from both regions. In an unpublished document, Day (1990) reported Leptobrachium sp. from Pulau Tioman, Pahang, West Malaysia based on five larvae collected in the Tengkuk Air Cave (Gua) at 1000 m elevation on Gunung Kajang. Lim and Lim (1999) referred these lar- vae to Leptolalax gracilis based on three additional lar- vae they collected at a lower elevation (ca. 400 m) on Gunung Kajang. More recently, Grismer et al. (2004) reported two adults from Gua Tengkuk Air. Examination of those adults indicates that they belong to the genus Leptolalax not Leptobrachium (but they are not L. gracilis). We therefore, refer these specimens and the larvae of Lim and Lim (1999) to the new species described herein. Materials and Methods The following measurements were made with sliding calipers to the nearest 0. 1 mm: adults; snout-vent length (SVL), tibial length (TL), head width (HW), head length (HL), and diameter of tympanum (TYM). Tadpoles (Table 1); interorbital distance (IOD), intemarial dis- tance (IND), tail length (TL), and tail height (TH). Tadpoles were preserved in 10% formalin and trans- ferred to 70% ethanol. Due to shrinking and wrinkling, head-body width (HBW), head-body height (HBH), and tail height (TH) are likely to be underestimated. Observations on external morphology were made with the use of a dissecting microscope. Specimens were compared to material listed in Appendix I and descrip- tions in Berry (1975), Inger (1985), Inger et al. (1997), Inger et al. (1995), Inger and Stuebing (1997), Malkmus (1992), and Matsui (1997). Leptolalax kajangensis , new species (Figs. 1-2) Holotype. - ZRC 1.7714; adult male, 34.0 mm SVL; found inside Gua Tengkuk Air, Gunung Kajang, Pulau Tioman, Pahang, West Malaysia at 1000 m in elevation. Collected by Jesse L. Grismer and Timothy M. Youmans on 21 July, 2001. Paratype. - ZRC 1.7715; adult male, 35.0 mm SVL. Same data as holotype. Diagnosis. - Leptolalax kajangensis differs from all other Malaysian species of Leptolalax in lacking distinct dark crossbands on the limbs and having a generally dark, unicolor to weekly patterned dorsum. It differs fur- ther from L. arayai, L. dringi, L. gracilis, L. hamidi, and L. maurus, in having smooth, as opposed to rough, warty skin. It differs further from L. dringi, L. gracilis, and L. heteropus in lacking dark spots on the ventrum. It dif- fers further from L. gracilis and L. hamidi in lacking a dark inguinal blotch and from L. gracilis in lacking a distinctly bicolored forelimb and dark markings on the ventral surface of the foreleg. Description of holotype. - Habitus moderately slender, head longer than wide; snout rounded in dorsal profile, weakly projecting beyond lower jaw; nostrils raised, directed dorsolaterally, and situated on canthi, near tip of snout; canthi rounded; lores slightly oblique; eye large, diameter slightly longer than length of snout; intemarial distance less than interorbital distance; interorbital width slightly greater than length of snout; tympanum visible, less than one-half diameter of eye, separated from eye by width of tympanum; vomerine teeth absent; tongue notched, without papillae. Forelimbs slender; fingers slender, unwebbed, tips rounded; first finger equal in length to second; no subar- ticular tubercles or elongate comified pads visible beneath fingers; large inner palmar tubercle present and © 2004 by Asiatic Herpetological Research Vol. 10, p.9 Asiatic Herpetological Research 2004 Figure 1. Photograph of Leptolalax kajangensis , new species, on forest leaf litter. much smaller outer palmar tubercle at base of fourth digit. Hindlimbs relatively short; heels overlap when limbs flexed; tibiotarsal articulation of adpressed limb reaches tip of snout; tips of toes rounded; third and fifth toe equal in length; slight webbing between first and second and second and third toes only; oval inner metatarsal tubercle present; subarticular tubercles absent. Skin on back smooth to faintly pebbled; flanks faintly pebbled; limbs smooth to faintly pebbled; promi- nant supratympanic fold extending from eyelid to shoul- der; ventral surfaces smooth. Coloration. - In life and ethanol, dorsal surfaces almost black with no visible pattern except for minute, faint, light-colored spots; limbs slightly lighter with faint dark mottling; faint dark mottling on flanks blending to gray- ish dusty ventral surfaces punctated by minute gray spots. Supratympanic fold darker than dorsum. Iris metallic gold in life. Measurements of holotype. - SVL 34.0 mm; TL 14.2 mm; HW 18.5 mm; HL 10.0 mm; TYM 1.6 mm. Variation. - The paratype closely approximates the holotype in all aspects of coloration and morphology. It differs in the following measurements: SVL 35.0 mm; TL 14.6 mm; HW 19.4 mm; HL 9.4 mm TYM 1.8 mm. Etymology. - This species is named after the type local- ity of Gunung Kajang, Pulau Tioman, West Malaysia Tadpoles. - Table 1 lists measurements, growth stages, and tooth row formulae of tadpoles. Based on specimens (ZRC 1.3339-41); stages 30, 36, and 37 (Gosner, 1960) from a small stream at 333 m below Gua Tengkuk Air on G. Kajang and specimens (0051-54 uncatalogued spec- imens in the British Museum reported by Day, 1990) stage 30 (Gosner, 1960) from Gua Tengkuk Air. Head and body relatively large and round; nostrils dorsally located, closer to tip of snout than eye; eyes dorsolater- al, not visible from below; spiracle sinistral, nearer to eye than vent; vent dextral, opening at margin of ventral fin; dorsal fin slightly deeper than ventral fin, not extending onto head-body; fins not deeper than caudal musculature; oral apparatus not emarginate; mouth sub- terminal; bordered by a single row of conical papillae; large submarginal papillae without denticles; jaw sheaths black, robust, strongly arched, and finely serrat- ed; upper jaw lacking medial notch; denticles small; rows A1 and P5 markedly shorter than adjacent rows. Tadpoles (ZRC. 1 .3339-41) from stream dusky brown above with faint darker mottling; venter cream colored, nearly immaculate; faint dark stippling on caudal fins, edges clear and immaculate; lateral line hash marks dis- tinct. Tadpoles from Gua Tengkuk Air (uncatalogued BM specimens) lack pigment and are white to transpar- ent in life. Comparisons with other species. - Leptolalax kajan- gensis differs from all other Malaysian species of Leptolalax on the basis of skin surface texture and col- oration. Leptolalax arayai , L. dringi , L. gracilis , L. hamaidi , L. m auras , and L. pe/odvtoides all have coarse- ly textured skin ranging from distinctive corrugations in Table 1. Selected measurements (mm), growth stage (GS), and labial tooth row formulae (LTRF) of Leptolalax kajangensis. Abbreviations follow Materials and Methods. * = Specimen damaged. Cat. no. HBW HBL HBH IOD IND TL TH GS LTRF 0051 13.5 23.3 8.4 4.8 3.2 39.9 10.0 30 6(2-6)/5(1-4) 0052 15.2 23.8 9.3 6.6 4.9 49.9 13.7 38 6(2-6)/5(1-4) 0053 13.5 23.5 10.0 5.0 4.3 ★ * 30 6(2-6)/5(1-4) 0054 14.5 25.5 10.0 5.4 3.9 45.5 11.0 30 6(2-6)/5(1-4) 0055 14.1 25.9 10.0 6.7 4.8 43.5 12.0 38 6(2-6)/5(1-4) ZRC. 1.3339 11.2 23.5 8.2 6.0 2.6 41.4 ★ 30 6(2-5)/5(2-4) ZRC. 1.3340 19.6 30.8 14.5 7.3 4.5 56.4 * 35 6(2-6)/1 (1-3) ZRC. 1.3341 18.0 31.9 14.2 7.8 4.6 51.9 ★ 36 5(4)/4(1 -3) 2004 Asiatic Herpetological Research Vol. 10, p. 10 Figure 2. Tadploe of Leptolalax kajangensis at type local- ity. L. dringi to isolated tubercles of varying sizes in L. gra- cilis (Inger and Stuebing, 1997; Inger et al. 1997; Matsui, 1997). Leptolalax kajangensis resembles L. pic- tus and L. pelodytoides in having smooth to weakly peb- bled skin on the dorsum (Inger et al. 1995) but differs from L. pictus in lacking small tubercles on its sides (Malkmus, 1992). Leptolalax kajangensis falls within the size range (SVL 34.0-35.0 mm) of L. dringi (SVL 30.0-38.8 mm), L. gracilis (SVL 31.0-48.0 mm), L. het- eropus (SVL 33.0-35.0 mm), L. pelodytoides (SVL 30.0- 42.0 mm), and L. pictus (SVL 3 1 .0-47.0 mm) but is larg- er than L. arayai (SVL 29.9) and L. mams (SVL 26.0- 32.0). Leptolalax kajangensis differs most notably from other species in coloration. Its generally unicolor black dorsal pattern contrasts sharply with the various mottled body patterns of L. dringi , L. hamidi, L. heteropus , L pelodytoides , and L. pictus. It does resemble L. maurus to some extent in that L. maurus has a black unicolored dorsum but differs in that the latter also has lighter col- ored limbs with darker crossbars and light colored spots on the flanks, patterns which are absent in L. kajangen- sis. Leptolalax gracilis also has a generally dark dorsal coloration with varying degress of dark mottling but is distinctive in having a light colored brachium. The sin- gle specimen of L. arayi has a light ground color with darker crossbands on the limbs. Natural history. - Both specimens of Leptolalax kajan- gensis were found in a cave (Gua Tengkuk Air) near the summit of Gunung Kajang at approximately 1000 m in elevation. This subterranean, obliquely oriented cavern is formed from the overhang of large boulders piled on top of one another. It contains a small pond (3 m x 4 in) drained by a small subterranean stream (1-3 m in width by 2-4 cm in depth) running for 3-4 m along the cave floor. Both specimens of L. kajangensis were observed in the afternoon sitting on top of large rocks adjacent to the stream approximately 10 m from the entrance to the cave and approximately 10 m below the outside ground level. It was from the subterranean pond that Day (1990) collected "large tadpoles" he referred to as Leptobrachium sp. We observed additional tadpoles and assume these to be larvae of Leptolalax kajangensis based on the fact that the type material were collected from the same stream and adults were observed calling from the edge of the pond. Three additional larvae examined here (ZRC. 1.3339-41) were collected from a lower stream on Gunung Kajang at approximately 400 m on 26 June 1996 (Lim and Lim, 1999). We tentatively assign these to Leptolalax kajangensis because they match the larvae from Tengkuk Air in morphology (Table 1). However, the Tengkuk Air specimens lack pigment and are white to transparent in life (Fig. 2). ZRC. 1 .3339-41 are coun- tershaded with dark pigment above and have minute dark flecks on a light venter. This indicates that either L. kajangensis is not confined to only the upper most ele- vations of Gunung Kajang or that there may be an addi- tional species of Leptolalax found lower down on the mountain. Biogeography. - Pulau Tioman had a land positive con- nection with Peninsular Malaysia as late as the Pleistocene (Voris, 2000). The presence of amphibian species on Pulau Tioman such as Leptolalax kajangen- sis, Megophrys nasuta , Rana hosii , R. picturata and oth- ers that require streams with moderate to strong currents for reproduction suggests these species are unlikely can- didates for long distance dispersal over flat, low-lying landscape (Inger and Voris, 2001). Pulau Tioman was part of a large granitic arc of mountains extending from what is currently peninsular Malaysia through the Kepulauan Anambas and Natunas across the Greater Sunda Shelf which provided a dispersal corridor for montane species (Inger and Voris, 2001) across the flat, low-lying exposed Sunda Shelf. Thus, the presence of stream-breeding species requiring moderate to fast flow- ing water on Pulau Tioman is most likely a result of vic- ariance. Acknowledgments We thank Mr. Sahir bin Othman, Director of Wildlife, PERHILITAN, for permission to conduct field work in the Seribuat Archipelago. We thank Peter Ng and Kelvin Lim of Raffles Museum of Biodiversity Research (ZRC) and Harold Voris and Alan Resetar of the Field Museum of Natural History (FMNH) for allowing us to examine specimens in their care. For comments on the manuscript we are grateful to R. F. Inger and for field assistance we thank Geoff Powells. Vol. 10, p. 11 Asiatic Herpetological Research 2004 Literature Cited Berry, P. Y, 1975. The Amphibian Fauna of Peninsular Malaysia. Tropical Press, Kuala Lumpur. Day, M. 1990. Zoological research. In: Day, M. and T. Mowbray (eds.). University of Bristol Tioman Archipelago Expedition, Peninsular Malaysia, 1988, Final Report. Unpublished Report pp. 25-43. Gosner, K. L. 1960. A simplified table for staging anu- ran embryos and larvae with notes on identification. Herpetologica 16:183-190. Grismer, J. L., L. L. Grismer, I. Das, N. S. Yaakob, L. B. Liat, L. T. Ming, T. M. Youmans, R. A. Sosa, and H. Kaiser. 2004. Species diversity and checklist of the herpetofauna of Pulau Tioman, peninsular Malaysia with a preliminary overview of habitat utilization. Asiatic Hereptological Reserch 10:247-279. Inger, R. F. 1985. Tadpoles of the forested regions of Borneo. Fieldiana Zoololgy. New Series No.26:l- 89. Inger, R. F., M. Lakim, A. Biun, and P. Yambun, 1997. A new species of Leptolalax (Anura:Megophryidae) from Borneo. Asiatic Herpetological Reserach 7:48- 50. Inger, R. F. and R. B. Stuebing, 1997. A Field Guide to Frogs of Borneo. Natural History Publications, Kota Kinabalau. Inger, R. F., R. B. Stuebing, and F. L. Tan. 1995. New species and new records of anurans from Borneo. Raffles Bulletin of Zoology 43 : 1 1 5- 1 3 1 . Inger, R. F. and H. K. Voris. 2001. The biogeographical relations of the frogs and snakes of Sundaland. Journal of Biogeography 28:863-891. Lim, K. K. P. and L. J. Lim. 1999. The terrestrial her- petofauna of Pulau Tioman, Peninsular Malaysia. Raffles Bulletin of Zoology. Supplement 6:131- 155. Malkmus, R. 1992. Leptolalax pictus, sp. nov. (Anura: Pelobatidae) vom Mount Kinabalu/Nord-Borneo. Sauria, Berlin 14:3-6. Mastsui, M. 1997. Call characteristics of Malaysian Leptolalax with a description of two new species (Anura: Pelodatidae). Copeia 1997:158-165. Voris, H. K. 2000. Maps of Pleistocene sea levels in Southeast Asia: shorelines, river systems, time dur- ations. Journal of Biogeography 27:1153-1167. 2004 Asiatic Herpetological Research Vol. 10, pp. 12-16] A New Species of Kukri Snake, Oligodon (Colubridae), from Pulau Tioman, West Malaysia. Tzi Ming Leong1 and L. Lee Grismer2 * Department of Biological Sciences, National University of Singapore, Singapore 119260 - Department of Biology, La Sierra University, Riverside, California 92515-8247, USA Abstract. - A unique species of Oligodon is described from the type locality of Pulau Tioman, West Malaysia. In terms of scalation, it is most comparable with the Bornean O. subcarinatus, but does not exhibit any feeble keeling of scales. In addition, its body color and patterns are unique in having a red dorsum, pink ventral surface, and indis- tinct pale bars on the nape and body. Key words. - Oligodon , kukri snake, Pulau Tioman, West Malaysia. Introduction The kukri snakes belonging to the genus Oligodon Boie, 1827 are so named because of the presence of unique posterior maxillary teeth, shaped like Ghurka kukri knives. In addition, members belonging to this genus are small to medium-sized ground dwelling species charac- terised by having a large slightly upturned rostral shield, short head, round pupil, and a cylindrical body with smooth scales (Tweedie, 1983; Cox et al., 1998). Many species possess a distinct dark chevron mark on the nape and a stripe across the anterior part of the head and down over/through the eye. Although Oligodon is well repre- sented in South and Southeast Asia, there are only three species on Peninsular Malaysia, namely O. octolineatus Schneider, O. purpurascens Schlegel, and O. signatus Gunther (Tweedie, 1983). In Borneo, eight species have been recorded [O. annulifer Boulenger, O. cinereus Gunther, O. everetti Boulenger, O. octolineatus Schneider, O. purpurascens Schlegel, O. signatus Gunther, O. subcarinatus Gunther, and O. vertebralis Gunther]. However, the occurrences of true O. annulifer and O. cinereus on Borneo remain to be verified (Stuebing and Inger, 1999). On Pulau Tioman, ca. 40 km from the southeast coast of the peninsula, one species ( O . purpurascens) has been reliably recorded thus far (Grismer et al., 2004; Hendrickson, 1966). The presence of O. octolineatus on the island, though possible, remains to be verified (Lim and Lim, 1999). Day (1990: 38) reported the presence of a distinct, new form of Oligodon from the cross-island (Tekek-Juara) trail, but did not provide any diagnostic characters. It was merely mentioned that this form resembled O. signatus , but had differences in terms of head scalation and dorsal colouration. We collected a specimen from the same locality on 16 July, 2001 that is different from all other nominal species of Malaysia and is herein named and described as new. Materials and Methods Prior to preservation in 10% formaldehyde, the speci- men was photographed and liver tissue sampled. Total length, tail length, and snout-vent length were obtained using a measuring tape (to nearest 1 mm). Additional measurements taken, using vernier callipers (to nearest 0.1 mm), include eye diameter (ED); head length (HL), taken from the union of the posteromedial comers of the parietals to the tip of the snout; head depth (HD), taken from the dorsal surface of the head to the ventral surface of the jaw immediately posterior to the eye; and snout length (SL), taken from the anterior margin of the eye to the tip of the snout. The scale counts included upper labials, number of upper labials in contact with eye, lower labials, preoculars, postoculars, ventrals, subcau- dals, and midbody scales. Comparative material was examined from the Zoological Reference Collection (ZRC) [Raffles Museum of Biodiversity Research (RMBR), National University of Singapore], the Department of Wildlife and National Parks, Peninsular Malaysia (DWNP) herpetological collection, the Bishop Museum, Hawaii (BPBM), and Museum fur Naturkunde, Humboldt-Universitat, Berlin, Germany (ZMB). Oligodon booliati, sp. nov. Holotype: ZRC.2.5153, adult female, collected on the night of 16 July, 2001, at 2130 hrs by T. M. Leong and K. M. Crane, while it was crawling on a concrete stair- case in primary forest along the Tekek-Juara trail, ca. 150 m ash, Pulau Tioman (Pahang, Peninsular Malaysia). Deposited at the Zoological Reference Collection (ZRC). Paratypes: (1) BPBM 13933, subadult male, collected on 17 April 1962 by J. R. Hendrickson, at Ulu Lalang, © 2004 by Asiatic Herpetological Research Asiatic Herpetological Research 2004 Vol. 10 p. 13 Figure 1. Dorsal view of Oligodon booliati holotype, ZRC.2.5153 (live coloration). ca. 700 m asl and (2) ZMB 64446, adult female, collect- ed in May 2001 by W. Grossmann and C. Scafer on the top of the Tekek-Juara Trail at 300 in asl. Diagnosis. - 6-7 upper labials, 2nd and 3 rd or 3rd and 4th touching eye, 7 lower labials, 17 mid-body scales, 143-153 ventrals, 54-60 subcaudals. Loreals present. Head scales without distinct patterns. No distinct stripe running through eye. In life, body deep maroon red dor- sally and along flanks, salmon pink on the ventrals and subcaudals. Ventrals without dark spots. Indistinct dark brown transverse bars (19-22) on body, starting from nape and fading increasingly towards tail. Thin, dark brown stripes on anterior sides of 5th and 6th upper labi- als, immediately posterior to eye. Description of Holotype. - Adult female, total length: 510 mm, tail: 121 mm, snout-vent: 389 mm, ED: 2.2 mm, HL: 10.8 mm, HD: 6.9 mm, SL: 5.0 mm; head stout (HD/HL 0.64), slightly broader than neck; snout moder- ate, oblique in dorsal profile, oval in lateral profile; eye 20% of head length, pupil round; rostral shield large, tri- angular, visible from above, width (3.5 mm) greater than height (3.0 mm), concave below; rostral in direct contact with first upper labials, nasals and internasals; trapezoid prefrontals posterior to intemasals, curving at canthus rostrals to meet triangular postnasals; single, almost tri- angular frontal, slightly longer (4.8 mm) than wide (4.2 mm), posterior point terminating 1.2 mm beyond poste- rior margin of eye; frontal flanked by prominent supraoculars, length of supraocular (3.0 mm) 1 .4 of ED; 7 upper labials, increasing in size to 6th, 3rd and 4th in contact with eye; a small (0.5 x 0.5 mm) loreal scale present on left, but absent on right, allowing contact between right prefrontal and 2nd upper labial; one preoc- ular and one postocular present on both sides; 7 lower labials, first pair elongated transversely, confining men- tal to meet medially, 4th pair largest; anterior chinsh ields longer than (almost double) posterior chinshields; 17 scale rows at neck and midbody, but 15 at one head length anterior to vent; 143 ventrals, lateral margins vis- ible from the sides; 59 subcaudals, all divided; anal shield single. Colour in life deep maroon red on dorsum and flanks (Fig. 1). Ventrals salmon pink, the colour becoming increasingly intense towards the tail tip (Fig. 2), without any melanistic pigmentation. Head scales without markings dorsally. Distinguishable grey-brown pigmentation visible in other head scales: dorsal portion of 4th upper labial (point of contact with eye), anterior margins of 5th and 6th upper labials, anterior margins of 1st to 4th lower labials, in between anterior chinshields towards the anterior, within mental, and lower surface of rostral. Crossbars on body faint. 21 crossbars from nape towards tail tip. Upon preservation, and storage in alco- 2004 Asiatic Herpetological Research Vol. 10, p. 14 Table 1 Measurements, scale counts, and number of crossbars among Oligodon booliati sp. nov. type materials (holo- type and two paratypes). L/R = Left/Right sides. Holotype ZRC.2.5153 Adult female Paratype #1 BPBM 13933 Subadult male Paratype #2 ZMB 64446 Adult female Snout-vent length (mm) 389 260 291 Total length (mm) 510 348 373 Upper labials (L/R) 7/7 7/7 6/6 Upp. labials touching eye 3rd & 4th 3rd & 4th 2nd & 3rd Lower labials (L/R) 7/7 7/7 7/7 Anterior nasal (L/R) 1/1 1/1 1/1 Posterior nasal (L/R) 1/1 1/1 1/1 Loreal (L/R) 1/0 1/1 1/1 Pre-ocular (L/R) 1/1 1/1 1/1 Post-ocular (L/R) 1/1 1/1 1/1 Supra-ocular (L/R) 1/1 1/1 1/1 Midbody 17 17 17 Ventrals 143 146 153 Subcaudals 59 60 54 # crossbars on body 21 22 19 Figure 2. Ventral view of Oligodon booliati holotype, ZRC.2.5153 (live colouration). hoi, the colours gradually faded to a lighter shade. A lat- eral view of the head is illustrated in Fig. 3. Description of paratypes. - Rostral shield large, visible from above. BPBM 13933 has 7 upper labials with the 3rd and 4th touching eye. ZMB 64446 has 6 upper labials with the 2nd and 3rd touching the eye. Loreal scale (0.5 x 0.5 mm) present on both sides in BPBM 13933 and fused to the postnasal on both sides in ZMB 64446 (1.0 x 1.0 mm). One preocular and one postocular. Seven lower labials, 17 midbody scales, 146 ventrals, 60 sub- caudals in BPMB 13933 and 153 ventrals and 54 sub- caudals in ZMB 64446. First pair of infralabials separat- ed medially by the mental in ZMB 64446. Total length (BPBM 13933): 348 mm; tail: 88 mm; snout-vent: 260 i i Figure 3. Lateral view of head (left side) of Oligodon boo- liati holotype, ZRC.2.5153. Scale bar = 5mm. mm. Total length (ZMB 64446): 373 mm; tail: 82 mm; snout-vent: 291 mm. Anal shield single, subcaudals paired. Colour in preservative faded to a cream colour. No distinctly striking markings on head, faint crossbars on body (22 in BPBM 13933, 19 in ZMB 64446). Measurements and scale counts of type materials are summarised in Table 1 . Etymology. - This new species is named in honor of Lim Boo Liat, of the Department of Wildlife and National Parks (Peninsular Malaysia), whose contribu- tions to our better understanding of Malaysia's natural history dates back to the 1950's. Flis publications include the description of a new snake ( Macrocalamus tweediei Li in, 1963) and the popular reference book Poisonous Snakes of Peninsular Malaysia (Lim, 1982). Vol. 10 p. 15 Asiatic Herpetological Research 2004 Table 2. Comparisons between the Oligodon species (arranged alphabetically) of Peninsular Malaysia and Borneo, including Oligodon booliati sp. nov. (measurements and scalations after Manthey & Grossmann, 1997). Lo = Loreal, UL - Upper Labials, MB = Midbody, V = Ventrals, SC = Subcaudals. Oligodon species Total length (cm) Lo UL UL touching eye MB V SC annulifer 43 1 7-8 3 & 4 15 148-162 40-58 booliati sp. nov. 51 1 6-7 2 & 3/3 & 4 17 143-153 54-60 cinereus 73 1 7-8 4 & 5 15/17 151-185 29-43 everetti 42 1 7 3 & 4 15 138-154 46-65 octolineatus 68 1 6 3 & 4 17 155-197 43-61 purpurascens 95 1 8 4 & 5/5 19/21 160-210 40-60 signatus 60 0 7 3 & 4 17 141-157 47-59 subcarinatus 40 1 7 3 & 4 17 155-166 50-54 taeniatus 34 1 8 3-5 17 146-169 30-47 vertebralis 35 1 7 3 & 4 15 136-154 35-54 Discussion The number of upper labials (6-7) in Oligodon booliati may be used to distinguish it from O. purpurascens (8) and O. taeniatus (8), although O. annulifer and O. cinerens may occasionally possess eight upper labials. O. booliati shares the same number of midbody scales (17) as O. octolineatus, O. signatus, and O. subcarina- tus (instead of 15 in O. annulifer , O. everetti and O. ver- tebral is) but may be distinguished from O. octolineatus by the absence of any longitudinal stripes along the body, from O. signatus by the presence of loreal scales, and from O. subcarinatus by the lack of feeble keels on its scales. This new species is assumed to be endemic to Pulau Tioman. Comparisons of measurements and scala- tion between O. booliati and the other species of Oligodon in Peninsular Malaysia and Borneo are sum- marised in Table 2. Comparative Material Examined Oligodon bitorquatus. - ZRC.2.3875, Bandung, West Java; ZRC.2.3876, Gunong Hedjo, Cheribon, West Java; ZRC.2.3957, Pengalengan, Java. Oligodon octolineatus. - ZRC.2.2295, 2399, 2559, 3161, 3850, 3853-3855, 3859-3860, 3865, 5058, Singapore; 3861, 3863-3864, Johor Bahru, Johor, Peninsular Malaysia; 3862, 3867, Selangor, Peninsular Malaysia. Oligodon purpurascens. - ZRC.2.3869, Sumatra, Indonesia; ZRC.2.3877-3879, Singapore; ZRC.2.3880- 3881, Pulau Gallang, Riau, Sumatra, Indonesia; ZRC.2.3882, 3884, 3885, 3887, Johor, Peninsular Malaysia; ZRC.2.3883, Forest Research Institute Malaysia, Kepong, Selangor, Peninsular Malaysia; ZRC.2.3886, 3888, Fraser's Hill, Pahang, Peninsular Malaysia; Bishop Mus. 14211, Pulau Tioman. Oligodon signatus. - DWNP.R.0005, Forest Research Institute Malaysia, Kepong, Selangor, Peninsular Malaysia; ZRC.2.4159, Bukit Asahan, Malacca, Peninsular Malaysia; ZRC.2.3203, 3388, 3400, 3871- 3873, 3958, 4842, Singapore. Oligodon taeniatus. - ZRC.2.4161, Bangkok, Thailand. Acknowledgments We are grateful to Sahir bin Othman (Perhilitan - Department of Wildlife and National Parks, Peninsular Malaysia), for permission to conduct research in the Seribuat Archipelago; Lim Boo Liat (DWNP), Norsham Yaakob (Forest Research Institute Malaysia), Karen M. Crane (La Sierra University) for their enthusiastic and active participation in the field; Karla H. Kishinami (BPBM) and Wolfgang Grossmann (ZMB) for the loan of specimens; Peter K. L. Ng and Kelvin K. P. Lim (RMBR) for granting access to literature and specimens; Robert B. Stuebing for examining the holotype and reviewing the manuscript with encouraging rigour; Robert F. Inger (FMNH) for critical and useful com- ments which greatly improved the manuscript. Literature Cited Boie, F. 1827. Bemerkungen uber Merrem's Versuch eines Systems der Amphibien, I. Lieferung: Ophidier. - Isis van Oken, Jena, 20:508-566. Cox, M. J., P. P. Van Dijk, J. Nabhitabhata and K. Thirakhupt. 1998. A photographic guide to snakes and other reptiles of Peninsular Malaysia, Singapore and Thailand. New Holland. 144 pp. 2004 Asiatic Herpetological Research Vol. 10, p. 16 Day, M. 1990. Zoological Research. In: Day, M. and T. Mowbray (eds.). University of Bristol Tioman Archipelago Expedition, Peninsular Malaysia, 1988, Final Report. Unpublished Report pp. 25-43. Hendrickson, J. R., 1966. Observations on the fauna of Pulau Tioman and Pulau Tulai. 5. The reptiles. Bulletin of Natural History 34:53-71. Grismer, J. L., L. L. Grismer, I. Das, N. S. Yaakob, L. B. Liat, T. M. Leong, T. M. Youmans, and H. Kaiser. 2004. Species diversity and checklist of the herpeto fauna of Pulau Tioman, Peninsular Malaysia, with a preliminary overview of habitat utilization. Asiatic Herpetological Research 10:247-279. Lim, B. L. 1963. Macrocalamus tweediei, a new species of Reed Snake from Malaya. Bulletin of the Nat. Mus., Singapore 32:99-102, 1 Fig. Lim, B. L. 1982. Poisonous snakes of Peninsular Malay- sia. Second Edition, Malayan Nature Society in association with Institute for Medical research, Kuala Lumpur, 73 pp. Lim, K. K. P. and L. J. Lim, 1999. The terrestrial her- petofauna of Pulau Tioman. Peninsular Malaysia. Raffles Bulletin of Zoology. Supplement No. 6:131- 155. Manthey, U. and W. Grossmann. 1997. Amphibien und Reptilien Siidostasiens. Natur und Tier-Verlag, Matthias Schmidt, Munster. 511 pp. Stuebing, R. B. and R. F. Inger, 1999. A field guide to the snakes of Borneo. Natural History Publications (Borneo), Kota Kinabalu, v + 254 pp. Tweedie, M. W. F., 1983. The snakes of Malaya. 3rd Edition. Singapore (National Printers Pte. Ltd.) 167 pp. 2004 Asiatic Herpetological Research Vol. 10, pp. 17-21 A New Philautus (Amphibia: Rhacophoridae) from Northern Laos Bryan L. Stuart1-’3’* and Harold F. Heatwole4 1 Field Museum oj Natural History, Department of Zoology, Division of Amphibians & Reptiles, MOOS. Lake Shore Drive, Chicago, Illinois, USA 60605-2496; •ji; Corresponding author: E-mail: bstuart@fieldmuseum.org -University of Illinois at Chicago, Department of Biological Sciences, 845 W. Taylor, Chicago, Illinois, USA 60607-7060 ^Wildlife Conservation Society, P.O. Box 6712, Vientiane, Laos 4 North Carolina State University, Department of Zoology, P.O. Box 7617, Raleigh, North Carolina, USA 27695-7617; E-mail: harold heatwole@ncsu.edu Abstract. - A new Philautus is described from Phou Dendin National Biodiversity Conservation Area in northern Laos. Philautus petilus sp. nov. is most remarkable by having a very slender, elongate habitus. Other distinguishing characteristics include having a tympanum diameter 80% of the eye diameter, white asperities on the dorsum, and distinctive coloration consisting of a soft yellow-beige dorsolateral surface with broken black stripes posteriorly, a lavender wash on dorsal surface of limbs, upper lip, and sides, a black stripe below edge of canthus extending from snout tip to flanks near level of mid-body, and a black spot equal in diameter to the tympanum located just anterior to the inguinal region. Key words. - Laos, new species, Philautus , Rhacophoridae. Introduction Bourret (1942) remains the major work on the amphib- ians of Laos, supplemented only recently by descrip- tions of new species (Inger and Kottelat 1998; Stuart and Papenfuss 2002). Consequently, the amphibian fauna of Laos is imperfectly known. From 6-26 October 1999, we conducted a herpeto- faunal survey of Phou Dendin National Biodiversity Conservation Area in eastern Phongsaly Province, northern Laos, near to the tri-border area of Laos, Vietnam, and China (Figure 1 A). The area surveyed was mostly covered in hilly evergreen forest, sometimes mixed with stands of natural bamboo, with small, rocky streams flowing down hillsides into the larger, swift Nam Ou and Nam Khang Rivers, at elevations from 600-1000 m. During the course of that work we found a single, adult female specimen of an enigmatic, rha- cophorid treefrog, which we describe here as a new species of Philautus. Materials and Methods The single specimen found was caught in the field by hand, preserved in 10% buffered formalin, and later transferred to 70% ethanol. A tissue sample was taken by preserving pieces of liver in 95% ethanol betoie the specimen was fixed in formalin. The specimen was deposited at the Field Museum of Natural History (FMNH). Measurements largely follow those of Bain et al. (2003) and were made with dial calipers to the nearest 0.1 mm. Abbreviations used are: SVL = snout-vent length; HDL = head length from tip of snout to the com- misure of the jaws; HDW = head width at the commisure of the jaws; SNT = snout length from tip of snout to the anterior comer of the eye; EYE = eye diameter; IOD = interorbital distance; TMP = horizontal diameter of tym- panum; TEY = tympanum-eye distance from anterior edge of tympanum to posterior comer of the eye; FPL = length of finger 111 disk from the base of the pad to its tip; FPW = width of finger 111 disc at the widest part of the pad; TPL = length of toe IV disk; TPW = width of toe IV disk. Measurement ratios are reported as percent- ages (%) rounded to the nearest integer. Philautus petilus. new species (Figure IB) Material examined. -Holotype: FMNH 257902, adult female, collected by the authors on 23 October 1999 in Phou Dendin National Biodiversity Conservation Area, Phongsaly District, Phongsaly Province, Laos, 22°05,44,'N 102°08'10,,E, at 600 m elevation. Diagnosis. - An elongate, slender Philautus having a head width only 27% of SVL: tympanum diameter 80% © 2004 by Asiatic Herpetological Research Vol. 10, p. 18 Asiatic Herpetological Research 2004 CHINA VIETNAM MYANMAR LAOS THAILAND CAMBODIA Figure 1 A-C. A. Map showing the type locality (black dot) of Philautus petilus sp. nov. in Phongsaly Province, north- ern Laos. B. The adult female holotype (FMNH 257902) of Philautus petilus sp. nov., anesthetized prior to preserva- tion. Photograph by Bryan L. Stuart. C. The hand of the adult female holotype (FMNH 257902) of Philautus petilus sp. nov. in preservation. Photograph by Nikolai L. Orlov. of eye diameter; white asperities on head, eyelids, back, dorsal surface of tibia and forelimbs, and anterior half of sides; no fringes, row of enlarged tubercles, or accesso- ry flaps of skin on outer margins of limbs; black stripe below edge of canthus extending from tip of snout to flanks near level of mid-body; black spot slightly anteri- or to inguinal region, equal in diameter to the tympa- num. Description of Holotype. - Habitus elongate, slender; head width 27% of SVL; head slightly longer than wide; snout obtusely pointed in dorsal view, projecting beyond lower jaw, round in profile, not depressed; nostril later- al, near tip of snout; canthus rounded but distinct, con- stricted behind nostrils; lores slightly concave, oblique; eye diameter less than snout length, interorbital distance wider than upper eyelid; tympanum visible, not depressed relative to skin of temporal region, tympanic rim slightly elevated relative to tympanum, tympanum diameter 80% of eye diameter; weak supratympanic fold from eye to shoulder; vomerine teeth very small, in oblique rows closer to choanae than to each other; tongue deeply notched posteriorly. Tips of all four fingers expanded, about two times the width of phalanges, with circummarginal grooves, width of finger I disc 60% the width of finger 111 disc, width of finger III disc 71% the diameter of tympanum; relative finger lengths I < II < IV < III; webbing absent; fingers III and IV with large middle subarticular tuber- cle and smaller palmar tubercle at base; fingers I and II with large palmar tubercle at base. Tips of toes expanded, width of toe IV disc slightly smaller than width of finger III disc; toe V longer than toe III; toe I webbing to midway between subarticular tubercle and disc, continuing only as narrow fringe to disc; toes II, III, and IV webbing to distal subarticular tubercle, continuing only as narrow fringe to disc; toe V webbing to midway between distal subarticular tubercle and base of disc, continuing only as narrow fringe to base of disc; inner metatarsal tubercle elongated, outer metatarsal tubercle very small, almost inconspicuous. Skin on dorsal and ventral surfaces smooth, except for distinct, white asperities on head, eyelids, back, dor- sal surface of tibia and forelimbs, and anterior half of sides; no fringes, row of enlarged tubercles, or accesso- ry flaps of skin on outer margins of limbs. Left ovary with fewer than 25, developing, creamy- white ova (color in preservative). In life, top of head and back light brown with dark brown reticulations and scattered black spotting; dorso- lateral surface of head and body soft yellow-beige, with short, broken black stripe on dorsolateral surface from level of midbody extending toward groin; lavender wash on dorsal surface of limbs, upper lip, and sides; black stripe below edge of canthus extending from tip of snout to anterior border of eye, and from posterior border of eye along supratympanic fold to flanks near level of mid-body; black spot slightly anterior to inguinal region. 2004 Asiatic Herpetological Research Vol. 10, p. 19 equal in diameter to the tympanum; black ‘M’-shaped marking over anus; black spot on tarsus closer to articu- lation than foot, narrow black crossbars on hindlimbs, some black flecking on forelimbs; venter creamy-white, dark spotting on chin and throat, pigmentation on under- side of hands, feet, and tibiotarsus. In preservative, yel- low-beige and lavender coloration lost. Measurements (mm) of holotype. - SVL 33.8; HDL 9.9; HDW 9.2; SNT 4.1; EYE 3.0; IOD 2.3; TMP 2.4; TEY 1.5; FPL 1.5; FPW 1.7; TPL 1.3; TPW 1.4. Comparisons. - The generic assignments of small, rha- cophorid (or rhacophorine) treefrogs are uncertain and debated (Bossuyt & Dubois 2001; Wilkinson et al. 2002). Therefore, we compare P petilus with all other species of small to medium-sized rhacophorid (or rha- cophorine) treefrogs having reduced finger webbing that are reported from the vicinity of northern Laos, regard- less of what genus they are currently referred to. These include Chirixalus doriae Boulenger, C. hansenae (Cochran), C. laevis (Smith), C. nongkhorensis (Cochran), C. palpebralis (Smith), C. vittatus (Boulenger), Philantus abditus Inger, Orlov, & Darevsky, P. albopunctatus Liu & Hu, P banaensis Bourret, P. carinensis (Boulenger), P. gracilipes Bourret, P. gryllus Smith, P. jinxiuensis Hu, P. longchua- nensis Yang & Li, P. maosonensis Bourret, P. menglaen- sis Kou, P. ocellatus Liu and Hu, P. odontotarsus Ye and Fei, P. parvulus (Boulenger), P. rhododiscus Liu and Hu, Rhacophorus appendiculatus (Gunther), R. baliogaster Inger, Orlov & Darevsky, R. bisacculus Taylor, R. verru- cosus Boulenger, Theloderma asperum (Boulenger), and T. stellatum Taylor (Bourret 1942; Taylor 1962; Inger et al. 1999; Fei 1999; Orlov et al. 2002). Philautus petilus differs from all species of Chirixalus Boulenger by lacking the two outer fingers appearing to be opposable to the two inner ones (present in Chirixalus). Philautus petilus further differs from C. doriae and C. nongkhorensis by lacking outer finger webbing (outer fingers 1/3 webbed in doriae and nongkhorensis), from C. hansenae, C. laevis and C. vit- tatus by lacking light-colored dorsolateral stripes (pres- ent in hansenae, laevis and vittatus), and from C. palpe- bralis by lacking a yellow streak from below eye to shoulder (present in palpebralis). Philautus petilus dif- fers from P. abditus by having a visible tympanum (hid- den in abditus) and lacking large black spots on the legs (present in abditus). Philautus petilus differs from P albopunctatus by having dorsal asperities (absent in albopunctatus) and lacking white blotches on the snout, dorsum and above anus (present in albopunctatus). Philautus petilus differs from P. carinensis by having the snout longer than the eye diameter (snout shorter than eye diameter in carinensis) and by having a slender habitus (stocky habitus in carinensis). Philautus petilus differs from P gracilipes by having a head width 27% of SVL (head width 45% of SVL in gracilipes), having the width of finger III disc 71% the diameter of tympanum (width of finger III disc equal to the diameter of tympa- num in gracilipes), and lacking mostly green coloration with dark-brown eyelids (present in gracilipes). Philautus petilus differs from P. jinxiuensis by lacking a large interorbital dark blotch extending posteriorly into two broad, dark dorsolateral stripes (present in jinxiuen- sis). Philautus petilus differs from P. longchuanensis by having a tympanum diameter larger than width of finger III disc (tympanum diameter smaller than width of fin- ger III disc in longchuanensis). Philautus petilus differs from P. maosonensis by having the head slightly longer than wide (head wider than long in maosonensis ), by having the snout projecting beyond the lower jaw (snout not projecting beyond lower jaw in maosonensis), by having the tympanum diameter 80% of eye diameter (tympanum diameter approximately 2/3 eye diameter in maosonensis), and by lacking a thin band between the eyelids, a large dark marking on the back, and a dark spot behind the axilla (present in maosonensis). Philautus petilus differs from P. menglaensis by having smooth skin with asperities on the dorsum (warty skin on dorsum in menglaensis) and by having a tympanum diameter larger than the width of finger III disc (tympa- num diameter smaller than or equal to width of finger III disc in menglaensis). Philautus petilus differs from P. ocellatus by lacking a round black blotch on the occiput (present in ocellatus). Philautus petilus differs from P parvulus by having the snout longer than the eye diam- eter (snout shorter than eye diameter in parvulus), hav- ing a visible tympanum (hidden in pamulus), and having the nostril close to the tip of snout (nostril midway between eye and tip of snout in parvulus). Philautus petilus differs from P. rhododiscus by lacking dark brown coloration with black spots, grayish-white ventral marbling, and reddish-orange finger and toe discs (pres- ent in rhododiscus). Philautus petilus differs from P. banaensis, P. gryllus , P odontotarsus, R. appendicula- tus, R. bisacculus, and R. verrucosus by lacking dermal fringes or tubercles on the limbs (present in banaensis, gryllus, odontotarsus, appendiculatus , bisacculus, and verrucosus). Philautus petilus differs from all species of Theloderma Tschudi by having smooth skin with dorsal asperities (skin rugose in Theloderma). Etymology. - The species name petilus (L.) means slen- der, referring to the distinct habitus of the holotype. Ecology. - The holotype was collected at 2215 h on a bamboo leaf 1 m above the ground in hilly evergreen Vol. 10, p. 20 Asiatic Herpetological Research 2004 torest mixed with bamboo, approximately 200 m from the bank of the Nam Ou River, at 600 m elevation. Remarks. - The generic assignments of small, rha- cophorid (or rhacophorine) treefrogs are uncertain, debated, and likely to be considerably revised in the near future (Bossuyt & Dubois 2001; Wilkinson et al. 2002). Many species have been moved among the genera Chirixalus Boulenger and Philautus Gistel (Frost 2002). Philautus petilus does not have the two outer fingers appearing to be opposable to the two inner ones (Figure 1C), which is diagnostic of the genus Chirixalus (Liem 1970). Historically, Philautus was diagnosed by the absence of vomerine teeth (Leim 1970; Bossuyt and Dubois 2001), but this character is known to vary7 with- in a species, and consequently Liem (1970) included species in the genus Philautus that sometimes have vomerine teeth. Philautus petilus has very small vomer- ine teeth. Bossuyt and Dubois (2001) proposed that only species having direct aerial development (lacking a free- swimming aquatic tadpole) be included in the genus Philautus. The mode of reproduction in P. petilus is unknown, but it does have a small clutch size (the left ovary of the holotype holds fewer than 25 ova). Dring (1987) reported total clutch sizes of only 2-26 eggs in five species of Philautus , including the type species, P. aurifasciatus. In the absence of a phylogeny and more substantial reproductive data, we recognize that our placement of petilus into the genus Philautus is tenta- tive. Acknowledgments The opportunity to work in Laos was made possible by the Wildlife Conservation Society / Division of Forest Resource Conservation Cooperative Program. The Ministry of Agriculture and Forestry (Vientiane, Laos) permitted export of specimens to the Field Museum of Natural History. Bee Thaovanseng assisted with field- work in Laos. Financial support was provided by The John D. and Catherine T. MacArthur Foundation (with Harold Voris and Robert Inger), the National Geographic Society (Grant no. 6247-98), and the Wildlife Conservation Society. Harold Voris, Alan Resetar, Jamie Ladonski, and Jennifer Mui facilitated examining specimens at the Field Museum of Natural History. Sophie Molia translated French descriptions and Tan Fui Lian translated Chinese descriptions. Robert Inger, Nikolai Orlov, and Jeff Wilkinson shared their taxonomic opinions on the specimen. Nikolai Orlov photographed the preserved specimen. Robert Inger, Raoul Bain, and an anonymous reviewer improved the manuscript. Literature Cited Bain, R. H., A. Lathrop, R. W. Murphy, N. L. Orlov, and Ho Thu Cue. 2003. Cryptic species of a cascade frog from Southeast Asia: taxonomic revisions and descriptions of six new species. American Museum Novitates 3417:1-60. Bossuyt, F. and A. Dubois. 2001. A review of the frog genus Philautus Gistel, 1848 (Amphibia, Anura, Ranidae, Rhacophorinae). Zeylanica 6(1): 1-1 12. Bourret, R. 1942. Les batraciens de l’lndochine. Memoires de Flnstitut Oceanographique de flndochine 6:1-547. Dring, J. 1987. Bornean treefrogs of the genus Philautus (Rhacophoridae). Amphibia-Reptilia 8(1987): 19- 47. Fei, L. 1999. Atlas of Amphibians of China. Zhengzhou: Henan Publishing House of Science and Technology. 432 pages, [in Chinese]. Frost, D. R. 2002. Amphibian Species of the World: an online reference. Version 2.21 (15 July 2002). Electronic database available at http://research.amnh.org/herpetology/amphibia/ index.html. Inger, R. F. and M. Kottelat. 1998. A new species of ranid frog from Laos. The Raffles Bulletin of Zoology 46(l):29-34. Inger, R. F., N. Orlov & I. Darevsky. 1999. Frogs of Vietnam: a report on new collections. Fieldiana, Zoology New Series, 92:1-46. Liem, S. S. 1970. The morphology, systematics, and evolution of the Old World treefrogs (Rhacophoridae and Hyperoliidae). Fieldiana Zoology 57:1-145. Orlov, N. L., R. W. Murphy, N. B. Ananjeva, S. A. Ryabov, & Ho Thu Cue. 2002. Herpetofauna of Vietnam, a checklist. Part I. Amphibia. Russian Journal of Herpetology 9(2):8 1-104. Stuart, B. L. and T. J. Papenfuss. 2002. A new salaman- der of the genus Paramesotriton (Caudata: Salamandridae) from Laos. Journal of Herpetology 36(2): 145-148. '004 Asiatic Herpetological Research Vol. 10, p. 21 raylor, E. H. 1962. The amphibian fauna of Thailand. University of Kansas Science Bulletin 63(8): 265- 599. Wilkinson, J. A., R. C. Drewes, and O. L. Tatum. 2002. A molecular phylogenetic analysis of the family Rhacophoridae with an emphasis on the Asian and African genera. Molecular Phylogenetics and Evolution 24: 265-273. 2004 Asiatic Herpetological Research Vol. 10, pp. 22-27~| Rediscovery of the Philippine Forest Turtle, Heosemys leytensis (Chelonia; Bataguridae), from Palawan Island, Philippines Arvin C. Diesmos1’2’3, Genevieve V. A. Gee3, Mae L. Diesmos3’ 4, Rafe M. Brown2’3’5, Peter J. Widmann3’6, and Judeline C. Dimalibot7 1 National Museum of the Philippines, Padre Burgos Avenue, Ermita 1000, Manila, Philippines; Current address: Department of Biological Sciences, National University of Singapore, Block S3 14 Science, Drive 4, Singapore 117543; E-mail: kaloula@i-manila.com. ph - Angelo King Center for Research and Environment Management; Marine Laboratory, Silliman University, Bantayan, Dumaguete City, Negros Oriental, Philippines 6200. J Wildlife Conservation Society of the Philippines, Room 106 Institute of Biology, University of the Philippines, Diliman 1101, Quezon City, Philippines; E-mail: jutisha@yahoo.com 4 Department of Biological Sciences, College of Science, University of Santo Tomas Espaha, Manila; E-mail: msleonida@lycos.com 5 Section of Integrative Biology, University of Texas, Austin Texas, 78712; Current address: Museum of Vertebrate Zoology, 3101 Valley Life Science Building, University of California, Berkeley, CA 94720; Email: rafe@mail.utexas.edu ^ KATALA Foundation, Jacana Road, Bancao-Bancao, PO. Box 390, Puerto Princesa City 5300, Palawan, Philippines; E-mail: widpeter@yahoo.com 7 Palawan Council for Sustainable Development, Sta Monica, Puerto Princesa City 5300, Palawan, Philippines. Abstract. - We report new observations from natural populations of the critically endangered Philippine forest turtle, Heosemys leytensis. Previously known from two cotypes (reportedly from Leyte Island) that were destroyed during World War II, a lone specimen in a U.S. collection, and a specimen purchased on Palawan Island in the late 1980s, its status in the wild has been uncertain since its discovery. Our recent surveys of Palawan and nearby Dumaran islands have documented natural populations that are under immediate threat due to over-harvesting and loss of habi- tat. Records of captive animals and interviews with residents from these islands suggest that this species is heavily exploited for food, pet trade, and ornamental fish pond curiosities. There is an urgent need to establish a conservation program to study and protect remaining natural populations. Heosemys leytensis, Asian freshwater turtles, turtle trade, Philippine forest turtle, Palawan Island, Key words. - Philippines. Introduction Taylor (1920) described the Philippine forest turtle, Heosemys leytensis, on the basis of two specimens that were collected by Gregorio Lopez. These specimens were reportedly collected from a swamp at the Municipality of Cabalian, southern Leyte Province, Leyte Island, Philippines (Fig. 1). The cotypes (a male and a female) were eventually deposited in the Philippine Bureau of Science (Taylor, 1944) but were destroyed during the World War II firebombing of Manila (Brown and Alcala, 1978; Buskirk, 1989). Between Taylor’s (1920) description and the late 1980s, no additional specimens or information became available for this species, although its status as a valid species has never been challenged (e.g., Pritchard, 1979; Ernst and Barbour, 1989; Iverson, 1992). In 1988, Timmerman and Auth reported on a specimen purchased from a local resident of the Municipality of Taytay, northern Palawan Island (Fig. 1). Buskirk (1989) described a neotype for the species (CAS 60930) based on a single specimen also reportedly from Cabalian, Leyte. Since these reports, numerous herpetologists, including us, have searched for H. leytensis at Cabalian, Leyte (Fig. 1) without success. The apparent rarity of the species formed the basis of its listing under CITES Appendix II and by IUCN as a Critically Endangered species (Hilton-Taylor, 2000). Chelonian biologists questioned whether the species was really rare or just unstudied, extinct or extirpated, and whether the speci- men reported by Timmerman and Auth (1988) was from © 2004 by Asiatic Herpetological Research Vol. 10, p. 23 Asiatic Herpetological Research 2004 Figure 1 . - Map of Palawan Island group in relation to the Philippines (inset) and Leyte Island. The type locality (Taylor, 1920) of H. leytensis is indicated with a star; recent trade or captive animal locations include (1) Brooke’s Point, (2) Rizal, (3) Aborlan, and (4) Puerto Princesa and known natural populations include (5) Taytay, and (6) Dumaran Island. a natural population on Palawan or the result of interis- land trade (Ernst and Barbour, 1989; Iverson, 1992; Das, 1995; Gaulke, 1995). The question remained whether H. leytensis occurred on Leyte Island or whether the origi- nal type locality data were in error. In late 2001, as part of a comprehensive status assessment of Palawan’s endemic amphibians and rep- tiles, we began a survey of forested sites throughout the island. We soon became aware of three nonmarine tur- tle species present in some local wet markets and in the possession of local wildlife traders. Two species Cuora amboinensis and Cyclemys dental a , are common on Palawan (Taylor, 1920; Alcala, 1986; Gaulke and Fritz, 1998; Widmann, 1998; ACD and RMB, pers. obs.). A third species, frequent in the wildlife and food trade, was identified as Taylor’s (1920) Heosemys leytensis. New observations. - The live specimens we examined match published descriptions of H. leytensis (Taylor, 1920; Buskirk, 1989; Ernst and Barbour, 1989): cara- pace unkeeled except for posterior vertebrals; vertebrals broader than long; anterior marginals projecting beyond cervicals, rendering anterior rim from slightly to strong- ly serrated; plastron much smaller than carapace, nar- Figure 2. - Live H. leytensis from natural population on Dumaran Island, northern Palawan: (A) An individual of undetermined sex in a small stream on Dumaran Island; (B) close up of the head. rowing anteriorly and posteriorly; angular notch between gulars deep and distinct; notch between gulars and humerals present, less distinct; anal notch deep and circular; three to four enlarged transverse scales present on anterior side of each foreleg; coloration rusty brown with darker margins on anterior scutes; narrow7 white to pale yellow line crosses head just behind auricular open- ings, medially divided in some specimens (Figs. 2-5). A full technical redescription of the morphology of H. leytensis will be published elsewhere (Diesmos et al.. unpublished data). We located captive animals for sale in markets at the Municipalities of Brookes Point, Aborlan, Rizal. Puerto Princesa City, and Taytay (Fig. 1). The animals were for sale as pets, ornamental fish pond curiosities, and for food. Additionally, H. leytensis individuals were found in public restaurants in the capital city' of Puerto Princesa (Fig. 4b). In many areas, residents expressed the belief that the keeping of pet H. leytensis specimens brings the owner good luck. We found natural populations in the vicinity' of Lake Manguao, Municipality of Taytay, Palawan Island and on Dumaran Island (Fig. 2). Exact localities are not given to protect these populations. Several individuals of each natural population were observed in slow-mov- 2004 Asiatic Herpeto/ogical Research Vol. 10, p. 24 Figure 3. - (A) Dorsal view of carapace and (B) ventral view of plastron of a subadult H. leytensis of underter- mined sex (captive pet, reportedly wild-caught locally) from Dumaran Island, N. Palawan. ing streams, quiet side pools, and nearby disturbed gallery forests (Fig. 4a), at most a few meters from the water’s edge. Residents in these localities reported to us that turtles are always located in the general vicinity of water, but that they can be found many meters away from water as well. Residents also report that H. leyten- sis burrows in stream banks and retreats under large nearby limestone boulders in the dry season when streambeds run dry. Interviews with Tagbanwa tribe members in the Municipality of Taytay suggest that in some areas this species is fairly common. Reports of natural popula- tions in the southern localities of Rizal and Brookes Point will need to be confirmed. In these areas inter- viewed persons claimed that H. leytensis was present in nearby forests but we were unable to locate wild animals ourselves. Discussion Our recent field observations confirm that H. leytensis occurs naturally on Palawan and at least on one of its northern satellite islands. Despite numerous surveys of suitable habitat at Cabalian, Leyte conducted by E. Figure 4. - (A) Preferred stream habitat of H. leytensis from Dumaran Island, northern Palawan; (B) Heosemys leytensis , Cuora amboinensis , and Cyclemys dentata specimens alive in captivity in restaurant of Puerto Princesa City, Palawan Island. Taylor, A. Alcala, and ourselves, no additional specimens of H. leytensis have been collected there. Interviews with residents in the vicinity of Cabalian, have failed to find verbal accounts of fresh-water turtles that fit the description of H. leytensis. We suspect that the species does not and never has naturally occurred on Leyte. We prefer the use of the common name “Philippine forest turtle” given that we have only observed animals in rem- nant old-growth forests and our sense is that this species is forest dependent. It is possible that Taylor or Lopez mislabeled or oth- erwise confused locality information assigned to the original co-types on Leyte and the third specimen at CAS (Buskirk, 1989). Taylor (1920) also reported Cyclemys dentata from Cabalian, Leyte (see also Iverson, 1992). This species has not since been reported from Leyte and is otherwise restricted in the Philippines to Palawan and the Sulu archipelago (Fig. 1; Taylor. 1920; Gaulke, 1995; Gaulke and Fritz, 1998). The fact that another conspicuous Palawan turtle species was reported at the same time and from the same site on Leyte (Taylor, 1920) suggests that a group of specimens from Palawan were mixed into collections from Leyte or Vol. 10, p. 25 Asiatic Herpetological Research 2004 Figure 5. - A live Heosemys leytensis from the Municipality of Taytay, northern Palawan Island, Philippines. Watercolors by Mr. Rene Aquino. were mislabeled. Based on information from the CAS herpetological registry, it is clear that G. Lopez also col- lected on Coron and Busuanga (Fig. 1) which would appear to be a likely source of the presumably erroneous “Leyte” specimens of C. dentata and H. leytensis. Thus, we suspect that a locality error is the basis of the specif- ic epithet and the long-held belief that H. leytensis natu- rally inhabits the island of Leyte. Whether H. leytensis has ever been introduced outside of Palawan or the country, remains to be documented. Finally, given the geological history and the Pleistocene formation of isolated paleoislands in the Philippines (Heaney, 1985; Hall, 1996, 1998) it is not surprising that H. leytensis may be restricted to Palawan and satellite islands. Based on available information from other groups of Philippine endemics, it is some- what rare for a species to be shared between both the Palawan (Palawan + Busuanga + Coron + Culion + Dumaran) and the Mindanao (Mindanao+BohoI+Leyte+ Samar) Pleistocene Aggregate Island Complexes (PAlCs). That is, based on previously-elucidated pat- terns of biogeography (Brown and Alcala, 1970; Brown and Diesmos, 2001; Brown and Guttman, 2002; Evans et al., 2003), we would expect to find Philippine endemics with restricted distributions on the Palawan PAIC or the Mindanao PAIC, but not necessarily both. There are some exceptions to these apparent trends, but they appear to be rare and limited to non-endemic wide- spread species that are also shared with the islands of the Sunda Shelf (Borneo, Java, Sumatra, etc.), or wide- spread Philippine endemics that are also found through- out the rest of the archipelago (Inger, 1954; Alcala and Brown, 1998; Brown and Alcala, 1970, 1978. 1980) Recommendations. - We recommend that an immediate exhaustive survey of the Palawan PAIC (including Balabac, Coron, Busuanga, Culion, and Dumaran) be undertaken to determine the status of natural H. leyten- sis populations. Basic knowledge of the species' distri- bution. habitat requirements, and natural population size will be a necessary requirement for designing effective conservation strategies. To combat illegal hobbyist, con- sumptive, and/or medicinal trade, wildlife managers will need to have reasonable estimates of numbers of animals 2004 Asiatic Herpeto/ogical Research Vol. 10, p. 26 Table 1 . - Standard measurements of H. leytensis speci- mens from captivity (Nos. 1-20) and a natural population (Nos. 21-24; Dumaran Isl ). Carapace Length and Width are straight-line distances; Carapace Width measured at widest point; Tail Length measured from posterior edge of cloaca to tip of tail. Sex undetermined; all measure- ments are in mm. Number Carapace Carapace Length Width Plastron Length Tail Length 1. 177.0 134.3 153.8 15.9 2. 183.8 139.6 151.6 17.6 3. 189.6 144.5 157.2 19.8 4. 192.5 142.4 158.1 18.3 5. 192.7 148.4 150.8 19.1 6. 196.4 152.2 151.4 20.4 7. 200.1 148.1 165.9 22.2 8. 203.6 151.8 171.4 19.2 9. 210.2 157.3 118.1 35.6 10. 215.9 160.8 179.7 19.9 11. 222.3 171.2 186.6 35.1 12. 231.2 172.2 185.2 22.8 13. 248.5 191.5 206.8 18.6 14. 261.7 191.0 210.3 25.5 15. 266.3 195.5 201.5 26.2 16. 269.9 196.4 213.0 23.4 17. 271.7 198.3 206.7 21.7 18. 275.3 200.5 205.6 19.7 19. 278.0 200.4 208.9 27.1 20. 280.0 201.6 205.2 29.1 21. 290.6 207.8 216.3 27.7 22. 297.8 211.4 208.5 30.9 23. 299.6 212.4 212.1 28.2 24. 299.9 212.9 213.0 35.0 being illegally harvested. Legislative protection of the species will need to be adjusted to recognize its current known distribution on Palawan and not Leyte. We expect that a specific conservation strategy will be nec- essary to protect this species from unchecked exploita- tion. The fact that the entirety of Palawan is officially designated a national protected area provides some assurance, but we suspect additional measures will need to be undertaken to protect this species while promoting its study. The legal “Protected Area” status of Palawan Island clearly is not deterring local exploitation of this species. Local education programs and public awareness campaigns targeting both the general public and local environmental authorities may be the key to insuring that H. leytensis does not become another casualty of the “Asian turtle crisis” (van Dijk et al., 2000). Many basic questions regarding the distribution, demography, ecolo- gy, reproductive biology, and phylogenetic affinities of H. leytensis remain to be answered. Acknowledgments This publication is a contribution of HerpWatch Palawan 2001, a project funded by the BP Conservation Programme (Silver Award No. 1554). ACD, MLD, GVAG, and JCD wish to thank their former institutions for supporting this work: De La Salle University- Dasmarinas, Haribon Foundation, and Palawan State University. The Protected Areas and Wildlife Bureau of the Philippine Department of Environment and Natural Resources and the Palawan Council for Sustainable Development facilitated research permits. We especially thank Althea Lota, Anson Tagtag, Marlynn Mendoza, Carlo Custodio, Mundita Lim, Wilfrido Pollisco, Linda Bacosa, Joselito Alisuag, Adelwisa Sandalo, and Rene Villegas. Indira Widmann, Siegfred Diaz, Deborah Villafuerte, Rolito Dumalag (Philippine Cockatoo Conservation Project), and Sabine Schoppe (State Polytechnic College of Palawan) extended much appre- ciated assistance and shared relevant information. We thank C. R. Infante for help during initial field survey work on Palawan and I. Das and M. Gaulke for provid- ing some critical reference material. We thank James Parham (University of California, Berkeley) and Bryan Stuart (Field Museum, Chicago) for providing refer- ences and for applying just the right amount of pressure to help us finish this report, and we thank Greg Pauly, Jen Weghorst, George Zug, and Bryan Stuart for critical reviews of earlier drafts of the manuscript. Special thanks are due to Rene Aquino (National Museum of the Philippines) for use of his painting of H. leytensis in life. Literature Cited Alcala, A. C. 1986. Guide to Philippine Flora and Fauna. Vol. X, Amphibians and Reptiles. Natural Resource Management Center, Ministry of Natural Resources and the University of the Philippines, Manila, Philippines. Alcala, A. C., and W. C. Brown. 1998. Philippine Amphibians: an Illustrated Field Guide. Bookmark Press, Makati City, Philippines. Brown, R. M„ and A. C. Diesmos. 2001. Application of lineage-based species concepts to oceanic island frog populations: the effects of differing taxonomic philosophies on the estimation of Philippine biodi- veristy. The Silliman Journal 42:133-162. Brown, R. M., and S. I. Guttman. 2002. Phylogenetic systematic of the Rana signata complex of Philippine and Bornean stream frogs; reconsidera- Vol. 10, p. 27 Asiatic Herpetological Research 2004 tion ot Huxley’s modification of Wallace’s Lme at the Oriental-Australian faunal zone interface. Biological Journal of the Linnean Society 76:393- 461. Brown, W. C., and A. C. Alcala. 1970. The zoogeogra- phy of the Philippine Islands, a fringing archipela- go. Proceedings of the California Academy of Science 38:105-130. Brown, W. C., and A. C. Alcala. 1978. Philippine Lizards of the Family Gekkonidae. Silliman University Press, Dumaguete City, Philippines. Brown, W. C., and A. C. Alcala. 1980. Philippine Lizards of the Family Scincidae. Silliman University Press, Dumaguete City, Philippines. Buskirk, J. R. 1989. A third specimen and neotype of Heosemys leytensis (Chelonia: Emydidae). Copeia 1989:224-227. Das, I. 1995. Status of knowledge on the biology and conservation of non-marine turtles of the Philippines. International Congress of Chelonian Conservation, Gonfaron, France. Ernst, C., H., and R. W. Barbour. 1989. Turtles of the world. Smithsonian Institution Press, Washington, DC. Evans, B. J., R. M. Brown, J. A. McGuire, J. Supriatna, N. Andayani, A. C. Diesmos, D. Iskandar, D. J. Melnick, and D. C. Cannatella. 2003. Phylogenetics of fanged frogs: testing biogeograph- ical hypotheses at the interface of the Asian and Australian faunal zones. Systematic Biology 52:794-819. Gaulke, M., 1995. On the distribution of emydid turtles and the anuran genus Microhyla in the Philippines. Asiatic Herpetological Research 6: 49-52. Gaulke, M. and U. Fritz. 1998. Distribution patterns of batagurid turtles in the Philippines. Herpetozoa 11:3-12. Hall, R. 1996. Reconstructing Cenozoic SE Asia. Pp. 153-184 In: Tectonic evolution of southeast Asia. Hall, R., and D. Blundell (eds). Geological Society, London. Hall, R. 1998. The plate tectonics of Cenozoic SE Asia and the distribution of land and sea. Pp 99-132 In: Biogeography and geological evolution of southeast Asia Hall, R., and J. D. Holloway (eds). Brackhuys, Leiden. Heaney, L. R. 1985. Zoogeographic evidence for middle and late Pleistocene land bridges to the Philippines. Modern Quaternary Research of SE Asia 9:127- 143. Hilton-Taylor, C. (Compiler) 2000. 2000 1UCN Red List of Threatened Species. IUCN, Gland, Switzerland and Cambridge. Inger, R. F. 1954. Systematics and zoogeography of Philippine Amphibia. Fieldiana 33:181 -53 1 . Iverson, J. B. 1992. A revised checklist with distribution maps of the turtles of the world. Privately pub- lished, Richmond, IN. Pritchard, P. C. H. 1979. Encyclopedia of turtles. TFH Publications, Neptune, NJ. Taylor, E. H. 1920. Philippine turtles. Philippine Journal of Science 16:111-144. Taylor, E. H. 1944. Present location of certain herpeto- logical and other type specimens. University of Kansas Science Bulletin 30, No. 11:160 Timmerman, W. W., and D. L. Auth. 1988. Geographic distribution: Heosemys leytensis. Herpetological Review 19:21. van Dijk, P. P., B. L. Stuart, and A. G. J. Rhodin. 2000. Asian Turtle Trade. Proceedings of a Workshop on Conservation and Trade of Freshwater Turtles and Tortoises in Asia. Chelonian Research Monographs, 2. Widmann, P. 1998. A Guide to the Ecosystems of Palawan Philippines. ViSCA-GTZ and Times Edition, Singapore. 2004 Asiatic Herpetological Research Vol. 10, pp. 28-37 Molecular Systematics of Old World Stripe-Necked Turtles (Testudines: Mauremys) Chris R. Feldman1’* and James F. Parham2’3 1 Department of Biology, Utah State University, Logan, UT, 84322-5305, USA, * Corresponding Author E-mail: elgaria@biology.usu.edu. "Evolutionary Genomics Department, Joint Genome Institute, 2800 Mitchell Drive, Walnut Creek, CA, 94598, USA. University of California Museum of Paleontology, University of California, Berkeley, CA, 94720-3140, USA, E-mail: parham@socrates. berkeley. edu Abstract. - Nine extant species of Mauremys (including Ocadia and Chinemys) represent a geographically widespread yet morphologically and ecologically conservative group of batagurid turtles. Here we examine the evolutionary rela- tionships of Mauremys using 1539 base pairs of mitochondrial DNA encoding portions of COI, ND4, and three adja- cent tRNA genes. These data contain 246 parsimony informative characters that we use to erect hypotheses of rela- tionships for Mauremys. Both maximum parsimony and Bayesian methods suggest that Mauremys japonica, M. sinensis , M. nigricans , and M. reevesii form a well-supported monophyletic clade, as do M. mutica and M. annamen- sis. Furthermore, our analyses show that M. mutica is paraphyletic with respect to M. annamensis. The western taxa M. leprosa, M. caspica, and M. rivulata remain problematic and do not form a monophyletic group sister to the Asian taxa. Nevertheless, an east-west biogeographic hypothesis cannot be discounted with our molecular genetic data. Key words. - Turtles, Bataguridae, Mauremys, molecular phylogenetics, mitochondrial DNA Introduction The Old World turtle genus Mauremys is represented by morphologically and ecologically conservative species that are diagnosed by a rigid plastron and a striped head and neck. These semi-aquatic, batagurid (= geoemydid, see Joyce et al., in press) turtles occupy lotic and lentic environments in both forested and arid habitats through- out Asia and the Mediterranean. The genus contains some of the most commercially important freshwater turtles in Asia. For example, M. mutica is one of the most commonly reared and highly traded chelonians in Asia (Lau and Shi, 2000). Other Mauremys species have been at the center of a conserva- tion and systematics controversy. In fact, two newly described Mauremys may be polyphyletic hybrids (Parham et al., 2001; Wink et al., 2001; Spinks et al., 2004). Given the mounting conservation interest in the tur- tle fauna of Asia (van Dijk et al., 2000), understanding the extant diversity and phylogenetic relationships among the Bataguridae are areas of active research (Wu et al., 1999; Honda et al., 2002b; Barth et al., 2004; Spinks et al., 2004). The genus Mauremys has received particular attention because of this recent conservation crisis and taxonomic confusion. The first examination of evolutionary relationships within Mauremys was a mor- phological treatment of the genus based on shell and scute measurements (Iverson and McCord, 1994). Consistent with the disjunct distribution of Mauremys, Iverson and McCord (1994) suggested that East Asian taxa form a monophyletic group, sister to a Mediterranean and Middle Eastern clade. A subsequent study used 12S and 16S ribosomal genes to resolve the phylogenetic relationships among species of Mauremys (Honda et al., 2002a). In contrast to the east-west hypothesis of Iverson and McCord (1994), Honda et al. (2002a) suggested that the deepest phylogenetic splits within Mauremys occur between Asian taxa. The ribo- somal mtDNA data also cast doubt on the monophyly of traditional Mauremys by including the east Asian species, Chinemys reevesii, as the sister taxon to M. japonica. Two recent studies examined more extensive sequence data, predominantly cyt b mtDNA, as well as a more comprehensive sampling of batagurids (Barth et al., 2004; Spinks et al., 2004). Both studies firmly estab- lished the placement of Mauremys within the Bataguridae and show that the Chinemys and Ocadia are phylogenetically nested within Mauremys (Barth et al., 2004; Spinks et al., 2004). Barth et al. (2004) offer two possible solutions to reconcile the paraphyly of Mauremys: 1) split the species of Mauremys into four genera; 2) lump Chinemys and Ocadia into an expanded Mauremys. While Barth et al. (2004) refrain from a tax- onomic decision, Spinks et al. (2004) adopt an expand- ed Mauremys. We also endorse an inclusive Mauremys © 2004 by Asiatic Herpetological Research Vol. 10, p. 29 Asiatic Herpetological Research 2004 because we consider expanding genera to well-support- ed clades of species functionally preferable to proliferat- ing monotypic genera based on subjective, typological ideas of uniqueness (Feldman and Parham, 2002; Parham and Feldman, 2002; Spinks et al., 2004). Our objective here is to provide an independent esti- mate of Mauremys phylogeny using different molecular markers from other recent systematic investigations and separate museum voucher specimens (Barth et al., 2004; Spinks et al., 2004). We hope that our data help resolve areas of uncertainty in the emerging consensus on Mauremys systematics. In addition, our study will add to the growing body of information on the evolutionary history and diversity of Asia’s threatened batagurid fauna (Wu et al., 1999; Honda et al., 2002b; Barth et al., 2004; Spinks et al., 2004). Materials and methods Taxon sampling and laboratory protocols. - We obtained liver tissue from 17 museum specimens repre- senting nine currently recognized species of Mauremys and three species of Cuora (Appendix 1). The nine species of Mauremys used in our study include: M. annamensis, M. caspica, M. japonica, M. leprosa, M. mutica, M. nigricans , M. reevesii, M. rivulata, and M. sinensis. We do not consider “M iversoni ”, “M pritchardF, “O. glyphistoma ” or “0. philippenF to be valid taxa because specimens matching these species (all described from the pet trade) are likely hybrids (Parham et al., 2001; Wink et al., 2001; Spinks et al., 2004). In addition, we also excluded “M megalocephald’\ which is probably a diet-induced variant of M. reevesii (Iverson et al., 1989; Barth et al., 2002). However, we do include a “M iversonF-Uke hybrid specimen described in Parham and Shi (2001) because mtDNA from this hybrid specimen is demonstrably Mauremys (Parham et al., 2001). All vouchers correspond to well-documented reference material and original species descriptions. We isolated genomic DNA from tissue samples by standard proteinase K digestion and phenol/chloroform purification (Maniatis et al., 1982). We amplified 700 bp of mtDNA encoding a section of COI via PCR (Saiki et al., 1988) using primers HCO-2193 and LCO-1490 (Folmer et al., 1994). We amplified an additional 900 bp region of mtDNA encoding a portion of ND4 and flank- ing tRNA histidine (tRNAhis), serine (tRNAser), and part of leucine (tRNAIeu) using primers ND4 and Leu (Arevalo et al., 1994). We used the following thermal cycle parameters for 50pl amplification reactions: 35 cycles of lmin denature at 94°C, lmin anneal at 45°C (COI) or 52°C (ND4), and 2min extension at 72°C. We purified PCR products using the Wizard Prep Mini Column Purification Kit (Promega, Inc.) and used puri- fied template in lOpl dideoxy chain-termination reac- tions (Sanger et al., 1 977) using ABI Big Dye chemistry (Applied Biosystems, Inc.) and the primers listed above. Following an isopropanol/ethanol precipitation, we ran cycle-sequenced products on a 4.8% Page Plus (Ameresco) acrylamide gel using an ABI 377 automated sequencer (Applied Biosystems, Inc.). We sequenced all samples in both directions. Sequence analyses. - We aligned DNA sequences with the program Sequencher™ 4.1 (Gene Codes Corp.), and translated protein coding nucleotide sequences into amino acid sequences using MacClade 4.0 (Maddison and Maddison, 2000). We identified tRNA genes by manually reconstructing their secondary structures using the criteria of Kumazawa and Nishida (1993). We deposited all mitochondrial DNA sequences in GenBank (Appendix 1). We performed a partition homogeneity test (PH), similar to the incongruence length differences test (ILD; Farris et al., 1994), to determine whether the ND4 and COI data could be combined. We used PAUP* 4.0b 10 (Swofford, 2002) to generate a null distribution of length differences using 1000 same-sized, randomly generated partitions from the original data with replacement. To evaluate base substitution saturation at first, sec- ond, and third codon positions, we plotted the uncorrect- ed percent sequence divergence of transitions and trans- versions versus the corrected maximum likelihood esti- mates of divergence for each codon position. Phylogenetic analyses. - We used maximum parsimony (MP; Farris, 1983) and maximum likelihood-based Bayesian (Larget and Simon, 1999) phylogenetic meth- ods to infer evolutionary relationships among batagurid species. We conducted MP analyses in PAUP* and Bayesian analyses with MrBayes 3.0b4 (Huelsenbeck and Ronquist, 2001). We polarized the phylogeny via outgroup comparison (Maddison et al., 1984) using the Asian box turtles Cuora mouhotii, Cuora picturata , and Cuora trifasciata. Other molecular phylogenetic studies suggest these turtles are appropriate outgroup taxa (Wu et al., 1999; Honda et al., 2002b; Barth et al., 2004; Spinks et al., 2004). We executed MP analyses with the branch-and- bound search algorithm (Hendy and Penny, 1982) using equally weighted, unordered characters. To assess nodal support, we used the bootstrap resampling method (BP; Felsenstein, 1985) employing 1000 pseudoreplicates of branch-and-bound searches in PAUP*. Additionally, we calculated branch support (DI; Bremer, 1994) for all nodes using the program Tree Rot 2c (Sorenson, 1999). We performed Bayesian analyses to estimate branch lengths and search for additional tree topologies. To 2004 Asiatic Herpetological Research Vol. 10, p. 30 determine the most appropriate model of DNA substitu- tion tor reconstructing Mauremys relationships under the Bayesian method, we executed hierarchical likeli- hood ratio tests (LRT; Felsenstein, 1993; Goldman, 1993; Yang, 1996) in the program Modeltest 3.06 (Posada and Crandall, 1998). Because MrBayes 3.0b4 can perform singular phylogenetic analyses using differ- ent models of evolution, we performed two separate LRTs on the two mtDNA regions. The model of nucleotide substitution that best fit the COI data was the HKY model (Hasegawa et al., 1985) in conjunction with r (Yang, 1994a; 1994b), and I (Gu et al., 1995), while the slightly less complex HKY + T model of DNA evo- lution best fit the ND4 data. We then performed Bayesian tree searches, allowing separate parameter estimates under the two models of DNA substitution for the COI and ND4 data partitions. We did not specify a topology or nucleotide substitution model parameters a priori. We ran Bayesian analyses for 3 x 10^ genera- tions using the Metropolis-coupled Markov chain Monte Carlo (MCMCMC) algorithm with four heated Markov chains per generation, sampling trees every 100 genera- tions. To determine when the Markov chains had con- verged on stable likelihood values, we plotted the -lnl scores against the number of generations (Huelsenbeck and Ronquist, 2001). We then computed a 50 % major- ity rule consensus tree after excluding those trees sam- pled prior to the stable equilibrium. Nodal support is given by the frequency of the recovered clade, which corresponds to the posterior probability of that clade under the given models of sequence evolution (PP; Rannala and Yang, 1996; Huelsenbeck and Ronquist, 2001). Lastly, we performed three Bayesian runs to be sure that independent analyses converged on similar log- likelihood scores (Leache and Reeder, 2002). Results Genetic variation. - Sequences from the protein coding regions appear functional and there are no gene rearrangements in the data (Kumazawa and Nishida, 1995; Kumazawa et al., 1996; Macey and Verma, 1997; Macey et al., 1997). However, ND4 in the batagurids studied here appears truncated relative to that of emydid turtles, which have three additional residues: Phenylalanine, Tyrosine, and Cysteine (Feldman and Parham, 2002). Instead, these batagurids possess a stop codon, followed by a 12 bp stretch of highly polymor- phic DNA between ND4 and tRNAhis. Additionally, tRNAser has a short D-stem, instead of a D-arm replace- ment loop like that of most metazoan taxa (Kumazawa and Nishida, 1993). This unusual tRNA condition is also seen in emydid turtles (Feldman and Parham, 2002). The PH test shows that length difference between the sum of the COI and ND4 trees and the combined COI and ND4 trees is not significantly different from the randomly generated test statistic ( P = 0.93). Therefore, we combined the aligned DNA sequences for subse- quent phylogenetic analyses. Of the 1539 aligned nucleotides, 369 are variable and 246 are parsimony informative. Among ingroup taxa, 289 sites are variable and 205 parsimony informa- tive. Of the 369 variable characters, 60 occur at 1st codon positions, 15 at 2nd positions, 261 at 3rd positions, and 33 in tRNAs. The scatter diagrams are linear and show no evidence of multiple hit problems for transi- tions or transversions (data not shown). Phylogenetic relationships. - The branch-and-bound equally weighted MP analysis produces a single most parsimonious tree (L = 661; Cl = 0.626; RI = 0.683) that is consistent with the model-based Bayesian analyses (Fig. 1). All three Bayesian analyses converge on the same topology and nearly identical mean log-likelihood values, parameter estimates, and nodal support. Thus we simply present results from the final search. The parti- tioned HKY + T+ I and HKY + TBayesian analysis (3 x 10^ generations) attains stable log-likelihood values within the first 15,000 generations, but we were conser- vative and discarded the first 20,000 generations. Because we sampled trees every 100 generations, we discarded the first 200 trees and retained 29,800 Bayesian trees, which we used to generate a 50% major- ity rule tree, and for which consensus values represent a group’s posterior probability (Huelsenbeck and Ronquist, 2001). The summary topology of the nearly 30,000 Bayesian trees (mean -lnl = 5205.5110, G2 = 24.5038; mean ti/tv (COI) = 10.8360; G2 = 10.6335; mean a(COI) =0.5479, a2 = 0.8874; mean Pinvar (COI) = 0.4163, O2 = 0.0291; mean ti/tv (ND4) = 12.3499; G2 = 9.0505; mean a(ND4) =0.2431, G2 = 0.0009) differs from the MP tree in the placement of only one taxon (Fig. 1). In both analyses, species of Cuora unambiguously group to the exclusion of Mauremys , (BP = 100%; DI = 19; PP = 100%). Mauremys japonica is a member of a clade containing M. nigricans, M. reevesii and M. sinen- sis (BP = 100%; DI = 13; PP = 100%), yet relationships among these taxa are not well resolved, as indicated by the low nodal support and conflict between MP and Bayesian reconstructions. The MP tree places M. sinen- sis sister to a group linking M. japonica , M. nigricans, and M. reevesii (DI = 1), wherein M. nigricans and M. reevesii form an additional clade (BP = 86%; DI = 5). Alternatively, the Bayesian tree connects M. japonica to M. sinensis (PP = 59%), sister to the M. nigricans the M. reevesii clade (PP = 99%). The M. japonica , M. nigri- cans, M. reevesii, and M. sinensis clade is sister to a Mauremys leprosa (Morocco) I Mauremys leprosa (Morocco) Vol. 10, p. 31 Asiatic Herpetological Research 2004 .2 'S3 cl cn a to S 5- CO S' si c cd <3 S' 2 co f 2 si a ■a jE3 cd cq i S' <3 o Co S' Si si I CO CO 5! 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Cb CD CO e a o d o > 8 a ■§ &S 5 Q3 > CD .O 5 CN 3 M. rivulata 4 M. sinensis — -J q Q. 2 o 3 CD O co 45 S 8 o -iS o ■c o 5 8 o =3 5 5 S' O T- O O CO CD o Is- T- CN CO co CO CO 03 T — t — T — T_ poorly supported M. mutica, M. armamensis, “M iver- sonf\ M. leprosa, and M. caspica assemblage (DI = 2; PP = 61%). Within this large group, M. mutica , M. anna- mensis and “M iversonf form a strongly supported clade (BP = 99; DI = 15; PP = 100%). In fact, inclusion of both M. armamensis and “M iversoni ’ render M. mutica paraphyletic; two M. mutica (ROM 25613, 25614) are more closely related to M. annamensis and “M iverson F than they are to a Chinese M. mutica (MVZ 230476) (BP = 100%; DI = 36; PP = 100%). Within this “ mutica complex”, M. annamensis and “M iversonr are weakly allied (BP = 60%; DI = 1 ; PP = 86). This entire Umutica complex” is then sister to a weakly supported M. leprosa and M. caspica clade (DI = 1). Finally, both MP and Bayesian analyses suggest that M rivulata is sister to a monophyletic clade containing the rest of Mauremys, but this phylogenetic arrangement receives almost no statistical support (DI = 2; PP = 60%). Phylogenetic relationships. - Both MP and Bayesian phylogenetic methods show that M. japonica is a mem- ber of a clade containing M. nigricans, M. reevesii, and M. sinensis, exclusive of other Mauremys. The M. japonica, M. nigricans, M. reevesii, and M. sinensis clade is joined to a poorly supported M. mutica, M. annamensis, M. leprosa, and M. caspica assemblage. Within this grouping, M. mutica and M. annamensis form a solid clade, congruent with shell and scute data (Iverson and McCord, 1994), other molecular data (Barth et al., 2004; Spinks et al., 2004) but not 12S and 16S mtDNA data (Honda et ah, 2002a). Our analyses further suggest that M. mutica is paraphyletic. Two M. mutica (ROM 25613, 25614) purchased in Vietnam are more closely related to M. annamensis than they are to topotypic M. mutica (MVZ 230476) from China. We tested the paraphyly of M. mutica by constraining the MP searches to recover only those trees that produce a monophyletic M. mutica. The shortest two trees generat- ed by the constraint search are 697 steps long (Cl = 0.594; RI = 0.636), 36 steps longer than the uncon- strained MP tree. The two-tailed Wilcoxon signed-ranks test (Templeton, 1983) fails to support (P < 0.0001) the monophyly of M. mutica. The mutica complex is linked to a tenuous M. leprosa and M. caspica group. Lastly, both MP and Bayesian phylogenetic analyses tentatively place M. rivulata sister to a monophyletic clade contain- ing the remaining ingroup taxa. Genetic Variation. - Our samples of M. leprosa from Spain and Morocco, and M. caspica from Iran and Bahrain, show no intraspecific haplotype diversity Discussion Vol. 10, p. 33 Asiatic Herpetological Research 2004 (Table 1), yet exhibit sizeable morphological variation (Busack and Ernst, 1980). This discrepancy between intraspecific mtDNA diversity and geographic variation seems to be common among turtles (e.g., Lenk et al., 1999; Starkey et ah, 2003) and may be related to exten- sive phenotypic plasticity or the slow rate of molecular evolution in turtles (Avise et ah, 1992; Lamb et ah, 1994). In contrast, most interspecific mtDNA variation appears extensive, with uncorrected sequence diver- gences higher than 8% between a number of ingroup taxa (Table 1). Additionally, the mitochondrial sequence divergences between M. rivulata and M. caspica (Table 1), formerly considered conspecifics (Fritz and Wischuf, 1997), are equivalent to or greater than the genetic dis- tances observed between other congeneric emydid and batagurid turtles (e.g., Feldman and Parham, 2002; Starkey et ah, 2003; Stuart and Parham, 2004). Hence, these mtDNA data, together with the differing shell mor- phologies, distinct color patterns, and unique habitat preferences of M. rivulata and M. caspica (Busack and Ernst, 1980), support the recent elevation of M. rivulata as a distinct evolutionary lineage independent of M. caspica (Fritz and Wischuf, 1997). Mauremys annamensis, a robust batagurid endemic to central Vietnam, is characterized by extensive axillary buttresses, a massive bridge, a slightly tricarinate and high-domed shell, a vividly striped head and neck, and reverse sexual size-dimorphism (McDowell, 1964; Iverson and McCord, 1994). The taxon is so distinctive it was once placed into its own genus, Annamemys Bourret 1939. McDowell (1964) originally demonstrat- ed that M annamensis and M. mutica share a number of derived features and Iverson and McCord (1994) subse- quently confirmed a close kinship between these taxa with shell measurements. Hence, the close relationship revealed by our mitochondrial genes is not novel. What is surprising, however, is that M. annamensis differs from Vietnamese M. mutica and our “M iverson?'- like hybrid by only two transitions. Furthermore, this clade shows a roughly 6% uncorrected sequence divergence from topotypic M. mutica from Zoushan Island, Zheijung Province, eastern China. In contrast, distantly collected samples of M. leprosa and M. caspica show no such intraspecific mtDNA variation (Table 1). These data question our ideas of species limits within Mauremys. Is M. annamensis a distinct species? Does M. mutica represent multiple species? Several potential hypotheses might account for these unexpected results. M annamensis may simply represent a recent species, derived from M. mutica , or even a geographical variant of M. mutica. The dramatic morphological differences exhibited by M. annamensis could reflect intense selection and rapid phenotypic evo- lution while the minute mitochondrial divergences and paraphyly represent the nature of speciation and unsort- ed polymorphism. Alternatively, there may be historical or ongoing introgression between M. annamensis and Vietnamese M. mutica , perhaps facilitated by selection. Two additional hypotheses involve the possibility of hybridization. While our specimen of M. annamensis conforms to the species description, it was acquired from a Chinese turtle farm (Appendix 1) where M. anna- mensis and M. mutica are reared together in large num- bers (J.F. Parham, pers. obs.). Hence, our M. annamen- sis could be a captive hybrid between M. annamensis and M. mutica , though we find no morphological char- acters supporting this notion. Ideally, we would examine the morphology and compare the sequences of a wild- caught M. annamensis to our sample, but to our knowl- edge, no tissued, field-collected vouchers of M. anna- mensis exist in collections; all modern museum speci- mens of M. annamensis have been obtained from either animal markets or the pet trade. Another possibility is that the Vietnamese M. muti- ca could be hybrid offspring of female M. annamensis and male M. mutica, accounting for the scant mtDNA differences between Vietnamese M. mutica and M. annamensis and the sizeable divergences between these samples and topotypic M. mutica. Although the “Vietnamese M. mutica ” are phenotypically similar to typical M. mutica, their darker coloration is evocative of M. annamensis. Both Barth et al. (2004) and Spinks et al. (2004) found substantial mitochondrial variation between M. mutica and M. annamensis, but we do not know the provenance or morphology of their samples. The hybridization of batagurid turtles has lead to other cases of taxonomic confusion (Parham and Shi, 2001; Parham et al., 2001; Shi and Parham, 2001; Wink et al., 2001; Spinks et al., 2004) and cannot be discount- ed here. Unfortunately, our small sample size prohibits us from effectively evaluating these hypotheses. Clearly a more detailed genetic study is needed to unravel this problem. With our present knowledge, any change in conservation policies for M. annamensis, one of the world’s most poorly known turtles, would be premature. Biogeography. - The distribution of Mauremys is char- acterized by a major break between the Zagros Mountains of western Iran (easternmost M. caspica ) and the Annamite Mountains of central Vietnam (range of M. annamensis). This disjunction includes the entire Indian subcontinent (home to a diverse, endemic batagurid fauna), and the inhospitable Tibetan plateau. We suggest that the collision of India into Asia may be the vicariant event responsible for the current distribution of Mauremys, as proposed for anguine lizards (Macey et al., 1999). Molecular data are ambiguous on this point. 2004 Asiatic Herpetological Research Vol. 10, p. 34 Given that neither eastern nor western species assem- blages appear monophyletic (though a Wilcoxon signed ranks test topology test cannot discount this hypothesis [P = 0.35]), the current divergences between the living species may have occurred before the development of the Indo-Tibetan gap. The collision and subsequent uplift of the Tibetan plateau took place in multiple stages between 50 and 10 MYBP (Shackleton and Chang, 1988; Dewey et al., 1989; Windley, 1988). Hervet (2004) attributed some Paleogene (>50 MYBP) European fossils to the stem of Mauremys , but did not investigate their relations to east Asian Mauremys. In addition to employing additional molecular markers to vouchered museum specimens, the integration of all extant Mauremys into analyses of morphological charac- ters and fossil taxa will be necessary to unravel the his- torical biogeography of this clade of turtles. Acknowledgments We thank T. J. Papenfuss, C. Cicero, and D. B. Wake (MVZ), and R. W. Murphy (ROM) for kindly contribut- ing specimens and tissues essential to this project. We are grateful to M. E. Pffender and P. G. Wolf for gener- ously providing laboratory space, and D. G. Mulcahy and W. B. Simison for much needed lab assistance. We appreciate helpful discussions about phylogenetic meth- ods from E. M. O’Neill, A. D. Leache, and F. T. Burbrink, and useful comments on this manuscript from M. D. Matocq, J. R. Mendelson III, E. D. Brodie Jr., H. B. Shaffer, and two anonymous reviewers. We also thank P. Q. Spinks and U. Fritz for sharing unpublished data. Finally, we thank K. Padian and T. J. Papenfuss for funding, space, advice, and encouragement. This is University of California Museum of Paleontology Contribution #1826 and LBNL #54656. This research was performed under the auspices of the U.S. Department of Energy, Office of Biological and Environmental Research. References Arevalo, E., S. K. Davis and J. W. Sites. 1994. Mitochondrial DNA sequence divergence and phy- logenetic relationships among eight chromosome races of the Sceloporus grammicus complex (Phrynosomatidae) in central Mexico. Systematic Biology 43: 387-418. Avise, J. C., B. W. Bowen, T. Lamb, A. B. Meylan and E. Bermingham. 1992. Mitochondrial DNA evolu- tion at a turtle’s pace: evidence for low genetic vari- ability and reduced microevolutionary rate in the Testudines. Molecular Biology and Evolution 9: 457-473. Barth, D., D. Bernhard, D. Guicking, D. 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Maximum likelihood phylogenetic esti- mation from DNA sequences with variable rates over sites: approximate methods. Journal of Molecular Evolution 39: 306-314. Yang, Z. 1996. Maximum likelihood models for com- bined analyses of multiple sequence data. Journal of Molecular Evolution 42: 587-596. Appendix 1. Specimens used and GenBank Accession numbers for DNA sequence data. Acronyms are: MVZ = Museum of Vertebrate Zoology, Berkeley, California; ROM = Royal Ontario Museum, Toronto, Ontario; AF or AY = GenBank (http://www.ncbi.nlm.gov). Mauremys annamensis - Purchased in turtle farm in Hainan Province, China, no real locality data; MVZ 238937; AY337338, AY337346. Mauremys caspica - Field collected on Bahrain Island, Bahrain; MVZ 230971; AY337339, AY337347. Mauremys caspica - Field collected in West Azarbaijan Province, Iran; MVZ 234281; AY337340, AY337348. “ Mauremys iversoni ” - Purchased in turtle farm in Hainan Province, China, no real locality data; MVZ 230475; AF348275, AF34281. Mauremys japonica - Pet trade specimen, no locality data; MVZ 234647; AY337341, AY337349. Mauremys leprosa - Field collected in Tetouan Province, Morocco; MVZ 178059; AY337342, AY337350. Mauremys lep- rosa - Field collected in Cadiz Province, Spain; MVZ 231989; AY337343, AY337351. Mauremys mutica - Field collected in Zoushan Island, Zhejiang Province, China; MVZ 230476; AF348262, AF348278. Mauremys mutica - Purchased from a turtle trader in Yen Bai Province, Vietnam; ROM 25613; AF348260, AF348279. Mauremys mutica - Purchased from a turtle trader in Yen Bai Province, Vietnam; ROM 25614; AF348261, AF348280. Mauremys rivulata - Field-col- lected in Bursa Province, Turkey; MVZ 230212; AY337344, AY337352. Mauremys ( =Ocadia ) sinensis - Field-collected in Hainan Province, China; MVZ 230479; AY337345, AY337353. Mauremys nigricans - Pet trade specimen, no locality data, MVZ 130463; AF348264, AF348289. Mauremys reevesii - Pet trade specimen, no locality data, MVZ 230533; AF348263, AF348288. Cuora picturata - Purchased from a turtle trader in Dong Nai Province, Vietnam, ROM 37067; AF348265, AF348292. Cuora trifasciata - Pet trade specimen, no locality, MVZ 230636; AF348270, AF348297. Cuora mouhotii - Purchased from a turtle trader in Bac Thai Province, ROM 35003; AF348273, AF348286. 2004 Asiatic Herpetological Research Vol. 10, pp. 38-52 A Preliminary Report on Southeast Asia’s Oldest Cenozoic Turtle Fauna from the Late Middle Eocene Pondaung Formation, Myanmar. J. Howard Hutchison1, Patricia A. Holroyd1, and Russell L. Ciochon2 1 Museum of Paleontology, 1101 Valley Life Sciences Building, University of California, Berkeley, California, 94720, U.S.A. J - Departments of Anthropology and Pediatric Dentistry, University of Iowa, Iowa City, Iowa, 52242, U.S.A. Abstract. - Late middle Eocene fossils from the Pondaung Lormation of central Myanmar document Southeast Asia’s oldest Cenozoic turtle fauna. Although the material is fragmentary, seven distinct turtle taxa are recognized. These include a podocnemid pleurodire, anosteirine and carettochelyine carettochelyids, two or more trionychine triony- chids, and a testudinid. Of these, only the carettochelyine carettochelyid is complete enough to recognize as a new taxon, Burmemys magnifica, gen. et sp. nov. The Pondaung turtle fauna is one of the best known of its age from Southeast Asia but comparisons with the limited literature of the Eocene faunas from China, Mongolia, and the Indian subcontinent indicate it is probably biogeographically unique. Among the recognized genera, only Anosteira is known from other Eocene Asian localities, and the presence of pleurodires is unusual. Key words. - Reptilia, Testudines, Carettochelyidae, Burmemys , Myanmar, Pondaung Lormation, Paleontology, Eocene. Introduction The origins of Southeast Asia’s herpetofauna are poorly understood, as there are few fossils that document the origin of the major groups inhabiting the region. The oldest known herpetofauna from this region is from the Pondaung Lormation, a late middle Eocene (approx. 37 Ma) set of rocks exposed in the Chindwin-Irrawaddy Basin of Myanmar (formerly Burma). The Pondaung fauna is best known for its mammalian fauna (e.g., Colbert, 1938; Tsubamoto et al., 2000), and little atten- tion has been devoted to the remainder of the fauna. In prior reports, Buffetaut (1978) noted the presence of both unidentified crocodilians and dyrosaurids; Sahni (1984) and Rage (1987) noted unidentified Lacertilia. These reports were based primarily on a rather limited collection made by Bamum Brown in 1922 and housed in the American Museum of Natural History, New York. Savage and Russell (1983) and Broin (1987) list “Pelomedusid/Emydidae”, “Carettochelyoidea”, and tri- onychids from the Pondaung. Outside the Pondaung region, the only other report of turtles from Southeast Asia is Ducrocq et ah’s (1992) mention of two types of ?Emydidae from the late Eocene site of Krabi, Thailand. Here we present a preliminary description of the turtles based on a more thorough study of these collections, and additional collections in the University of California Museum of Paleontology, Berkeley, California. Localities and age. - fossils occur in a number of local- ities occurring in the upper 100+ meters of the otherwise marine Pondaung formation. The majority of the speci- mens discussed here come from localities to the west and northwest of Mogaung village, Myaing Township, central Myanmar (fig. 1), that have been collected inter- mittently over the past 80 years. As a consequence, most specimens have limited, descriptive locality data that provides locations based on distances from known vil- lages. Recent fieldwork has provided detailed, GPS based mapping of the most productive outcrops and per- mit us to place most of the historic localities in a more accurate and stratigraphically detailed framework. Those localities we can place with confidence are shown in figure 1. Localities whose positions are approximate are shown with dashed lines. Concordances for locali- ties that have been published under more than one name or number are provided in the caption of figure 1 and are based on Colbert (1938), maps on file at the American Museum of Natural History, field notes of J. Wyatt Durham and Donald E. Savage on file at the University of California Museum of Paleontology, data contained in Tsubamoto et al. (2000, 2002) and Gunnell et al. (2002), and field observations by PAH and RLC. fossils occur in place and as erosional lag coming out of reddish to purplish mudstones (Pig. 2 A-C). fossil wood is also commonly found (Pig. 2D), attesting to the presence of the ancient forest. Soe et al. (2002) interpreted sediments including these localities as swale- fills and/or paleosols deposited in an ancient floodplain; stratigraphic sections for these localities are contained in Gunnell et al. (2002). Based on comparisons of tempo- ral distribution and faunal resemblance data of the © 2004 by Asiatic Herpetological Research Vol. 10, p. 39 Asiatic Herpetological Research 2004 28° N 24 “N 20° N ia°N Figure 1. Locality Map of Pondaung Formation localities. V78090=Thandaung kyitchaung and possibly AMNH locali- ties A14-16, 18-19; V83106, 3.5 mi SW of Mogaung = AMNH A31; V83111 "1 .25 mi NW Paukkaung" probably equals Pk2; V83116 probably equals Yarshe kyitchaung. V96001-V96002 = AMNH A22 and Lema kyitchaung; V98019 "Thidon or near Bahin", possibly equal to Pkl or Pk2. Pondaung mammalian fauna with other Asian and North American mammal faunas, as well as additional con- straining evidence from marine invertebrates, Holroyd and Ciochon (1994) concluded that the Pondaung fauna is best considered latest middle Eocene in age and broadly contemporaneous with Asian faunas assigned to the Sharamurunian Land Mammal Age, a finding con- firmed by recent fission-track dates that provide a date of 37.2 +/- 1.2 Ma (Tsubamoto et al., 2002). Abbreviations. - AMNH, American Museum of Natural History, New York, New York, U.S.A.; UCMP. University of California Museum of Paleontology, Berkeley, California, U.S.A. Systematic Paleontology Testudines Batsch, 1788 Pleurodira Cope, 1865 Pelomedusoides Cope, 1868 Podocnemididae Cope, 1868 ?Podocneniididae unident. Referred Material. - UCMP locality V83108; UCMP 153798, right peripheral 3. UCMP locality V 83 113; UCMP 147052, partial left hypo-xiphiplastron. UCMP locality V96002: UCMP 142245, left incomplete epi- p I astro n. MAP 1 MAP 2 2004 Asiatic Herpetological Research Vol. 10, p. 40 Figure 2. Fossil localities of the Pondaung Formation. A. UCMP locality V96001, Lema kyitchaung; B. UCMP locali- ty V96002, Lema kyitchaung; C-D. UCMP locality V96007, near Mogaung, showing the common occurrence as float of both turtle bone (C) and petrified wood (D) on the surface. Description. - The epiplastron (UCMP 142245, Fig. 3A) lacks the posterolateral part but is otherwise well preserved. The scale covered surfaces are very finely textured with delicate but well-defined sulci. Faint growth corrugations are present on the gular scale (extragular of Hutchison and Bramble, 1981). There is a prominent anteriorly-projecting gular spur, and the epi- plastron margin is distinctly concave between the mid- line and the gular spur. There is an intergular scale (gular of Hutchison and Bramble, 1981) spanning the midline that projects anteriorly into the anterior embayment. The scales overlap extensively onto the dorsal surface with little exposure of the visceral surface. The intergular expands slightly posteriorly on the ventral surface and Vol. 10, p. 41 Asiatic Herpetological Research 2004 Figure 3. A-C. Podocnemidae? indet. A. UCMP 142245, incomplete left epiplastron, dorsal, medial suture and ven- tral views. B. 153798, right peripheral 3, external and visceral views. C.UCMP 147052, hypo-xiphiplastron fragment, dorsal view, spot indicates center of bump. D-E. Testudinidae. D. UCMP 142226, partial right epiplastron, cross-sec- tion (as indicated) and dorsal views. E. UCMP 149166, partial right xiphiplastron, dorsal view. Scale bars equal 1 cm. extends onto the entoplastron. Dorsally the intergular extends slightly more than one-half the length of the inter-epiplastral suture and is parallel-sided. On the ven- tral surface, the gular is triangular with the lateral mar- gins converging to a point at the entoplastron margin. The hypo-xiphiplastron fragment, UCMP 147052 (Fig. 3C), is broken on all the edges except the free mar- gin. It exhibits a narrow overlap of the femoral and anal scales onto the dorsal surface (less than one-fifth the transverse length as preserved). The swelling at the anterolateral comer indicates an ascendant hypoplastral buttress. The sutures are fused. A short expanse of the femoral-anal scale sulcus is preserved at the extreme posterior end. Medial to the scale margins on the dorsal side is a large elliptical swelling that has a smooth sur- face and may have been divided by the hypo-xiphiplas- tron suture. The peripheral 3 (UCMP 153798, Fig. 3B) lacks the dorsal margin. The body of the peripheral is robust and without a change in plane between the pleural and mar- ginal surfaces. The surface is smooth and unsculptured. The sulci are shallow but well defined. The free margin is acutely angled. On the visceral side, the marginal scales rise up from only about one-third of the peripher- al depth. There is no indication of an axillary scale. The finely dentate suture for the hyoplastron buttress rises anteriorly and may have overlapped peripheral 2-3 suture. There is a gap in the hyoplastral suture near the posterior margin, for passage of the musk duct. The length between the anterior and posterior sutures along the free margin of the peripheral is 37.9 mm. Discussion. - The dorsal scale overlap, truncated anteri- or margin, undivided intergular, and relatively thick epi- plastra resemble selected extant or fossil Pelomedusoides (Bothremydiae, Podocnemididae, and Pelomedusidae). The prominent epiplastral spurs resem- ble those of the pelomedusid Kenyemys Wood, 1983, from the Pliocene of Kenya. Flowever, the Pondaung form differs in the greater excavation of the gular embayment, intergular extending onto the entoplastron, and restriction of the gular scales to the epiplastra (i.e., not reaching the midline). The scale arrangement is sim- ilar to that of the podocnemidid Neochelys Bergounioux, 1954 (Broin, 1977; Jimenez et al., 1994) from the Eocene of Europe. Neochelys may also possess a rela- tively prominent gular spur (Broin, 1977, fig. 59), but differs in the less extensive dorsal overlap of the scales and lesser development of an epiplastral embayment. 2004 Asiatic Herpetological Research Vol. 10, p. 42 Figure 4. Anosteira sp., A. UCMP 131736, right hypoplastron, ventral view. B. UCMP 131737, lateral fragment of left hypoplastron, ventral view. C. UCMP 147030, left peripheral 6, external, posterior, visceral, and anterior views. Scale bars equals 1 cm. The presence of a prominent musk duct on the peripheral 3, absence of an axillary scale, and strong indication of a hyoplastral buttress rising onto the first costal is consistent with Neochelys- like pleurodires. The hypoplastron fragment may be referable to the same taxon, but the area of the pelvic sutures is broken off. The general similarity to at least some Neochelys favors a placement of the Pondaung Formation speci- mens in the Podocnemididae. Cryptodira Cope, 1868 Testudinidae Gray, 1825 Testudinidae undet. Referred Material. - UCMP locality V6204: UCMP 149166, right xiphiplastron fragment. UCMP locality V96009: UCMP 142226, partial right epiplatron. Description. - The epiplastron (UCMP 142226, Fig. 3D) lacks the gular region. The remaining part of the free margin is greatly thickened along the anterior edge of the dorsal scale covered portion. The posterior rim of this thickened gular area overhangs the visceral surface. The ventral surface is longitudinally convex. The sutures are moderately thick and dentate. The anterolateral part of a right xiphiplastron (UCMP 149166, Fig. 3E) is referred to the Testudinidae on the basis of the strong overlap of the femoral scale dorsally, its inflated appearance, and fairly porous sur- face texture. Discussion. - The morphology of the epiplastron is typ- ical of testudinids and a few batagurids. The rather porous bone, inflation of the gular area, and general nature of the sutures and surface texture agrees best with that of a testudinid. The overhang of the posterior gular rim is derived in testudinids and absent or poorly devel- oped in such tortoises as Hadrianus Cope, 1872, Stylemys Leidy, 1851, Sharemys Gilmore, 1931, Kansuchelys Yeh, 1963, and Ergilemys Ckhikvadze, 1972. The epiplastron thus resembles more derived tor- toises such as Testudo Linnaeus, 1858. Testudinoidea Fitzinger, 1826, indet. Referred material. - UCMP locality V96019: UCMP 147051, posterior part of left hypoplastron. UCMP locality V78090: UCMP 170495, partial neural. UCMP locality V98109: UCMP 170522, shell fragments. Description. - The hypoplastron is represented by a fragment (UCMP 147052) that preserves the portion Vol. 10, p. 43 Asiatic Herpetological Research 2004 Figure 5. Burmemys magnifica gen. et sp. n. A. UCMP 61212, adult left hypoplastron (type), ventral view. B. UCMP 131745. juvenile left hypoplastron, ventral view. C. UCMP 154993, posterior part of juvenile left epiplastron, ventral view. D. UCMP 131747, anterior part of juvenile left xiphiplastron, dorsal and ventral view. E. UCMP 157444, juvenile left costal 2, external view. F. UCMP 147022, neural, external view. G. UCMP 157442, suprapygal, external view. Scale bars equals 1 cm. 2004 Asiatic Herpelological Research Vol. 10, p. 44 S§t8t»g \ *’.•.■** j?- % : Figure 6. Burmemys magnifies gen. et sp. n. A. AMNH 14196, left peripheral 1, anterior suture, external and visceral views. B. UCMP 147021, left peripheral 1, external, posterior suture and visceral views. C. UCMP 147027, left periph- eral 2, external and visceral views. D. UCMP 147002, left peripheral 3, external, visceral and posterior suture views. E. UCMP 61211, left peripheral 4, external and visceral views. F. UCMP 147001, left peripheral 4 fragment, visceral and posterior suture views (arrow points to flat hyoplastral suture). G. UCMP 131756, juvenile left peripheral 5, suture view. H. UCMP 61218, left peripheral 6, external, posterior suture, visceral and anterior suture views. I. UCMP 142223, right peripheral 7, external, anterior suture, visceral, posterior suture views. J. UCMP 157445, juvenile left peripheral 7 external, visceral, and anterior suture views. K. UCMP 142244, right peripheral 8, external, anterior suture, viscer- al and posterior views. L. AMNH 1911, left peripheral 10 and pygal, external view. Scale bars equal 1 cm. Vol. 10, p. 45 Asiatic Herpetological Research 2004 posterior to the buttress. The free margin is slightlv con- vex. The femoral scale distinctly but narrowly overlaps the dorsal side. The margin dorsal margin of the scale is marked by a shallow sulcus and the bone continues to thicken medially before thing nearer the midline. A par- tial neural (UCMP 170495) has a distinct carina with a rounded top. Discussion. - The referred fragmentary specimens do not appear to belong to other known taxa in the fauna and agree in general morphology with testudinoids, probably testudinids or batagurids. The neural resembles those of carinate batagurids. Carettochelyidae Boulenger, 1887 Anosteirinae Lydekker, 1889 Anosteira Leidy, 1 87 1 Anosteira sp. Referred material. - UCMP locality UCMP V78090: UCMP 131752, peripheral 9 or 10; UCMP 131754, hypoplastron fragment; UCMP 131755, posterior frag- ment of nuchal; UCMP 147115, neural. UCMP locality V83106: UCMP 131736, medial right hypoplastron fragment; UCMP 131737, lateral hypoplastron frag- ment; UCMP 131741, peripheral 7; UCMP 131742, peripheral 8; UCMP 131744, costal fragments; UCMP 131746, anterior fragment of a right peripheral 6. UCMP locality V96001: UCMP 147005, hypoplastron frag- ment; UCMP 147011, left peripheral 7. UCMP locality V96002: UCMP 147030, left peripheral 6. UCMP local- ity V96008: UCMP 147024, right peripheral 2. UCMP locality V96009: UCMP 142225, peripheral 9 or 10. Description: The hyoplastron resembles those seen in typical Anosteira and Pseudanosteira Clark, 1932, and lacks the truncated anteromedial articulation of the new genus described below. This specimen differs from Allaeochelys Noulet, 1867, in having a narrower poste- rior lobe and narrower bridge area. The peripherals are referred to Anosteira on the basis of their small size and well-formed sutures. Most also show the presence of weekly-defined sulci on the external surface. All the peripherals have sharp margin- al carina, and the surface is finely pustulate. The gom- photic pits for reception of the plastron on peripheral 6 (UCMP 147030, Fig. 4C) lie within a longitudinal trough that traverses the peripheral. The latter is 12.6 mm along the free margin carina and 12.5 mm from the carina to the costal suture. A partial peripheral 6 (UCMP 131746) has the trough on the plastral suture filled with 8-9 vertically elongated pits and a shaip lateral carina. The two gomphotic pits on peripheral 7 also occur with- in a trough, but on peripheral 7 the trough is only approximately two-thirds the length of the bone. The peripheral 7 (UCMP 147011) is 12.0 mm along the cari- na. The specimen tentatively identified as peripheral 9 or 10 (UCMP 131752) is deeper than long (16 mm along the margin, 18 mm in depth). The posterior nuchal fragment has the typical caret- tochelyid nuchal pedicle. A faint transverse sulcus is present, and another faint longitudinal sulcus near the midline is visible. A small neural (UCMP 147115) is also referred to Anosteira on the basis of the small size and patterned surface, narrow length to width ratio, and low and broad central carina. Discussion. - Anosteira is known from both Asia (5 species) and North America (1 species) in the Eocene. The closely related genus Pseudanosteira is limited to North America and distinguishable from Anosteira only by details of the top of the carapace. No elements in the Pondaung collection resemble Pseudanosteira. The presence of sulci on the peripherals, nuchal, and costal fragments indicates it should be assigned to Anosteira. The Pondaung specimen is most parsimoniously referred to Anosteira in the absence of any evidence that Pseudanosteira occurs anywhere in Asia. Previous records of Anosteira are confined to China and Mongolia. Carettochelyinae Boulenger, 1887 Burmemys magnified gen. et sp. nov. Holotype. - UCMP 61212, adult left hypoplastron (Fig. 5 A) from UCMP Locality V6204 (near Myaing), found by J. Wyatt Durham, late Professor of Paleontology at the University of California, Berkeley. Paratypes. - AMNH locality “1 mile northeast of Gyat, Magwe Province”: AMNH 1911, pygal, peripheral 10 fragment, and costal fragment. AMNH locality “1 mile north of Koniwa”: AMNH 1919, left first peripheral; AMNH 1928, distal half of right first peripheral; AMNH 14196, partial left peripheral 1; AMNH 14197, plastron fragment. UCMP locality V6204: UCMP 61211, left peripheral 4; UCMP 61218, left peripheral 6. UCMP locality V78090: UCMP 131750, juvenile lateral hypoplastron fragment, UCMP 131751, juvenile periph- eral fragment; UCMP 131753 juvenile xiphiplastron fragment; UCMP 154994, proximal costal fragment. UCMP locality V83106: UCMP 13 1738, juvenile right hypoplastron; UCMP 131739, juvenile hypoplastron fragment; UCMP 131745, juvenile left hypoplastron. UCMP locality V 83 111: UCMP 128406, right peripher- al 2. UCMP locality V83 1 1 6: UCMP 1 3 1 748, hypoplas- tron fragment. UCMP locality V83143: UCMP 131747, Vol. 10, p. 46 Asiatic Herpetological Research 2004 anterior part of left xiphiplastron. UCMP locality V96001: UCMP 147001, left peripheral 4 fragment; UCMP 147002, partial left peripheral 3; UCMP 147003, anterior peripheral fragment; UCMP 147009, neural; UCMP 147010, peripheral fragment; UCMP 147012, juvenile medial hypoplastron fragment. UCMP locality V96002: UCMP 142244, right peripheral 8; UCMP 154984, anterior peripheral fragment. UCMP locality V96008: UCMP 147021, left first peripheral; UCMP 147023, posterior peripheral fragment; UCMP 147027, left peripheral 2; UCMP 147028, partial right peripheral 1; UCMP 147029 distal fragment of a costal. UCMP locality V96009: UCMP 142223, right peripheral 7. UCMP locality V99498: UCMP 157443, neural; UCMP 157446, shell fragments. Referred material. - UCMP locality V96001: UCMP 147004, plastron fragment. UCMP locality V83106: UCMP 131740, hyoplastron fragment. UCMP locality V78090: UCMP 131756, juvenile left peripheral 5; UCMP 154993, posterior fragment of left epiplastron. UCMP locality V99498: UCMP 157442, suprapygal; UCMP 157444, left costal 2; UCMP 157445, left periph- eral 7. Diagnosis. - Burmemys is distinguished from other carettochelyines by the combination of asymmetrical articulation of the hyo-hypoplastra, narrow hypoplastral bridge, and large size (estimated carapace length greater than 1000 mm). Description. - The holotype hypoplastron (UCMP 61212, Fig. 5 A) is massive. The anterior suture of the left hypoplastron consists of two sutures. The suture with the left hyoplastron is sinusoidal, curving antero- medially, and joins a distinct, straight and anteromedial- ly-facing suture, presumably for articulation with the right hyoplastron. The ventral sculpture consists of a pattern of irregular, closely-spaced tubercles that radiate from a focal point lateral to the middle of the medial moiety. Laterally, the tubercles coalesce into ridges radi- ating laterally. The sutures are finely dentate and thick (13 mm). The lateral margin and posterior half of the medial part is broken away in the type, but these are pre- served in the juvenile specimen (UCMP 131745, Fig. 5B). The width of the medial part of the hypoplastron measured from the apex of the inguinal notch to the plastral midline is only one-half or less of the maximum hypoplastral width. The inguinal notch is open and not confined as in Carettochelys Ramsey, 1887. The anteri- or-posterior width of the bridge area is one-half or less the width of the xiphiplastral lobe of the hypoplastron. The referred juvenile specimens exhibit the same sutur- al shapes as the adult (type) but the inguinal notches are shallower, sculpture less organized, and lateral extent of the lateral arm of the bridges are shorter. The posterior part of a juvenile epiplastron (UCMP 154993, Fig. 5C) is referred to Burmemys on the basis of the convex curvature of the lateral margin that indicates a short and rounded anterior lobe, and an obtuse angle between the entoplastral and hyoplastral sutures indicat- ing a short and broad entoplastron. Two xiphiplastra (UCMP 131747, Fig. 5D; UCMP 131753) are referred to Burmemys on the basis of rela- tively larger size, converging (non-parallel) medial and lateral margins of the anterior moiety, and thinning rather than thickening toward the midline suture. Both specimens are small (proximal width of UCMP 131747 is 20 mm) and thus considered as juveniles. The juvenile left costal (UCMP 157444, Fig. 5E) is nearly uniformly thin, parallel sided, and sculptured with a subdued and random pattern of low pustules and short ridges. The distal margin forms about a 45 degree angle to the sides. There are no sulci. The parallel sides and high angle of the distal margin indicate a second costal. The distal end of an adult costal (UCMP 147029) is subtlety sculptured with longitudinal irregular ridges. The distal suture is weakly dentate but patent except above the rib. The rib ends protrudes prominently. Although damaged, the distal width is about 70 mm. A large neural (UCMP 147009, Figs. 5F) is relative- ly narrow, lacks a midline carina, and has subtle sculp- ture of very shallow dimples. It has a midline length of 51 mm, maximum width of 32 mm, and maximum thickness of the lateral side of 15.4 mm. The suprapygal (UCMP 157442, Fig. 5G) is trian- gular with a distinct medial carina. The surface sculpture consists of irregular vermiform ridges that radiate from the central area of the posterior margin. It is longer than wide (32.2 mm long, 31.5 mm wide). At least nine peripheral positions are represented. The sculpture is variable consisting of distinct tubercles at one extreme to anastomosing pits and ridges at the other. The free margins of adult specimens are rounded but may be acute in juveniles. There are no indications of scale sulci. The first peripheral exhibits distinct sutures with the first costal, nuchal and second peripheral. The free mar- gin perimeter is asymmetrically curved. The largest specimens (AMNH 1919, Fig. 6A; UCMP 147021, Fig. 6B) have perimeter lengths of 114 and 119 mm, maxi- mum depths of 90 and 81 mm, maximum thicknesses at the posterior suture of 33 and 27 mm, and maximum thicknesses at anterior suture of 28 and 25 mm respec- tively. 2004 Asiatic Herpetological Research Vol. 10, p. 47 The two second peripherals differ in size. UCMP 147026 (Fig. 6C) is massive with a free margin length of 82 mm, maximum depth of 67 mm, and maximum thick- ness of the anterior suture of 20 mm. Comparable meas- urements of UCMP 128406 are 45, 40, and 11 mm respectively. The second peripheral is roughly rectangu- lar in external view. The only specimen referred to the third peripheral (UCMP 147002, Fig. 6D) is lacking the anteroventral and posterodorsal corners. The anterior part of the dor- sal suture is a semi-scarf joint — probably for the rib end of the first costal. The peripheral thickens noticeably towards the posterior suture and reaches a thickness of 32 mm at the suture. The fourth peripheral (UCMP 61211, Fig. 6E) is damaged anteriorly and dorsally and locally abraded. Its length along the lateral carina is 75 mm. The free mar- gin curves posteromedially on the posterior moiety to form a plastral articulation. The plastral articular surface is relatively flat but deep (up to 16 mm) and without pits for the hyoplastral buttress or normal dentations, thus indicating a weakly ligamental and kinetic joint. The lat- eral carina is broadly rounded. The peripheral 4 frag- ment (UCMP 147001, Fig. 6F) also shows this rather flat and deep (19 mm) hyoplastral suture. A small peripheral, probably a left peripheral 5 (UCMP 131756, Fig. 6G) is considered a juvenile of this species. The plastral arm is very short with a longitudi- nal trough enclosing a series of gomphotic pits. The lat- eral carina is slightly rounded and broadly upturned. The length of the lateral carina is 22 mm and has a posterior thickness of about 9 mm. A relatively complete left peripheral 6 (UCMP 61218, Fig. 6H) has a damaged plastral margin and lacks the dorsal suture. The plastral and costal arms converge posteriorly. The plastral articulation is broken anteriorly but posteriorly has a longitudinal trough indicating inter- digitation with the hypoplastron. The lateral carina is rounded and slightly upturned. The length along the lat- eral carina is 82 mm. A peripheral 7 (UCMP 142223, Fig. 61) of an adult measures 99 mm along the marginal carina, 100 mm from the carina to costal margin, posterior thickness of 29 mm and an anterior thickness of more than 45 mm. The hypoplastral suture is damaged but trough-like, extends about half-way along the medial side, and appears to have housed one or two recessed pits. The isolated peripheral 7 (UCMP 157445, Fig. 6J), a pre- sumed juvenile, closely resembles Carettochelys with the hypoplastral buttress rising up the central part of the medial side. The free margin is sharp and broadly upturned. The length of the free margin is about 34 mm. The adult peripheral 8 (UCMP 142244, Fig. 6K) is massive and slightly shorter than deep (97 mm along the lateral carina and 105 mm from the carina to costal suture). An anteriorly-deepening trough divides the medial surface into dorsal and ventral arms anteriorly. The pygal (AMNH 1911, Fig. 6L) is distinctly trapezoidal with a short anterior side and a low but sharp medial crest. The sculpture consists of widely spaced irregular tubercles that fade out near the medial crest and free margin. An associated posterior peripheral 10 frag- ment has a sculpture of irregular ridges and tubercles that radial from a central focus. Discussion. - The absence of scales and large size place Burmemys in the Carettochelyinae. Of the three Eocene genera of Carettochelyinae, Burmemys differs from all in the presence of two distinct anterior articular sutures on one of the hypoplastra. This most likely represents an asymmetrical articulation with the hyoplastra, with one of the hyoplastra extending well across the midline to form an angled articulation with the opposite hypoplas- tron. Even where the hyo-and hypoplastra are not mirror images with one of the hypoplastra contacting the oppo- site hyoplastron (e.g., Anosteira in Hay, 1908, Fig. 353), the midline suture remains straight as in other Paleogene carettochelyids and the plastral midline suture usually exhibits some limited kinesis. This asymmetry is not unusual in turtles, but within carettochelyids was known only to a lesser degree in some Carettochelys. The hypoplastron in extant Carettochelys insculpta Ramsey, 1887 may cross the midline to form a short angled suture with the opposite hyoplastron (AMNH 84212, and Rooij, 1915, fig. 123a). This occurs on the left hyoplas- tra on both of these. Burmemys resembles anosteirines and differs from extant Carettochelys, Hemichelys Lydekker, 1887, (pi. XII, fig. 2) from the Eocene of the Punjab, and Chorlakkichelys Broin, 1987 (pi. 1, fig. 2) from the Eocene of Pakistan in the relatively broad inguinal notch. Burmemys additionally differs from Chorlakkichelys and Carettochelys in having a distinct- ly narrow bridge area. The general proportions of the hypoplastron resemble those of Allaeochelys from the Eocene of Europe (Broin, 1977, PI. XVI, Fig. 3). The suprapygal differs from Allaeochelys, Carettochelys, Hemichelys and probably Chorlakkichelys in being longer than wide. Burmemys is also the largest carettochelyid described to date. Based on scaling up of the elements in comparison to other carettochelyids, we can estimate that the shell length of Burmemys exceeded 1000 mm. This estimate suggests that Burmemys is among the largest turtles known, but is smaller than estimates Head et al. (1999) provided for Eocene trionychids from Pakistan, which may have reached more than 2000 mm in length, and is smaller than the giant Bridgerian triony- chid from Wyoming (Gaffney, 1979). Vol. 10, p. 48 Asiatic Herpetological Research 2004 Figure 7. Trionychinae. A. UCMP 61213, right hypoplastron, ventral view. B. UCMP 1537993, fragment of right hypoplastron, ventral view. C. UCMP 147022, neural, external view. D. UCMP 170520, costal fragment, external view. Scale bars equals 1 cm. Trionychidae Gray, 1825 Trionychinae Gray, 1825 Trionychinae genus indet. Trionychinae, large form Referred material. - UCMP locality V6204: UCMP 61213, right hypoplastron. UCMP locality V78090: UCMP 170497, costal fragments. UCMP locality V83116: UCMP 153799, fragment of right hypoplas- tron. UCMP locality V83143: UCMP 173809, plastron fragment. UCMP locality V96001: UCMP 147020, costal fragment. UCMP locality V96002: UCMP 154983, costal proximal fragment. UCMP locality V96008: UCMP 147022, neural. UCMP locality V96009: UCMP 142222, plastron fragment. Description. - The hypoplastron (UCMP 61213, Fig. 7A) lacks the projecting spines (present in UCMP 153799, Fig. 7B) but is otherwise relatively complete. The calloused area has well defined edges and covers most of the ventral surface, except for the long offset shelf on the medial edge. The calloused area is sculp- tured with distinct pits and ridges, while the shelf is amorphously roughened. The buttress is composed of two protruding spikes. The posteromost of these is bro- ken off at the base, but the larger anterior one is divided into three fluted points at its tip. The isolated neural (UCMP 147022, Fig. 7C), prob- ably 6 or 7, is hexagonal and narrows distinctly posteri- orly. The dorsal surface is weakly sculptured, lack sulci, and is flat anteriorly but is formed into a central carina posteriorly. The neural is 46 mm long and 50 mm wide. Discussion. - The large size of the hypoplastron and general conformation indicates a trionychine and gener- ally resembles Pelochelys Gray 1864, Chitra Gray 1 844, and Pelodiscus Fitzinger 1835 in these features, but dif- fers in having a wide, unsculptured medial shelf. Additional material would be needed to refine identifi- cation. Trionychinae, small form Referred material. - UCMP locality V6204: UCMP 61210 plastron fragment. UCMP locality V83106: UCMP 147116, two costal fragments. Description. - Two distal costal fragments exhibit a well-defined sculpture of pits and ridges with indica- tions of longitudinal welts. The pattern extends to the free margin with only a slight sculpture-free zone at the free margin, suggesting an adult turtle of relatively small 2004 Asiatic Herpetological Research Vol. 10, p. 49 size in comparison with the preceding taxon. The sculp- ture resembles that of a trionychine rather than a cyclanorbine such as Lissemys Smith, 1931. Trionychinae, ornate form Referred material. - UCMP locality V98109: UCMP 170520, two costal fragments. Description. - The costal fragments (UCMP 170520, Fig. 7D) exhibit a striking sculpture of large, elongate tubercles rising above a surface composed of a general- ly organized pattern of longitudinal rows of shallow pits and low rides. The longitudinal axes of the raised tuber- cles vary from anterior-posterior to medial-lateral and are large (11-15 mm) relative to the overall size of the larger costal fragment (maximum preserved width of 32 mm). Carapace sculpturing varies between individuals and also ontogenetically, but the peculiar sculpture shows some resemblance to that seen in some extant Aspideretes Hay, 1904. Discussion In addition to the turtles, a variety of other lower verte- brates are present in the Pondaung Formation including a carcharhinid shark, Galeocerdo Muller and Henle 1837 (UCMP 142238), a clariid catfish (UCMP 128411), at least four species of agamid lizards (UCMP 128410, 130290, 142227, 142232), paleophid and colu- broid snakes (see Head et al, in prep.), and a minimum of two crocodilians, including a pristichampsine croco- dylian (UCMP 147127) and a dyrosaurid (Buffetaut, 1978). Unfortunately, reports on Asian lower vertebrates of comparable age (Sharamurunian Asian Land Mammal Age or late middle Eocene) are few. Thus, the limited lit- erature, combined with the fragmentary nature of the Pondaung fossils themselves, make detailed compar- isons with other faunas difficult. Nonetheless, compar- isons with known Sharamurunian lower vertebrate fau- nas reveal only a few similarities between the Pondaung and any other locality. Pleurodires are previously unde- scribed from Asia, although Broin (1987) notes the pres- ence of “Pelomedusidae and / or Emydidae” from the middle Eocene of Pakistan and Oligocene of India. An adocid was described by Gilmore (1931) from the late middle Eocene of Mongolia, but none were identified in the Pondaung assemblage. The carettochelyid genus, Anosteira , has been reported from age-equivalents in Manchuria (Zangerl, 1947) and Guangdong, China (Sun Ailing et al., 1992) and from slightly younger sediments in Shandong and Guangdong provinces (Yeh, 1963). The only other carettochelyines described from Asia are Chorlakkichelys and Hemichelys from the early middle Eocene of Pakistan (Lydekker, 1887; Broin, 1987), and Burmemys is the most easterly and southerly Eocene record of the subfamily. Trionychids, as elsewhere, are an important part of the fauna, but our material is not sufficiently diagnostic to make any meaningful biogeo- graphic comparisons. Testudinids are widely reported in Chinese and Mongolian Eocene faunas (Gilmore, 1931; Ye, 1963), but all of these appear to be more generalized forms similar to Hadrianus or Kansuchelys. The Pondaung form appears to be more like the modern Testudo, and thus distinct from contemporaneous Chinese and Mongolian taxa. Among other reptiles, the only other agamid lizard known in the Asian Sharamurunian is Tinosaurus yuan- quensis from the Heti Formation (Li, 1991), but it is a diminutive form that bears no resemblance to the Pondaung agamids. Crocodilians are known elsewhere, but not in detail. In overall diversity, the Pondaung fauna shares more general resemblances to the better- known Irdinmanhan faunas, especially that of the Kuldana Formation of Pakistan (Broin, 1987). Based on the fragmentary evidence available to date, several observations can be made regarding the Pondaung lower vertebrate fauna. Faunal endemicity is supported by the number of unique taxa, and the compo- sition of the turtle fauna is unusual with trionychoids (especially carettochelyids) dominating. Faunal compo- sition and the large size of these turtles are consistent with an interpretation of these sites as representing a warm, tropical floodplain environment, deposited fairly near shore. The aquatic habits of most of the lower ver- tebrates suggest that during late middle Eocene time the Pondaung region was a well-drained floodplain environ- ment, a finding consistent with previous geological interpretations (e.g., Bender, 1983, Soe et al., 2002). Acknowledgments We are grateful to Donald E. Savage, Ba Maw, and Thaw Tint for reinitiating fieldwork in the Pondaung region in the late 1970’s, and Brig. -Gen Than Tun and Major Bo Bo from the Office of Strategic Studies, Ministry of Defense for arranging our visits to Myanmar. Tin Thein, Aye Ko Aung, Aung Naing Soe, and other members of the Pondaung Fossil Expedition team are thanked for their assistance during our stays and their help while in the field. Eugene Gaffney, Charlotte Hotton, and Mark Norell kindly provided access to and assistance in use of the AMNH collections and associated records. Walter Joyce reviewed the man- uscript and provided constructive criticisms. Funding for fieldwork in Myanmar has been provided by the Smithsonian Foreign Currency Program (from Public Vol. 10, p. 50 Asiatic Herpetological Research 2004 Law 480 funds), the L.S.B. Leakey Foundation, the University of California Museum of Paleontology, the University of North Carolina at Charlotte Foundation, the University of Iowa Center for Pacific and Asian Studies, the Office of the Vice President for Research and the Human Evolution Research Fund of the University of Iowa Foundation. This is UCMP contribu- tion no. 1836. Literature Cited Batsch, A. J. G. C. 1788. 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Fission-track zircon age of the Eocene Pondaung Formation, Myanmar. Journal of Human Evolution 42: 361-369. Wood, R. C. 1983. Kenyemys williamsi, a fossil pelome- dusid turtle from the Pliocene of Kenya. Pp. 74-85. In: A. G. Rhodin et al. (eds). Advances in Herpetology and Evolutionary Biology. Ye (Yeh), X. 1963. Fossil turtles of China. Palaeontologica Sinica, new Series C (18): 112 pp. Zangerl, R. 1947. A new anosteirine turtle from Manchuria. Fieldiana, Geology 10:13-21. 2004 Asiatic Herpetological Research Vol. 10, pp. 53-109 A Review of the Comparative Morphology of Extant Testudinoid Turtles (Reptilia: Testudines) Walter G. Joyce1 and Christopher J. Bell2 1 Department of Geology and Geophysics, Yale University, New Haven, CT 06511 E-mail: walter.joyce@yale.edu Department of Geological Sciences, The University of Texas at Austin, Austin, TX 78712 E-mail: cjbell@mail. utexas. edu Abstract. - With an expansive geographic distribution, an excellent fossil record, and over 140 recognized extant species, testudinoid turtles constitute one of the most diverse and widespread clades of turtles. The current under- standing of the distribution of morphological characters among testudinoid turtles is poor. Improved knowledge will help to facilitate accurate identification of fossil remains, and to provide a reliable morphological data set for phylo- genetic analyses. We provide a critical review of skeletal and scute characters commonly utilized in previous system- atic analyses of Testudinoidea. Description and illustration of character states, discussion of their distribution within Testudinoidea, and polarity determinations for 93 characters are provided. Our preliminary results indicate that onto- genetic changes in skeletal structure are an important source of variation within Testudinoidea. Sexual variation, onto- genetic variation, and intra- and inter-population variation are inadequately documented for most testudinoid taxa. Furthermore, data matrices of morphologic characters in the existing literature must be carefully reconsidered. Previously published morphologic data provide reasonably strong support for the monophyly of 'Testudinidae.' Strong morphologic support for a monophyletic 'Emydidae' is lacking, and 'batagurid' monophyly has not been rigorously tested in the literature. Because a new research cycle centered on testudinoid phylogeny is now under way, it is essen- tial to critically re-examine the underlying assumptions and working hypotheses that have governed this field of study over the last 20 years. Key words. - Testudines, Testudinoidea, Testudinidae, Emydidae, Bataguridae, Geoemydidae, morphology, systemat- ics. Introduction Pond turtles and land tortoises (collectively, Testudinoidea) form one of the largest and most wide- spread clades of living turtles, with more than 140 extant species and an almost worldwide distribution. The dis- covery and description of many new fossil testudinoids in the last half century, combined with the emergence and ascendancy of molecular techniques in systematics, provide new opportunities to explore the evolutionary history of the group in unprecedented detail. Concomitant with the appearance of these new data sets and analytical techniques comes an increasing apprecia- tion for conservation efforts to preserve these turtle lin- eages and help to secure their future in the face of increasing human predation and habitat encroachment. This is true especially for the Asian representatives of this clade (e.g., van Dijk et al., 2000) but also is relevant at a more generalized and inclusive level (e.g., Rhodin, 2000). Our recent attempts to diagnose fossil testudinoids reliably and to place them within a phylogenetic context led to the recognition that a critical re-evaluation of mor- phological data and purported synapomorphies for the subclades of testudinoid turtles is desirable. A more thorough understanding of morphological data sets will provide not only a means by which molecular trees may be independently assessed, but also will form an essen- tial foundation for diagnosing and interpreting fossil specimens. This in turn will facilitate the integration of fossil taxa into future systematic analyses, and will enhance our understanding of the paleobiogeography and divergence times of extant lineages. The recent flurry of published works appears to rep- resent the beginning of a new research cycle ( sensu Kluge, 1991) in testudinoid systematics. We suggest that an important part of this cycle will be a critical re-exam- ination of the working hypotheses that have governed testudinoid systematics since the publication of McDowell’s (1964) seminal work on the group. A key component of this will be the assessment of fundamen- tal, often unstated, assumptions that underlie current hypotheses of relationship. Our contribution to this research cycle is the first critical reappraisal of morpho- logical characters applied to testudinoid systematics since the work of Hirayama (1985). The emerging improvement in our understanding of testudinoid rela- tionships based on molecular sequence data will certain- © 2004 by Asiatic Herpetological Research Vol. 10, p. 54 Asiatic Herpetological Research 2004 ly result in numerous new questions (e.g., regarding paleobiogeography, the timing of sequence and evolu- tionary divergences, and the evolution of morphological adaptations) that will demand a clearer understanding of testudinoid morphology. The purpose of this paper is to present a preliminary revision and discussion of the morphological characters previously utilized in investigations of testudinoid sys- tematics. Our goal here is not to produce a phylogenetic hypothesis (indeed, we deliberately eschew such a pro- duction), but rather to evaluate the morphological data that have been, and will be, used to generate such hypotheses. To enhance our discussion and facilitate improved communication about testudinoid morpholo- gy, we provide illustrations of all characters states we discuss. Abbreviations. - Institution and collection abbrevia- tions: CAS, California Academy of Sciences, San Francisco, California; CM, Carnegie Museum of Natural History, Pittsburgh, Pennsylvania; FMNH, Field Museum of Natural History, Chicago, Illinois; IVPP, Institute of Vertebrate Paleontology and Paleoanthropology, Beijing, China; KU, The University of Kansas Natural History Museum, Lawrence, Kansas; LMNH, Louisiana Museum of Natural History, Baton Rouge, Louisiana; MCZ, Museum of Comparative Zoology, Harvard University, Cambridge, Massachusetts; TNHC, Texas Natural History Collections, Texas Memorial Museum, Austin, Texas; TUMNH, Tulane University Museum of Natural History, New Orleans, Louisiana; YPM, Yale Peabody Museum, New Haven, Connecticut. Abbreviations used in figures: AB, abdominal scute; af, articular facet; an, angular; bo, basioccipital; CE, cervical scute; co, costal bone; ent, entoplastron; epi, epipterygoid; fdm, foramen dentofaciale majus; fr, frontal; fpp, foramen palatinum posterius; HU, humeral scute; hyo, hyoplastron; hypo, hypoplastron; ju, jugal; MA, marginal scute; mx, maxilla; ne, neural bone; pa, parietal; pal, palatine; PEC, pectoral scute; pf, pre- frontal; PL, pleural scute; pm, premaxilla; po, postor- bital; pt, pterygoid; qj, quadratojugal; qu, quadrate; VE, vertebral scute; vf, vomerine foramen; vo, vomer. Material and Methods We examined 309 testudinoid specimens representing 93 species, but focused our efforts on 46 representative species. The list of specimens examined is provided in Appendix 1. Turtle shell nomenclature follows Zangerl (1969) and cranial nomenclature follows Gaffney (1972). Of the 46 focal species, most were recognized as valid species by Ernst and Barbour (1989), with the exception of texana, which they placed within ‘ concin - na .’ Generic allocations for testudinoid species varied widely over the last 50 years and are subject to differing opinions today, particularly because the monophyly of many testudinoid genera remains untested. We conse- quently suppress the use of generic names wherever pos- sible and use species epithets only. This procedure also has the advantage of precisely associating observations with species only instead of higher taxonomic cate- gories. Most extant turtles have distinct species names, but among those turtles discussed in this review the species epithets insculpta, nelsoni , oculifera , ornata, and platynota each appear twice ( insculpta under Glyptemys and Carettochelys; nelsoni under Pseudemys and Terrapene\ oculifera under Graptemys and Psammobates\ ornata under Pseudemys and Terrapene; platynota under Geochelone and Notochelys ). For clari- ty in these instances, we indicate our usage with a sin- gle-letter generic abbreviation. A complete list of all cur- rently recognized testudinoid species and all outgroup species used herein is provided in Appendix 2 together with a list of their various generic assignments used in the last 50 years. Our use of the classic higher categories is always restricted to their phylogentic crown. ‘Emydinae’ (sensu McDowell, 1964) are also referred to as ‘Emydidae,’ ‘emydids,’ or North American pond turtles; ‘Batagurinae’ (sensu McDowell, 1964) as ‘Bataguridae,’ ‘batagurids’ or Asian pond turtles; and ‘Testudinidae’ (sensu McDowell, 1964) as ‘testudinids’ or (land) tor- toises. We make no a priori assumptions of monophyly for any of these categories, and retain single quotations around these names throughout the text to emphasize our uncertainty. We attempted to examine most significant morpho- logical characters commonly utilized in systematic stud- ies of testudinoids, but the majority of our observations concern the skeletal system and scute characters. Almost all characters were derived from the literature. Major sources for each category were: ‘Batagurinae’ (Hirayama, 1985; McCord et al., 1995; Yasukawa et ah, 2001); ‘Testudinidae’ (Crumly, 1982, 1985, 1994); and ‘Emydinae’ (Gaffney and Meylan, 1988; Burke et ah, 1996). Additional characters were also found in Mlynarski (1976), Shaffer et ah (1997), and other sources cited in the character discussions. With few exceptions, morphological features were examined on specimens themselves; evaluations based on previously published literature are indicated where applicable. Sexual dimorphism, ontogenetically influenced poly- morphisms, and geographic variation in morphology are not well explored in testudinoid turtles. These areas are in need of much more research. A full exploration of 2004 Asiatic Herpetological Research Vol. 10, p. 55 such variation is beyond the scope of this work, but we aie able to make some preliminary observations regard- ing morphological change through ontogeny in some anatomical systems. Polarizing characters with the help of outgroups proved to be a difficult task, mostly because all relevant extant sister taxa are highly specialized after more than 65 million years of independent evolution. Furthermore, hypotheses of the systematic relationships of the major groups of cryptodires reveal a highly unstable picture (e.g., Bickham, 1981; Gaffney, 1975, 1985; Gaffney et al., 1991; Shaffer et al., 1997) making it impossible to make any a priori decisions regarding the succession of outgroups. We consequently assessed polarity for most characters by examining select outgroup taxa and the ingroup taxa. Where polarity is not clear from outgroup comparison, we sometimes relied upon ingroup com- monality. To allow full transparency, we discuss every polarity decision at the end of each character descrip- tion. Outgroup taxa include the cryptodires caretta , odoratus, serpentina , and spinifera, and the pleurodires gibba , siebenrocki, subglobosa, and subrufa. For a num- ber of characters, especially of the shell, neither ingroup nor outgroup analysis of extant taxa proved useful. In these instances, polarity was based on literature descrip- tions of the “lindholmemydid” taxa Gravemys, Lindholmemys , and Mongolemys . These Cretaceous, Asian, fossil taxa are not well described in the literature, but sufficient material and description exists to use these taxa to help polarize character states (e.g., Khosatzky and Mlynarski, 1971; Sukhanov, 2000; Danilov and Sukhanov, 2001). The group may not be monophyletic, but putative members currently are hypothesized to sit along the phylogentic stem of Testudinoidea (Danilov and Sukhanov, 2001). We purposefully did not use the fossil taxon ‘ Echmatemys’ as an outgroup taxon (Hirayama, 1985), because its phylogenetic position out- side of Testudinoidea or even ‘Batagurinae’ is not suffi- ciently demonstrated. All figures were produced using digital photogra- phy and processed using Adobe Photoshop. Images were digitally enhanced using the burn and burnish tools and the unsharp mask filter option. Taxonomic and Systematic Background. - Despite the increased attention directed towards testudinoids by sci- entists, hobbyists, and nonprofessional enthusiasts in the last thirty-five years, our collective conceptualization of the higher-level (beyond the specific and generic) sys- tematics within this clade remained virtually unchanged since the work of McDowell (1964). The various tax- onomies in current use owe their existence in large part to historical contexts that are not well appieciated by many authors. A brief summary is given here. During the second half of the 19th century, a num- ber of attempts were made to work out higher-level tes- tudinoid relationships and to apply taxonomic conven- tions that were designed (to a greater or lesser extent) to communicate conceptualizations of these relationships. In his synopsis on the turtles of North America, Agassiz (1857) united all pond turtles into the Emydoidae and subdivided this group into a three monotypic subfami- lies (Deirochelyoidae for reticularia, Evemydoidae for blandingii, Cistudinina for T. ornata and Carolina ) fol- lowed by the subfamilies Clemmydoidae (for G. insculp- ta, guttata, marmorata, and muhlenbergii ) and Nectemydoidae (for those species currently placed in the genera Pseudemys, Trachemys, Graptemys , Malaclemys and Chrysemys). Most land tortoises were placed in Testudinidae by Theobald (1868); he also included all ‘leaf turtles and tortoises’ (e.g., amboinensis, emys, den- tata, grandis, tricarinata ) in Geoemydidae, and an eclectic group of aquatic turtles, including mega- cephalum, serpentina, kinostemids, and all remaining testudinoids, in Emydidae. Subsequently, all land tor- toises (including emys and impressa) were united in the Testudinidae by Gray (1870). Those species currently placed in Pseudemys and Trachemys were assigned by Gray (1870) to the Pseudemydae; the Asian taxa baska, borneoensis, thurjii, kachuga, and ocellata were assigned to the Bataguridae, and all hinged pond turtles, ‘true terrapins,’ and ‘snail-eating pond turtles’ to the Holarctic families Cistudinidae, Emydidae, and Malaclemmydae, respectively. Despite these early attempts, most subsequent authors (e.g., Boulenger, 1889; Siebenrock, 1909; Lindholm, 1929; Smith, 1931; Bourret, 1941) ignored (or were unaware of) these works and simply divided all testudinoid turtles into two speciose subgroups: tortois- es (Testudinidae or Testudininae) and pond turtles (Emydidae or Emydinae). This situation remained static for nearly 100 years until the comprehensive and influ- ential work of McDowell (1964). He not only divided all known pond turtles into several species complexes (‘ Hardella,' ‘ Batagur ,’ ‘ Orlitia ,’ ‘ Geoemyda ,’ ‘ Chrysemys ,’ ‘ Deirochelys,' and ‘ Emys ’ complexes), but also concluded that pond turtles can be divided clearly into two subgroups, the predominantly North American ‘Emydinae’ and the Asian and central American ‘Batagurinae.’ Furthermore, McDowell (1964) reasoned that tortoises are not the sister group of pond turtles, but rather were likely derived from a ‘batagurine’ ancestor. These conclusions were later corroborated by the first, and to date only, comprehensive morphological cladistic analysis of ‘batagurine’ systematics (Hirayama, 1985). The influence of McDowell’s (1964) work is best understood when considering its continuous impact on Vol. 10, p. 56 Asiatic Herpetological Research 2004 subsequently proposed phylogenies. Despite differences of opinion regarding generic- and species-level system- atic arrangements, virtually all major synthetic works in the last thirty years followed McDowell’s (1964) subdi- vision of pond turtles into the ‘Batagurinae’ and ‘Emydinae' (e.g., Mlynarski, 1976; Pritchard, 1979; Ernst and Barbour, 1989), even though 'Batagurinae’ may best be regarded as a paraphyletic taxon (McDowell, 1964; Hirayama, 1985). The fundamental division proposed by McDowell is also reflected in more recent studies centered on using various molecular tech- niques to elucidate phylogeny, the majority of which dealt with treatments of in-group relationships within one or the other of McDowell’s groups (Sites et al., 1984; Bickham et al., 1996; Carr and Bickham, 1986; Wu et al., 1999; McCord et al., 2000; Feldman and Parham, 2002; Honda et al., 2002; Iverson et al., 2002; Stephens and Wiens, 2003). Admittedly, the list of autapomorphic characters compiled by Crumly (1985) for the ‘Testudininae’ com- pellingly corroborates the hypothesis of tortoise mono- phyly. However, most characters that currently unite ‘Emydinae’ or ‘Batagurinae + Testudininae’ seem to support these groupings weakly, because the derived states typically are not found within all species of the ingroup and commonly also are observed in species of the alleged sister group (e.g., Hirayama, 1985; Gaffney and Meylan, 1988). In addition, several of the characters that purportedly distinguish ‘Batagurinae + Testudininae’ from the ‘Emydinae’ probably should be considered primitive for the entire group (Gaffney and Meylan, 1988). Even if some characters do successfully unite a group, monophyly is not established until the involved characters are demonstrated to be derived with- in Testudinoidea. Furthermore, the simple demonstra- tion of monophyly for a given group does not automati- cally imply that it must be the sister to the remaining taxa. For instance, ‘Emydinae’ may be monophyletic, but monophyly does not necessarily demand that ‘Emydinae’ be regarded as the sister to ‘Batagurinae + Testudininae.’ It is at least plausible that ‘Emydinae’ is situated within ‘Batagurinae,’ a possibility that is not adequately explored and tested in the literature. Similarly, most of the groupings considered by Gray (1855, 1870) and Agassiz (1857) were not dis- cussed in recent literature, even though they might be valid. For instance, given the considerable list of mor- phological similarities that are shared by hinged turtles of the New and Old World (e.g., development of a plas- tral hinge, reduction of posterior neural elements, fusion of the femoral trochanter, great reduction of the tempo- ral arch) perhaps Gray (1870) was truly visionary in uniting these turtles as the ‘Cistudinidae.’ Only a global cladistic analysis with no a priori assumption regarding internal relationships can evaluate these alternatives and produce testable results. It is toward this end that we offer our critical reappraisal of morphological characters in testudinoids. Results and Discussion Cranium (1) Shape of the fissura ethmoidalis; 0 = narrow or closed, keyhole-shaped, Fig. 1; l = very wide, Fig. 2 (modified from Crumly, 1982, 13; Hirayama, 1985, 1; McCord et al., 1995, 5). The general configuration of the fissura in ‘emy- dids’ and ‘batagurids’ is keyhole-shaped (McDowell, 1964). Different proportions and widths are apparent (especially in the ventral part of the fissura), and were scored by Hirayama (1985) and Crumly (1982) as dis- crete character states. Our survey of many taxa reveals morphological intermediates, and the expression of var- ious states appears to have an ontogenetic component in which younger individuals exhibit a relatively larger fis- sura, which corresponds to a less-ossified nasal cavity. However, a rather significant morphological gap can be observed between tortoises and pond turtles. For the purpose of this review, we lumped Hirayama’s (1985) states into our state 0, and Crumly ’s (1982) into our state 1. The scoring presented by Hirayama (1985) and McCord et al. (1995) permitted phylogenetic resolution within ‘batagurids’, and that of Crumly (1982) within tortoises. Our revised scoring permits support only for the hypothesis of a monophyletic ‘Testudinidae.’ Polarity: Pleurodires lack a defined fissura eth- moidalis. A keyhole-shaped fissura ethmoidalis is pres- ent in spinifera, odoratus, caretta, serpentina , and Mongolemys, and this condition is considered primitive for testudinoids. (2) Medial inflection of the inferior descending process- es of the frontal; 0 = absent, or very small, Fig. 3; 1 = present, well-developed, medial contact present or almost present, Fig. 4 (modified from Hirayama, 1985, 2). In most turtles, a gutter (the sulcus olfactorius) is formed along the ventral surface of the ffontals. This gutter transmits the olfactory nerve. The lateral rims of the sulcus sometimes form processes that descend ven- tromedially to surround the nerve from below (McDowell, 1964). According to Hirayama (1985) these processes are well-developed, or are in contact medially, in ocellata and hamiltonii. We confirm the presence of well-developed processes in these taxa and add petersi, N. platynota, and all sampled tortoises to the list. We recommend that this character not be subdivided into additional character states, because the descending processes of the ffontals grow larger through ontogeny. 2004 Asiatic Herpetological Research Vol. 10, p. 57 Polarity: A medial inflection is absent in all out- groups and the vast majority of the ingroup. We consid- er its presence to be derived. (j) Frontal contribution to the orbital rim; 0 — present, no prefrontal/postorbital contact on dorsal surface, Fig. 5; 1 = absent, frontal excluded from orbital rim by pre- frontal/postorbital contact, Fig. 6 (modified from Crumly, 1982, 17; Hirayama, 1985, 3; Shaffer et al., 1997, 97; Yasukawa et al, 2001, 1). Three states for this character were scored by Hirayama (1985) and Yasukawa et al. (2001): frontal contribution always or usually present, frontal some- times excluded from orbital rim, and frontal always excluded from orbital rim. Our sample size for many taxa does not permit a reliable assessment of intraspecif- ic variation in this character, and thus our initial scores differed for some taxa from those of Hirayama (1985). We add peter si and N. platynota to the list of taxa in which the frontal appears always to be excluded. For those taxa that sometimes exclude the frontals, we con- firm this polymorphic condition in crassicollis, and add agassizii and annulata. Our sample was too small to confirm the reported polymorphic condition in amboinensis , and pulcherrima by Hirayama (1985) and Yasukawa et al. (2001), but we scored these taxa as polymorphic based on their observations. We also fol- lowed Crumly (1982) by coding pardalis as polymor- phic, even though we were not able to observe this in our sample. Our coding differs from that of Shaffer et al. (1997) for Heosemys and reeves ii. In their analysis, they used spinosa (in which the frontal contributes to the orbital rim), but we used grandis (in which it does not). In both specimens of reevesii available to us, the frontal clearly does not participate in the orbital margin. Given the contrary statement by Shaffer et al. (1997), reevesii may be polymorphic for this character. Polarity: The frontal participates in the orbital rim in pleurodires and spinifera , it is excluded in odoratus and caretta, and it is polymorphic in serpentina. No pat- tern is apparent within the ingroup. Given that the frontal clearly contributes to the orbital rim in Mongolemys, we consider its absence to be derived. (4) Contact between jugal and pterygoid; 0 = present, medial process of jugal well-developed and touching the pterygoid, Fig. 7; 1 = absent, medial process reduced, Fig. 8 (modified from Hirayama, 1985, 11, 12; McCord et al., 1995, 3; Burke et al, 1996, 23; Yasukawa et al, 2001, 4, 5). The jugal of most testudinoid turtles is expanded at its ventral end to form a medial process that contacts the pterygoid medially (McDowell, 1964). Presence or absence of the medial process, and presence or absence of a medial contact with the pterygoid were treated as two characters by Hirayama (1985) and Yasukawa et al. (2001). The scoring for the two characters appears to be redundant and we followed the recommendation of Gaffney and Meylan (1988) by combining them. We confirm the loss of a medial contact between the jugal and the pterygoid in galbinifrons, flavomarginata, and mouhotii (Hirayama, 1985; Yasukawa et al., 2001), but we found this condition to be polymorphic in speng- leri (also reported by McCord et al., 1995), and in triju- ga. Our observations are concordant with those of Burke et al. (1996). Polarity: A contact between the jugal and pterygoid in present in spinifera , odoratus, and serpentina , but is absent (although the two bones closely approach one another) in caretta. We conclude that the contact between the two bones is the primitive condition for tes- tudinoids and that their separation is derived, a conclu- sion also reached by Hirayama (1985). Our polarity determination is opposite that used by McCord et al. (1995), who mistakenly claimed to have derived their polarity assessment from Hirayama (1985). (5) Contact between jugal and palatine; 0 = absent, Fig. 9; 1 = present, Fig. 10 (Gaffney and Meylan, 1988, F5.4). The presence of a contact between the medial process of the jugal and the palatine was used previous- ly in support of a monophyletic Deirochelyinae (Gaffney and Meylan, 1988). We confirm the formerly observed distribution of this character within ‘emydids’ with the exception of reticularia, which does not exhib- it a contact. A contact is present in numerous ‘batagurids,’ such as borneoensis, reevesii, and hamil- tonii, but was absent in all examined members of ‘Testudinidae.’ Polarity: A contact between the jugal and the pala- tine is present in caretta, odoratus, and pleurodires, but is absent in spinifera and serpentina. A contact is absent in Mongolemys. We consequently consider its presence to be derived. (6) Contact of the epipterygoid with the jugal; 0 = clear- ly absent, Fig. 11; 1 = present, or almost present, epipterygoid forms a long lateral process that approach- es the jugal, Fig. 12 (Gaffney and Meylan, 1988, F8.1; Shaffer et al., 1997, 106). According to Gaffney and Meylan (1988) the epipterygoid and the medial process of the jugal approach one another or are in contact in reticularia and the various species they included in Pseudemys and Trachemys. They also noted a contact between these two elements in species they classified in Graptemys, but the condition in those taxa was interpreted to be a result of Vol. 10, p. 58 Asiatic Herpetological Research 2004 the medial expansion of the jugal and not a lateral expansion of the epipterygoid, and consequently was regarded as non-homologous (Gaffney and Meylan, 1988). We confirm the contact or near contact of these two elements in reticularia, decor ata, scripta, alaba- mensis, P. nelsoni, rubriventris, texana , flavimaculata, geographica , kohnii, nigrinoda , G. ocidifera, ouachiten- sis, and versa , and also report it in picta and terrapin. Contact was clearly absent in the specimens of harbour i, ernsti, and gibbonsi we examined. We made no assess- ments of homology, but accept any contact between these two elements as the derived state (as was done by Shaffer et al., 1997). Among ‘batagurids,’ we also found a close approach in reevesii. Polarity: A contact, or near contact, between the epipterygoid and the jugal is absent in all outgroups and the vast majority of the ingroup. We consider its pres- ence to be derived. (7) Contact of the inferior process of the parietal with the medial process of the jugal; 0 = absent, Fig. 13; 1 = present, Fig. 14 (Hirayama, 1985, 13). Our coding differs significantly from that of Hirayama (1985). In reevesii , N. platynota, and bealei we found no contact between the parietal and jugal, although these were the only three taxa in which Hirayama (1985: table 2) scored it to be present. However, we found a pronounced contact between these two elements in subtrijuga, a species scored by Hirayama (1985: table 2) as lacking such a contact, but shown on his tree (Hirayama, 1985: fig. 2) as a unique ‘batagurid’ feature convergent with some ‘emydids.’ Among ‘emydids,’ a well-developed contact occurs in barbouri and other broad-headed species currently clas- sified in Graptemys. Polarity: There is no contact between the inferior process of the parietal and the medial process of the jugal in all outgroups and the vast majority of the ingroup. A contact is considered to be the derived condi- tion. (8) Contact of the inferior process of the parietal with the maxilla; 0 = absent, Fig. 13; 1 = present, Fig. 14 ( Hirayama , 1985, 14). Our coding differs from that of Hirayama (1985). According to his character matrix (table 2) a contact should be present between the inferior process of the parietal and the maxilla in reevesii and mouhotii, but his cladogram (fig. 2) indicated that the presence of a con- tact should be regarded as a uniquely derived autapo- morphy of subtrijuga. We found no contact in reevesii or mouhotii. Of the testudinoid species we examined, sub- trijuga is the only one that shows this feature. Polarity: There is no contact between the inferior process of the parietal and the maxilla in all outgroups. Its presence is considered to be derived. (9) Extent of quadratojugal; 0 = quadratojugal well developed, firmly attached to jugal, Fig. 15; 1 = quadra- tojugal present, contact lost with jugal, Fig. 16; 2 = quadratojugal so heavily reduced that it appears to be absent in many skeletal specimens, Figs. 17, 18 (modi- fied from Hirayama, 1985, 16; Shaffer et al., 1997, 47; Burke et al., 1996, 21; McCord et al., 1995, 6; Yasukawa et al., 2001, 7, 8). Variation in the structure of the temporal region of turtles was discussed in detail by Zdansky (1924) and comments specific to testudinoids were provided by Zangerl (1948) and McDowell (1964). We originally scored the reduction of the quadratojugal as three differ- ent characters: loss of contact with the jugal, loss of con- tact with the squamosal, and the apparent loss of the quadratojugal. All five logically possible combinations were observed, but in most testudinoid turtles the tem- poral arch is so slender that the contact between the quadratojugal and squamosal is commonly reduced to a sliver that would have to be scored as ‘just barely pres- ent’ or ‘just barely absent.’ We therefore abandoned our efforts to evaluate the contact between the quadratojugal and squamosal. Our observations generally agree with those of McDowell (1964), Hirayama (1985), Burke et al., (1996), McCord et al. (1995), and Yasukawa et al. (2001). We purposefully avoid addressing the apparent lack of a quadratojugal in many species as an absence, because previous work by Zdansky (1924) showed that the quadratojugal of some ‘batagurids’ is so poorly ossi- fied and connected to the surrounding elements that it tends to be lost in skeletal specimens (Figs. 17, 18). An example of this problem can be found among the many conflicting statements made regarding the presence of this element in N. platynota (e.g., Smith, 1931; Bourret, 1941; McDowell, 1964; Ernst and Barbour, 1989). Polarity: The quadratojugal is present and firmly attached to the jugal in all outgroups, with the exception of chelids. Its reduction is considered to be derived. (10) Contribution of jugal to the rim of upper temporal emargination (Hirayama, 1985, 15); 0 = absent, Figs. 19, 20; 1 = present, Fig. 21. Participation of the jugal in the rim of the upper temporal emargination was reported previously in hamiltonii and ocellata (Hirayama, 1985). We confirm its presence in both species, but in one of the hamiltonii specimens we examined (MCZ 120333) the jugal forms a significant part of the rim only on one side of the skull; on the other side, which appears abnormal and likely represents a teratology, it does not. In all other species 2004 Asiatic Herpetological Research Vol. 10, p. 59 available to us, the jugal does not participate in the rim. In subtrijuga, the jugal is excluded from the upper tem- poral emargination by narrow extensions of the postor- bital and quadratojugal (Fig. 20). Polarity: The jugal participates in the upper tempo- ral rim ot spinifera , but it is excluded in odoratus, caret- ta , serpentina , Mongolemys , and most pleurodires. We consider the participation of the jugal in the rim of the upper temporal emargination to be the derived condition within testudinoids. (11) Contact between the quadratojugal and the articu- lar facet of the quadrate; 0 = absent, Fig. 22; l = pres- ent, quadratojugal sends a process ventrally along the rim of the cavum tympani and touches the lateral edge of the articular facet, Fig. 23 (modified from Hirayama, 1985, 17). The original character definition (Hirayama, 1985, character 1 7) is inappropriate, because the jugal does not contact the articular surface of the quadrate in any turtle except for madagascariensis and dumerilianus (Gaffney and Meylan, 1988). However, because Hirayama indi- cated in his tree that the only ‘batagurid’ taxon to exhib- it this character is subtrijuga , we assume that he was referring to a contact between the quadratojugal and the articular facet of the quadrate, a characteristic of subtri- juga only among testudinoids. A contact between the two elements was reported previously for reevesii in the character matrix published by Hirayama (1985), but we conclude that this must be a publishing error, because it stands in conflict with his tree. In the specimens of reevesii available to us, there is no contact. It is possible that the scoring for subtrijuga and reevesii were flipped, at least in part, in the Hirayama (1985) matrix (in which the taxa were listed next to one another). Polarity: A contact between the quadratojugal and the articular surface of the quadrate is absent in spinifera , but present in odoratus , caretta, and serpenti- na, and consequently could be considered primitive. However, based on ingroup commonality and the absence of a contact in Mongolemys , we consider a con- tact to be derived for Testudinoidea. (12) Contact between quadratojugal and maxilla; 0 = absent, Fig. 22; 1 = present, Figs. 23, 24 (Hirayama, 1985, 18). According to Hirayama (1985), among ‘batagurids’ a contact between the quadratojugal and maxilla is only present in subtrijuga and reevesii. We did not find a contact in our specimens of reevesii , but confirm its presence in subtrijuga. Gaffney and Meylan (1988) listed a contact between the quadratojugal and maxilla as a synapomor- phy for Platystemina {megacephalum + ‘ Chelydropsis'} and as an independently evolved synapomorphy for Kinostemidae, whereas Shaffer et al. (1997) noted a contact to be present in Kinostemidae, C. insculpta, and megacephalum. Our observations confirm the presence of a contact in all of these extant groups. Among testudi- noids, subtrijuga is unique in having an extensive con- tact in lateral view (Fig. 23). In several ‘emydids,’ a con- tact is present on the inside of the temporal arch ( bar - bouri, and nigrinoda; polymorphic in geographica , G. oculifera, and texana; Fig. 24). For now, we scored all taxa as present, regardless of whether the contact is vis- ible in lateral view, medial view, or both. In several other ‘emydid’ taxa, the bones closely approach one another, but do not actually meet, on the inside of the temporal arch ( alabamensis , ernsti, flavimaculata, gibbonsi, kohnii, P. nelsoni ). Polarity: A contact is present between the quadrato- jugal and maxilla in odoratus, but absent in spinifera, caretta, serpentina, and Mongolemys. We consider the presence of a contact to be derived for Testudinoids. (13) Medial contact of the maxillae along the anterior margin of the jaw; 0 = absent, Figs. 25, 26; 1 = present, Fig. 27 (modified from Hirayama, 1985, 20; McCord et al., 1995, 2; Yasukawa et al., 2001, 10). In most testudinoids, the anteromedial ends of the maxillae are separated medially by the premaxillae along the anterior margin of the jaw (Fig. 25). Hirayama (1985) noted that the maxillae have a medial contact in some ‘batagurids,’ which was confirmed by McCord et al. (1995) and Yasukawa et al. (2001) for spengleri and several other species that they included in the genus Geoemyda. We found a broad medial contact of the max- illae in spengleri and annulata (Fig. 27). In some species, the maxillae approach one another along the ventral rim of the nasal opening (e.g., amboinensis, mouhotii, pulcherrima, crassicollis ), but a well-devel- oped contact is never present (Fig. 26). Polarity: The maxillae do not meet medially along the anterior margin of the jaw in odoratus, caretta , ser- pentina, and Mongolemys. A medial contact is present in spinifera but only along the ventral border of the exter- nal nares. We consider a medial contact along the ante- rior margin of the jaw to be the derived condition with- in Testudinoidea. (14) Size of the foramen orbito-nasale; 0 = small, less than 1/6 of orbit length, Figs. 28, 29; 1 = large, more than 1/6 of orbit length, Fig. 30 (modified from Hirayama, 1985, 33; Gaffney and Meylan, 1988, F9.3, FI 0.2, G10.3, HI 1.1, HI 6.3; Crumly, 1982, 25; Crumly, 1994, 12). We were cautious when first approaching this char- acter due to the inconsistent usage and definition of Vol. 10, p. 60 Asiatic Herpetological Research 2004 small and ‘large’ by various authors. However, after assessing the size ot this foramen based on its size rela- tive to the length of the orbit, we were surprised to see that we were able to reproduce Hirayama’s (1985) scor- ing for the ‘batagurids’ without too many difficulties. In contrast, our initial observations of tortoises were in stark contrast to those of Crumly (1982, 1985, 1994) and Gaffney and Meylan (1988). This may be due to the thin nature of the palatine of many tortoises, and the relative ease with which that part of the palate can be damaged during skeletal preparation and handling. Furthermore, the foramen becomes progressively more closed with increased ontogenetic age (Crumly, 1982). We encoun- tered similar problems in attempting to score reticularia and blandingii. Because we deem this character to be potentially useful for helping to resolve phylogeny with- in ‘batagurids’ and ‘emydids,’ we decided to score all testudinoids with delicate palatines (i.e., all tortoises, reticularia , and blandingii ) as ‘unknown.’ We acknowl- edge that our redefinition of the character is still subjec- tive and somewhat problematic, but using this definition we were able to unambiguously score all the ingroup taxa we examined. Polarity: The foramen orbito-nasale is large in ser- pentina, odoratus, and spinifera, but small in caretta. We consider presence of a large foramen orbito-nasale to be the derived condition within testudinoids, because the foramen is small in Mongolemys. (15) Contact between maxilla and vomer; 0 = present, Fig. 31; 1 = absent, vomer separated from the maxilla by the premaxilla, Fig. 32 (Hirayama, 1985, 31; Crumly, 1982, 21; Yasukawa et al., 2001, 14). We generally agree with previous scorings for this character (Hirayama, 1985, Crumly, 1982, Yasukawa et al., 2001). We confirm the absence of a contact in amboinensis and pulcherrima, but our scoring differs slightly for those taxa that Hirayama (1985) coded as ‘intermediate apomorphic,’ a character state that we interpret as polymorphism. Of those taxa that Hirayama (1985) and Yasukawa et al. (2001) scored as intermedi- ate (flavomarginata, caspica, annulata ), our sample size is too small to confirm whether both character states are present. We consequently follow these authors by scor- ing those taxa as polymorphic. Polarity: A contact between the maxilla and vomer is present in all outgroups. The loss of this contact is derived for testudinoids. (16) Size of the foramen palatinum poster ius; 0 = large, Fig. 33; 1 = small, Fig. 34 (modified from Hirayama, 1985, 22; Gaffney and Meylan, 1988, F2.2, F6.1; McCord et al., 1995, 4; Yasukawa et al., 2001, 12). Our characters 16 and 17 were published originally by Hirayama (1985) as one character that combined two morphological features: the size of the foramen palat- inum posterius (f.p.p.) and participation of the pterygoid in the margin of the f.p.p. Although four possible com- binations of these features are logically possible, only two were originally included (participation present, f.p.p. large; participation absent, f.p.p. small). Gaffney and Meylan (1988) also used this character within ‘emy- dids,’ but their character applied only to the exclusion of the pterygoid from the f.p.p. We decided to subdivide Hirayama’s (1985) character into one character that describes the size of the f.p.p. and a second that address- es the position of the pterygoid relative to the f.p.p. We found no difficulty in identifying the f.p.p. as ‘large’ or ‘small,’ (Figs. 33 and 34, respectively), and no ambiguous condition was encountered. Because our character definition only includes two character states, our scorings do not reflect those of other workers with a more limited target group (e.g., McCord et al., 1995). In juveniles the f.p.p. tends to be larger, but during later ontogenetic stages the f.p.p. is slowly reduced in size. Polarity: The f.p.p. is small in odoratus, spinifera, and most pleurodires; it is absent in caretta, but is large in serpentina and Mongolemys. We consider a small f.p.p. to be the derived state. (17) Position of the pterygoid relative to foramen palat- inum posterius (f.p.p.); 0 = pterygoid situated posterior to the f.p.p., Fig. 33; 1 = pterygoid situated posterior to the f.p.p., but sends a process anterior and lateral to the f.p.p., Fig. 34. Our survey of testudinoids indicated that reliable assessment of participation of the pterygoid in the f.p.p. may be difficult because many species show an ontoge- netic change in configuration of this part of the palate. In juveniles, the f.p.p. typically includes the pterygoid in its posterior margin. During later ontogenetic stages the pterygoid is excluded. In spite of this, it appears that the relative position of the pterygoid tends to stay constant during ontogeny. Polarity: The anterior end of the pterygoid is situat- ed posterior to the f.p.p. in odoratus, serpentina, most pleurodires, and Mongolemys, but is situated lateral to the f.p.p. in spinifera. We consider a posterior position to be primitive for Testudinoidea. (18) Epipterygoid participation in the trigeminal fora- men; 0 = absent, Fig. 35; 1 = present, epipterygoid clearly separates the parietal and pterygoid in lateral view, Fig. 36. The anteroventral rim of the trigeminal foramen of most testudinoid turtles is formed by the parietal and pterygoid. The epipterygoid commonly comes close to the foramen, but does not form part of it. In spengleri 2004 Asiatic Herpetological Research Vol. 10, p. 61 and mouhotii , the epipterygoid consistently participates in the margin of the trigeminal foramen thus separating the parietal and pterygoid, at least in lateral view. Polarity: The epipterygoid forms part of the anteroventral rim of the trigeminal foramen in the majority of outgroups, with the exception of pleurodires that lack a definitive ossified epipterygoid (Gaffney, 1979). The condition is unclear for Mongolemys. However, within the ingroup we found this character only in the seemingly rather specialized turtles spengleri and mouhotii. We consequently consider its presence to be secondarily derived. (19) Vomerine foramen ; 0 = absent, Fig. 37; 1 = pres- ent, Fig. 38 (Gaffney and Meylan, 1988, H4.1; Crumly, 1994, 15). The vomerine foramen (= anteromedial vomerine aperture of Crumly, 1982 and 1994 [in part]) is a small opening that pierces the vomer along the midline just posterior to the foramen praepalatinum (Bramble, 1971). The presence of a vomerine foramen was noted in agas- sizii and berlandieri by Bramble (1971), and was used by Gaffney and Meylan (1988) to unite various species currently placed in Gopherus as a clade. Its irregular presence in elegans, elongata, chilensis, and radiata was reported by Crumly (1982). Specimens in our sam- ple enable us to confirm its presence in agassizii, berlandieri , and chilensis. Polarity: The vomerine foramen occurs in only a few ‘testudinids’ and it is absent in all outgroups. Its presence is considered derived for Testudinoidea. (20) Development of the foramen praepalatinum as a canal (canalis praepalatinum) that is concealed by a bony secondary palate in ventral view; 0 = absent, Fig. 39; 1 = present, Fig. 40 (modified from Hirayama, 1985, 24). In most testudinoids, the foramen praepalatinum is a small opening that connects the nasal cavity with the roof of the oral cavity (Fig. 39). However, in a number of taxa with extensively developed secondary palates, the anterior nasal artery passes through an elongated canal that is concealed in ventral view by the bony sec- ondary palate (e.g., baska , tentoria , petersi: ; Fig. 40). We refer to this structure as the canalis praepalatinum. Our scorings are fully consistent with those of Hirayama (1985). Polarity: The foramen praepalatinum is absent in spinifera (Gaffney, 1979) and caretta (Nick, 1912), and is developed as a true foramen that is exposed in ventral view in odoratus, serpentina , and Mongolemys. The development of a canalis praepalatinum is considered the derived condition within Testudinoidea. (21) Contact between pterygoid and basioccipital; 0 = present, Fig. 41; 1 = absent, Fig. 42 (modified from Gaffney and Meylan, 1988, FI. I, F10.3, H18.3; Crumly, 1994; Shaffer et al., 1997, 103). Two of the most often-cited characters that purport- edly help to distinguish the ‘Emydidae’ from the ‘Bataguridae’ are the batagurine process and the contact between the pterygoid and the basioccipital (McDowell, 1964). Both traits are commonly combined as one char- acter (e.g., Gaffney and Meylan, 1988) and even seem to have been confused with one another (e.g., Mlynarski, 1976; Shaffer et al., 1997). The batagurine process is a poorly-defined feature that, in McDowell’s original usage (1964) appears to consist of a lateral process of the basioccipital that floors the recessus scalae tympani. Many testudinoid species (including non-‘batagurines’) have a laterally-projecting process of the basioccipital; it may or may not floor the recessus scalae tympani, but it is often obscured from view in articulated specimens. When disarticulated material is examined, a broader dis- tribution of this feature (assuming it is interpreted as we have done above) across testudinoids is revealed. Within Testudinoidea, the pterygoid commonly sends a process posteriorly and contacts the basioccipi- tal just lateral to the basisphenoid. This character appears to be absent in most ‘emydids,’ but is present in terrapin and those species that are currently attributed to Graptemys (Gaffney and Meylan, 1988). We noticed a strong ontogenetic component to this character within ‘emydids.’ The pterygoid is commonly rather short dur- ing younger ontogenetic stages, but finally reaches the basioccipital in later stages. For instance, among our specimens of terrapin, orbicularis , and texana, the pterygoid does not contact the basioccipital in younger individuals, but a clear contact is present in adults. We score such species as polymorphic, but note that ontoge- netically influenced polymorphisms are not well explored in turtles. Among ‘batagurids,’ we noted a similar pattern. Contrary to general belief we were not able to observe a contact between the pterygoid and the basioccipital in all taxa traditionally classified in this group (e.g., trijuga , pulcherrima, sinensis). As with the ‘emydids,’ there seems to be an ontogenetic effect in which younger specimens tend not to have a contact. Where recognized, we scored these species as polymorphic. We found much conflicting data regarding the dis- tribution of this character among tortoises (Crumly, 1982, 1985, 1994; Gaffney and Meylan, 1988). Among the specimens we examined, we note the complete absence of a contact only in graeca\ polymorphisms were observed in polyphemus and horsfieldi. Again, an ontogenetic component is apparent. Polarity; A contact between the pterygoid and Vol. 10, p. 62 Asiatic Herpetological Research 2004 basioccipital is present in all outgroups. Its absence is considered to be derived. (22) Contact of the pterygoid with the articular facet of the quadrate ; 0 = absent, Fig. 43; 1 = present, Fig. 44 (Hirayama, 1985, 38). According to the data matrix published by Hirayama (1985) he only observed this contact in reeves ii, however, in his tree (fig. 2) the contact is mapped as an autapomorphy of subtrijuga. We confirm the presence of a contact between the posterior process of the pterygoid and the articular surface of the quadrate in subtrijuga. It is the only taxon we examined that dis- plays the derived condition. Polarity: A contact between the pterygoid and the articular surface of the quadrate is absent in all out- groups and the vast majority of the ingroup. We consid- er its presence to be derived. (23) Closure and depth of the incisura columella auris; 0 = absent, incisura is open, Fig. 44; 1 = present, incisura closed, Fig. 45 (Crumly, 1985; Gaffney and Meylan, 1988, HI. 3). The incisura columella auris is a notch that is formed by the quadrate and that holds the stapes and eustachian tube (Gaffney, 1972). In a number of turtles, the incisura closes to fully surround the stapedial shaft (Gaffney and Meylan, 1988). Within testudinoids, the incisura evidently is closed in most tortoises (Crumly, 1985). We are able to confirm the presence of such a closed incisura in all ‘testudinids’ we examined with the exception of one specimen of kleinmanni (CAS 228431), the smallest of the species now classified in Testudo. In a number of ‘batagurids’ and ‘emydids’ the incisura commonly is very narrow and even appears to be closed, however, a closer look under the microscope combined with a probing needle reveals that this appar- ent closure is produced by dry tissues that remain in this area in many articulated skulls. The only ‘batagurid’ for which we sometimes found a closed incisura is N. platynota; in that species the quadrate does not fuse together forming a solid ring behind the incisura, but this is also the case for many tortoises (e.g., some belliana, emys, some homeana, some kleinmanni). In some cases, the polymorphism we noted (e.g., in belliana and home- ana) appears to be a result of ontogenetic age, with older individuals displaying a greater degree of fusion at the posterior part of the incisura. Polarity: The polarity of this character is somewhat ambiguous, because the incisura columella auris is closed in serpentina and spinifera , but open in caretta and odoratus. We conclude that its presence is derived within testudinoids because it is absent in Mongolemys. Mandible (24) Angular contribution to the sulcus cartilaginis Meckelii; 0 = present, the angular contributes to the sul- cus and is as long or longer than the prearticular, Fig. 47; 1 = absent, the angular is shorter than the preartic- ular, Fig. 48 (modified from Gaffney and Meylan, 1988, FI. 4). A broad contact of the angular with Meckel’s carti- lage was used by McDowell (1964) to characterize the ‘Emydinae’ and later used by Gaffney and Meylan (1988) as a synapomorphy to unite the same grouping. As it was originally worded, this character is difficult to observe in its literal sense for most museum specimens, because the Meckel’s cartilage usually is not present in modern and fossil skeletal specimens. We suggest that the spirit of McDowell’s (1964) character can be evalu- ated by examining the participation of the angular in the sulcus cartilaginis Meckelii. We confirm that a small to broad angular contribution is present in all ‘emydids’ with the exception of rubriventris. In most ‘batagurids,’ the angular is a short bone that does not participate in the sulcus and barely spans half the distance the prearticular does. However, a small but clear contribution to the sul- cus is present in an eclectic group comprised of baska , dentata, thurjii , punctularia, and some grandis. We were not able to carefully evaluate potential polymorphisms in these taxa. Polarity: An angular contribution to the sulcus car- tilaginis Meckelii is present in all cryptodiran outgroup taxa. We consider is absence to be derived. (25) Contact between surangular and dentary; 0 =sim- ple contact, Fig. 49; 1 = strongly interdigitated suture, Fig. 50 (Crumly, 1982, 12; Crumly, 1985; Gaffney and Meylan, 1988, H6.1). In most testudinoids, the surangular and dentary meet along the lateral side of the mandible in a simple, overlapping contact. According to Crumly (1982, 1985), this contact is stabilized through a finger-like process of the surangular that interdigitates with the dentary in all tortoises except emys , impressa, and those species he classified in Gopherus. We confirm the absence of this character in agassizii, berlandieri, emys, impressa, and polyphemus, but also did not observe it in areolatus. Polarity: An interdigitated contact between the surangular and dentary is absent in all outgroups and the majority of the ingroup. We consider its presence to be derived. (26) Height of the processus coronoideus; 0 = as high as dentary, Fig. 51; 1 = rising significantly above the den- tary, Fig. 52 (modified from Hirayama, 1985, 45). The coronoid of most turtles is a small bone that produces a minor knobby projection that rises only little 2004 Asiatic Herpetological Research Vol. 10, p. 63 above the adjacent dentary, if at all. According to McDowell (1964) and Hirayama (1985) the coronoid is larger and rises moderately above the dentary in borneensis and crassicollis, and in reeves ii and subtriju- ga the coronoid is very large and produces a robust process that sits high above the dentary. We confirm these observations, however, we were also able to observe moderately developed coronoid processes in kachuga and tentoria. Among ‘emydids,’ we also observed moderately developed coronoids in barbouri, ernsti, jlavimaculata, geographica , gibbonsi, kohnii , and terrapin. Unlike Hirayama (1985), we only utilize one derived character state, because it is difficult to objectively measure and discretize the relative height of the coronoid among turtles. Polarity: The coronoids of caretta, serpentina , and odoratus are small and do not rise above the dentary, but the coronoid of spinifera is well developed and forms a moderate process. The lower jaw is not described for Mongolemys. We consider well-developed coronoids to be derived within testudinoids. (27) Foramen dentofaciale majus; 0 = small, Fig. 53; 1 = large and situated within a large lateral fossa, Fig. 54 (Hirayama, 1985, 47). The foramen dentofaciale majus of most testudi- noids is a small opening that is situated on the lateral side of the mandible, just ventral and slightly anterior to the coronoid. The foramen dentofaciale majus is greatly enlarged in thurjii and ocellata and is situated at the anterior end of an expanded lateral fossa (Hirayama. 1985). We confirm the enlargement in those taxa, and further note its presence in petersi. Polarity: The foramen dentofaciale majus is small in all outgroups and the vast majority of the ingroup. Its presence is considered to be derived. Triturating Surfaces (28) Participation of palatine in the triturating surface of the upper jaw; 0 = absent, Fig. 55; 1 = present, Figs. 56, 57 (Hirayama, 1985, 26; Gaffney and Meylan, 1988, F2. 1). In some testudinoids, the palatine has a ventrolater- al maxillary process that participates in the triturating surface of the upper jaw. The degree of participation varies among taxa, and within some species. A clear and extensive participation is present in barbouri , ernsti , geographica , gibbonsi , petersi , rubriventris, script a, terrapin, texana , and versa. It is weakly developed in hamiltonii, ocellata, G. oculifera, ouachitensis, reevesii, subtrijuga, and some individuals of baska. Participation was used by Gaffney and Meylan (1988) to unite Terrapene spp., blandingii , guttata, G. insculpta, marmorata, muhlenbergii, and orbicularis as a clade within the ‘Emydidae.’ In our observations, how- ever, this participation also is absent among other ‘ emy- dids ’ such as jlavimaculata, kohnii, nigrinoda, picta, and reticularia (we were not able to evaluate adequately the potential for polymorphism in these taxa). In addition, it appears that the absence of a palatine participation rep- resents the plesiomorphic state for testudinoids, thus eliminating its value for diagnosing monophyletic groups. Polarity: The palatine does not participate in the trit- urating surface of spinifera, but does so in odoratus, caretta, and serpentina. The palatine does not participate in the triturating surface of Mongolemys and only occurs in Testudinoids with highly derived secondary palates. We consequently consider the participation of the pala- tine in the triturating surface to be derived within tes- tudinoids. (29) Participation of the vomer in the triturating surface of the upper jaw; 0 = absent, Figs. 56, 57, 58; 1 - pres- ent, Fig. 59 (Hiray>ama, 1985, 25). The triturating surface is the grinding surface of the jaw. In most turtles, it is formed on the upper jaw pre- dominantly by the maxilla and premaxilla. However, in turtles with extensive secondary palates the vomer may also participate. In texana, the vomer may have a ventral projection that barely separates the maxillae in the mid- line, but it does not participate in the triturating surface proper because it sits in a dorsal concavity within the palate (Fig. 56). Our scorings differ from those of Hirayama (1985) for mouhotii (which Hirayama scored as “intermediate apomorphic” and we score as absent because it does not have a secondary palate) and subtri- juga (in our specimens the vomer does not descend to the palatal surface, but this species may be polymor- phic). This character is polymorphic in barbouri. Polarity: Because the vomer does not participate in the triturating surface of odoratus, spinifera, caretta, serpentina, and Mongolemys, its participation is consid- ered to be the derived condition for Testudinoidea. (30) Presence and number of lingual ridges of the tritu- rating surfaces of the upper and lower jaws; 0 = no lin- gual ridges present, Fig. 60; 1 = one lingual ridge pres- ent, Figs. 61-62; 2 = two lingual ridges present (modi- fied from Hirayama, 1985, 29, 44; Gaffney and Mey4 an, 1988, F7.2, F9.1) Most turtles lack lingual ridges on their triturating surfaces (Fig. 60), but one or two such ridges are devel- oped in a number of testudinoids (Figs. 61-62). These ridges run parallel to the labial surface of the maxilla and dentary, and typically do not meet their counterpart on the midline. They are not necessarily a continuous structure (Fig. 61), and may be divided into several com- Vol. 10, p. 64 Asiatic Herpetological Research 2004 ponents. In some cases, an extensive ridge-like structure can create the appearance of an additional ridge at the extreme lingual margin of the maxilla bordering the internal nares; however, we consider these to be the thickened rim ot the internal nares rather than an addi- tional ridge. Among ‘batagurids’ and ‘emydids,’ one lin- gual ridge is present in alabamensis , borneoensis , deco- rata, kachuga , P. nelsoni, ocellata, petersi, rnbriventris, scripta, sinensis, tentoria, texana, and thnrjii. Two lin- gual ridges are developed in baska. We found lingual ridges in all tortoises we examined except erosa, bel- liana , and homeana. Hirayama (1985) originally scored this character as two separate characters, one for the mandible and one for the maxilla. In our observations, the triturating sur- face of the lower jaw closely mimics that of the upper jaw, creating an occlusal surface that closely reproduces the function of cusps in mammalian cheek teeth. Both characters were scored in parallel in Hirayama’s matrix, and we see no reason to consider them independent. Polarity: All outgroups lack lingual ridges on the triturating surfaces. We consequently consider their presence to be derived. (31) Well- dev el oped serrations on labial or lingual ridges of the triturating surfaces of the upper and lower jaws; 0 = absent, Fig. 60; 1 = present, Fig. 61 (modified from Hirayama, 1985, 21, 27, 41, 43, 46; Gaffney and Meylan, 1988, F9.2; Yasukawa et al., 2001, 11). Well-developed serrations on the lingual and labial ridges of the upper and lower jaws are developed in sev- eral ‘batagurids’ and ‘emydids’. A number of tortoises and other ‘batagurids’ (e.g., carbonaria, pardalis, sulca- ta, annulata, and areolata) exhibit serrations on their ramphothecae, but unlike the bony, tooth-like serrations of borneoensis, thurjii, petersi, or texana, these crenula- tions are weakly developed, leaving very little or no trace of serrations on the underlying bone. In compari- son to those taxa with well-developed serrations, it very difficult to establish a consistent scoring system for taxa with fine crenulations, because many specimens will not exhibit any serrations, probably due to wear of the ram- phothecae. Unlike Hirayama (1985) and Yasukawa et al. (2001) we scored all taxa with such weak serrations as absent. Unfortunately, even in those taxa with well-devel- oped serrations, the serrations are not always evenly developed on all ridges. We consequently combined all of Hirayama’s (1985) characters relating to serrations into one character. Because serrations commonly occur on all available ridges, this treatment will also help to avoid unconsciously weighting the presence of serra- tions with up to five characters. In those taxa that have them, the ridges themselves often have very different morphologies; this character needs to be critically reevaluated with adequate sample sizes for the relevant taxa. Polarity: All of our outgroup taxa and the majority of the ingroup taxa lack strong serrations. We interpret their presence to be derived. (32) Median ridge or sulcus of the triturating surface of the upper jaw; 0 = both structures absent, Fig. 60; l = median ridge present, Fig. 62; 2 = median sulcus pres- ent, Fig. 63 (modified from Hirayama, 1985, 30; Crumly, 1985, 1994, 4; Gaffney and Meylan, 1988, H3.1). In a number of testudinoid turtles with partially developed secondary palates and lingual ridges, addi- tional structures are formed along the midline of the upper jaw that typically correspond to reciprocal struc- tures of the lower jaw. The upper jaw of petersi is char- acterized by a narrow sulcus (Fig. 63) and the mandible exhibits a low median ridge. On the other hand, in baska, borneoensis, thurjii, kachuga, agassizii, berlandieri, and polyphemus, a ridge runs along the mid- line (Fig. 62), which typically corresponds to a sulcus in the lower jaw. An incipient ridge also was reported in emys (Crumly, 1994), but we were not able to confirm this on the specimen available to us. Polarity: A median ridge is absent in all outgroups and the vast majority of the ingroup. Its presence is con- sidered to be derived. (33) Posterior extension of the lower triturating surface behind the symphysis of the dentary; 0 = absent, Fig. 64; 1 = present, Fig. 65 (Hirayama, 1985, 42; Gaffney and Meylan, 1988, G5.2). In several ‘batagurids,’ the triturating surface of the dentary forms a shelf along the midline that extends so far posteriorly that the symphysis cannot be seen when the mandible is observed in dorsal view (McDowell, 1964). Our scorings fully agree with those of Hirayama (1985) for the ‘Bataguridae,’ but we disagree with Gaffney and Meylan (1988) who asserted that this char- acter also occurs in some ‘Emydidae.’ Admittedly, sev- eral species currently placed in the genera Graptemys, Pseudemys, and Trachemys have greatly expanded tritu- rating surfaces of the dentary, but in all of the specimens available to us, the symphysis is always visible in dorsal view. Polarity: An extended triturating surface of the den- tary does not occur in any outgroup taxon. We consider its presence to be derived. Carapace (34) Carapace strongly tricarinate in adult; 0 = absent, Figs. 66, 67; 1 = present, Fig. 68 (modified from 2004 Asiatic Herpetological Research Vol. 10, p. 65 Hirayama, 1985, F; McCord et al., 1995, 10; Yasukawa et al., 2001, 24). Three distinct carapacial ridges are present in the adults of reevesii, hamiltonii, spengleri , subtrijuga, tri- juga, and mouhotii. We cannot replicate Hirayama’s (1985) placement of this character as an autapomorphy in hamiltonii. In our observations, the carinae in hamil- tonii are not better developed than in some other taxa. In fact, they are more weakly developed than those in mouhotii (Fig. 68). Because keels are present in the young and subadults of such taxa as crassicollis (Fig. 67), mutica, and sinensis, but disappear with age, and because we were not able to observe the juveniles of most species, we restricted this character to those species that exhibit well-developed tricarinae as adults. Three keels were reported to be present in the adults of dentata (McCord et al., 1995), but we cannot confirm this (tricarinae are not present on our younger speci- mens). Polarity: Tricarinae are absent in our outgroup species ( caretta , odoratus, serpentina , spinifera, gibba, siebenrocki, subglobosa, and subrufa), but do appear occasionally in some of their close relatives, such as scorpioides, temminckii, and fimbriatus. We consider the presence of tricarinae to be derived within Testudinoidea. (35) Significant serration of the posterior peripherals; 0 = absent, Fig. 66; 1 = present, Fig. 68 (modified from Hirayama, 1985, D; McCord et al., 1995, 11; Yasukawa et al., 2001, 23). We generally agree with previous observations reported for this character (Hirayama, 1985). However, because the carapace rim is at least slightly serrated in almost all turtles, we rephrase the character definition to include only significantly serrated posterior peripherals as found, for example, in crassicollis, dentata, grandis, mouhotii, N. platynota, and spengleri. Among the ‘emy- dids’ and ‘testudinids,’ the peripherals of barbouri, erosa, fiavimaculata, geographica, homeana, kohnii, nigrinoda, oculifer, G. oculifera, pseudogeographica, and versa also are serrated. It is important to note that our scores are based on the peripheral bones; the amount of carapacial serration greatly depends on the presence or absence of the marginal scutes in the specimens used, because the scutes greatly accentuate the amount of ser- ration, if present. We find no conflict with the codings of McCord et al. (1995) and Yasukawa et al. (2001). Polarity: Serrated posterior peripherals are present in caretta, but absent in odoratus, serpentina , and most pleurodires. However, due to ingroup commonality and its absence in taxa placed within “Lindholmemydidae” we conclude that its presence is derived. (36) Carapace of adult tectiform in cross-section with a strong posterior projection on the third vertebral scute; 0 = absent, Fig. 69; 1 = present, Fig. 70 (Hirayama, 1985, N). According to Hirayama (1985), this character only occurs in tecta and tentoria. For our sample, we were able to confirm its presence in tecta and tentoria and also observed it in barbouri ( barbouri and other species now classified in Graptemys may be sexually dimorphic for this character). The descriptive term ‘tectiform’ is somewhat problematic, because any turtle shell can be considered ‘roofed’ (Fig. 69). We regard a carapace as tectiform if its sides are more-or-less flat and meet along the midline at a rather sharp angle (Fig. 70). Many tes- tudinoids, and notably those ‘emydids’ currently classi- fied within Graptemys, have a somewhat tectiform cara- pace as juveniles, but that morphology typically is lost in the adults. Polarity: Because all outgroups and the majority of the ingroup do not have a tectiform carapace, we consid- er its presence to be derived. (37) Shape and orientation of the second neural; 0 = second neural hexagonal, short sides positioned anteri- orly, Fig. 71; 1 = second neural hexagonal, short sides positioned posteriorly, Fig. 72; 2 = second neural octagonal, Fig. 73 (modified from Hirayama, 1985, G; Yasukawa et al., 2001, 25). (38) Shape and orientation of the third neural; 0 = third neural hexagonal, short sides positioned anteriorly, Fig. 71; 1 = third neural hexagonal, short sides positioned posteriorly, Fig. 72; 2 = third neural square, Fig. 74; 3 = third neural octagonal, Fig. 75 (modified from Hirayama, 1985, G; Yasukawa et al, 2001, 25). Originally, Hirayama (1985) only discussed the ori- entation of the neurals in general, which is fully suffi- cient for his ‘batagurid’ ingroup, because almost all indi- viduals exhibit his two suggested character states. However, in most tortoises the second and/or third neu- rals are not hexagonal, but rather are square or octago- nal, making it impossible to assign them to one of Hirayama’s (1985) character states. We consequently split Hirayama’s original character into two characters, restricted their application to the second and third neu- ral, and added additional character states. Our observations generally agree with those of Hirayama (1985) and Yasukawa et al. (2001) for ‘batagurids,’ with the exception of annandalei in which we found the short side of the second and third neurals to be positioned anteriorly, and not posteriorly as was indicated by Hirayama (1985). Polarity: The short side of the second and third neu- ral bones faces posteriorly in odoratus and spinifera but Vol. 10, p. 66 Asiatic Herpetological Research 2004 anteriorly in caretta. The shape of the second and third neurals is extremely variable in serpentina. However, in Lindholmemys and Mongolemys the short side of the second and third neurals is positioned anteriorly. We consider that condition to be primitive for Testudinoidea. (39) Medial contact of the seventh and/or eighth costal bones; 0 = absent, Fig. 76; 1 = present, Fig. 77 (Hirayama, 1985, V; Yasukawa et al., 2001, 26). In some testudinoid turtles, the posterior costal bones meet along the midline due to the reduction of the posterior neural elements. The original character defini- tion provided by Hirayama (1985) and Yasukawa et al. (2001) was worded to indicate a contact between the seventh and eighth costal bones among some ’batagurids.’ We are unable to reproduce their results if the character definition is taken literally. For example, in all our specimens of amboinensis and galbinifrons, the eighth costals meet on the midline, but the seventh costals do not. However, if the character definition is modified to include any contact of the seventh or eighth costals, our results are concordant with those of Hirayama (1985) and Yasukawa et al. (2001). In addition to the Asian box turtles, we report a medial contact of the posterior costals in baska, Carolina, coahuila, T. nel- soni, T. ornata and rnbida. Polarity: A medial contact of the seventh and/or eighth costals is absent in serpentina, but present in spinifera, caretta, many kinostemids, and many pleu- rodires. Although a composite reconstruction of a puta- tive “lindholmemydid” from the Early Cretaceous of Japan was illustrated with the seventh costals in contact at the midline (Hirayama et al., 2000, fig. 11), such a contact is absent in other specimens of Lindholmemys and Mongolemys. Its presence within testudinoids is pre- dominantly in the highly derived box turtles, and we conclude that its presence is derived for Testudinoidea. (40) Cervical scute; 0 = present, Fig. 78; 1 = absent, Fig. 79 (modified from Crumly, 1985, 1994, 34; Gaffney and Meylan, 1988, H5.2, FI 10.1; Shaffer et al., 1997, 41). The presence and shape of the cervical scute is used commonly to determine phylogenetic relationships within tortoises. According to Crumly (1985) the cervi- cal scute is very narrow or absent in all tortoises except agassizii, berlandieri, emys, flavomarginatus, impressa, and polyphemus. We generally agree with these observa- tions, but when this character is applied to all testudi- noids intraspecific variation is so great that the character becomes essentially useless. We consequently limit our scoring to the mere presence or absence of the cervical scute. We confirm the observations of Gaffney and Meylan (1988) that this scute is absent in carbonaria, chilensis, elegans, nigra, par dalis, and sulcata and addi- tionally code homeana and erosa as polymorphic. Polarity: The cervical scute is present in all cryp- todiran outgroups that have scutes on their carapace. We consider its absence to be derived. (41) Number of vertebral scutes; 0 = five, Fig. 80; 1 = six or more, Fig. 81 (Hirayama, 1985, P). We confirm Hirayama’s (1985) observation that there are at least six vertebral scutes in N. platynota. Additional scutes occasionally occur in other species, but are best considered abnormalities; they typically lack the symmetrical associations with adjacent pleural scutes seen in N. platynota. Polarity: All testudinoids, except for N. platynota, have five vertebral scutes. We consider the presence of six scutes to be derived. (42) Position of the anterior sulcus of the fourth verte- bral scute; 0 = sulcus lies on the fifth neural, Fig. 82; 1 = sulcus lies on fourth neural, or on the suture between the fourth and fifth neural, Fig. 83; 2 = sulcus lies on the sixth neural, or on the suture between the fifth and sixth neural, Fig. 84 (modified from Hirayama, 1985, L+M). (43) Position of the posterior sulcus of the fourth verte- bral scute; 0 = sulcus lies on the eighth neural, or on the homologue of the eighth neural, if the seventh is reduced (e.g., in most tortoises), Fig. 85; 1 = sulcus lies on the seventh neural, or on the suture between the seventh and eight neural, Fig. 86; 2= eighth neural absent, sulcus overlies costals that meet at the midline, Fig. 87 (modi- fied from Hirayama, 1985, L+M). The size of the fourth vertebral scute was addressed with two characters by Hirayama (1985) but the total range of morphological variability in testudinoids is not encompassed by his character definitions. In most tes- tudinoids, the fourth vertebral scute covers the posterior half of the fifth neural bone, the sixth and seventh neu- rals, and the anterior half of the eighth neural. A number of variations are known, and simply counting the num- ber of neural bones covered by this scute results in prob- lems by creating a false perception of homology. For instance, a fourth scute that partially overlies the fourth and seventh neurals and fully covers the fifth and sixth, can strictly be said also to cover four neurals, but the ele- ments involved are only partially homologous with the common condition. We attempt to resolve these issues by establishing two new characters that preserve what we think was Hirayama’s (1985) original intent, but that permit a more accurate representation of the association between the fourth vertebral scute and the underlying neural bones. 2004 Asiatic Herpetological Research Vol. 10, p. 67 Some problems that are associated with scoring this character include the prevalence of scute abnormalities among testudinoids (e.g., Coker, 1905, 1910; Newman, 1906; Zangerl and Johnson, 1957). Specimens exhibit- ing such abnormalities were scored as unknown. The notable exception to this is N. platynota, in which a sixth, or even a seventh, vertebral scute is always pres- ent. Polarity: Determining the polarity through outgroup relationship is somewhat difficult, because almost every outgroup exhibits a different condition, especially for the posterior sulcus. Based on ingroup commonality, and the presence of our zero state in both Lindholmemys and Mongolemys, we consider the sulci of the fourth verte- bral scute to be primitively situated on the fifth and eighth neural bones. (44) Posterior margin of first vertebral scute significant- ly narrower than its anterior margin; 0 = absent, Fig. 88; 1 = present, Fig. 89 (modified from Hir ay ama, 1985, C). When originally proposed, this character was applied to a posterior constriction of all vertebral scutes (Hirayama, 1985). If strictly applied, this character is absent in all taxa, because the fifth vertebral scute never is constricted along its posterior edge relative to the anterior edge. If each scute is viewed by itself, it becomes apparent that especially the fourth vertebral scute tends to be narrowed posteriorly, as can be observed in all species now classified in the genera Graptemys, Heosemys, Trachemys, and Testudo among others. According to Hirayama (1985), posterior narrow- ing is limited to crassicollis and borneensis and unites them as a synapomorphy. We were able to replicate this distribution only if the character definition was restrict- ed to the first vertebral scute, in which the posterior mar- gin is significantly narrower than its anterior margin in those two species only. In making this change, however, this character becomes at least partly redundant with characters 45 and 47. Polarity: Outgroup analysis reveals that posterior narrowing of the first vertebral scute is present only in odoratus\ it is absent in Lindholmemys and Mongolemys. We regard the presence of a posterior nar- rowing of the first vertebral scute to be derived. (45) Anterior half of the first vertebral scute much nar- rower than posterior half especially in adults; 0 absent, Fig. 90; 1 = present, Fig. 91 (modified from Hirayama, 1985, R). We confirm the clear presence of an anteriorly nar- rowed first vertebral scute in dentata and spinosa as reported by Hirayama (1985), and note that grandis is polymorphic. Because the anterior sulcus of the first ver- tebral scute commonly is restricted to the nuchal bone in several other taxa, but the scute shows no anterior nar- rowing, we limit the original character definition to the shape of the first vertebral scute only. This peculiar mor- phology seems to be the result of growth that is limited to the anterior edge and the posterior half of the lateral edge of the first vertebral scute. As a consequence, this character is not apparent in juveniles, but becomes increasingly accentuated in adults. Polarity: Anterior narrowing of the first vertebral scute is absent in all outgroups and within the large majority of the ingroup. We consider its presence to be derived. (46) Significant contact of the tenth marginal scute with the fifth vertebral scute; 0 = absent, Fig. 92; 1 = pres- ent, Fig. 93 (modified from Hirayama, 1985, K). Contact of the tenth marginal scute with the fifth vertebral scute was reported previously only in baska, smithii, tecta, and tentoria (Hirayama, 1985). We are able to confirm the presence of a very well developed contact in all but smithii (not seen), and we add spinosa to the list of species in which this contact may occur (it is polymorphic for spinosa ; contact is present in CAS 228368, but absent in the smaller CAS 228459, so onto- genetic differences may explain the polymorphism). We also note slight contacts in some specimens of other species (e.g., agassizii, borneoensis, carbonaria, home- ana, orbicularis, pardalis, and polyphemus ), but by rewording Hirayama’s (1985) original character to include only significant contact, we are able to retain what we believe was his original intent. Polarity: Due to the absence of contact in all out- groups in which it is applicable and the predominance within the ingroup, we consider its presence to be derived. (47) Contact of the second marginal scute with the first vertebral scute; 0 = absent, Fig. 94; 1 = present, Fig. 95 (Hirayama, 1985, O; see also Tinkle, 1962, table 1, ‘Seam A ’). According to Hirayama (1985), the first vertebral scute usually (>90%) contacts the second marginal scute in japonica, leprosa, and caspica. He also noted that the scutes are sometimes in contact in N. platynota and bealei. For our sample, we are able to confirm this con- tact as a polymorphism for bealei , caspica , japonica , and leprosa , but the contact is clearly absent in all our specimens of N. platynota. We also note that these scutes are sometimes in contact in picta, amboinensis, orbicu- laris, and terrapin. Together with all of the above, these taxa were scored as polymorphic. The only taxa to exhibit a consistently well-developed contact are reticu- laria and blandingii. Vol. 10, p. 68 Asiatic Herpetological Research 2004 Polarity: A clear contact between the second mar- ginal scute and the first vertebral scute does not exist in caretta or serpentina , but both morphologies occur in kinostemids and pleurodires. The scutes are not in con- tact in Lindholmemys and Gravemys , but they are in con- tact in Mongolemys. The polarity for this character is ambiguous. (48) Contact of the sixth marginal scute with the third pleural scute; 0 = absent, Fig. 96; 1 = present, Fig. 97 (modified from Hirayama, 1985, B; see also Tinkle, 1962, table 3, ‘Seam C’). The contact between the sixth marginal and third pleural scutes is easily enough rendered as a simple "presence or absence’ character, but this hides the range of possible morphological variation. The degree of con- tact can range from extensive to a condition where the two scutes just barely contact at their comers. Several taxa exhibit a condition where these scutes either barely touch or do not touch one another at their comers, but whenever several specimens were available to us, they typically turned out to be polymorphic. For this reason Gaffney and Meylan (1988) called this character ‘dubi- ous.’ We scored such borderline cases as polymorphic, even if not enough specimens were available to corrob- orate this. We confirm Hirayama’s (1985) observations regarding the absence of a contact between these scutes in baska, ocellata, tecta, tentoria, and thurjii, and we add petersi and spinosa to that list. Taxa that we score as polymorphic include annandalei, borneensis, borneoen- sis, caspica, crassicollis, grandis, hamiltonii, japonica, punctularia , reevesii, sinensis, subtrijuga, and trijuga. Whereas all ‘emydids’ lack a contact, tortoises exhibit both character states. The presence of a contact between the sixth margin- al scute and the third pleural scute was considered by Hirayama (1985) to unite crown group ‘batagurids’ and ‘testudinids’ as a synapomorphy. Given the patchy dis- tribution of this character, and widespread polymor- phism, it seems to be of little use. Polarity: A contact between the sixth marginal scute and the third pleural scute is absent in most outgroups. The exception is caretta ; this is not surprising because caretta has five instead of four pleural scutes. We con- sider the presence of a contact to be derived. (49) Twelfth marginal scute; 0 = two present, their com- mon sulcus only partially subdivides the pygal bone, Fig. 98; 1 = two present, but their common sulcus fully subdivides the pygal bone, Fig. 98; 2 = both twelfth marginal scutes fused along the midline, Fig. 99 (modi- fied from Mlynarski, 1976; Crumly, 1985, 1994, 35; Gaffney and Meylan, 1988, H2.1). According to McDowell (1964, p. 240 and table 1 number 4) members of the ‘Emydinae’ can be distin- guished from the ‘Batagurinae’ based on an incomplete subdivision of the pygal bone by the median sulcus of the posterior-most marginals. We found exceptions with picta, N. platynota, pulcherrima, reevesii, ret icul aria, and spengleri, which do not always clearly exhibit the pattern that would be predicted by McDowell’s (1964) statement, but we note that the expression of this charac- ter will depend significantly on the shape of the pygal bone. In all tortoises except emys and impressa, the twelfth marginal scutes are fused to from a single supra- caudal scute that covers the posterior part of the cara- pace (Crumly, 1985). For this condition, we created a third character state. Polarity: The twelfth marginal scutes are fully sep- arated in all outgroups that have them, and their com- mon sulcus fully subdivides the pygal bone. The pygal bone in “lindholmemydids” is polymorphic, with the 0 state found in Lindholmemys and Mongolemys, and the 1 state in Gravemys. Either state 0 or state 1 is primitive for Testudinoidea; the midline fusion of the twelfth mar- ginals is a derived feature for ‘Testudinidae’. Bridge (50) Sutured contact between plastron and carapace; 0 = present, plastron and carapace are tightly connected by an osseous bridge, Fig. 100; 1 = absent, plastron is attached to carapace by connective tissue, Fig. 101 (modified from Hirayama, 1985, Q; Shaffer et al., 1997, 58; Yasukawa et al. 2001, 21a). (51) Presence and development of anterior buttresses; 0 = anterior buttresses absent, Fig. 102; 1 = anterior but- tresses present but small, and not in contact with the first costal bones, Fig. 103; 2 = anterior buttresses long and thin and just barely in contact with the costal bones, if at all, Fig. 104; 3 = anterior buttresses well developed and in clear contact with the first costal bones, Fig. 105; 4 = anterior buttresses very large and in direct contact with the first dorsal rib, Fig. 106 (modified from Hirayama, 1985, Q; Gaffney and Meylan, 1988, A 14.2; Yasukawa et al, 2001, 28). (52) Presence and development of posterior buttresses; 0 = posterior buttresses absent, Fig. 107; 1 = posterior buttresses present but small, and not in contact with the costal bones, Fig. 108; 2 = posterior buttresses long and thin and just barely in contact with the costal bones, if at all, Fig. 109; 3 = posterior buttresses well developed and in clear contact with costal bones V and VI, Fig. 110; 4 = posterior buttresses well developed but onlv in clear contact with costal bone V, Fig. Ill (modified from Hirayama, 1985, Q; Gaffney and Meylan, 1988, A 14.2, Shaffer et al, 1997, 55; Yasukawa et al, 2001, 29). 2004 Asiatic Herpetological Research Vol. 10, p. 69 (5j) Medially-directed pivoting process for plastral hinge developed on fifth peripheral bone; 0 = absent, Fig. 112; 1 = present, Fig. 113. (54) Complete or almost complete overlap of hyoplas- tron/hypoplastron suture by the pectoral/ abdominal sul- cus; 0 = absent, Fig. 114; 1 = present, Fig. 115 (modi- fied Gaffney and Meylan, 1988, F3.2; Burke et al., 1996, 16; McCord et al., 1995, 13; Yasukawa et al., 2001, 21b). In most testudinoid turtles, the plastron is attached to the carapace via a fully ossified bridge and variably developed plastral buttresses. In species with a kinetic plastron, the bridge is typically absent and the plastron is attached to the carapace via connective tissues (e.g., amboinensis, blandingii , Carolina , dentata, galbinifrons, mouhotii , orbicularis , T. ornata). The original configu- ration of this character tied the presence of plastral kine- sis to the reduction of the buttresses (Hirayama, 1985). However, within testudinoid turtles the morphology of the buttresses varies significantly and independently from plastral kinesis. We consequently split this charac- ter into three discrete characters concerned with the morphology of the bridge and the buttresses. We also developed two new characters that pertain to the morphology of the bridge region: the presence of medially-directed processes on the fifth peripherals that act as pivots for the plastral bones during shell closure (Bramble, 1974) and a revised plastral kinesis character (Gaffney and Meylan, 1988; McCord et al., 1995) that considers plastral-kinesis to be well developed only in those taxa in which the pectoral/abdominal sulcus fully overlaps the hyoplastron/hypoplastron suture, allowing optimal movement between the two plastral lobes. Most taxa with plastral kinesis also have well-developed piv- oting processes on the fifth peripherals, but notable exceptions are orbicularis and N. platynota. In blandingii , the process is modified into an anteroposte- riorly-elongated, flattened process that extends along most or all of the length of the fifth peripheral (Fig. 113). In some specimens of blandingii , a similar structure is developed on the sixth peripheral as well. Polarity: Reconstructing the basal condition for these characters within testudinoids is difficult, because all living cryptodiran outgroups do not have plastral but- tresses and commonly lack osseous bridges. However, the bridge of Gravemys, Mongolemys, and Lindholmemys is osseous, shows no signs of kinesis, and (at least in Lindholmemys) the anterior and posterior but- tresses are well developed and touch the costal bones. We consider that morphology to be primitive for Testudinoids. (55) Contact between inguinal and femoral scutes; 0 = absent, Fig. 116; 1 = present, Fig. 117 (Crumly, 1985, 1994, 42; Gaffney and Meylan, 1988, H3.3, HI 5.2). Within tortoises the complete or frequent absence of a contact between the inguinal scute and the femoral scute was used previously to hypothesize the monophy- ly of several smaller clades, for example {graeca + her- manni + horsfieldi + kleinmanni + marginata + tornieri } (Crumly, 1985) and {agassizii + berlandieri + flavomar- ginatus + polyphemus } (Gaffney and Meylan 1988). We confirm the absence of a contact in representatives of the first group, but not in the second. Among the second group (traditionally classified together in Gopherus) the contact is strongly reduced, but still is present. Among ‘batagurids’ and ‘emydids,’ a contact is absent in all taxa with the noteworthy exception of hamiltonii. Polarity: Determining the polarity for this character is somewhat difficult because all living outgroups have an arrangement of plastral scutes that is rather different from testudinoids. However, based on ingroup common- ality and the absence of a contact in the “Lindholmemydidae,” we conclude that the presence of a contact between the inguinal and femoral scutes should be considered derived for Testudinoidea. (56) Presence of musk glands; 0 = inguinal and axillary gland present; 1 = axillary gland present only; 2 = musk glands absent (modified from Crumly, 1985). (57) Presence of anterior musk duct foramina; 0 = musk glands and their foramina present, Fig. 118; 1 = musk glands present, but foramina not developed; 2 = musk glands and foramina absent (modified from Hirayama, 1985, A; Gaffney and Meylan, 1988, FI. 2, F5.3; Burke et al., 1996, 20). (58) Presence of posterior musk duct foramina; 0 = musk glands and their foramina present, Fig. 119 ; 1 = musk glands present, but foramina not developed; 2 = musk glands and foramina absent (modified from Hirayama, 1985, A; Gaffney and Meylan, 1988, FI. 2, F5.3; Burke et al., 1996, 20). According to Hirayama (1985), the presence of musk duct foramina characterizes the paraphyletic assemblage ‘Batagurinae’ (sensu Hirayama, 1985, not Gaffney and Meylan, 1988). We believe the difference of opinion between Hirayama (1985) and Gaffney and Meylan (1988) regarding this character is based on fail- ure to make clear the distinction between the presence of musk glands and the presence of musk duct foramina. Musk glands are developed in almost all extant turtles (Waagen, 1972), and we consequently agree with Gaffney and Meylan (1988) that their presence should be considered primitive for all cryptodiran turtles. Vol. 10, p. 70 Asiatic Herpetological Research 2004 However, even though most turtles have musk glands, true musk duct foramina are developed only in some pleurodires (e.g., Chelodina, Emydura), some ‘emy- dids,’ and all “batagurids,’ making a monophyletic Testudinoidea (sensu Hirayama, 1985) possible. Distinct musk duct grooves are present on the anterior peripher- als of Kinostemidae, and tiny foramina are sometimes associated with these (Hutchison, 1991). Because the presence of musk glands does not nec- essarily result in the development of musk duct forami- na, we decided to score these two characters separately. We relied on an unpublished thesis on the musk glands of turtles (Waagen, 1972) to determine the presence of musk glands for most taxa. In scoring taxa not investi- gated by Waagen (1972) we only recorded presence of musk glands if musk duct foramina provided positive evidence for their presence (e.g., baska, bealei, borneen- sis, galbinifrons , kachuga, mouhotii , petersi, pulcherri- ma, spengleri, and tentoria). Many tortoises, conse- quently, had to be scored as unknown, because they were not analyzed by Waagen (1972) and do not exhibit musk duct foramina (e.g., elongata, homeana, horsfiel- di, and pardalis ). Our scoring of the musk duct foramina is derived from a combination of osteological observation and data provided by Waagen (1972). Taxa not reported to pos- sess musk glands (Waagen, 1972) were checked for musk duct foramina, but none were found. For those species that Waagen reported as having musk glands, we sought musk duct foramina on osteological specimens. Several taxa with musk glands, but only lightly ossified bridges, do not exhibit musk duct foramina (e.g., blandingii, dentata , flavomarginata, orbicularis , and pulcherrima) or show an asymmetry with foramina only visible anteriorly (e.g., N. platynota). In taxa that pos- sess them, the musk duct foramina are sometimes con- tained entirely within the peripherals (e.g., N. platyno- ta), and sometimes between the peripheral and the plas- tral buttress (e.g., reevesii). Polarity: Given the presence of musk glands in all extant outgroups (Waagen, 1972), their absence should be considered derived. Musk duct foramina are not described for “lindholmemydids” but J. H. Hutchison specifically searched for them in Mongolemys speci- mens housed at IVPP and found no trace of them. Because musk duct foramina are developed in the vast majority of the ingroup, we consider their absence to be derived for testudinoids. Plastron (59) Extensive overhanging lip of the epiplastra; 0 = absent, Fig. 120; 1 = present, Fig. 121 (Gaffney and Meylan 1988, H5.1, 9.2). In most testudinoid turtles, the epiplastra are rather flat with a slight increase in thickness along the anterior margin. In contrast, many tortoises have strongly thick- ened epiplastral margins that sometimes form an over- hanging lip along the interior rim of the plastron. According to Gaffney and Meylan (1988) the presence of such an overhang of the epiplastra unites all tortoises except those classified in the genera Manouria (Fig. 120) and Gopherus, with a reversal occurring in giant insular forms. We confirm this general pattern, but we note the absence of an overhang in tornieri. A interior overhang is absent also in all extant ‘emydids’ and ‘batagurids,’ but a small overhang is present in extinct Ptychogaster and Echmatemys (Mlynarski, 1976, figs. 78, 8 1), taxa generally considered to belong to either the ‘Emydidae’ or ‘Batagurinae.’ Polarity: An overhanging lip on the epiplastra is absent in all outgroups and the majority of the ingroup. We consider its presence to be derived. (60) Intersection of the entoplastron by the humeropec- toral sulcus; 0 = absent, Fig. 122; 1 = present, Fig. 123 (Hirayama, 1985, X; Crumly, 1985; Gaffney and Meylan, 1988, F5.1; McCord et al., 1995, 15). This character was used to help resolve relation- ships within ‘batagurids’ by Hirayama (1985) but we were unable to replicate his results in our analysis. We agree with Crumly (1985) that the sulcus crosses the entoplastron in at least one species classified in the genus Indotestudo (i.e., elongata ), but the sulcus is at the entoplastron/hyoplastron suture in our specimen of forsteni. The condition in species now commonly classi- fied in Testudo varies widely (e.g., the suture crosses the entoplastron in graeca and horsfieldi, but does not in hermanni or kleinmanni). We also agree with Gaffney and Meylan (1988) on their distribution of this character among the ‘Emydidae,’ however, our scoring for picta, orbicularis, and blandingii is polymorphic, because the sulcus generally runs along the suture between the ento- plastron and the hypoplastra, but may barely fall on either side. Polarity: The polarity is ambiguous if only extant taxa are considered. The plastron of most outgroups is too different from that of testudinoids to be of any use for polarizing this character. For instance, the plastron of spinifera lacks scutes, and that of serpentina, odoratus, and caretta is too heavily modified to enable a meaning- ful comparison. Both character states are commonly found in ‘batagurids,’ ‘emydids,’ and testudinids, mak- ing an ingroup analysis futile. The humeropectoral sul- cus is distinctly posterior to the epiplastron in Gravemys and Mongolemys, so we consider an intersection of this suture with the entoplastron to be derived. 2004 Asiatic Herpetological Research Vol. 10, p. 71 (61) Anal notch of the plastron; 0 = present, Fig. 124; 1 = greatly reduced, Fig 125; 2 = absent, Fig. 126 (mod- ified from Hirayama, 1985, W; Yasukawa et al, 2001, 22). The plastron of most testudinoid turtles has a signif- icant anal notch. The absence of such an anal notch for amboinensis, galbinifrons, and flavomarginata was reported by Hirayama (1985) and we confirm those observations. An anal notch also is absent in belliana, Carolina , coahuila , erosa , homeana, T. nelsoni , and T. ornata. To accommodate the presence of a reduced anal notch we modify Hirayama’s (1985) character by creat- ing a third character state. A reduced anal notch is found in blandingii, orbicularis, N. platynota, and reticularia. In at least one species (mouhotii), a distinct anal notch is present in larger individuals, but small specimens have a reduced notch (scored as polymorphic in our matrix); this suggests that development of an anal notch may be subject to ontogenetic variation in at least some testudi- noids. Polarity: In caretta and serpentina an anal notch is not present, however, their plastra are narrow and tapered posteriorly. The fleshy plastron of spinifera is smooth along its posterior margin, but this cannot be observed in osteological preparations. A notch is weak- ly developed in at least some Lindholmemys, but is absent in Mongolemys. A notch is present in kinostem- ids, pleurodires, most of the ingroup, and in Gravemys. We conclude that its absence should be considered derived for our ingroup. (62) Anal scutes fused; 0 = absent, Fig. 127; 1 = pres- ent, Fig. 127 (Hirayama, 1985, Z). The anal scutes of adult galbinifrons, and flavomar- ginata are at least slightly fused, especially along their posterior medial border. We fully agree with Hirayama’s (1985) treatment for this character. Anal scute fusion can be identified easily in macerated specimens (Fig. 127), because the anal scutes will not separate from one anoth- er, as will all other scutes. Polarity: Anal scute fusion is absent in the vast majority of turtles, and is considered to be the primitive condition. (63) Plastral scutes with vibrant, radiating color pat- tern; 0 = absent, Fig. 128; 1 = present, Fig. 129 (Hirayama, 1985, S; McCord et al, 1995, 16 Yasukawa et al, 2001, 32). Vibrant, radiating color patterns of the plastral scutes of dentata, grandis, and spinosa were noted by Hirayama (1985) and McCord et al. (1995). We add tcheponensis to this list, as well as the testudinids geo- metricus and P. oculifera. In our specimens, the pattern of dentata and tcheponensis is not as vibrant as in gran- dis and spinosa. Polarity: Vibrant, radiated color patterns are miss- ing in all outgroups and the majority of the ingroup. Their presence is derived. Postcranium and Soft Tissue (64) Development of a suprascapula; 0 = absent; 1 = present, Fig. 130 (Gaffney and Meylan, 1988, F3.1; Burke et al, 1996, 11). (65) Development of an episcapula; 0 = absent; 1 = present, Fig. 130 (Gaffney and Meylan, 1988, F4.1; Burke et al, 1996, 11). The presence of both a suprascapula and an epis- capula apparently is limited to blandingii and the species currently classified in Terrapene. A suprascapula is also present in orbicularis. Both structures are involved in the locking mechanism of the anterior plastral lobe dur- ing shell closure (Bramble, 1974). These structures are difficult to verify in most osteological preparations, because they may dissociate from the scapula and be dif- ficult to recognize, and because they may ossify only in older individuals. The specimen we dissected to illus- trate these features (TNHC 62532, a T. ornata with cara- pace length of 103 mm) has a completely cartilaginous episcapula, and a predominantly cartilaginous supras- capula (Fig. 130). It is therefore much easier to confirm their presence than verify their absence. We followed Bramble’s (1974) account of these structures and scored our matrix accordingly, as probably did Gaffney and Meylan (1988) and Burke et al. (1996). Polarity: Suprascapulae and episcapulae are absent in all outgroups and the majority of the ingroup. Their presence is considered to be derived. (66) Shape of coracoid blade; 0 = long and narrow, Fig. 131; 1 = short and very wide, Fig. 131 (Crumly, 1985, 1994; Gaffney and Meylan, 1988, HI. 7). The coracoid blade of all ‘emydids’ and ‘batagurids’ is an elongate bone with a narrow, short shaft and a long, wedge-shaped coracoid blade that is about two times wider than the base. In tortoises, this bone is still wedge-shaped, but relatively much shorter and with a blade that is considerably wider, typically four times the width of the base (Crumly, 1985, 1994; Gaffney and Meylan, 1988). We agree with previously published observations. Polarity: The coracoid blade of caretta, serpentina, and odoratus is long and narrow and that of spinifera is long, but not wedge-shaped. We consider a long and nar- row coracoid blade to be primitive for Testudinoidea. (67) Number of manual claws; 0 = five, Fig. 132; 1 = four, Fig. 133 (modified from Hirayama, 1985, J). Vol. 10, p. 72 Asiatic Herpetological Research 2004 Most testudinoid turtles have five manual claws with the exception of baska and horsfieldi, both of which have only four. We did not verify this character independently for all species, due to an overall lack of articulated skeletons and our limited access to pickled specimens. However, because the number of claws of the forelimbs is an easily determinable, discrete number that is regularly noted and described in the literature, we scored all remaining taxa from the comprehensive infor- mation provided by Ernst and Barbour (1989). Polarity: Five manual claws are present in serpenti- na, odoratns, and almost all pleurodires (excluding species currently classified in Chelodina and Hydromedusa ); three are present in spinifera, and two in caretta. The condition in “lindholmemydids” is unknown. Based on ingroup commonality, we consider five claws to be primitive for the ingroup. (68) Number of phalanges of manus and pes; 0 = digi- tal formula of2-3-3-3-3 or 2-3-3-3-2, Fig. 132; 1 = dig- ital formula with less than 2-3-3-3-2, Fig. 133 (Crumly, 1985; Gaffney and Meylan, 1988, Hl.l). The digital formula of most testudinoid turtles is 2- S-3-3-3 or 2-3-3-3-2. Among tortoises, the manus and pes are greatly shortened and the digital formula is typi- cally reduced to 2-2-2-2-2 or less (Auffenberg, 1974:135-136; Crumly, 1985). Due to the dissociated nature of most of the material we viewed, we were not able to verify the digital formulae of most of the turtle taxa we included. However, when articulated hands and feet were present, we never found anything to contradict the statements made above. We scored all tortoises based on information provided by Auffenberg (1974) and Crumly (1985). Polarity: All outgroups and the majority of the ingroup do not have a reduced digital formula. We con- sequently consider the reduced formula to be derived. (69) Webbing between digits; 0 = present, well devel- oped, Fig. 134; 1 = absent, or at least strongly reduced, Fig. 135 (Hirayama, 1985, b). Due to their semi-aquatic nature, most testudinoids have well-developed webbing between the digits of their hands and feet. In more terrestrial species, however, the webbing often is reduced. Unfortunately, there seems to be a gradient in the development of webbing, from extremely well developed (e.g., baska, reticularia) to moderately developed (e.g., dentata, guttata) to virtual- ly non-existent (e.g., spengleri ). We nevertheless were able to reproduce Hirayama’s (1985) distribution for the ‘batagurids’ with the exception of grandis and spinosa, which have reduced webbing (grandis is the only ‘bor- derline’ taxon we found, but its webbing is reduced rel- ative to those taxa we scored as having well-developed webbing). Among ‘emydids,’ we note that the webbing is heavily reduced in Carolina, T. nelsoni, and T. ornata. All tortoises lack webbing. Polarity: All outgroups and the majority of ingroup taxa have webbed hands and feet. We consider the absence of webbing to be derived. (70) Sexual size dimorphism; 0 = absent; 1 = present, female much larger than male (Gaffney and Meylan, 1988, F5.2; Burke et al., 1996, 37). In almost every species of turtle, there is some expression of sexual size dimorphism (Berry and Shine, 1980; Gibbons and Lovich, 1990). The difference in size between the sexes can be expressed as a ratio and typi- cally shows considerable variation depending on the population (Gibbons and Lovich, 1990). We initially tried to score this character with three character states, as done by Burke et al. (1996), differentiating between species with larger males, larger females, and equally sized sexes, but we abandoned that, because exact data are missing for most ‘batagurid’ taxa. We consequently only score taxa as being sexually dimorphic if females are at least 1 .4 times larger than the males. Our scores are derived from Gibbons and Lovich (1990) and Ernst and Barbour (1989). Polarity: Sexual size dimorphism is prevalent in most outgroups. In spinifera the female is much larger, in odoratus and serpentina the male tends to be slightly smaller, in caretta the sexes are of similar size. The out- group polarity is thus ambiguous, but in the majority of the ingroup pronounced sexual dimorphism is absent. Problematic Characters We encountered difficulties in evaluating a number of previously used characters, and we provide some sum- mary statements for those in this section. Most of these characters were not pursued thoroughly in our study because we were not able to understand the original descriptions, were unable recover discrete character states, or because at an early point in our investigation of the character we detected significant variation in expres- sion of character states within taxa. In the latter case greater sample sizes or new methodological techniques (e.g., Wiens, 1995; Smith and Gutberlet, 2001) will be required to tease out a phylogenetic signal. (A) Frontal contribution to the supratemporal rim (Hirayama, 1985, 4). The anterior extent of the upper temporal emargina- tion is difficult to define in many taxa, and is impossible to determine in those with a fully emarginated temporal region (e.g., T. ornata ). The result is a high degree of ambiguity and a general lack of discrete character states. 2004 Asiatic Herpetological Research Vol. 10, p. 73 (B) Contact between postorbital and quadrate (Hirayama, 1985, 10). In the vast majority of ingroup taxa, there is no con- tact between the postorbital and the quadrate. Such a contact was observed only in japonica and punctularia by Hirayama (1985). In specimens of japonica available to us, we were not able to confirm this contact. CAS 228348 is a skeleton from a diseased specimen of punc- tularia. On the right side of the skull there is a possible (but only slight) contact. It is possible that the contact is actually between the postorbital and the quadratojugal (Fig. 136). We also found a minimal contact in one spec- imen of annulata. Given these diverging observations and the minute contact that is present in our material, we regard (for now) any contact within the ingroup as an abnormality. (C) Absence of the “ posterior process of the postor- bital” (Hirayama, 1985, 8). We cannot determine unambiguously what Hirayama (1985) meant by this character. In our assess- ment of testudinoids, both a posterolateral and a postero- medial process of the postorbital can occur. In Hirayama’s (1985) analysis, only grandis and spinosa lack a “posterior process of the postorbital.” These species are also the only ‘batagurid’ taxa to fully lack a temporal arch. We suspect that this character may some- how be referring to a lack of a bony temporal arch. (D) Processus inferior parietalis ‘‘medially approximat- ing each other, cranial cavity ant eroventr ally narrow- ing” (Hirayama, 1985, 5; McCord et al., 1995, 7; Yasukawa et al, 2001, 2). We acknowledge the validity of this character as was originally worded by McDowell (1964). However, we find it difficult to determine how strongly the con- striction of the brain case must be before it can be con- sidered present. We were unable to develop unambigu- ous discrete character states for this feature. (E) Subdivision of the foramen nervi trigemini (Crumly, 1982; Hirayama, 1985, 6). This character was used originally by Crumly (1982) to infer phylogenetic relationships within ‘Testudinidae’. For his ingroup, Crumly (1982) observed a great amount of polymorphism, with no sin- gle species either completely lacking or always exhibit- ing a subdivision of the foramen. He also noted asym- metry for this character between the left and right side of some individuals. We confirm the common presence of a subdivided trigeminal foramen in representatives of ‘Testudinidae,’ and the occasional presence in individu- als of ‘Emydidae’ and ‘Bataguridae’ (e.g., areolata, den- tata, flavimaculata, N. platynota , and rubida). A signifi- cant amount of variation can be observed in two speci- mens of borneensis available to us, that exhibit left/right asymmetry and the full spectrum from a fully intact (Fig. 137), to partially subdivided (Fig. 138), to fully subdi- vided (Fig. 139) trigeminal foramen. Given that most taxa are represented by three or fewer skulls in our study, it is apparent that we are not able to fully docu- ment the amount of variation exhibited by testudinoids. (F) Contact between postorbital and squamosal (Hirayama, 1985, 9). Gaffney et al. (1991) noted that absence of this con- tact is associated with the upper temporal emargination and considered it informative at the level of their analy- sis. Within our ingroup, all turtles have substantial upper temporal emarginations, resulting in the contact being just barely present, or just barely absent, or polymorphic (e.g., picta, petersi , texana, crassicollis). See comments above under character 9. (G) Median premaxillary notch (Hirayama, 1985, 19; Yasukawa et al., 2001, 9) (H) Large cusps near the suture of the premaxillae and maxillae (Hirayama, 1985, 28). Initially, we were faced with the problem of defin- ing these two characters independently from one anoth- er, because the presence of two tightly spaced, opposing cusp-like structures along any margin will automatically result in the development of a median notch. An addi- tional problem relating to these characters is the ques- tion of whether these features should be observed on the ramphotheca or the maxilla. Large, tooth-like cusps are clearly present in a num- ber of taxa (e.g., thurjii ) but so is the full spectrum of smaller cusps, making it impossible to clearly define discrete character states. Furthermore, if all species were evaluated for medial notches that existed even if the cusps were removed, all taxa in our sample would show a medial notch. We were unable to develop a consistent method for scoring this character for all testudinoid species. (I) ‘‘Antero-medial portion of the upper triturating sur- face formed by premaxillae and maxillae” (Hirayama, 1985, 23; Yasukawa et al., 2001, 13). We are neither able to replicate the full meaning of this character nor formulate truly discrete character states. A connection with the development of the second- ary palate is evident, but the morphology of this region seems to be sufficiently covered by a number of other characters. Vol. 10, p. 74 Asiatic Herpetological Research 2004 (J) Participation of the vomer in the foramen praepalat- innm (Crumly, 1982, 10; Hirayama, 1985, 32; Yasukawa et al., 2001, 15). Within testudinoid turtles, the foramen praepalat- inum perforates the nasal cavity at the border between the premaxilla and the vomer. When the foramen is posi- tioned slightly more anteriorly, it is fully surrounded by the premaxilla, when it is minutely farther posterior it is surrounded by the vomer. Considering the impact of such minute changes, it is not surprising that our scoring for this character generally seems to be in conflict with that of Hirayama (1985) and Crumly (1982). This char- acter appears to be subject to great intraspecific varia- tion. (K) Foramen palatinum posterius enclosed within the brain cavity (Hirayama, 1985, 34). According to Hirayama (1985), in reevesii (only) the foramen palatinum posterius is enclosed within the region of the brain cavity due to a flared descending process of the parietal. We cannot confirm this observa- tion for any testudinoid turtles (including three speci- mens of reevesii). (L) Participating bones in the processus trochlearis oticum (Hirayama 1985, 37; Gaffney and Mey l an, 1988, Gaffney et al. 1991, 6; McCord et al., 1995, 8; Shaffer et al., 1997, 74, 258 Yasukawa et al., 2001, 18). The relative participation of the prootic, parietal, and quadrate to the processus trochlearis oticum was used previously by a number of authors to infer phylo- genetic relationships within turtles. Our observations confirm the great variety of morphologies that can be observed in this region. However, the amount of intraspecific variation is considerable and the full spec- trum of possible morphologies seems to be filled, mak- ing it difficult to discern discrete character states. Future research in the area may result in more clearly defined discrete character states. (M) Length of the crista supraoccipitalis (Hirayama, 1985, 40). A long crista supraoccipitalis was observed by Hirayama (1985) for borneensis. The character states he used are defined by relative length of the crista supraoc- cipitalis to the “condylo-basal length.” Unfortunately, we could not replicate this because it is not clear exact- ly how the length of the crista was measured. Furthermore, a true morphological gap seems to be missing between the admittedly very long crista of borneensis and other ‘batagurids’ with an elongated crista. This character is problematic, because it is poor- ly defined and lacks discrete character states. (N) Bony sutures and sulci lost in old adults (Hirayama, 1985, I). According to Hirayama (1985), loss of sutures and sulci occurs in baska, borneoensis, and borneensis only. We are able to confirm this, but we do not have individ- uals of all other species that are sufficiently old enough to positively confirm that they also do not exhibit this feature at old age. In subsequent treatments, loss of sutures and loss of sulci should be treated as separate characters. (O) Ossification of cornu branchiale II (Hirayama, 1985, 48; Yasukawa et al., 2001, 20). This character was used previously to unite tortois- es with a number of ‘batagurid’ taxa (Hirayama, 1985). The hyoid apparatus of turtles is often disarticulated in skeletal preparations, making is difficult to positively confirm if an ossified cornu branchiale is present or absent. However, for those taxa for which we were able to observe the hyoid apparatus, we were not able to con- firm Hirayama’s (1985) observation of a reduced cornu branchiale II in some ‘batagurids’ (e.g., galbinifrons, spengleri). Instead, these taxa exhibit a cornu branchiale II that is not significantly different from most other ‘batagurids.’ (P) Double articulation between the fifth and sixth cer- vical vertebrae (Hirayama, 1985; Gaffney and Mey lan, 1988, FI. 5). Most articular surfaces of the cervical column are rather homogenous within all testudinoid turtles (Williams, 1950). A double articulation between the fifth and sixth cervical previously was considered to be a unique character that unites the ‘Emydinae’ (McDowell, 1964). This character also was used by Hirayama (1985) and with reservations by Gaffney and Meylan (1988). Our observations generally confirm the presence of a more or less clear double articulation in most ‘emydids,’ however, this features is also present in a number of ‘batagurids’ confirming that this character is highly vari- able within the ingroup (Williams, 1950; Gaffney and Meylan, 1988). Unfortunately, discrete character states are lacking; we were able to observe the full morpholog- ical spectrum from a clear singular articulation to a clear double articulation. (Q) Scapular prong with lateral concavity (Hirayama, 1985, E). Hirayama (1985) reported this character as an autapomorphy for subtrijuga only. However, we cannot identify this morphology in any of our specimens of sub- trijuga. 2004 Asiatic Herpetological Research Vol. 10, p. 75 (R) Large facet of the ilium for the testoscapularis and testoiliacus (Hirayama, 1985, T; Yasukawa et al., 2001, 34) (S) Extensive development of both testoscapularis and testoiliacus (Hirayama, 1985, U). An extensive development of the testoscapularis and testoiliacus muscles together with an associated large scar on the ilium was reported by Bramble (1974) for Asian and North American box turtles. Whereas we have no reason to doubt his assessment of the develop- ment of these muscles for box turtles, we were not able to score this character for most of the remaining taxa. The shape of the ilium was explored and illustrated for some ‘emydids’ and ‘batagurids’ by Yasukawa et al. (2001:122-123). (T) Ossification of the epipubis (Gaffney and Meylan, 1988, F5.5). The identification (and confirmation of presence or absence) of an ossified epipubis (Fig. 141) is somewhat difficult for most species, because it seems to ossify rather late in ontogeny, and can fall off during prepara- tion. Our tentative observations confirm the presence of an ossified epipubis in numerous adult ‘emydids’ and ‘batagurids,’ typically terrestrial forms (e.g., T. ornata, G. insculpta , N. platynota, yuwonoi). An improved sam- ple of adult specimens of all taxa, however, is necessary to reveal the true distribution of this character. (U) Diploid Number of Chromosomes (Hirayama, 1985, 0; Shaffer et al., 1997, 43). The diploid number of chromosomes was used by Carr and Bickham (1986) to hypothesize a sister group relationship between the ‘Emydinae’ and subtrijuga, fol- lowed by borneensis and crassicollis and finally the rest of the ‘Batagurinae.’ Whereas most ‘batagurids’ alleged- ly have 52 chromosomes, subtrijuga, borneensis, crassi- collis, and ‘emydids’ are supposed to have 50 chromo- somes. We view these results with caution, because a brief review of the relevant literature reveals great dif- ferences in chromosomal counts for a variety of taxa. For instance, according to the work of Killebrew (1977) and Bickham (1981), amboinensis has 52 chromosomes, however, Gorman (1973) reported only 50. Similar con- flicts can also be found for dentata (Bickham, 1981; DeSmet, 1978; Gorman, 1973; Stock, 1972), subtrijuga (Bickham, 1981; Killebrew, 1977), trijuga (DeSmet, 1978; Carr and Bickham, 1986), sinensis (Bickham, 1981; Killebrew, 1977; Stock, 1972), crassicollis (Killebrew, 1977; Stock, 1972; Bickham and Baker, 1976), and some of the species currently placed in Graptemys (Killebrew, 1977; McKown 1972), Trachemys (DeSmet, 1978; Killebrew, 1977; Stock, 1972) and Mauremys (Killebrew, 1977; Stock, 1972). This conflict in primary data is probably best understood when considering the nature of testudinoid chromo- somes: whereas 14 pairs of chromosomes have a consid- erable size, all of the remaining ones are extremely small. Given these circumstances, it seems reasonable to hypothesize that one pair of chromosomes may be unrecognized during analysis. (V) Plica media spade-shaped (Gaffney and Meylan, 1988, F7.1). The penile soft anatomy of turtles was comprehen- sively reviewed by Zug (1966) and one of his characters, the shape of the plica media, was used by Gaffney and Meylan (1988) to unify species placed in Chrysemys, Deirochelys, Trachemys, and Pseudemys as a mono- phyletic group. In his detailed description of the plica media, Zug (1966) referred to the shape of this structure as being “spade-shaped” in those taxa, but made similar claims for other taxa too. Furthermore, based on the illustrations that were provided by Zug (1966) for other taxa, the plica media of species placed in Graptemys, Malaclemys, Rhinoclemmys, and Platysternon appear “spade-shaped” also, even though Zug (1966) did not explicitly use those descriptive words. This anatomical system should be carefully reevaluated for all testudi- noids, with special attention given to definition of dis- crete characters. (W) Ossifications within the fenestra postotica. In some taxa, portions of the fenestra postotica are closed or obscured by ossifications (noted, but without exemplars, by Gaffney, 1972). In our largest specimen of grandis (CAS 228443) a short, spike-like ossified process extends posterodorsally from the dorsal edge of the quadrate process of the pterygoid, and crosses the fenestra postotica. It is situated ventral to the stapes (col- umella auris), medial to the incisura columella auris, and lateral to the fenestra ovalis (Fig. 141). In some speci- mens, the dorsal tip of a similar structure approaches or meets a posterodorsally-inclined process that extends from the dorsal surface of the pterygoid, near the suture with the prootic. In our largest specimen of N. platynota (CAS 228342) the two processes meet to enclose the stapedial shaft in a ring of bone situated at approximate- ly the midpoint between the fenestra ovalis and the medial opening of the incisura columella auris. Our other, smaller, specimen of N. platynota shows no devel- opment of these processes. It seems likely that there is an ontogenetic component in the expression of this fea- ture. It does not appear to have any systematic signifi- cance, but in any case it is not widespread within Testudinoidea. Vol. 10, p. 76 Asiatic Herpetological Research 2004 Conclusions Our observations can be used to draw some tenta- tive conclusions regarding the current level of under- standing about morphological variation within testudi- noid turtles. In addition, we provide some cautionary statements about the quality of morphological data now in use for assessing systematic relationships among these turtles. It is clear from a perusal of the relevant lit- erature, and from our data, that there is reasonably strong morphological support for a monophyletic ‘Testudinidae.’ Support for monophyly of ‘emydines’ and ‘batagurines’ is not as impressive. The paraphyly of ‘Batagurinae’ (with respect to ‘Testudinidae’) was explicitly proposed by Hirayama (1985) and has been generally accepted since that time, although some strides have been made towards resolving relationships among some ‘batagurine’ taxa. The monophyly of ‘Emydinae’ seems to have been implicitly assumed by many workers, but remains to be established in the con- text of a rigorous phylogenetic analysis of all relevant taxa. The interpretation of several morphological fea- tures shared between some ‘emydines’ and some ‘batagurines’ as either convergence or synapomorphy remains an important and interesting challenge. For example, there are intriguing morphological similarities between subtrijuga and some species classified in the genus Graptemys (e.g., contact of the jugal and descend- ing process of the parietal; contact of the quadratojugal with the articular facet of the quadrate; contact between the quadratojugal and the maxilla; ventral process of the pterygoid approaching the articular surface of the quadrate). These similarities may be due to functional convergence as a result of a molluscivorous diet, but they raise questions about the propriety of utilizing sub- trijuga as an outgroup for systematic studies of ‘emy- dines’ (e.g., Burke et al., 1996). Additional similarities are reported for chromosome numbers in subtrijuga and ‘emydines’ (see ‘Problematic Character’ U, above). A seriously deficient understanding of morphologi- cal variation is one of the greatest inadequacies of cur- rent perspectives on morphological data in turtles gener- ally, and testudinoids specifically. Few published studies have been conducted to evaluate the range and causes of sexual, ontogenetic, intra- and inter-population variation in morphological characters within testudinoids. Our preliminary considerations of ontogenetic variation, combined with reports of sexual variation (e.g., Berry and Shine 1980; Gibbons and Lovich, 1990; Stephens, 1998) and new studies exploring the complex interac- tions of morphological evolution with behavioral char- acteristics and environmental conditions in turtles (e.g., Lindeman, 2000; Herrel et al., 2002; Joyce and Gauthier, 2003; Claude et al., 2003) emphasize the importance of pursuing these questions further. Our decision not to produce a phylogenetic hypothesis in this paper was based primarily on two considerations. The first is the relatively small sample size we used for many taxa in this study (although it is comparable to sample sizes from other, previously published, studies), and the fact that several taxa are not represented in our work. The second consideration is our sense that the current under- standing of morphological variation in testudinoid tur- tles is insufficiently mature to permit reliable phyloge- netic hypotheses based solely on morphological data. The most expedient way to address the need for greater documentation of variation within testudinoid species is to utilize existing museum collections to the greatest extent possible, and secondarily to develop responsible collecting programs that are designed with this need in mind. Our results also indicate that morphological data matrices currently in the literature should not be taken at face value. We had particular difficulties replicating some of the scoring in the Hirayama (1985) matrix. That seminal analysis (completed prior to the widespread use of computer-assisted analytical methods in systematics) laid the foundation for nearly all subsequent work on ‘batagurine’ morphology (including our own), and its importance in shaping our current conceptualization of ‘batagurine’ phylogeny cannot be overstated. The work of pathfinders in all fields of inquiry is often subjected to the greatest scrutiny by the next generation of researchers. Our statements and contradictory observa- tions in this paper in no way denigrate Hirayama’s work; instead, we view our efforts as minor attempts to correct the few inconsistencies in his analysis, and to contribute our observations to the body of knowledge that he began to synthesize 20 years ago. The accurate interpretation and understanding of the morphological descriptions of previous authors were among the great challenges we faced when we began our studies of testudinoid skeletal morphology. Much of our confusion could have been averted if adequate illustra- tions accompanied all published character descriptions, but such documentation often is an expensive undertak- ing. Our photographs of character states discussed in this paper are intended to facilitate communication among turtle enthusiasts, and to provide a baseline for future comparisons and discussions about testudinoid morphology. We hope that adequate illustration of all newly proposed characters will become standard prac- tice among turtle systematists. It seems likely that our interpretations of characters will differ in some respects from those of our colleagues, and we anticipate that our decisions regarding ‘problematic characters’ (discussed above), and our choices with respect to ‘lumping’ or ‘splitting’ previously published character states, will generate much spirited discussion in the years ahead. 2004 Asiatic Herpetological Research Vol. 10, p. 77 Figure 1. Character 1(0): CAS 228437, texana, Figure 2. Character 1(1): CAS 228404, belliana, anterior view. anterior view. Figure 3. Character 2(0): CAS 228458, texana , Figure 4. Character 2(1): CAS 228404, belliana , left lateral view. left lateral view. Figure 5. Character 3(0): CAS 228444, mouhotii , left ventrolateral view of orbit. Figure 6. Character 3(1): CAS 228443, grandis, left ventrolateral view of orbit. HM Vol. 10, p. 78 2004 Figure 7. Character 4(0): CAS 228443, grandis , right anterolateral view of orbit. Figure 8. Character 4(1): CAS 228420, mouhotii, right anterolateral view of orbit. Figure 9. Character 5(0): CAS 228444, mouhotii, left anterolateral view of orbit. Figure 10. Character 5(1): CAS 228438, texana , left anterolateral view of orbit. Figure 11. Character 6(0): CAS 228443, grandis, right posterolateral view. Figure 12. Character 6(1): CAS 228439, texana, right posterolateral view, postorbital removed. 2004 Asiatic Herpetological Research Vol. 10, p. 79 Figure 13. Character 7(0) and 8(0): CAS 228438, Figure 14. Character 7(1) and 8(1): CAS 228445, texana, left anterolateral view of orbit. subtrijuga, left anterolateral view of orbit. Figure 15. Character 9(0): CAS 228447, orbicularis, left lateral view. Figure 16. Character 9(1): CAS 228444, mouhotii, left lateral view. Figure 17. Character 9(2): YPM 14074, galbinifrons, left lateral view. Figure 18. Character 9(2): YPM 14080, galbinifrons, left lateral view. Vol. 10, p. 80 Asiatic Herpetological Research 2004 Figure 19. Character 10(0): CAS 228444, mouhotii, left lateral view. Figure 20. Character 10(0): CAS 228446, subtrijuga, left lateral view. Figure 21. Character 10(1): YPM 10339, hamiltonii, left lateral view of orbit. Figure 22. Character 11(0) and 12(0): CAS 228447, orbicularis, left lateral view. Figure 23. Character 11(1) and 12(1): CAS 228446, subtrijuga, left lateral view. Figure 24. Character 12(1): CAS 228438, texana, right posteroventral view of lower temporal fossa. 2004 Asiatic Herpetological Research Vol. 10, p. 81 Figure 25. Character 13(0): CAS 228361, reevesii, anterior view. Figure 26. Character 13(0): CAS 228419, amboinensis , anterior view. Figure 27. Character 13(1): CAS 228371, spengleri , anterior view. Figure 29. Character 14(0): CAS 228443, grandis , left posterolateral view of orbit. Figure 28. Character 14(0): CAS 228444, mouhotii, left posterolateral view of orbit. Figure 30. Character 14(1): CAS 228438, texana , left posterolateral view of orbit. Vol. 10, p. 82 Asiatic Herpetological Research 2004 Figure 31. Character 15(0): CAS 228342, N. platynota , ventral view of palate. Figure 32. Character 15(1): CAS 228419, amboinensis, ventral view of palate. Figure 33. Character 16(0) and 17(0): ' Figure 34. Character 16(1) and 17(1): CAS 228447, orbicularis , ventral view of palate. CAS 228420, mouhotii, ventral view of palate. Figure 35. Character 18(0): CAS 228443, Figure 36. Character 18(1): CAS 228444, grandis, right lateral view of trigeminal foramen. mouhotii, right dorsolateral view of trigeminal foramen. 2004 Asiatic Herpetological Research Vol. 10, p. 83 Figure 37. Character 19(0): CAS 228447, orbicularis, ventral view of palate. Figure 38. Character 19(1): TMM 2813, berlandieri, ventral view of palate. Figure 39. Character 20(0): CAS 228335, crassicollis, ventral view of palate. Figure 40. Character 20(1): FMNH 259430, tentoria, posteroventral view of palate. Figure 41. Character 21(0): CAS 228443, grandis, ventral view of brain case. Figure 42. Character 21(1): CAS 228338, reticularia, ventral view of brain case. Vol. 10, p. 84 Asiatic Herpetological Research 2004 Figure 43. Character 22(0): CAS 228443, grandis , ventromedial view of right basicranium. Figure 45. Character 23(0): CAS 228437, texana , right lateral view of quadrate. Figure 44. Character 22(1): CAS 228445, subtrijuga , ventromedial view of right basicranium. Figure 46. Character 23(1): CAS 228342, N. platynota, right lateral view of quadrate. Figure 47. Character 24(0): CAS 228447, orbicularis , right lateral view of mandible. Figure 48. Character 24(1): CAS 228335, crassicollis , right lateral view of mandible. 2004 Asiatic Herpetological Research Vol. 10, p. 85 Figure 49. Character 25(0): CAS 228447, orbicularis, left lateral view of mandible. Figure 50. Character 25(1): CAS 228411, carbonaria, left lateral view of mandible. V Figure 51. Character 26(0): CAS 228404, belliana , left lateral view of mandible. Figure 52. Character 26(1): CAS 228361, reevesii, left lateral view of mandible. Figure 53. Character 27(0): CAS 228361, reevesii, left posterolateral view of mandible. Figure 54. Character 27(1): YPM 10861, thurjii, left posterolateral view of mandible. Vol. 10, p. 86 2004 Figure 55. Character 28(0): CAS 228342, N. platynota, ventral view of palate. Figure 56. Character 28(1) and 29(0): CAS 228437, texana , ventral view of palate. Figure 57. Character 28(1) and 29(0): CAS 228445, subtrijuga , ventral view of palate. Figure 58. Character 29(0): CAS 228419, amboinensis , ventral view of palate. Figure 59. Character 29(1 ): CAS 228361 , reevesii , ventral view of palate. 2004 Asiatic Herpetological Research Vol. 10, p. 87 Figure 60. Character 30(0), 31(0), and 32(0): CAS 228447, orbicularis , oblique ventral view of palate. Figure 62. Character 30(1) and 32(1): TMM 2813, berlandieri, ventral view of palate. Figure 61. Character 30(1) and 31(1): CAS 228437, texana , oblique ventral view of palate. Figure 63. Character 32(2): CM 124246, petersi, ventral view of palate. Figure 64. Character 33(0): CAS 228443, grandis, dorsal view of mandible. Figure 65. Character 33(1): CAS 228361, reevesii, dorsal view of mandible. Vol. 10, p. 88 Asiatic Herpetological Research 2004 Figure 66. Character 34(0) and 35(0): CAS 228448, blandingii, dorsal view of carapace. Figure 67. Character 34: CAS 228451, crassicollis , dorsal view of juvenile carapace showing tricarinae. Figure 68. Character 34(1) and 35(1): CAS 228444, mouhotii, dorsal view of carapace. Figure 69. Character 36(0): CAS 228376, Figure 70. Character 36(1): CM 259430, tentoria galbinifrons, posterior view of shell. anterior view of shell. 2004 Asiatic Herpetological Research Vol. 10, p. 89 Figure 7 1 . Character 37(0) and 38(0): CAS 228346, blandingii, dorsal view of carapace. Figure 72. Character 37(1) and 38(1): CAS 228343, spengleri, dorsal view of carapace. Figure 73. Character 37(2): CAS 228408, elongata, dorsal view of carapace. Figure 74. Character 38(2): CAS 228399, horsfieldi , dorsal view of carapace. Figure 75. Character 38(3): CAS 228445, subtrijuga, dorsal view of neurals II - IV. Vol. 10, p. 90 Asiatic Herpetological Research 2004 Figure 76. Character 39(0): CAS 228399, Figure 77. Character 39(1): CAS 228375, horsfieldi, posterodorsal view of carapace. Carolina, posterodorsal view of carapace. Figure 78. Character 40(0): CAS 228371, spengleri, dorsal view of carapace. Figure 79. Character 40(1): CAS 228430, carbonaria, anterodorsal view of carapace. Figure 80. Character 41(0): CAS 228368, spinosa, dorsal view of carapace. Figure 81. Character 41(1): CAS 228450, N. platynota, dorsal view of carapace. 2004 Asiatic Herpetological Research Vol. 10, p. 91 Figure 82. Character 42(0): CAS 228338, Figure 83. Character 42(1): FMNH 259430, reticularia , posterodorsal view of carapace. tentoria, posterodorsal view of carapace. Figure 84. Character 42(2): CAS 228344, emys, Figure 85. Character 43(0): CAS 228413, posterodorsal view of carapace. insculpta, posterodorsal view of carapace. Figure 86. Character 43(1): YPM 14678, platynota, posterodorsal view of carapace. Figure 87. Character 43(2): CAS 228345, amboinensis, posterodorsal view of carapace. Vol. 10, p. 92 Asiatic Herpetological Research 2004 Figure 88. Character 44(0): CAS 228368, spinosa , dorsal view of carapace. Figure 89. Character 44(1): CAS 228335, crassicollis , dorsal view of carapace. Figure 90. Character 45(0): CAS 228430, carbonaria , anterodorsal view of carapace. Figure 92. Character 46(0): YPM 11653, spengleri, posterodorsal view of carapace. Figure 91. Character 45(1): CAS 228341, grandis, dorsal view of first vertebral scute, anterior to top. Figure 93. Character 46(1): CAS 228368, spinosa , posterodorsal view of carapace. 2004 Asiatic Herpetological Research Vol. 10, p. 93 Figure 94. Character 47(0): CAS 228371, spengleri, dorsal view of carapace. Figure 95. Character 47(1): YPM 10382, blandingii , dorsal view of carapace. Figure 96. Character 48(0): CAS 228450, Figure 97. Character 48(1): CAS 228430, N. platynota, left dorsolateral view of carapace. carbonaria, left dorsolateral view of carapace. Figure 98. Character 49(0): CAS 228458, texana (left); 49(1): CAS 228341, grandis (right); dorsal view of pygals. Figure 99. Character 49(2): CAS 228449, pardalis , posterior view of carapace. Vol. 10, p. 94 Asiatic Herpetological Research 2004 Figure 100. Character 50(0): CAS 228399, horsfieldi, lateral view of carapace. Figure 101. Character 50(1): CAS 228375, Carolina , lateral view of carapace. Figure 102. Character 51(0): CAS 228342, platynota, ventral view of anterior plastral lobe. Figure 104. Character 51(2): CAS 228408, elongata, internal view of shell from posterior. Figure 103. Character 51(1): CAS 228419, amboinensis, ventral view of anterior plastral lobe. Figure 105. Character 51(3): CAS 228406, insculpta , internal view of shell from posterior. 2004 Asiatic Herpetological Research Vol. 10, p. 95 Figure 106. Character 51(4): YPM 14073, thurjii, Figure 107. Character 52(0): YPM 691, Carolina , left ventral view of first thoracic rib. ventral view of posterior plastral lobe. Figure 108. Character 52(1): CAS 228434, blandingii, ventral view of posterior plastral lobe. Figure 110. Character 52(3): CAS 228361, reevesii, medial view of partial right shell. Figure 109. Character 52(2): CAS 228349, pardalis, medial view of partial right shell. Figure 111. Character 52(4): CAS 228341, grandis , medial view of partial right shell. Vol. 10, p. 96 Asiatic Herpetological Research 2004 Figure 112. Character 53(0): CAS 228447, orbicularis, ventral view of carapace. Figure 114. Character 54(0): CAS 228399, horsfieldi, ventral view of shell. Figure 113. Character 53(1): CAS 228345, amboinensis (left); CAS 228373, blandingii (right); ventral view of carapaces. Figure 115. Character 54(1): CAS 228345, amboinensis, ventral view of shell. Figure 117. Character 55(1): YPM 12653, erosa, right posterolateral view of carapace. Figure 116. Character 55(0): CAS 228403, tornieri, right posterolateral view of carapace. 2004 Asiatic Herpetological Research Vol. 10, p. 97 Figure 120. Character 59(0): CAS 228416, impressa , dorsal view of epiplastra. Figure 121. Character 59(1): CAS 228397, carbonaria , dorsal view of epiplastra. Figure 118. Character 57(0): CAS 228335, crassicollis , anterior view of shell. Figure 119. Character 58(0): CAS 228335, crassicollis, posterior view of shell. Figure 122. Character 60(0): CAS 228437, texana, ventral view of anterior plastral lobe. Figure 123. Character 60(1): CAS 228345, amboinensis, ventral view of anterior plastral lobe. Vol. 10, p. 98 Asiatic Herpetological Research 2004 Figure 124. Character 61(0): CAS 228437, texana , ventral view of posterior plastral lobe. Figure 126. Character 61(2): CAS 228376, galbinifrons , ventral view of posterior plastral lobe. Figure 128. Character 63(0): CAS 228450, platynota, ventral view of plastron. Figure 125. Character 61(1): YPM 14678, N. platynota, ventral view of posterior plastral lobe. Figure 127. Character 62(0): CAS 228345, amboinensis (left); 62(1): CAS 228376, galbinifrons (right); ventral view of anal scutes. Figure 129. Character 63(1): CAS 228368, spinosa, ventral view of plastron. 2004 Asiatic Herpetological Research Vol. 10, p. 99 Figure 130. Character 64(1) and 65(1): TNHC 62532, ornata, lateral view of right scapulacoracoid. Figure 132. Character 67(0) and 68(0): YPM 14677, blandingii, top view of lower arm and manus. Figure 134. Character 69(0): YPM 2983, terrapin, top view of lower arm and manus. Figure 131. Character 66(0): CAS 228345, amboinensis (left); 66(1): CAS 228397, carbonaria (right); coracoids. Figure 133. Character 67(1) and 68(1): YPM 16450, horsfieldi, top view of lower arm and manus. Figure 135. Character 69(1): YPM 14445, spengleri, top view of lower arm and manus. Vol. 10, p. 100 Asiatic Herpetological Research 2004 Figure 136. CAS 228348, punctularia, right lateral view. Figure 137. FMNH 224107, borneoensis, left anterolateral view of trigeminal foramen. Figure 138. FMNH 224107, borneoensis, right anterolateral view of trigeminal foramen. Figure 139. FMNH 224122, borneoensis , right anterolateral view of trigeminal foramen. Figure 140. CAS 228378, Carolina, dorsal view Figure 141. CAS 228443, grandis, of pelvis. posteroventral view of otic region. 2004 Asiatic Herpetological Research Vol. 10, p. 101 As practicing vertebrate paleontologists, we also hope that our efforts here will stimulate additional inves- tigation and publication of the extensive fossil record of testudinoid turtles. Morphology is, of course, of para- mount importance for the interpretation of fossils. Our visits to many museums in the last several years revealed an abundance of unpublished testudinoid fossil material. Although a phylogenetic analysis was not a goal of this research, we adopt the convenient and now familiar means of summarizing morphological data by providing a character matrix that summarizes some of our observations. Although character data can be used to assist paleontologists in diagnosing fossil testudinoid specimens, we also encourage paleontologists to utilize our study as a starting point for basic morphologic descriptions. Description of differential diagnostic char- acters generally is an adequate minimum for the erection of a new taxon. However, such a diagnosis is, in itself, not particularly helpful to systematists trying to score a matrix and place a fossil into a broader phylogenetic context. Descriptions of new fossil specimens (and taxa) will be most useful if they include discussions of char- acter state data for all preserved anatomical regions. This current summary of morphological characters that traditionally are used in systematic treatments of testudi- noids can be used as a preliminary guide to the anatom- ical regions and features that would be most useful when included as part of a description of new fossil material. Acknowledgements We are greatly indebted to a number of institutions and their staff for providing generous access to skeletal and wet specimens: Jens Vindum (CAS), Stephen Rogers and John Wiens (CM), Alan Resetar and Maureen Kearney (FMNH), John Simmons and Linda Trueb (KU), Jim McGuire (LMNH), Jose Rosado (MCZ), Travis LaDuc and David Cannatella (TMM), Harold Dundee (TUMNH), and Gregory Caldwell- Watkins and Jacques Gauthier (YPM). We extend our thanks to Rick Van Dyke for the gift of specimens to support this research. Gabe Bever, Julien Claude, Jason Downs, Jacques Gauthier, Jason Head, Pat Holroyd, Howard Hutchison, Lyndon K. Murray, Jim Parham, and Krister Smith (in alphabetical order) provided useful comments and criticisms regarding this project. All images containing YPM specimens were reproduced with courtesy of the Peabody Museum of Natural History, Yale University, New Haven, CT. Special thanks go to Ted Papenfuss and Jim Parham for provid- ing us with the unique opportunity to publish the results of this study, and to Julien Claude and Howard Hutchison for their detailed and insightful reviews. The Brett Stems Award for Chelonian Research (CAS), the Visiting Scholar Fund (FMNH), and Yale University Graduate Fellowships provided funding to WGJ. Additional financial support was provided by the Geology Foundation of The University of Texas at Austin. Literature Cited Agassiz, L. 1857. Contributions to the Natural History of the United States of America. First Monograph. Part II. North American Testudinata. Pp. 235-452, Plates 1-7. Little, Brown and Company, Boston. Auffenberg, W. 1974. 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Occasional Papers of the Museum of Zoology, University of Michigan 647:1-24. 2004 Asiatic Herpetological Research Vol. 10, p. 105 Appendix 1 List of specimens used. agassizii, CAS 33867, FMNH 216746, FMNH 250843; alabamensis , CM 95968, CM 95991; amboinensis, CAS 153872, CAS 228345, CAS 228369, CAS 228412, CAS 228419, FMNH 224009; annandalei , FMNH 258876, FMNH 258879, FMNH Moll3036; annulata, CAS SUR7425, CM 87903, YPM 15410; arachnoides, MCZ 54050; areolatus , MCZ 42214; areolata , CM 47957, CM 87904; barbouri, CAS SUR12063, TUMNH 15400, TUMNH 15428, TUMNH 15429, TUMNH 16899; basket, FMNH 224095, FMNH 224097, FMNH 224213, FMNH 224124, FMNH 224226; bealei, CM 118554, FMNH 255270, FMNH 226542; belliana, CAS 228394, CAS 228404; berlandieri , TNHC 2813; blandingii, CAS 12837, CAS 228346, CAS 228373, CAS 228434, CAS 228448, YPM 14677, YPM 10382; borneensis , FMNH 224001, FMNH 224003, FMNH 224004, FMNH 224005, FMHH 224140; borneoensis, FMNH 224107, FMNH 224122, FMNH 224129, FMNH 251499, MCZ 42198; carbonaria , CAS 228397, CAS 228411, CAS 228427, CAS 228430; caretta, CAS 8383, FMNH 31021; Carolina , CAS 228375, CAS 228378; caspica, CAS 141118, CM 118517, FMNH 19714, FMNH 74505, FMNH 98764; chilensis, CM 112252; coahuila, KU 46929, KU 92623, MCZ 120335; crassi- collis, CAS 228335, CAS 228451, FMNH 11091, MCZ 7821, MCZ 134451; decor ata, CM 118590; dentata, CAS 134332, KU 47170, MCZ 29567, CAS 228333, CAS 228362, CAS 228414; elegans, CAS 228396; elon- gata , CAS 228408, FMNH 183740, FMNH 257382; emys, CAS 228344, FMNH 63749, FMNH 224034; ern- sti, TUMNH 13460, TUMNH 13462, TUMNH 16899; erosa, YPM 12653; flavimaculata , TUMNH 15375, TUMNH 15404, TUMNH 15747, TUMNH 15787; flavomarginata, CAS 18040, CAS 228356, CAS 228357, CAS 228359, FMNH 21651 5\flavomarginatus, MCZ 1 64926 \forsteni, CAS 228433; galbinifrons, CAS 228358, CAS 228360, CAS 228367, CAS 228376, YPM 14074, YPM 14080; geographica , CAS 12809, TUMNH 15391, TUMNH 15392, TUMNH 15410, TUMNH 15430; geometricus, MCZ 32184; gibba, CAS 228392; graeca, CAS 217732, CAS 228435, LMNH 54855, LMNH 54861; grandis , CAS 228341, CAS 228443, FMNH 224038, YPM 15431; guttata , CAS 8696, CAS 228372, CAS 228386, FMNH 211573; hamiltonii , MCZ 120333, YPM 10399; hermanni, CAS 228400, CAS 228401, CAS 228402; homeana , CAS 228409, CAS 228423, CAS 228428, CAS 228429, CAS 228455, FMNH 19794; horsfieldi , CAS 120707, CAS 228398, CAS 228399, CAS 228421, CAS 228425, YPM 16450; impressa, CAS 228416; G. insculpta, CAS 228406, CAS 228407, CAS 228413; japonica, YPM 15482, YPM 15486; kachuga, FMNH 224128, FMNH 224152; kleinmanni, CAS 228422, CAS 228426, CAS 228431; kohnii, TUMNH 10237, TUMNH 12121, TUMNH 14544, TUMNH 15678; leprosa, CM 137031; marmorata, CAS no number, CAS 188533, CAS 220052, FMNH 22076; mouhotii, CAS 228365, CAS 228374, CAS 228420, CAS 228444; muhlenbergii, LMNH 55352, MCZ 52248; mutica, LMNH 54883; P. nelsoni, CM 67311; nigra, CAS 8125, CAS 8289; nigrinoda , TUMNH 15147, TUMNH 15317, TUMNH 15408, TUMNH 15750; ocellata, CAS 228336; G. oculifera, TUMNH 26, TUMNH 3359, TUMNH 7548, TUMNH 12402, TUMNH 16928; P. oculifera , CAS 165598, CAS 220645, CAS 220646; odoratus, YPM 13622, CAS 228351, CAS 228352, CAS 228353; orbic- ularis, CAS 173223, CAS 228347, CAS 228415, CAS 228447, LMNH 10347, YPM 15479; T. ornata, CAS 228381, CAS 228382, CAS 228383, CAS 228384, TNHC 62532; ouachitensis, CM 61656, CM 84696; pardalis, CAS 148630, CAS 228349, CAS 228410, CAS 228418, CAS 228432, CAS 228449; petersi, CAS 8608, CM 124246, CM 124247; picta, CAS 13889, CAS 228379, CAS 228380, CAS 228385; N. platynota, CAS 228342, CAS 228450, CM 118586, FMNH 224216, YPM 14678; polyphemus, CAS 14090, LMNH 43344, LMNH 43354, LMNH 59320; pulcherrima, CAS 11754, CAS 228355, CAS 228366, CAS 228377; pul- chra, LMNH 48092; punctularia, YPM 465, CAS 228348; reevesii, CAS 31437, CAS 228361, CAS 228364; reticularia, CAS 228338, CAS 228388, LMNH 14565, LMNH 14569, LMNH 49145, LMNH 54856, LMNH 78580; rubida, CM 87907, CM 87908; rubriventris, CM 34409, CM 45188; scripta, CAS SUR8642, CAS 228436, CAS 228442, LMNH 15684; serpentina, CAS 228452, YPM 6369, CAS 228457; siebenrocki, CAS 228393; signatus, CAS 228405, MCZ 42217, MCZ 42218; sinensis, CAS 18031, CAS 228339; spengleri, CAS 21008, CAS 228331, CAS 228332, CAS 228343, CAS 228371, YPM 11653, YPM 14445; spinifera, CAS 65705, CAS 228350, CAS 228354, YPM 656; spinosa, CAS 228368. CAS 228459; subglobosa, CAS 228334; subtrijuga, CAS 16996, CAS 228445, CAS 228446, CAS 228453, CAS 228454; sub- rufa, CAS 228389, CAS 228390, CAS 228391; tchepo- nensis, CAS 228363, CAS 228370; tecta, CM 89923; tentoria, FMNH Moll3026, FMNH Moll3028, FMNH Moll3032, FMNH 259430; tentorius, MCZ 41944, MCZ 46604; terrapin, CAS 43640, CAS 228340, CAS 228387, YPM 2983; texana , CAS 30965, CAS 228437, CAS 228438, CAS 228439, CAS 228440, CAS 228441, CAS 228456, CAS 228458; thurjii, FMNH 224135, FMNH 224153, MCZ 62523, MCZ 62524, YPM 14072, YPM 14073, YPM 14074; tornieri, CAS 139704, CAS 228395, CAS 228403, CAS 228417, CAS 228424; tri- Vol. 10, p. 106 Asiatic Herpetological Research 2004 fasciata, CAS 228337, MCZ 5218; trijuga, CAS 12463, dussumieri CM 89921, CM 124227, YPM 15453; versa, TUMNH 4484, TUMNH 10510, TUMNH 16192 -yuwonoi, YPM elegans 12626. elongata emolli emys Appendix 2 ernsti Generic synonymies for all currently recognized testudi- erosa noid species and all outgroup species used herein. fern oral is Synonymies are based only on common usage during flavimaculata the last 50 years. flavomarginata flavomarginatus Ingroup floridana adiutrix Chrysemys, Pseudemys, Trachemys forsteni agassizii Gopherus, Xerobates funerea alabamensis Chrysemys, Pseudemys gaigeae amboinensis Cuora galbinifrons angulata Chersina, Goniochersus, Neotestudo, geographica Testudo geometricus annamensis Annamemys, Mauremys gibbonsi annandalii Hieremys graeca annulata Callopsis, Geoemyda, Nicoria, grandis Rhinoclemmys guttata arachnoides Pyxis hamiltonii areolata Callopsis, Geoemyda, Nicoria, hermanni Rhinoclemmys home an a areolatus Homopus horsfieldi aurocapitata Cuora impressa barbouri Graptemys, Malaclemys G. insculpta baska Batagur japonica bealei Clemmys, Sacalia kachuga belliana Kinixys kleinmanni berlandieri Gopherus kohnii blandingii Emydoidea, Emys, Neoemys leprosa borneensis Orlitia leytensis borneoensis Callagur lobatsiana boulengeri Homopus marginata caglei Graptemys m arm or at a callirostris Chrysemys, Pseudemys, Trachemys melanosterna carbonaria Chelonoidis, Geochelone mouhotii Carolina Terrapene muhlenbergii caspica Clemmys, Mauremys mutica chilensis Chelonoidis, Geochelone nasuta coahuila Terrapene natalensis concinna Chrysemys, Pseudemys nebulosa crassicollis Siebenrockiella P. nelsoni decorata Chrysemys, Pseudemys, Trachemys T. nelsoni decussata Chrysemys, Pseudemys, Trachemys nigra dentata Cyclemys, Geoemyda nigricans denticulata Chelonoidis, Geochelone nigrinoda depressa Geoemyda, Heosemys ocellata dhongoka Kachuga G. oculifera diademata Geoemyda, Rhinoclemmys P. oculifera dorbigni Chrysemys, Pseudemys, Trachemys oldhamii Aldabrachelys, Dipsochelys, Geochelone, Megalochelys Geochelone Geochelone, Indotestudo Chrysemys , Pseudemys, Trachemys Geochelone, Manouria Graptemys Kinixys Homopus Graptemys, Malaclemys Cistoclemmys, Cuora, Geoemyda Gopherus Chrysemys, Pseudemys Geochelone, Indotestudo, Manouria Callopsis, Geoemyda, Rhinoclemmys Chrysemys, Pseudemys, Trachemys Cistoclemmys, Cuora Graptemys, Malaclemys Psammobates Graptemys Testudo Geoemyda, Heosemys Clemmys Geoclemys Testudo Kinixys Agrionemys, Testudo Geochelone, Manouria Calemys, Clemmys, Glyptemys Clemmys, Mauremys Kachuga Pseudotestudo, Testudo Graptemys Clemmys, Mauremys Geoemyda, Heosemys Kinixys Testudo Clemmys, Emys Geoemyda, Rhinoclemmys Cuora, Cyclemys, Geoemyda, Pyxidea Calemys, Clemmys, Glyptemys Clemmys, Mauremys Callopsis, Geoemyda, Rhinoclemmys Kinixys Chrysemys, Pseudemys, Trachemys Chrysemys, Pseudemys Terrapene Chelonoidis, Geochelone Chinemys Graptemys, Malaclemys Morenia Graptemys, Malaclemys Psammobates Cyclemys 2004 Asiatic Herpetological Research Vol. 10, p. 107 orbicularis Emys yaquia Chrysemys, Pseudemys, Trachemys T. ornata Terrapene yniphora Asterochelys, Geochelone P. ornata Chrysemys, Pseudemys, Trachemys yunnanensis Cuora ouachitensis Graptemys yuwonoi Geoemyda , Leucocephalon, pani Cuora Notochelys par dal is Geochelone zhoui Cuora peter si Morenia pi eta Chrysemys Outgroup planicauda Acinixys, Pyxis caretta Caretta G. platynota Geochelone dumerilianus Peltocephalus , Podocnemis N. platynota Notochelys C. insculpta Carettochelys polyphemus Gopherus, Xerobates fimbriatus Chelus pseudogeographica Graptemys, Malaclemys gibba Phrynops, Mesoclemmys pulcherrima Callopsis, Geoemyda, Rhinoclemmys madagascariensis Erymnochelys, Podocnemis pidchra Graptemys, Malaclemys megacephalum Platysternon pulchristriata Cy cl emys odoratus Kinosternon, Sternotherus, punctularia Geoemyda, Rhinoclemmys Sternothaerus quadriocellata Clemmys, Sacalia scorpioides Kinosternon radiata Asterochelys, Geochelone serpentina Chelydra reeves ii Chinemys siebenrocki Chelodina, Macrochelodina reticularia Deirochelys spiniferaApalone, Trionyx rubida Callopsis, Geoemyda, Nicoria, subglobosa Emydura, Tropicochelymys Rhinoclemmys subrufa Pelomedusa rubriventris Chrysemys, Pseudemys temminckii Macroclemys, Macrochelys scripta Chrysemys, Pseudemys, Trachemys signatus Homopus silvatica Geoemyda, Heosemys Appendix 3 sinensis Ocadia Distribution of character states for 70 characters among smithii Kachuga, Pangshura 46 species of testudinoid turtles; a = 0/1 ; b = < 0/2; c = 0/3; spekii Kinixys d = 0/1/2; e = 1/2; f = 1/3; g = 2/3. spengleri Geoemyda spinosa Geoemyda, Heosemys agassizii stejnegeri Chrysemys, Pseudemys, Trachemys llalO 00000 000?0 11010 00110 00001 subtrijuga Malayemys 01000 03200 00000 00120 22001 22200 sulcata Geochelone 00000 10110 sylhetensis Kachuga, Pangshura amboinensis taylori Chrysemys, Pseudemys, Trachemys OOaOO 00010 00001 00000 00010 00000 tcheponensis Cy cl emys, Geoemyda o o o o o o o 00200 OaOll 11110 00001 tecta Kachuga, Pangshura 20000 00000 tentoria Kachuga, Pangshura annandalei tentorius Psammobates 0010? 00020 00011 10001 00010 00000 terrapen Chrysemys, Pseudemys, Trachemys 00100 01100 00000 00a 10 34000 00001 terrapin Malaclemys 00000 0000? texana Chrysemys, Pseudemys annulata thurjii Hardella OOaOO 00010 0010a 00000 00010 00000 tornieri Malacochersus 00000 03100 00000 000?0 11000 0??01 travancorica Geochelone, Indotestudo 00000 00010 tricarinata Geoemyda, Melanochelys barbouri trifasciata Cistoclemmys, Cuora 00001 11000 01010 10000 10000 101 aO trijuga Geoemyda, Melanochelys 00001 10000 00000 00000 34000 22200 trivittata Kachuga 00000 00001 venusta Chrysemys, Pseudemys, Trachemys baska versa Graptemys, Malaclemys 00001 00000 00010 10001 00000 00a02 werneri Testudo 11100 00010 00000 10110 43000 00000 Vol. 10, p. 108 Asiatic Herpetological Research 2004 00000 01000 00100 00010 bealei guttata 00000 00000 00010 00000 10000 00010 00000 00010 00000 00000 00000 10000 00000 00000 0??00 OaOlO 34000 0000? 00000 00000 00000 00000 34000 10201 00000 00000 00000 00000 blandingii hamiltonii 00110 00000 00000 00070 00000 10000 00000 01101 00001 00000 10000 00010 00000 OcOOO 00000 01001 11110 1120a 00111 00000 00000 OOalO 33001 0000a 10011 00000 00000 00000 borneensis homeana 00100 00000 00010 00000 00010 10000 11010 00000 00071 11000 00111 00000 00000 00000 00010 00a 10 33000 00000 00001 0110a 00000 00020 22001 77? la 00000 00000 20000 10110 borneoensis horsfieldi 00001 00000 00010 10001 00010 00001 11010 00000 00070 11000 aOlll 00001 11100 00000 00100 OOalO 43000 00000 00000 03200 00000 00020 22000 77711 00000 00000 00000 11110 carbonaria kachuga 11010 00000 000?a 11000 00111 00001 00101 00000 00010 10001 00010 10001 00000 01201 00000 00120 22001 22210 11100 00000 01000 10110 43000 00000 00000 10110 00000 0000? caspica mouhotii 00000 00010 0000a 00000 00010 00000 00010 00010 00000 11100 00010 00000 00000 00000 00000 OaalO 34000 00001 00011 01100 00000 00011 11110 00001 00000 00000 aOOOO 00010 crassicollis P. oculifera OOaOO 00000 00000 10000 00010 10000 11010 00000 0007? 11000 00111 00001 00001 00000 00010 OOalO 33000 00001 00001 01200 00000 00120 22001 77710 00000 00000 00000 10110 elongata orbicularis 11010 00000 0007a 11000 00111 00001 00000 00000 00000 00000 aOOOO 00000 00000 03200 00000 00020 22001 77711 00000 00000 00000 OaOOl 11010 1120a 00000 10110 10010 00000 dentata T. ornata 00000 00010 00000 00000 00000 00000 00010 00010 00000 00000 10000 00000 00001 01100 00001 00011 11000 01101 00000 00010 00200 00001 00110 ed201 00100 00000 20011 00010 emys par dal is 11010 00000 000?a 11000 00110 00001 11010 00000 0007a 11000 00111 00001 00000 01000 02000 OOaaO 22001 77700 00000 0f201 00000 00120 22001 77710 00000 10110 00000 10110 flavomarginata peter si 00010 00020 0000a 00000 00010 00000 01100 00000 00010 10001 00010 01111 00000 oino 00100 00011 11110 71101 12100 02000 00100 00110 34000 00000 21000 00010 00000 00001 graeca picta 11010 00000 00071 11000 10111 00001 00001 10000 00010 00000 10000 00000 00000 03200 00000 00a20 22000 22211 00000 00000 00000 OaOaO 34000 2220a 00000 10110 00000 0000a grandis N. platynota 0010a 00020 00000 00000 OOOaO 00000 01101 00010 00000 00000 OOalO 00000 00001 01100 0000a OOalO 34000 00001 00001 00000 10100 00001 03010 00101 2004 Asiatic Herpetological Research Vol. 10, p. 109 10000 00000 polyphemus 11010 00000 00070 11000 aOUO 00001 01000 0a200 00000 00120 22001 22200 00000 10110 pulcherrima OOaOO 00010 00001 00000 aOOlO 00000 00000 01100 00000 OOOaO 14000 01701 00000 00000 reeves ii 00101 10000 00000 10000 00010 10110 00110 00000 00000 OOaaO 33000 00001 00000 00000 reticularia 00000 10000 00070 00000 10000 00000 00000 00000 00000 OlOaO 34000 22200 10000 00001 scripta 00001 10000 00010 10000 10000 00101 10000 00000 00000 00000 34000 22200 00000 0000a sinensis 00001 00000 00010 10001 10010 00001 00000 00000 00000 OOaaO 33000 00001 00000 00000 spengleri OOOaO 00000 00100 10100 00010 00000 00011 01100 00000 00000 34000 00001 00000 00010 subtrijuga 00100 01100 11000 10000 01010 10100 00110 OOgOO 00000 00a 10 33000 00000 00000 00000 tentoria 00001 00000 00010 10001 00010 10001 10100 10000 01000 10110 43000 00000 00000 0000? terrapin 00001 10000 00010 10000 aOOOO 10100 00000 00000 00000 OaOOO 34000 ed200 00000 00001 texana 00001 10000 OaOlO 10000 aOOOO 00101 10000 00000 00000 00000 34000 22200 00000 0000a thurjii 00001 00000 00010 10001 00000 01011 11100 00000 00100 00110 43000 00000 00000 tornieri 00001 11010 00000 00071 11000 00111 00001 00000 0??00 00000 00020 22000 2220a 10110 2004 Asiatic Herpetological Research Vol. 10, pp. 110-113 Trade Data and Some Comments On the Distribution of Mauremys annamensis (Siebenrock, 1903) Minh Le1’*, Thang Hoang2, and Due Le3 1 American Museum of Natural History, Department of Herpetology, Central Park West at 79th Street New York, NY 10024 -19th Le Thanh Tong St., Hanoi, Vietnam 3 WWF Indochina Programme, 53-Tran Phu St., Ba Dinh, Hanoi, Vietnam; IPO Box 151, Hanoi, Vietnam * E-mail: minhl@amnh.org Abstract. - This trade survey of Annam Pond Turtle reveals that this species is likely to have larger distribution than previously thought. The records in the trade in Quy Nhon and Ho Chi Minh City suggest its range could extend much further south. In addition, given the one way south-north trade route, the absence of Mauremys mutica in the trade south of Hai Van Pass and the reported absence of M. annamensis in the trade north of the Pass support the hypoth- esis that the Pass is the natural barrier for the two species ranges. This hypothesis combined with the long existence of the Pass might indicate that the speciation between the two species happened when their ancestors dispersed across the Pass, and were subsequently isolated, by the means of rafting or walking through narrow land strip emerged dur- ing the low sea level period. In terms of conservation, M. annamensis has become much rarer even in the trade, sug- gesting immediate conservation measures to protect it. Key words. - Mauremys annamensis, Bataguridae, Hai Van Pass, Truong Son Range, distribution, biogeography. Distribution of Mauremys annamensis. - The Annam Pond Turtle, Mauremys annamensis, was first described by Siebenrock in 1903 based on a specimen collected from Phuc Son or Phuoc Son (15° 33' 00" N; 108° 04' 00" E) (southwest of Tourane, now known as the city of Da Nang) in Central Vietnam. Another specimen was collected by Bourret from Fai Fo (Hoi An), an ancient city about 50 km from Da Nang (Bourret, 1941). Since then, it seems that little effort has been made to record the distribution of this species in the wild. Iverson (1992) and Iverson et al. (1999) cite only the above records from Bourret for their maps of global turtle dis- tribution. According to Bourret (1941), this species was very abundant in the marshes and slow-moving water bodies in the lowland areas of the cities of Hoi An and Da Nang. Both Hoi An and Da Nang, however, are now very populated cities surrounded by rice paddies, which are unlikely to be suitable habitats for this species. This is because the intensive use of chemicals, such as herbi- cides and pesticides, in rice paddies through out Vietnam makes it difficult for turtles to survive in this environ- ment. To better understand its distribution, we did a 6-day trade survey in the August of 1996. The survey covered three cities, namely Quy Nhon (13° 46’ 00” N; 109° 14’ 00” E), Da Nang, and Hue (16° 28' 00" N; 107° 36’ 00" E), and their surrounding areas. We interviewed turtle dealers at the collecting points, where turtles were bought from collectors and awaited to be shifted to China. We used the book Turtles of the World (Ernst and Barbour, 1989) for identification key. In addition, since the initial purpose of the survey is to determine the trade status of M. annamensis, we identified, but did not record the availability of other turtle species in the col- lecting points. We observed that this species was still common in the trade in Quy Nhon and Da Nang. In Quy Nhon and its outskirt, we visited three collecting points. In the first one, we identified 2 adult M. annamensis. In the other two collecting points, we found 3 and 4 juveniles, respectively. In Da Nang and its surrounding areas, we visited four sites with three to four specimens in each sites. They were all young and juvenile turtles. From the interviews with local people in Quy Nhon and Da Nang, it was apparent that this species could well survive in the human-modified environment, such as lakes and fish- ponds, if there were no collecting activities by local peo- ple. Local trade dealers and collectors, encountered in collecting points in Quy Nhon and Da Nang, suggested that this species still existed in the water bodies in the nearby region. Le and Trinh (2001) also indicated that the species could occur in Tra My, Tien Phuoc, and probably Hiep Due Districts, Quang Nam Province. The occurrence of M. annamensis in Quy Nhon is very interesting since this species had been believed to have very restricted distribution. More remarkably, Le and Broad (1995) reported that M. annamensis even extended far south to Ca Mau, Minh Hai Province inhab- iting Melaleuca forests (around 9° 29' 00" N; 105° 20' © 2004 by Asiatic Herpetological Research Vol. 10, p. Ill Asiatic Herpeto/ogical Research 2004 Map scale: 1:15,000,000 Figure 1. Topographic map of Vietnam. 00" E;) situated at the southern tip of Vietnam. It is pos- sible that Le and Broad misidentified this species in their survey (Jenkins, 1995). However, Peter Paul van Dijk, Le Trong Dat, and Douglas Hendrie on May 30, 2000 found this species in Ben Chuong Duong Street shops in Ho Chi Minh City (Hendrie, 2000a). This evidence com- bined with the one-way trafficking of turtles (from south to north) (Le and Broad, 1995) indicates that M. anna- mensis may have much wider distribution than previous- ly thought. This hypothesis is also supported by Le and Trinh (2001). Their interviews with local dealers, in May 2001, in Quang Nam Province revealed that one dealer in Thang Binh District bought this species from the ship- ments transported from the south. Le and Trinh (2001) also suggested that A/, annamensis is naturally distrib- uted in Tra My, Tien Phuoc, and probably Hiep Due Districts, Quang Nam Province. These authors also indi- cated an interesting fact that turtle hunters only sell their animals to the local dealers. This manner of trade demonstrates that the local trade data can be informative in determining the limit of the natural ranges of some species. It is noted that we did not encounter any M. mutica in the areas in Quy Nhon and Da Nang. Le and Trinh (2001) also did not find any M. mutica in their trade investigation, which covered seven districts and one town in Quang Nam Province, and the city of Da Nang. According the pattern of turtle trade in Vietnam plotted by Le and Broad (1995), turtles have been collected from the south and transported to the north. They are finally destined in the border between and Vietnam and China, where they are traded in an enormous volume. The one-way south north trafficking leads to the conclu- sion that M. mutica does not occur in Da Nang, located in the southern side of Hai Van Pass, or southern areas of the city. In the survey, we found no evidence for M. anna- mensis distributed in the northern side of the Hai Van Pass. Interviews with turtle dealers from two collecting points in the city of Hue (north of Hai Van Pass) revealed that this species was only transported to the city' from the south and there was no record of this species in the surrounding areas. It is also interesting that the deal- ers called this species Rua Dep Nam (Beautiful Southern Turtle) as compared to Rua Dep Bac (Beautiful Northern Turtle), here referred to Mauremys mutica. According to them, M. mutica came only from areas north of Hai Van Pass. In the house of a trader in Hue, we observed about 20 M. mutica from 1 to 2 kg, but no M. annmensis. In the other site, we did not find any M. mutica or M anna- mensis. Given the fact that Hue and Da Nang is only 100 km apart, it is very likely that Hai Van Pass (at around 16°N) forms the natural boundary of these two species since the Pass stands in between two cities. Records from pre- vious studies also support this hypothesis (Iverson. 1992; Nguyen and Ho, 1996). Thus, the mountain range, which cuts through the country, is most certainly the northern boundary of M. annamensis' range and south- ern boundary of M. mutica's range. Because they are the lowland inhabitants, the Pass (1712m above sea level at the summit) seems to be a significant barrier. Some studies have suggested that the Pass is the border between two zoological regions. Northern Central Vietnam (Northern Truong Son) and Central Vietnam (Central Truong Son) for such groups as rodent, bird, fish, and lizard (Bobrov, 1993; Dao, 1978. Cao, 1989). In addition, Fooden (1996) showed in Figli that the distribution two closely related gibbon taxa, Hylobates gabriellae siki and H. gabriellae gabriellae , has been separated by the Pass. Even though primate is more likely to possess higher dispersal ability, the barri- er seems significant enough to block their expansion. In 2004 Asiatic Herpeto/ogical Research Vol. 10, p. M2 fact, the Truong Son Mountain Range in general has established dispersal barriers for Cuora galbinifrons species complex (Stuart and Parham, 2004). Hai Van Pass is a part of Truong Son Mountain Range (Annamite Mountains), which runs throughout most of the country’s length. The Pass meets the South China Sea and effectively divides the country into two different sections. It is formed by aluminous granite, and probably emerged about 250 Mya in the early Triassic (Lepvrier et al., 1997). If the Pass is actually the natu- ral boundary of these two species’ ranges, it can be hypothesized that the speciation between M annamensis and M mutica was occurred when their ancestors dis- persed across the Hai Van Pass and then were subse- quently isolated. Since these turtles are good swimmers, one possible scenario is that they rafted on the sea to get to the other side of the Pass. Another possibility is that they traveled south through a narrow land belt exposed during the low sea level period. According to Prentice and Denton (1988), before from 6 Mya to 0.9 Mya the sea level fluctuated at an average of 70m below the pres- ent sea level. At this level, a few kilometers of the con- tinental shelf could be opened to the east of the Pass (Voris, 2000) and well served as a travel route for these turtles. However, these hypotheses are very preliminary and, therefore, should be carefully tested in a much more comprehensive study in the future. Trend in the trade of M. annamensis. - In recent years, turtles in the Southeast Asian region, especially in Vietnam, have been critically threatened by the trade with China. The trade has been driven by the Chinese long tradition of using turtles as food and medicines. Many species might go extinct in very near future unless urgent protection measures are implemented (van Dijk, 2000; Hendrie, 2000). In Vietnam, species such as Cuora trifasciata , believed to be able to cure cancer, is at considerable risk due to its significant economic value (about 1 000 USD per individual or even more (Lovich et al ., 2000; Le and Trinh, 2001)). For M. annamensis , in addition to the general demand from China, there are also interests in keeping them as pets in countries such as the U.S. The United States Fish and Wildlife Service indicated that from 1996 to 1999 small numbers were imported to the U.S. from Vietnam (Consideration of Proposals for Amendment of Appendices I and II). Weissgold (2002) even maintained that the imports increased during 1999-2001. Due to the risk posed by the trade and habitat destruction, this species has been listed in the Appendix II and in the critically endangered category by CITES and IUCN, respectively. It is clear that the number of M. annamensis has declined dramatically in recent years. The market value of this species, approximately $5 to $7 per kg (Le and Broad, 1995; Le and Trinh, 2001; and this survey), can generate substantial interests among poor local people. In our survey in 1996, this species was still pretty com- mon in the trade. My personal observation in Dong Xuan market, Hanoi, in 1998 also confirmed the com- monness of this species. More recently, Hendrie (2000b) reported that the occurrence of this species in turtle con- fiscated shipments is less frequent compared to the pre- vious years. Le and Trinh (2001) reported that this species was very rare in the trade compared to other species — only second to Golden Turtle ( Cuora trifasci- ata). In fact, they only encountered only one juvenile in the whole period of the survey. Thus, this species should be given the highest priority in conservation programs in the near future. Acknowledgments We would like to thank Drs. Ross Kiester and James Juvik for their support of the field trip. Bryan Stuart encouraged and convinced ML that the information in this paper is important. Comments from two anonymous reviewers and Jim Parham help improve the paper. ML is supported by a NASA grant (No. NAG5-8543) to the Center for Biodiversityand Conservation at the American Museum of Natural History and a Columbia University/CERC Faculty Fellowship. Literature Cited Bourret, R. 1941. Les tortues de l’lndochine. Bulletin de Flnstitut Oceanographique de FIndochine 38:1-235 Bobrov V. V. 1993. Zoogeographic analysis of the lizard fauna (Reptilia, Sauria) of Vietnam. Zoologicheskii-Zhumal 72(8): 70-79 (in Russian, English summary) Cao, V. S. 1989. On the problem of zoogeographical division of the rodent fauna of Vietnam (Mammalia, Rodentia). Vertebrata-Hungarica 23:57-66 Consideration of Proposals for Amendment of Appendices I and II. WWW at http://www.cites.org/eng/cop/12/prop/E12-P21.pdf Dao, V.T. 1978. An experience of zoogeographical regionalization of Vietnam. Zoologicheskii-Zhurnal 57:582-586 (in Russian, English summary) Ernst, C. H. and R. W. Barbour. 1989. Turtles of the World. Smithsonian Institution Press, Washington D.C. and London. 313 pp. Vol. 10, p. 113 Asiatic Herpetological Research 2004 Fooden, J. 1996. Zoogeography of Vietnamese primates. International Journal of Primatology 17(5): 845- 899 Hendrie, D. B. 2000a. Compiled notes on the wildlife trade in Vietnam - January-May 30, 2000. Report to TRAFFIC Southeast Asia. Hendrie, D. B. 2000b. Status and conservation of tortoi- ses and freshwater turtles in Vietnam. Pp. 63-73. In P. P. van Dijk, B. L. Stuart, and A. G. J. Rhodin (eds.), Asian Turtle Trade: Proceedings of A Workshop on Conservation and Trade of Freshwater Turtles and Tortoises in Asia. Chelonian Research Monographs, Number 2. Iverson, J. B. 1992. A revised checklist with distribution maps of the turtles of the world. Privately pub- lished. 374 pp. Iverson, J. B., A. J. Kimerling, A. R. Kiester., L. E. Hughes, and J. Nicolello. 1999. World turtle databa- se. Website at http://emys.geo.orst.edu Le, D. D. and S. Broad. 1995. Investigations into Tortoise and Freshwater Turtle Trade in Vietnam. IUCN Species Survival Commission, IUCN, Gland, Switzerland and Cambridge, UK. 34pp. Le, T. D. and N. L. Trinh. 200 1 . Status of the Vietnamese turtle ( Mauremys annamensis Siebenrock, 1903) in the wild and in the trade in Quang Nam and Da Nang. Cue Phuong Conservation Project. (Unpublished Report). Lepvrier, C., H. Maluski, V. V. Nguyen, D. Roques, V. Axente, and C. Rangin. 1997. Indosinian NW- trending shear zones within Truong Son belt (Vietnam): 40Ar-39Ar Triassic ages and Cretaceous to Cenozoic overprints. Tectonophysics 283:105- 127. Lovich, J. E., R. A. Mittermeier, P. C. H. Pritchard, A. G. J. Rhodin, and J. W. Gibbons. 2000. Powdermill conference: Trouble for the world’s turtles. Turtle and Tortoise Newsletter 1:16-17 Nguyen, V. S. and C. T. Ho. 1996. [Checklist of Herpetofauna of Vietnam]. Scientific and Technical Publishing House, Hanoi. 264pp. (In Vietnamese). Prentice, M. L. and G. H. Denton. 1988. Deep-sea oxy- gen isotope record, the global ice sheet system, and hominid evolution. Pp 383-403. In F. Grine (ed), The Evolutionary History of the Robust Australopithecines. De Gruyter, New York. Stuart, B. L. and J. F. Parham. 2004. Molecular phyloge- ny of the critically endangered Indochinese box tur- tle ( Cuora galbinifrons). Molecular Phylogenetics and Evolution 31:164-177. Siebenrock, F. 1903. Schildkroten des ostlichen Hinterindien. Anzeiger der Kaiserlichen Akademie der Wissenschaften in Wien 1 12(1 ):333-353 . Van Dijk, P. P. 2000. The status of turtles in Asia. Pp. 1 5- 23. In P. P. van Dijk, B. L. Stuart, and A. G. J. Rhodin (eds.), Asian Turtle Trade: Proceedings of A Workshop on Conservation and Trade of Freshwater Turtles and Tortoises in Asia. Chelonian Research Monographs, Number 2. Voris, H. K. 2000. Maps of Pleistocene sea levels in Southeast Asia: shorelines, river systems and time durations. Journal of Biogeography 27:1153-1167 Weissgold, B. J. 2002. Turtle trade in North America: Legal requirements and trade trends. Report and presentation presented at the technical workshop on conservation and trade in freshwater turtles and tor- toises in Asia, Running, Yunnan Province, China. 2004 Asiatic Herpetological Research Vol. 10, pp. 1 14-1 1 9 1 Neotype of Testudo terrestris Forsskal, 1775 (Testudines, Testudinidae) Jarmo PeralA1 and Roger Bour2 ^ Department of Biological and Environmental Sciences, PO Box 65 (Biocenter 3, Viikinkaari 1), FIN-00014 University of Helsinki, Finland; E-mail: jarmo.perala@helsinki.fi ^ Laboratoire des Reptiles et Amphibiens, Museum national d’Histoire naturelle, 25 rue Cuvier, 75005 Paris, France; E-mail: bour@mnhn.fr Abstract. - We discuss and clarify the nomenclatural status of Testudo terrestris Forsskal, 1775. Testudo terrestris, a taxon based on a syntype series, was until now a valid name for tortoises from parts of Egypt, Arabia and the Levant, according to the International Code of Zoological Nomenclature. On the contrary, all type locality restrictions from the 20th century regarding the name Testudo terrestris Forsskal, done without the fixation of a name-bearing specimen (lectotype or neotype), are invalid. Because the name is valid, and because the syntypes, listed but not identified in the original description, are untraceable, and to permanently fix the name and its type locality, we designate a neotype for Testudo terrestris Forsskal based on a specimen from Aleppo, Syria. Key words. - Taxonomy, Testudines, Testudinidae, Testudo terrestris Forsskal, 1775, nomenclatural validity, neotype. Forsskal and Mediterranean tortoises. - During his fatal travel (1761-1763) in the eastern Mediterranean countries, the Finn Petter Forsskal (1732-1763) encoun- tered tortoises, briefly described by the name Testudo terrestris in his diary, which was published as a posthu- mous work by Carsten Niebuhr (1733-1815), a German traveller and surveyor, in 1775 [= Forsskal, 1775]. Strauch (1862: 69) remarks in a footnote that Forsskal (1775) discusses a tortoise, Testudo terrestris, which occurs in Aleppo and Lebanon, without describing it, and Anderson (1896: 68) writes that “it is useless attempting to identify this animal” “from Loheia, north of Hodeida,” with reference to Forsskal (1775). Forsskal ’s itinerary took him through the following localities: Malta, Smyrna (Izmir), Constantinople, Alex- andria, Rosetta, Cairo, Suez, Jedda, ending up in Yarim, Arabia Felix (Yemen), where Forsskal died of malaria in 1763. Tortoises were reported from A1 Lufyayyah (Yemen), Cairo (Egypt), Lebanon, Lattakia (A1 Ladhiq -Tyah) and Aleppo (both in Syria) (Fig. 1). Forsskal (1775) indicated that the chelonians are called “Zol- hafae” by the Arabs (Forsskal, 1775); as for their distri- bution, he specified: “Kahirae non frequens vivit, Aleppo autem & ad Libanon copiosor,” translated by Daudin (1801: 225) as: “It is rare in Cairo, but can be found rather abundantly near Aleppo and towards Mount Lebanon”. At best, Forsskal himself was thus able to see tortoises in situ in northern Egypt (Nile Delta and Suez area) only, that is those which are actually called either Testudo kleinmanni Lortet, 1883 (the Egyp- tian tortoise) or the vicariant T. werneri Perala, 2001. This interpretation is in accordance with the vernacular name of the Egyptian tortoise, Anderson (1898: 30; “Sohlafa” pronounced “Zihlifa”) and Flower (1933: 742; “Educated people in Egypt employ the word ‘Zal- heefah’”). Niebuhr’s subsequent route back home took him via Basra, Baghdad, Mosul, Diyarbakir, Aleppo, Lebanon and Palestine (Hansen, 1962; Niebuhr, 1772, 1973). Niebuhr, the only surviving member on the expe- dition to Arabia Felix, continued to Bombay (Mumbai), India, from where he shipped the natural history notes (Niebuhr, 1772: xix) and specimens (Niebuhr, 1973: 120) collected by Forsskal via London to Copenhagen. A considerable amount of material in the Forsskal col- lection in Copenhagen was destroyed in bombings by the British in 1807 (Nielsen, 1993). The type of T. ter- restris is said to be "apparently lost" (Webb in Iverson, 1992). The name is not fixed to a single name-bearing specimen. Nomenclatural validity of Testudo terrestris. - Testudo terrestris Forsskal, 1775, one of the oldest species group names in the genus Testudo Linnaeus, 1758 sensu lato (in the sense of Lapparent de Broin, 2001; Perala 2002a), is nomenclaturally valid, according to the present International Code of Zoological Nomenclature (ICZN, 1999): it was published before 1931 and thus does not need to have a full diagnosis by which it can be identified. Only “a description or a definition” per se is needed (ICZN, 1999: Article 12). In this respect the remark by Forsskal (1775) pertaining to the tortoises’ length (“...foot-long...”), as well as that made of the shapes of plastra of both sexes, is enough, even if these characters were incorrect. Moreover, although the name © 2004 by Asiatic Herpetological Research Vol.10, p. 1 15 Asiatic Herpeto/ogical Research 2004 Figure 1. Map of geographical localities mentioned in Forsskal's (1775) description, which together constituted the type locality of Testudo terrestris Forsskal until the present work. is accompanied by the remark “Obs.” (Forsskal, 1775: 12), it is also listed under the headings “Descripta” (p. viii) as well as “Nominata” (p. ix) alongside with valid names such as Testudo triunguis Forsskal, 1775, that is, current Trionyx triunguis. The book contains a multitude of other descriptions of valid taxa which cannot be rejected based on the arguments (see below) about the use of the Latin language, a contemporary standard, alone. Based on Niebuhr’s subsequent route back home, Gasperetti et al. (1993) suggested that it may actually be Niebuhr who is responsible for the name T. terrestris Forsskal. As Niebuhr’s role is not explicit in the original publication, the authorship of the name remains with Forsskal (ICZN, 1 999: Art. 50.1.1 ). Testudo terrestris overlooked, rejected, then revali- dated. - After its publication, and during about 180 years, the nominal species Testudo terrestris Forsskal, 1775, although nomenclaturally valid, was nevertheless overlooked by most authors (except Strauch, 1 862 and Anderson, 1896), some of them (Gray, 1831; Dumeril and Bibron, 1835) only mentioning Testudo zolkafa and Testudo zohalfa , respectively, based on Forsskal’s (1775) vernacular Zolhafae, and which nornina nuda appear in the above works as synonyms of Testudo graeca Linnaeus, 1758 and Testudo mauritanica Dumeril and Bibron, 1835, respectively. On the other hand, at least one scientist worked to revive the nominal species. A brief history of the case is presented by Bour in David (1994: 86). The name Testudo terrestris Forsskal was resurrected, and therefore revalidated, by Wermuth (1956: 402), who improperly (without speci- men fixation) designated “Arabia” as type locality. Simultaneously, Wermuth considered to ask the Interna- tional Commission on Zoological Nomenclature to invalidate the nominal species Testudo terrestris Forsskal; unfortunately, his attitude changed radically (1958: 149-153) and he used this name to designate the Near Eastern population of Testudo (as Testudo graeca terrestris Forsskal), restricting the type locality to “Libanon-Gebirge, Israel” [sic], and mistakenly extend- ing its range to Libya. It must be outlined that there is a gap in the range of Testudo graeca ( sensu lato) complex tortoises between Israel and Libya, the latter containing the range of the recently described species Testudo cyrenaica Pieh & Perala, 2002. Wermuth had at his dis- posal one Libyan specimen (SMF 36127; paratype of T. cyrenaica ), from Dernah, which he thought to be identi- cal with Middle-Eastern tortoises (Pieh & Perala, 2002). Wermuth’s validation was disapproved by Buskirk (in Ernst et al., 2000; unpublished manuscript from the early 1990s; and pers. comm, to both authors); H igh- field (in Ernst et al., 2000; and: http: //www.tor- toisetrust.org/articles/newfloweri.html); J. F. Parham (pers. comm.); and Perala (1996); among others. Their opposition is based on several arguments such as: a description is lacking with reference to the remark “Obs.” in Forsskal (1775); the species cannot be identi- fied from the “description”; there is no type specimen: a “false” type locality; the name “ Testudo terrestris ” is just Latin for a terrestrial chelonian, used without inten- tion to describe a new species. Need of a neotype. - Rather to resurrect Testudo terres- tris Forsskal (Wermuth, 1956). it would have been pref- erable: (1) either to suppress this imprecise name (and which is a more recent homonym of the nominal species Testudo terrestris Fermin, 1 765, for which Wermuth had to successfully request the invalidation by the Interna- tional Commission on Zoological Nomenclature; ICZN, 1963: Opinion 660); (2) either to use it to name Testudo kleinmanni , a name revalidated only in the 1950s by Mertens & Wermuth (1955), and independently by Lov- eridge & Williams (1957). It could still be possible to ask the ICZN to officially suppress the name Testudo 2004 Figure 2. Neotype of Testudo terrestris Forsskal, 1775, specimen n° NMW 18674: 2, sub-adult female. Upper left: lat- eral view (right side); Upper right: dorsal view; Lower left: ventral view. Lower right: living specimen of Testudo terres- tris, adult female, from Aleppo (topotype); CL = 185 mm, Ml = 143 mm, HE = 99 mm. terrestris Forsskal, 1775. However, we reject this option because: (1) the name is presently widely used (in all recent check-lists, with more than fifty references), although with vagueness about the identity of the con- cerned population; (2) there is no valid name to desig- nate the species of Testudo living in the Middle-East, more precisely in the area of the upper Euphrates - Tigris drainage; (3) the name Testudo terrestris Forsskal, 1775 became available by the very ruling of the International Commission on Zoological Nomencla- ture (ICZN, 1963), and it is unlikely that the Commis- sion would reverse its opinion. Accepting the nomenclature proposed by Wemuith, one of us (RB) proposed to emend the type locality of Testudo terrestris Forsskal to the vicinity of Aleppo (= Halab), Syria (Bour, 1989: 14), in the interest of clarifying the status of this taxon and as a first step towards the description of a neotype. However, such restriction of the type locality, as well as the earlier restrictions proposed by Wermuth (1956, 1958), are invalid according to the Code (ICZN, 1999; Art. 76.3), because these actions were not done in connection with the selection of a lec- totype or neotype. Therefore we here propose the formal description of a neotype of Testudo terrestris Forsskal, 1775. All the animals listed (although not identified; therefore untraceable) by Forsskal (from Al-Luljayyah, Cairo, Lebanon, Lattakia and Aleppo) actually represent syntypes, according to the Code (ICZN, 1999; Art. 72. 1 . 1 ), and thus the type locality of T terrestris encom- passes the region containing all those localities (ICZN, 1999; Art. 73.2.3). A neotype could legitimately be selected from any of those per se. Besides sea turtles, only Centrochelys sulcata (Miller, 1779), a land tortoise (possibly introduced), and Pelomedusa subrufa (Lacepede, 1788), a fresh-water turtle, are known to occur in Yemen (Obst & Wranik, 1987; Gasperetti et al., 1993; Al Safadi, 1997); therefore no Testudo sp. could have been observed in Al-Luljayyah by the Forsskal - Niebuhr expedition. Following the earlier choice, made in accordance with the available data, and also with the current taxonomical practice (e.g., Perala, 2002b). in order to preserve the stability of the nomenclature, and Vol.10, p. 117 Asiatic Herpetological Research 2004 to objectively delimit Testudo terrestris Forsskal from all other species in the Testudo graeca ( sd .) complex, we choose a specimen collected in Aleppo (and which locality has by chance a historical background in tor- toise literature: cf. Siebenrock, 1913). The neotype of Testudo terrestris. - To fix the name and type locality, we hereby designate a specimen from the Vienna Natural History Museum No. NMW 18674:2, collected in Aleppo by Viktor Pietschmann in March 1910, as the neotype of Testudo terrestris Forsskal, 1775, according to Articles 75 and 76 (ICZN, 1999). The neotype is a subadult female with a straight-line carapace length (CL) of 136.8 millimeters. Note: all morphometric characters are according to the standards published in Perala (2001). As a result of our neotype designation (ICZN, 1999; Art. 76.3), the type locality of Testudo terrestris Forsskal, 1775 is restricted to Aleppo (Alep, Halab; 36°12’ N, 37°09’ E), Syria (Syrian Arab Republic). (For the range of T. terrestris , see Perala, 2002b.) Description. - Sub-adult female. Most scutes marked with about a dozen of conspicuous growth ridges and grooves; areolae of the carapace feebly bumped, neatly displaced caudally, and also dorsally on the costals. Longitudinal profile high, regularly domed, highest at third vertebral, slightly behind the middle of the shell (Fig. 2, top left). The outline of the shell short, squarish, anterior and posterior free borders only very feebly cut out. Vertebrals wide in dorsal view, the fourth the small- est; the first one with almost straight lateral borders, moderately wider anteriorly than posteriorly (Fig. 2, top right). Cervical (= nuchal) four-sided, very wide; supra- caudal distally as wide as vertebrals, regularly convex in its middle; all common sutures of marginals nearly sub equal (= height of lateral ones, and length of anterior and posterior ones); only marginals 9-11 are slightly flaring. Gulars well prominent but short and narrow, their common suture very short; a pair of rather large axillaries on each side; pectorals with a very short com- mon (medial) suture, included about four times in the medial suture of humerals; inguinals small, separated in a larger distal part and a very small proximal part con- tacting femorals; rear lobe of plastron hardly mobile, short, anals both long and wide, with parallel anterior and posterior borders: their particular shape at first sight resembles the usual shape observed in males (Fig. 2, bottom left). Head covered above with two large and roughly pentagonal scutes, the frontal and the prefron- tal, symmetrical about their common suture. Five nails at each hand, the inner one smaller but well developed; four at each foot. Anterior side of the fore-arm covered between the elbow and the wrist by about fifteen large scutes, the four largest being triangular, neatly distinct from the background of the scaly skin; outer border of this area covered by a row of six triangular, overlapping scutes. Tail relatively long and regularly tapering, also giving a rather masculine appearance, ending with slightly enlarged but discrete flat scutes; a small isolated spur on each thigh. General color greenish yellow, often lighter close to the sutures, with large darker, grayish areas apparently deep in the scutes. Blackish marks reduced on the cara- pace, limited to incomplete and irregular narrow lines along the anterior sutures of the scutes (marginals, cos- tals), also on the lateral borders of vertebrals; areolae or areolar areas slightly and irregularly flecked with black, on costals 1-3 and on vertebrals 1-3. Dark patches wider on the plastron, issued from the areolae, roughly extend- ing along the rear third of each scute (from pectorals to anals), restricted to a narrow band on the humerals; lim- its of the patches are inconspicuous, with a gradual shading, delimiting few lighter or darker radiating lines. Soft parts mostly yellowish, in places with a brownish tinge, with well contrasting blackish flecks on the homy beak (upper and lower, forming a ‘moustache’), on pre- frontal, on the large triangular scutes of the fore-arms, and on the heels; all nails are also heavily pigmented, from dark brown to black. Additional morphometric data derived from the neotype are presented in the following (all measure- ments in mm): maximum plastron length (PL) = 122.7, midline plastron length (PL-m) = 1 1 1.8, maximum mid- body width (MI) = 100.2, maximum width of shell at posterior marginals (MA) - 104.4, maximum gular scute length (GU-1) = 15.3, maximum gular scute width (GU-w) = 27.3, gular scute height (GU-h) = 13.2, maxi- mum shell height (HE) = 70.0, maximum width of ante- rior shell opening (ASO-w) = 69.9, maximum height of anterior shell opening (ASO-h) = 21.9, left minimum bridge length (BR) = 59.9, maximum humeral scute width (HUM-w) = 63.7, maximum pectoral scute width (PEC-w) = 86.6, maximum abdominal scute width (ABD-w) = 89.9, maximum femoral scute width (FEM- w) = 64.1, maximum anal scute width (AN-w) = 51.9, maximum nuchal scute length (NU-1) = 1 1.4, maximum nuchal scute width (NU-w) = 10.0, intergular length (GU-m) = 13.4, interhumeral length (HUM-m) = 22.8, interpectoral length (PEC-m) = 7.7, interabdominal length (ABD-m) = 42.1, interfemoral length (FEM-m) = 10.8, interanal length (AN-m) = 18.7, maximum width of first vertebral scute (Vl-w) = 35.2, maximum width of second vertebral scute (V2-w) = 36.4, maximum width of third vertebral scute (V3-w) = 41.0, maximum width of fourth vertebral scute (V4-w) = 33.8, maxi- mum width of fifth vertebral scute (V5-w) = 41.3, maxi- mum length of first vertebral scute (V 1 -1) = 26.9, 2004 maximum length of second vertebral scute (V2-1) = 28.0, maximum length of third vertebral scute (V3-1) = 25.6, maximum length of fourth vertebral scute (V4-1) = 23.3, maximum length of fifth vertebral scute (V5-1) = 3 1 .8, first costal length (C 1) = 43.0, second costal length (C2) = 28.7, third costal length (C3) = 28.1, fourth cos- tal length (C4) = 22.9, maximum dorsal width of supra- caudal (SUP-d) = 23.4, maximum ventral width of supracaudal (SUP-v) = 40.4, maximum median length of supracaudal (SUP-1) = 21.7, maximum head width (HEAD) = 21.6, minimum distance between right eye and tympanum (EYE-TY) = 6.9, minimum distance between right eye and nostril (EYE-NO) = 6.5. Figure 2 (lower right) depicts a living female (topo- type) Testudo terrestris Forsskal from the type locality, Aleppo, Syria. Acknowledgments We warmly thank James Buskirk (Oakland, CA) and James Parham (University of California, Berkeley) for personal communications, as well as two anonymous reviewers for comments, which had a positive impact on the manuscript, Richard Gemel, Heinz Grillitsch and Franz Tiedemann (all from the Naturhistorisches Museum, Vienna) for the loan of specimens, the Aca- demic Kippis Society (AKS) for spiritual guidance, and Annemarie Ohler (Museum national d’Histoire naturelle, Paris) for logistical help. Literature Cited A1 Safadi, M. M. 1997. Yemeni Turtles & Tortoises. Yemen Times, vol.VII, 51 (last page), and http:// www.yementimes.com/97/iss51/lastpage.htm. Anderson, J. 1896. Sketch of the literature bearing on the reptilian and batrachian fauna of Arabia. Part IV. Pp. 68-76. 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Vol. 10, PP- 120-125 An Ocadia sinensis x Cyclemys shanensis hybrid (Testudines: Geoemydidae) Maik Schilde1, Dana Barth2 and Uwe Fritz3 1 Opalstr. 31, D-043 19 Leipzig, Germany; E-mail: maik.schilde@ufz.de -University of Leipzig, Institute of Zoology, Molecular Evolution & Animal Systematics, Talstr. 33, D-04103 Leipzig, Germany; E-mail: dbarth@rz.uni-leipzig.de 3 Zoological Museum (Museum fur Tierkunde), Natural History State Collections Dresden, A. B. Meyer Building, Konigsbriicker Landstr. 159, D-0 11 09 Dresden, Germany; E-mail: uwe.fritz@snsd.smwk.sachsen.de Abstract. - A captive bred Ocadia sinensis x Cyclemys shanensis hybrid is described. Its hybrid status was confirmed by a comparison of a 1036 bp fragment of the mitochondrial cytochrome b gene with the putative mother (C. shanen- sis) and genomic ISSR fingerprinting. This is the first report of an intergeneric hybrid between very distantly related geoemydid turtles. All previous geoemydid intergeneric hybrids have been crossings within or between two sister clades containing the currently accepted genera ( Chinemys , Mauremys, Ocadia ) and ( Cuora , Pyxidea). Key words. - Cyclemys, Ocadia, testudines, intergeneric hybrid. Introduction Recently several new cases of intergeneric chelonian hybrids became known to science (reviewed in Galgon and Fritz, 2002). Most of them belong to the Southeast Asian family Geoemydidae, long known under its junior synonym Bataguridae. However, current research on the molecular phylogeny of geoemydids has shown that some species traditionally attributed to different genera are more closely related than previously thought (Wu et al., 1999; McCord et ah, 2000; Honda et ah, 2002a, b; Barth et ah, in press; Stuart and Parham, in press), sug- gesting that they should be better lumped in the same genus. Thus, some of the hybrids may be in fact not intergeneric. In this paper we report a captive bred hybrid between two distantly related Southeast Asian geoemydids, representing an undoubtedly intergeneric cross. According to the cited studies, there are several major clades among geoemydids. One clade contains the currently recognized genera Chinemys, Mauremys, Ocadia, Cuora, and Pyxidea (McCord et ah, 2000; Honda et ah, 2002a, b; Barth et ah, in press), and anoth- er one, being the sister clade, Cyclemys, Sacalia, Heosemys, Hieremys, Notochelys, and Leucocephalon (McCord et ah, 2000; Honda et ah, 2002b). The turtle described herein is the result of a hybridization of an Ocadia sinensis male and a Cyclemys shanensis female, representatives of two of the major clades of the Geoemydidae. This hybrid demonstrates that very distantly related geoemydids are capable of hybridizing successfully. It underlines the possibility that some recently described Southeast Asian chelonians ( Ocadia glyphistoma, O. philippeni ), which are only known from few pet trade specimens, might also be hybrids. The specimen. - The turtle described below hatched in the live collection of M. Schilde from an egg of a Cyclemys shanensis, laid August 13, 2002. The second egg of the same clutch did not develop. The mother was a long term captive, and kept with a Cyclemys shanensis male and two Ocadia sinensis males. The elongated eggs measured 56.5 x 20.0 mm. One quickly developed a white band as typical for fertile eggs. It was incubated constantly at 28°C on Vermiculite. On October 26, 2002 a healthy turtle with a straight line shell length of 33 mm hatched (Figs. 1-4). Its color pattern resembled Ocadia sinensis but the general form was more similar to Cyclemys (roofed, distinctly tricarinate shell, serrated posterior marginal scutes), suggesting that it might be a hybrid. We decided to use two molecular methods to test this hypothesis. Materials and Methods We sequenced a 1036 bp portion of the mitochondrial cytochrome b gene (cyt b) of the captive bred turtle for comparison with the putative mother. Because mito- chondrial DNA is inherited in the maternal line, the sequence of the putative hybrid should be identical with the mother ( Cyclemys shanensis ). Species identification of the Cyclemys was done by morphological means and comparison with the mitochondrial cyt b sequences of Guicking et al. (2002); the Ocadia sinensis males were determined morphologically. Blood samples were obtained by coccygeal vein puncture. Samples were stored as described in Haskell © 2004 by Asiatic Herpetological Research Vol. 10, p. 121 Asiatic Herpetological Research 2004 Figure 1 . Figure 2. Figure 3. Figure 4. Fig. 1-4. The captive bred Ocadia sinensis x Cyclemys shanensis hybrid, September 2003 (11 months old). The roofed, distinctly tricarinate shell and the serrated posterior marginals resemble Cyclemys spp. The neck and facial stripes as well as the plastral pattern are similar to O. sinensis. The plastral pattern was more contrasting as a hatchling and has faded during growth. Photos: F. Hohler. and Pokras (1994) and Arctander (1988). Additional blood samples and photographs (dorsal and ventral aspects) of the captive bred turtle (MTD T1262), the Cyclemys shanensis female (MTD T816), and the two Ocadia sinensis males (MTD T8 1 7-8 1 8) are in the tissue collection of the Zoological Museum Dresden. DNA extraction was carried out using the QIAamp Blood Mini Kit (Qiagen). Primers mt-A (Lenk and Wink, 1997) and HI 5909 (Lenk et al., 1999) were used to amplify a DNA fragment containing 1036 bp of cyt b. PCR and sequencing conditions were as described in Barth et al. (in press). Sequencing reactions were performed on an ABI 3100 Genetic Analyzer. Alignment was carried out with CLUSTAL X, v. 1.8 (Thompson et ah, 1997) with default parameters. To demonstrate the considerable genetic difference between Cyclemys and Ocadia com- pared to other hybridizing geoemydids, Mega 2.1 (Kumar et ah, 2001) was used for estimation of genetic distances. Cyt b sequence data for calculating genetic distances are from Barth et ah (in press) and Guicking et ah (2002). To exclude the possibility of a gynogenetic or parthenogenetic origin of the specimen and to identify the putative father, we conducted genomic fingerprint- ing with Inter Simple Sequence Repeats (1SSR) for the captive bred specimen, the Cyclemys female and both Ocadia males. ISSR PCR produces species-specific genomic fingerprints (Gupta et ah, 1994; Zietkiewics et ah, 1994; Wink et ah, 1998. 2001; Nagy et ah, 2003) useful in detecting turtle hybrids (Wink et ah, 2001). Hybrid specimens share about 50% of their ISSR PCR products with the respective paternal and maternal species (Wolfe et ah, 1998: Wink et ah, 2001; Storch et ah, 2001 ). ISSR PCR is a simple and cheap method, and the results are easily reproducible (Bornet and Branchard, 2001). Gynogenesis or pseudogamy, the development of unfertilized eggs by activation through sperm of another species, as well as parthenogenesis is not known in turtles. However, if the captive bred spec- imen should be of such origin, the ISSR profiles should be identical with its biological mother. The primer 5 -GACAGACAGACAGACA-3' was used to generate ISSR fingerprints for the captive bred specimen, the putative mother ( Cyclemys shanensis ), and both Ocadia sinensis males. Each reaction mix con- tained 100 ng of genomic DNA, 20 pmol primer, 1 U 2004 Asiatic Herpetological Research Vol. 10, p. 122 Table 1. ISSR fingerprints of the Ocadia sinensis x Cyclemys shanensis hybrid and parental species (bioogica mother and father plus the second 0. sinensis male kept with the mother). Only polymorphic and diagnostic products shown. MTD T = Museum fur Tierkunde Dresden Tissue Collection; + = PCR product present, - = PCR product lacking. Fragment Length Cyclemys shanensis MTD T816 Hybrid MTD T1262 Ocadia sinensis MTD T817 Ocadia sinensis MTD T818 100 bp + + - - 480 bp - + + + 590 bp - + + + 810 bp + + - - 940 bp + + - - 1100 bp - + + - 1450 bp - + + + 1500 bp + + - - 1550 bp - + + - 1700 bp + + - - 2400 bp - + + + 2700 bp - + + + 2900 bp + + - - Taq-polymerase (SIGMA), 2.5 pi lOx PCR buffer (SIGMA) and 2.5 pi of 200 pM dNTPs in a total volume of 25 pi. Amplification conditions were 4 min initial denaturation (94°C), followed by 31 cycles of 1 min at 94°C, 1 min at 54°C, and 2 min at 72°C, final extension of 7 min (72°C). PCR reactions were performed on an Eppendorf thermocycler. 15 pi of each PCR reaction was separated on 2% agarose gels (25 cm), stained in ethidium bromide solu- tion (0.5 pg/ml) and visualized under UV light. The 100 bp DNA ladder Plus (MBI Fermentas) was used as a size standard. PCR was repeated under identical conditions to test reproducibility of results. DNA fragments were scored manually. Band sharing coefficients were calcu- lated according to Storch et al. (2001). Results and Discussion As expected, the cyt b sequence of the captive bred tur- tle and the putative mother ( Cyclemys shanensis ; EMBL acc. no. AJ604513) proved to be identical. ISSR finger- printing produced highly variable profiles which were species-specific and permitted individual identification of both Ocadia sinensis males (Table 1). The captive bred turtle shared 6 of its 13 bands with the mother (band sharing coefficient 0.5) and 7 bands with one of the O. sinensis males (band sharing coefficient 0.52). Because the captive bred turtle and this O. sinensis male exclusively share some fragments, we identified this tur- tle as the biological father. Thus, both methods con- firmed the hybrid origin of the captive bred turtle. Except for an unconfirmed, anecdotal newspaper record of natural hybrids between Cuora flavomargina- ta and Geoemyda japonica in Japan (Anonymous, 1995), all previous geoemydid intergeneric hybrids have been crossings within or between two sister clades con- taining the currently accepted genera ( Chinemys , Mauremys, Ocadia) and {Cuora, Pyxidea ); Chinemys reevesii x Cuora amboinensis kamaroma (Galgon and Fritz, 2002), Chinemys reevesii x Mauremys japonica (Yasukawa et al., 1992), Chinemys reevesii x Mauremys mutica (= "Mauremys pritchardi" , Wink et al., 2001), Cuora amboinensis kamaroma x Mauremys annamensis (Fritz and Mendau, 2002), Cuora bourreti x Pyxidea mouhotii (= "Cuora serrata ", Parham et al., 200 1 ; Stuart and Parham, in press), Cuora galbinifrons x Pyxidea mouhotii (= "Cuora serrata ", Parham et al., 2001; Stuart and Parham, in press), and Cuora trifasciata x Mauremys mutica (= " Mauremys iversoni", Parham et al., 2001 ; Wink et al., 200 1 ). Cyclemys belongs to anoth- er major clade, comprising the genera Cyclemys , Sacalia , Heosemys, Hieremys , Notochelys , and Leucocephalon (McCord et al., 2000). Cyclemys is sep- arated by a considerable genetic distance from Ocadia (Table 2), surpassing the genetic distances of the other hybridizing geoemydids. Superficially our hybrid Ocadia sinensis x Cyclemys shanensis resembles O. sinensis due to its striped head and neck and the spotted plastral pattern. This leads to the speculation that the morphologically similar Ocadia philippeni McCord and Iverson, 1992 and O. glyphistoma McCord and Iverson, 1994 might be also intergeneric hybrids, as earlier suggested by van Dijk (2000), Fau and Shi (2000), Parham and Shi (2001), and Galgon and Fritz (2002). Both species were described on the basis of only a few pet trade turtles (McCord and Iverson, 1992, 1994), and until now no Vol. 10, p. 123 Asiatic Herpetological Research 2004 Table 2. Pairwise genetic distances (cyt b) between hybridizing geoemydid species. Cyt b Geomydid species pairwise distances Chinemys reevesii - Cuora amboinensis 0.104 Chinemys reevesii - Mauremys japonica 0.050 Chinemys reevesii - Mauremys m utica 0.070 Cuora amboinensis - Mauremys annamensis 0.098 Cuora galbinifrons - Pyxidea mouhotii 0.059 Cuora trifasciata - Mauremys mutica 0.104 Ocadia sinensis - Cyclemys shanensis 0.118 additional specimens became known to science. For some individuals of three other pet trade taxa a hybrid status has been unambiguously demonstrated: Two tur- tles identified as Mauremys pritchardi McCord, 1997 proved to be hybrids of Chinemys reevesii and Mauremys mutica (Wink et al., 2001). Three Mauremys iversoni Pritchard and McCord, 1991 originated from crossing Cuora trifasciata and Mauremys mutica (Parham et al., 2001; Wink et al., 2001), and several Cuora serrata Iverson and McCord, 1992, have been demonstrated to be hybrids between Cuora galbinifrons and Pyxidea mouhotii and of Cuora bourreti and Pyxidea mouhotii (Parham et al., 2001; Stuart and Parham, in press). Until now it is unknown whether all specimens of these taxa are of hybrid origin, and if so, whether the crosses occurred in the wild, in captivity, or whether one or the other form might represent a natural, stable hybrid taxon (Parham et al., 2001; Wink et al., 2001). Many Southeast Asian chelonians are facing extinction due to overexploitation (van Dijk et al., 2000). Therefore, many conservation efforts are established around the globe, including CITES listing and captive breeding programs for several species. A correct taxonomy is the prerequisite for any conservation measure. Hence, it is crucial to determine whether the mentioned taxa repre- sent real evolutionary entities and deserve high priority in conservation, this includes also natural hybrid taxa (Allendorf et al., 2001), or whether they are only inci- dentally occurring hybrids, without any conservation relevance. Acknowledgments Special thanks go to Michael Wink and Daniela Guicking, Heidelberg, for sharing unpublished geoemy- did sequences with us and to Thomas U. Berendonk, Leipzig, for technical advice. James R. Buskirk, Oakland, provided the newspaper report on Cuora flavo- marginata x Geoemyda japonica hybrids. Literature Cited Allendorf, F. W., R. F. Leary, P. Spruell and J. K. Wenburg. 2001. The problems with hybrids: setting conservation guidelines. TREE 16:613-622. Anonymous. 1995. Hybrids between two protected tur- tles increase. Okinawa Times 15 August 1995. Arctander, P. 1988. Comparative studies of Avian DNA by restriction fragment polymorphism analysis. 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L and J. F. Parham, in press. Molecular phy- togeny of the critically endangered Indochinese box turtle ( Cuora galbinifrons). Molecular Phylogenet- ics and Evolution. Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin and D. G. Higgins. 1997. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 24:4876-4882. van Dijk, P. P. 2000. The status of turtles in Asia. Chelonian Research Monographs 2:15-23. van Dijk, P. P, B. L. Stuart and A. G. J. Rhodin (eds.). 2000. Asian Turtle Trade. In Proceedings of a Workshop on Conservation and Trade of Freshwater Turtles and Tortoises in Asia. Chelonian Research Monographs 2:1-164. Wink, M., H. Sauer-Giirth, F. Martinez, G. Doval, G. Blanco and O. Hatzofe. 1998. Use of GACA-PCR for molecular sexing of Old World vultures (Aves: Accipitridae). Molecular Ecology 7:779-782. Wink, M., D. Guicking and U. Fritz. 2001. Molecular evidence for hybrid origin of Mauremys iversoni Pritchard et McCord, 1991, and Mauremys pritchardi McCord, 1997. Zoologische Abhandlungen, Staatliches Museum fur Tierkunde Dresden 51:41-49. Wolfe, A. D., Q.-Y. Xiang and S. R. Kephart. 1998. Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable intersimple sequence repeat (ISSR) bands. Molecular Ecology 7:1107-1125. Wu, P, K. Zhou and Q. Yang. 1999. Phylogeny of Asian freshwater and terrestrial turtles based on sequence analysis of 12S rRNA gene fragment. Acta Zoologica Sinica 45:260-267. Vol. 10, p. 125 Asiatic Herpetological Research 2004 Yasukawa, Y., N. Kamezaki and N. Ichikawa. 1992. On hybrids between Mauremys japonica and Chinemys reevesii. Japanese Journal of Herpetology 14:206- 207. Zietkiewicz, E., A. Rafalski and D. Labuda. 1994. Genome fingerprinting by simple sequence repeat (ISSR)-anchored polymerase chain reaction ampli- fication. Genomics 20:176-183. 2004 Asiatic Herpetological Research Vol. 10, pp. 126-128 New Data on the Trade and Captive Breeding of Turtles in Guangxi Province, South China Haitao Shi1, Zhiyong Fan2, Feng Yin3, and Zhigang Yuan4 ' Department of Biology, Hainan Normal University, Haikou , 571158, Hainan Province, P. R. China. E-mail:haitao-shi@263.net; Fax: 0898-65890520; Tel: 0898-66752479; 65883521. -Fauna Division, The Endangered Species Import and Export Management Office of China (CNMA) #18 Hepingli Dongjie, Beijing 100714, P. R. China. J Department of Science and Education, China Wildlife Conservation Association, #18 Hepingli Dongjie, Beijing 100714, P. R. China. ^Department of Biology, Guangxi Medicine University, 530021, Nanning, PR. China. Abstract. - New data on the captive breeding and trade of turtles in Guangxi Province, China, are presented. These data are from four turtle farms and three markets surveyed in May 2002. The scale of captive breeding in Guangxi is larger than previously known. At the same time, the number of wild turtles in the markets may be decreasing. Issues concerning the licensing of turtle farms and the effectiveness of enforcement are discussed. Key words. - China, Guangxi, turtles, trade, farming. Introduction The People’s Republic of China is Asia’s most signif- icant importer of tortoises and freshwater turtles and more comprehensive studies on its wildlife trade are urgently needed (Li and Li, 1997; van Dijk, et al., 2000). There is a long history of wildlife consumption in Guangxi, and traditional Chinese medicine is as popular as in neighboring Guangdong Province. Li and Li( 1 997) reported that there are at least 91 species of animals involved in the wildlife trade in Guangxi, mostly turtles and snakes. The sheer volume of the wildlife trade on the border between China and Vietnam is astonishing, and may be unprecedented in the history of internation- al wildlife trade. Guangxi is one of the main corridor for the import and export of wildlife from land and sea into mainland China. Although China has recently increased its level of protection for imported and exported turtles (Meng et al., 2002), the effectiveness of these new meas- ures is not well demonstrated. We report data on four turtle farms and three markets surveyed in late May 2002. Turtle farms Shifu turtle farm (Shifu Town, Nanping City). - The turtle farm was founded in 1996, its area is 65 ha. At it’s height it included -70,000 turtles: -60,000 Trachemys sripta elegans, -7,000 Pelodiscus sinensis , and -600 Ocadia sinensis. However during a very heavy flood on July 5, 2001 in the area, almost all the turtles escaped or drowned. The farm is rebuilding, but now includes only -700 Mauremys mutica, -200 Palea steindachneri and -50 Cuora trifaciata. Quanming wildlife farm (Located at Quanming Town, Daxin County). - The farm began to raise turtles in 1996. It has -80 Mauremys mutica and 47 Chelydra serpentina. Their 36 female Chelydra lay 30-35 eggs three times every year. So they get more than 3000 eggs every year. They are expanding to include Macroclemys temminckii. Besides turtles, the wildlife farm also breed successfully 1800 Tokay Geckos ( Gekko gecko ) and for several decades has also bred the Masked Palm Civet (Paguma larvata ). Qingzhou turtle farm 1 (Qingzhou City). - This farm has been raising turtles since 1981, but was not very suc- cessful until 1997. In 2001, the farm had -1000 Mauremys mutica and -300 Cuora trifaciata. In the past they also bred Trachemys scripta elegans , but switched to Cuora trifaciata and Mauremys mutica due to the higher price for the two species. The owner of this farm claims that there are dozens more farms in Qingzhou that were founded based stock from his farm. The phe- nomena of Mauremys mutica mixed with Cuora trifaci- ta was found here, mirroring the conditions of a farm in Tunchang, Hainan Province (Shi & Parham, 2001). It was impossible for us to visit the farm without the local officials of Forestry Department forcing the farmer to accept us. The owner prefers to remain secret in order to avoid theft, taxes, and people wanting to borrow money. © 2004 by Asiatic Herpetological Research Vol. 10, p. 127 Asiatic Herpetological Research 2004 Qingzhou turtle farm 2 (Qingzhou city). - This farm was founded in 1986. It encompasses 4 hectares, and includes —7,000 Mauremys mutica , —6,000 Trachemys scripte elegans , -2700 Pelodicus sinensis, -800 Cuora trifaciata, 200 Cuora flavomarginata and 3 Pelochelys bibroni. Approximately, 30,000 hatchlings are bred every year. Unlike the owner of the previously reported farm, the owner of this farm, Wusong Ma, is friendly, generous, and open to visitors. Many famous people have visited his turtle farm, including a former Chinese national vice-premier, the Governor of Guangxi, and the Minister of Agriculture Department. Unfortunately, all this attention may have led to a large theft of Mauremys mutica in April of 2001. According to the owner of the farm, the value of these turtles was 50,000 USD. Despite this, Mr. Ma remains non-secretive and has even set up at least 20 additional farms. Remarks on turtle farms. - The senior author (HS) looked over the licenses for captive breeding of turtles at the Forestry Department of Guangxi Province. More than 600 farms (of various sizes, including small breed- ing operations) were licensed in Guangxi Province. Zou Yi, an official in the Guangxi Forestry Despartment, informed us that another governmental department (Agriculture Department) issues even more licenses for turtle farming, but we do not have these numbers. Moreover, most breeding operations are illegal and not licensed. Consequently, determining the actual number of commercial, often secretive, turtle breeders will require intensive survey and investigation. Turtle Markets Nanning Road Trade Market in Nanning City. - Five stalls with 14 species and 194 turtles were found in this market. The species included 58 Pelodiscus sinensis, 24 Pyxidea mouhotii, 21 Platysternon megacephalum, 17 Mauremys mutica, 15 Trachmys script a elegans, 1 3 Cyclemys dent at a, 13 Palea steindachneri, 12 Sac alia quadriocellata, 8 Indotestudo elongata, 4 Cuora amboinensis, 4 Ocadia sinensis, 2 Heosemys grandis, 1 Hieremys annandalei, andl Orlitia borneensis. Dongfeng Market in Qingzhou City. - Three stalls with 9 species and 64 were found in Dongfeng Market in Qingzhou. The species included 19 Trachmys scripta elegans, 1 8 Indotestudo elongata, 9 Manouria impressa, 5 Malayemys subtrijuga, 4 Cuora amboinensis, 4 Pyxidea mouhotii, 2 Geoemyda spengleri, 2 Heosemys grandis, and 1 Cuora galbinifrons. Danqing Wholesale Market in Nanning City. - 5 stalls with 4 species and 42 turtles were found in this market. Sixty (71%) of the turtles were Platysternon mega- cephalum. Platysternon was sold at every stall. I was told that they came from turtle farms, but they refused to tell where these turtle farms were. Remarks on turtles. - According to Lu Qi, the vice director of the Wildlife Management Section of the Guangxi Forestry Department, the past three years have seen a sharp decrease in the numbers of turtles in the markets and he attributes this to increased enforcement. This increased enforcement coincides with an ever- growing commercial effort to breed these turtles. For example, Manying Huang (professor of biology at Guangxi Medicine University) states that there are over 3000 families that raise Cuora trifasciata in Nanning City. However, the increased level of enforcement has led to many captive-bred turtles also being confiscated. If indiscriminant confiscations continue it could depress the development of captive breeding of turtles in China. Acknowledgments We would like to thank Manying Huang, professor at Guangxi Medicine University, Qi Lu, Zou Yi and others, officials in Forestry Deaprtment of Guangxi Province, for their assistance with the survey. This work was fund- ed by National Natural Science Foundation of China (No. 30260019), Foundation for University Key Teacher by the Ministry of Education and Wildlife Conservation Association of China. James Parham helped to correct the English and provided helpful comments. Literature Cited Li, Y. and D. Li. 1997. The investigation on wildlife trade across Guangxi borders between China and Vietnam. Conserving China’s Biodiversity. Reports of the Biodiversity Working group, China Council for International Cooperation on Environment and Development: 118-127. Meng, X., Z. Zhou and B. L. Stuart. 2002. Recent actions by the People’s Republic of China to better control international trade of turtles. Turtle and Tortoise Newsletter 5:15-16. Shi, H. and J. F. Parham. 2001. Preliminary Observations of a large turtle farm in Hainan Province, People’s Republic of China. Turtle and Tortoise Newsletter, issue 3: 4-6. 2004 Asiatic Herpetological Research Vol. 10, p. 128 van Dijk, P. P., B. L. Stuart and A. G. J. Rhodin (Eds.). 2000. Asian Turtle Trade: Proceedings of a Workshop on Conservation and Trade of Freshwater Turtles and Tortoises in Asia. Chelonian Research Monographs 2:1-164. 2004 Asiatic Herpetological Research Vol. 10, pp. 129-150 Recent Records of Turtles and Tortoises from Laos, Cambodia, and Vietnam Bryan L. Stuart1’2’3’4’* and Steven G. Platt4’5 1 Field Museum of Natural History, Department of Zoology, Division of Amphibians & Reptiles, 1400 S. Lake Shore Drive, Chicago, Illinois 60605 USA. E-mail: bstuart@fieldmuseum.org * Corresponding author and address z University of Illinois at Chicago, Department of Biological Sciences (M/C 066), 845 W. Taylor, Chicago, Illinois 60607 USA 3 Wildlife Conservation Society, P.O Box 6712, Vientiane, Lao PDR 4 Wildlife Conservation Society, P.O. Box 1620, Phnom Penh, Cambodia 5 Present address: Department of Math and Science, Oglala Lakota College, P.O. Box 490, Kyle, South Dakota 57752-0490 USA Abstract. - The chelonian fauna of Laos, Cambodia, and Vietnam remains poorly known and is currently threatened by widespread and intensive exploitation for food and traditional Chinese medicine. The distributions of many species are uncertain owing to a paucity of records. Because turtles are so extensively traded in the region, most records now come from animals in trade. We emphasize that authors must be explicit about how their records were obtained to allow other workers the ability to critically evaluate the accuracy of the distribution record. We here present detailed information on recent (1993-2002), vouchered records of 19 species of freshwater turtles, tortoises, and marine tur- tles collected in the field or obtained from hunters, abandoned hunting camps, villages, or markets in Laos, Cambodia, and Vietnam. Key words. - Testudines, turtles, tortoises, Laos, Cambodia, Vietnam, distribution. Introduction Although mainland Southeast Asia has long been regarded as a hotspot of chelonian diversity (van Dijk et al., 2000), the turtle and tortoise fauna of Laos, Cambodia, and Vietnam (formerly known as French Indochina) remains poorly known. Biological investiga- tion was limited prior to World War II, and since then decades of civil unrest, political instability, and military conflict have largely prevented fieldwork. Consequently few museum records exist (summarized by Iverson, 1992) and with the exceptions of Smith (1931) and Bourret (1941), little information is available on the occurrence and distribution of chelonians in former French Indochina. A recent photographic identification guide to the region (Stuart et al., 2001) and country reviews of Laos (Stuart and Timmins, 2000), Cambodia (Touch et al., 2000), and Vietnam (Flendrie, 2000) sum- marized information on chelonian distributions, but pro- vided few details concerning new records on which these accounts are based. Exploitation of chelonians for food and medicinal markets is widespread in Laos, Cambodia and Vietnam (Jenkins, 1995; Le Dien Due and Broad, 1995, Lehr, 1997; Timmins and Khounboline, 1999; van Dijk et al., 2000; Ziegler, 2002; Holloway 2003). Hunters in rural villages capture turtles and tortoises for local consump- tion or to sell to traders who periodically visit villages to purchase wildlife. Although turtles and tortoises are locally consumed and domestically traded in Laos, Cambodia, and Vietnam, most are exported to markets in southern China (Stuart et al., 2000; van Dijk et al., 2000). Chelonians from Laos and Cambodia are usually transported to Vietnam, where they join with Vietnamese turtles on northward routes to China (Stuart et al., 2000). Because Laos and Cambodia are source rather than destination or transfer countries, specimens obtained from markets in Laos or Cambodia usually originated from that country. However, trade specimens in Vietnam may have originated from Vietnam, Laos, Cambodia, or beyond. The volume of this trade is believed to pose a serious threat to the continued viabil- ity of wild chelonian populations throughout Southeast Asia (van Dijk et al., 2000). Because turtles and tortoises are extensively and visibly traded in Southeast Asia, most recent distribution records are based on animals observed in trade rather than collected from the wild. The geographic origin of many trade specimens can be difficult if not impossible to determine, especially those obtained in urban mar- kets. Uncritical acceptance of these records by workers has led to inaccurate characterization of species distribu- tions, with serious biological, conservation, legal, and regulatory implications. Additional confusion has result- © 2004 by Asiatic Herpetological Research Vol. 10, p. 130 Asiatic Herpetological Research 2004 ed from the realization that some species of Asian turtles described during the last two decades were based on type specimens obtained from Hong Kong animal deal- ers who provided inaccurate or fabricated locality data, leaving the geographic origin of many in doubt (Dalton, 2003; Parham et al., 2001). Distribution records that explicitly state how the turtles were obtained are there- fore clearly important, given the historical paucity of information, uncertainties in recent literature, and the serious conservation threats faced by these taxa. We here report recent distribution records of chelo- nians from Laos, Cambodia, and Vietnam that can be verified with voucher specimens or photographs. These records were obtained by (1) us during herpetological surveys conducted from February 1998 through May 2001, (2) other workers in the region between 1993 and June 2002, and provided to us, or (3) other workers who deposited specimens at the Field Museum of Natural History, Chicago, USA since 1993. For each record we note whether the specimen was collected in the field, found in abandoned hunting camps, obtained from hunters or residents in rural villages, or purchased from markets. It should be emphasized that our collecting activi- ties had little if any detrimental impact on populations of wild chelonians. The number of collected specimens on which we report is insignificant when compared to the millions of chelonians annually consumed by the wildlife markets of southern China (Lau et al., 2000). Furthermore, the majority of specimens we and others collected in the field were shells of animals consumed by rural villagers. Chelonian shells are commonplace and easily obtained in villages; shells are retained by hunters as trophies, sold or kept for medicinal purposes, and used as food containers for domestic animals and rice scoops. Our collecting activities certainly provided no stimulus for the additional harvesting of wild chelo- nians. Finally, we believe that further scientific collect- ing is warranted in the region, as most species remain under-represented in museum collections, and taxonom- ic study can affect conservation priorities (Parham and Shi, 2001; Stuart and Thorbjamarson, 2003). Methods and Conventions Measurements were taken to the nearest 0.1 cm with 80 cm sliding calipers. We use the following abbreviations: CL = maximum straight carapace length, including spines or projections (i.e. not necessarily the mid-line); CW = maximum carapace width, including spines or projections; PL = maximum plastron length, including spines or projections; BD = maximum depth of complete specimen (head and neck extended in trionychid speci- mens). Measurements were not reported if shell damage precluded accurate measurement. Records are presented under each species account in the following format where applicable: Field Museum of Natural History (FMNH) or figure number, type of record, measurements (defined above), locality includ- ing coordinates if available, approximate elevation and brief habitat description if field-collected, circumstances of origin if not field-collected by collector or photogra- pher, date specimen was collected or photographed, and name of collector or photographer. In the case of records obtained from hunters or villages, the name of collector refers to the person who preserved the specimen or pro- vided the record to us rather than to the name of the per- son who actually captured it from the wild. In the same cases, the date of collection refers to the date the collec- tor (as previously defined) obtained the specimen or took the photograph rather than to the date it was removed from the wild. GPS coordinates are presented only if the original collector provided them, and in the same format as orig- inally provided. Coordinates that we generated for the purposes of mapping the records are not presented. Marine turtle records are not mapped. Species Accounts Platysternidae Platysternon megacep/ialum Gray, 1831 [Map 1] Laos. - Fig. 1, photograph only, Huaphahn Province, Vieng Tong District, Ban Sa Kok Village, 20° IF N 103° 12' E, captured by resident of Ban Sa Kok, 29 April 1998, B. L. Stuart. FMNH 258749, complete specimen, CL = 9.3, CW = 7.3, PL = 6.9, BD = 2.6, Bolikhamxay Province, Khamkeut District, Nape border area, stream in wet evergreen forest, 19 March 1997, D. Davenport. Fig. 2 (and Fig. 4 in Stuart and Timmins, 2000), photo- graph only, Khammouan Province, Nakai District, Nakai-Nam Theun National Biodiversity Conservation Area, Ban Xiangthong Village, 17° 54' 05" N 105° 23' 50" E, one of eleven individuals in the possession of a Vietnamese trader leaving Ban Xiangthong, 17 November 1998, B. L. Stuart. Fig. 3, photograph only, Xe Kong Province, Dakchung District, Ban Daklan Village, 15° 21.61' N 107° 01.70' E, captured by resi- dents of Ban Daklan, D. Showier, December 1997. Vietnam. - FMNH 252164, complete specimen, CL = 1 5.5, C W — 1 1 .6, PL =12.7, BD = 5.2, Gia-Lai Province, Ankhe District, Buon Loi Village, 20 km northwest of Kannack town, Annamite Mountains, 14° 20' N 108° 36' E, 700-750 m, found in burrow under overhanging stream bank, 31 March 1995, I. Darevsky and N& L. Orlov. 2004 Asiatic Herpetological Research Vol. 10, p. 131 Remarks. - Ziegler (2002) reported the species in local trade in Ha Tinh Province, Vietnam. Geoemydidae Batagur baska (Gray, 1831 “1830-35”) [Map 2] Cambodia. - Platt et al. (2003) reviewed the status of B. baska in Cambodia and reported on a breeding popula- tion in the Sre Ambel River System of Koh Kong Province. Cuora amboinensis (Daudin, 1 802) [Map 3] Cambodia. - Fig. 4, photograph only, Battambang Province, Ek Phnom District, Koh Chivang Commune, Prek Toal Village on Tonle Sap Lake, 13° 14' 28" N 103° 39' 32" E, captured by residents of Prek Toal, 27 August 1999, B. L. Stuart, J. Smith, and K. Davey. FMNH 259411, broken carapace and plastron, Kandal Province, Trayo Village, 11° 19’ 02" N 105°09’ 47" E, obtained from hunter in Trayo, 05 July 2000, S. G. Platt. Fig. 5, photograph only, two living turtles, CL = 19.3 and PL = 17.5, CL = 20.6 and PL = 18.9, Kampong Thom Province, Sary Village, 12° 48.48' N 104° 44.19’ E, col- lected by Sary residents in Tonle Sap, 21 June 2000, S. G. Platt, Heng Sovannara, and Long Kheng. Fig. 6, pho- tograph only, CL = 20.4 cm, PL = 18.5, Koh Kong Province, Sre Ambel Town, in house of wildlife trader, 1 1 ° 07.30’ N; 1 03° 44.73’ E, 27 August 2000, S. G. Platt, B. L. Stuart, and Vuthy Monyrath. Fig. 7, photograph only, CL = 11.6, PL = 11.0, Koh Kong Province, Koh Kong Town near municipal airport, 11° 37.11’ N; 103° 00.98’ E, crossing road in open grassland bordered by Melaleuca and Rhizophora swamp, 07 February 2001, S. G. Platt, Heng Sovannara, and Long Kheng. Laos. - FMNH 255262, complete specimen, CL = 16.7, CW = 11.7, PL = 15.2, BD = 7.5, Champasak Province, Mounlapamok District, Dong Khanthung Proposed National Biodiversity Conservation Area, Ban Khiem Village, 14° 14’ N 105° 20’ E, captured by residents of Ban Khiem for food, 24 July 1998, B. L. Stuart. Vietnam. - Fig. 8, photograph only, Kien Giang Province, An Minh District, photographed in a reptile trade shop, 09° 45’ 04" N 104° 59’ 35" E, 31 October 2000, B. L. Stuart. Cuora galbinifrons Bourret, 1939 [Map 4] Laos. - FMNH 256544, complete specimen, CL = 16.6, C W = 12.2, PL = 1 6.2, BD = 8.0, Khammouan Province, Nakai District, Nakai-Nam Theun National Biodiversity Conservation Area, 17° 50’ N 105° 35’ E, 900 m, wet evergreen forest, found under brush in leaf litter on hill- side 500 m from nearest stream, 13 December 1998, B. L. Stuart. FMNH 255273, carapace only, CL = 18.1, CW = 12.9, Khammouan Province, Yommalat District, Khammouan Limestone (= Phou Hin Poun) National Biodiversity Conservation Area, Ban That Mouang Khai Village, 17° 32’ N 105° 04’ E, consumed by residents of Ban That Mouang Khai, 01 April 1998, B. L. Stuart. Vietnam. - FMNH 255694, complete specimen, CL = 19.2, CW = 13.0, PL = 18.0, BD = 8.5, Nghe An Province, Tuong Duong District, Pu Mat Nature Reserve, 19° 03’ N 104° 37’ E, 600 m, wet evergreen hill forest, in leaf litter along Khe Mat Stream, 14 September 1998, B. L. Stuart. FMNH 255695, complete specimen, repatriated to Vietnam before measurements could be taken, collecting information same as FMNH 255694. Remarks. - The subspecies C. galbinifrons galbinifrons Bourret, 1939 is treated here as a full species following the recommendation of Stuart and Parham (2004). Ziegler (2002) reported the species in local trade in Ha Tinh Province, Vietnam, and Fritz et al. (2002) reported hybrids of C. galbinifrons and C. bourreti in local trade in Ha Tinh and Quang Binh Provinces, Vietnam. Cuora mouhotii (Gray, 1862) [Map 5] Laos. - FMNH 258880, carapace only, CL = 16.1, CW = 11.0, Khammouan Province, Boualapha District, Hin Nam No National Biodiversity Conservation Area, Ban Tasang Village, eaten by residents of Ban Tasang, 30 December 1995, R. J. Timmins. FMNH 258881 , cara- pace only, CL= 17.3, CW = 13.0, collecting information same as FMNH 258880. FMNH 258887, plastron only, PL = 16.8, Bolikhamxay Province, Nam Kading National Biodiversity Conservation Area, eaten by vil- lagers living in Nam Kading, 01 May 1995, R. J. Timmins. Remarks. - The species mouhotii was previously placed in the monotypic genus Pyxidea Gray, 1863, but we allo- cate it to the genus Cuora following Honda et al. (2002) and Stuart and Parham (2004). Ziegler (2002) reported the species in local trade in Ha Tinh Province, Vietnam. Cyclemys atripons Iverson and McCord, 1997 [Map 6] Cambodia. - FMNH 259050, complete specimen, CL = 21.4, CW = 16.2, PL = 20.7, BD = 8.4, Mondolkiri Province, Pichrada District, Phnom Nam Lyr Wildlife Vol. 10, p. 132 Asiatic Herpetological Research 2004 Sanctuary, near 12° 32' 16" N 107° 32' 00" E, 600-700 m, evergreen gallery forest, found on sand bank at base of large boulder 1.5 m from swift, shallow stream, 21 June 2000, B. L. Stuart. FMNH 259051, complete spec- imen, CL = 22.7, CW = 17.0, PL = 21.2, BD = 8.0, Koh Kong Province, Sre Ambel District, Sre Ambel Town, 11° 07' 20" N 103° 44' 45" E, obtained from turtle trad- er who reported specimen came from Sophat Village, downstream from Sre Ambel Town, 27 August 2000, B. L. Stuart and S. G. Platt. FMNH 259052, complete spec- imen, CL = 18.0, CW = 15.0, PL =16.7, BD = 6.2, col- lecting information same as FMNH 259051. FMNH 259412, carapace and incomplete plastron, CL = 19.3, CW = 15.2, Koh Kong Province, Sre Ambel District, BoeungTradok Pong Village, 11° 31' 10" N 103° 46' 55" E, obtained from hunter in Boeung Tradok Pong, 24 August 2000, B. L. Stuart and S. G. Platt. FMNH 259414, plastron only, PL = 16.5, collecting information same as FMNH 259412. FMNH 259415, plastron only, PL = 16.2, collecting information same as FMNH 259412. FMNH 259416, plastron only, PL = 14.9, Koh Kong Province, Sre Ambel District, Chaouethail Pious Village on Sre Ambel River, 11° 18' 03' N, 103° 44' 56”E, obtained from hunter in Chaouethail Pious, 21 August 2000, B. L. Stuart and S. G. Platt. FMNH 259417, plastron only, PL = 19.2, collecting information same as FMNH 259416. FMNH 259422, plastron only, PL = 20.2, Koh Kong Province, Sre Ambel District, Chay Reap Village, west bank of Sre Ambel River, 11° 29' 10" N 103° 47' 00" E, <10 m, obtained from hunter in Chay Reap, 23 August 2000, B. L. Stuart and S. G. Platt. Fig. 9, photograph only, two living animals, CL = 13.7 cm and PL = 13.0 cm, CL = 20.8 cm and PL = 19.6 cm, Koh Kong Province, Kaoh Pao River, 11° 44.46' N; 103° 04.80' E, surrounding hills covered in dense ever- green forest with some mangrove along shoreline, obtained from fishermen, taken in crab traps set in river, 10 May 2001, S. G. Platt, Heng Sovannara, and Long Kheng. Remarks. - Fritz and Ziegler ( 1 999) reviewed records of Cyclemys from the region. Species boundaries within the genus Cyclemys remain uncertain (Fritz and Ziegler, 1999; Guicking et ah, 2002). The specimens we assigned to C. atripons have plastra that are largely yel- low with densely pigmented bridges; complete speci- mens exhibit nearly immaculate chins. These character- istics are typical of both C. atripons and C. pulchristri- ata Fritz, Gaulke & Lehr, 1997, two species that were described almost concurrently in 1997. Cyclemys atripons and C. pulchristriata have been considered the same taxon (Iverson in Guicking et ah, 2002). However, Fritz et ah (2001) concluded that C. atripons has more ventral neck stripes (7-8 light and 7-9 dark stripes when counted from one mouth comer to the other) than C. pul- christriata (5-7 light and 5-7 dark stripes). FMNH 259050 has 8 dark and 7 light ventral neck stripes, FMNH 259051 has 10 dark and 9 light ventral neck stripes, but in FMNH 259052 the ventral side of the neck is nearly immaculate like the chin and completely lacks striping. These few samples demonstrate that ventral neck stripes are more variable than stated by Fritz et ah (2001). In a phylogenetic analysis of a 982 bp fragment of the mitochondrial cytochrome b gene, Guicking et ah (2002) recovered two clades in the atripons-pulchristri- ata complex that differed by up to 4.5% sequence diver- gence. Samples referred to atripons and pulchristriata appeared in both clades, but the authors assigned these names according to whether the sample originated from Cambodia {atripons) or Vietnam {pulchristriata ), rather than based on their morphology. The findings of Guicking et ah (2002) suggest that more than one species of Cyclemys with mostly yellow plastra, densely pigmented bridges, and immaculate chins could exist, but it remains unclear whether the two clades corre- spond to what have been described as atripons and pul- christriata. We assign the name C. atripons rather than C. pulchristriata to our samples because the type locali- ty of C. atripons is geographically closer to most of our samples than to that of C. pulchristriata. Clearly, further studies into the morphological and genetic variation in Cyclemys are warranted, particularly with samples of certain provenance. Cyclemys tcheponensis (Bourret, 1939) [Map 7] Laos. - Fig. 10 (and Fig. 7d in Stuart et ah, 2001), pho- tograph only, Bolikhamxay Province, Thaphabat District, Phou Khao Khouay National Biodiversity Conservation Area, near That Xay Waterfall, 1 8° 27' N 103° 10' E, 300 m, dry evergreen forest mixed with bam- boo, sleeping on bottom of 4 x 4 m pool in forested stream, 26 June 1998, B. L. Stuart. FMNH 258870, complete specimen, CL = 9.7, CW = 8.0, PL = 8.7, BD = 3.6, Bolikhamxay Province, Khamkeut District, pur- chased in Lac Xao Market, 14 December 1996, D. Davenport. FMNH 258871, complete specimen, CL = 9.0, CW = 8.1, PL = 8.3, BD = 3.6, collecting informa- tion same as FMNH 258870. FMNH 258875, complete specimen, CL = 20.6, CW = 15.2, PL = 20.3, BD = 8.5, Khammouan Province, Nakai District, Houay Moey Stream (tributary of Nam Pheo River), Ban Na Meo Village, dry evergreen forest, 07 March 1997, D. Davenport and J. Chamberlain. FMNH 255263, com- plete specimen, CL= 17.1, CW = 13.6, PL= 16.1, BD = 6.5, Khammouan Province, Nakai District, Khammouan Limestone (= Phou Hin Poun) National Biodiversity 2004 Asiatic Herpctological Research Vol. 10, p. 133 Conservation Area, 17° 53' N 104° 52' E, 570 m, dry evergreen lorest mixed with deciduous trees and pine, caught on streambank by hunter using dog, 26 March 1998, B. L. Stuart and T. Chan-ard. Remarks. - Species boundaries within the genus Cycle my s remain uncertain (Fritz and Ziegler, 1999; Guicking et al., 2002). The specimens we assigned to C. tcheponensis have dark radiating patterns of the plastra, pigmented chins, head and neck stripes, and dorsal spot- ting on the crown of the head, as illustrated by Fritz and Ziegler (1999) and Fritz et al. (1997). Fritz and Ziegler (1999) reviewed records of Cyclemys from the region, and Ziegler (2002) reported the species in local trade in Ha Tinh Province, Vietnam. Cyclemys sp. [Map 8] Cambodia. - FMNH 259418, carapace only, CL = 19.5, CW = 14.7, Kampong Speu Province, Koh Kong Samling Logging Concession, 11° 24' 15" N 103° 49' 47" E, 200 m, recovered from hunter’s camp, mixed deciduous forest and grassland, 15 February 2000, J. Walston. FMNH 259419, carapace only, CL = 22.1, CW = 16.1, collecting information same as FMNH 259418. FMNH 259420, carapace only, CL = 19.1, CW = 15.9, collecting information same as FMNH 259418. FMNH 259421, plastron only, PL = 21.2, collecting information same as FMNH 259418. FMNH 259423, plastron only, PL = 21.0, Koh Kong Province, Sre Ambel District, Chay Reap Village, west bank of Prek Sre Ambel River, 11° 29' 10" N 103° 47' 00" E, <10 m, obtained from hunter in Chay Reap, 23 August 2000, B. L. Stuart and S. G. Platt. Laos. - FMNH 258893, carapace only, CL = 21.8, CW = 16.6, Champasak Province, Pakxong District, Ban Latsasin Village, near Xe Nam Noy River, 800 m, eaten by residents of Ban Latsasin Village, 02 April 1995, T. D. Evans. Remarks. - The condition of these shell fragments pre- cludes identifying them to species. They are not neces- sarily a species different from atripons or tcheponensis. Heosemys grand is (Gray, 1860) [Map 9] Cambodia. - FMNH 259409, carapace only, CL = 30.5, CW = 22.6, Phnom Penh, Oreussay Market, purchased in market, 17 May 1999, S. G. Platt. FMNH 259405, plastron only, PL = 26.5, Koh Kong Province, Sre Ambel District, Boeung Tradok Pong Village, 11° 31' 10' N 103° 46' 55" E, obtained from hunter in Boeung Tradok Pong, 24 August 2000, B. L. Stuart and S. G. Platt. FMNH 259406, plastron only, PL = 20.8, Koh Kong Province, Sre Ambel District, Chaouethail Pious Village on Sre Ambel River, 11° 18' 03" N 103° 44' 56" E, obtained from hunter in Chaouethail Pious, 21 August 2000, B. L. Stuart and S. G. Platt. Fig. 11, photograph only, CL = 3 1 .8, PL = 28.6, Koh Kong Province, Thmor Andart Village along Stoeng Metoek River, 11° 49.23' N, 102° 53.62' E, captured by residents of Thmor Andart, 10 May 2001, S. G. Platt, Heng Sovannara, and Long Kheng. FMNH 259407, plastron only, PL = 19.4, collecting information same as FMNH 259406. Laos. - FMNH 255271, carapace only, CL = 23.6, CW = 18.3, Khammouan Province, Thakhek District, Khammouan Limestone (= Phou Hin Poun) National Biodiversity Conservation Area, Ban Na Village, 1 7° 33' N, 104° 52' E, eaten by residents of Ban Na, 02 April 1998, B. L. Stuart. FMNH 258885, plastron only, PL = 27.0, Khammouan Province, Khammouan Limestone (= Phou Hin Poun) National Biodiversity Conservation Area, Ban Namphick Village, eaten by residents of Ban Namphick, 22 May 1994, R. J. Timmins. FMNH 258894, carapace only, CL = 36.3, CW = 24.7, collect- ing information same as FMNH 258885. FMNH 258889, plastron only, PL = 13.9, Laos, Khammouan Province, Khammouan Limestone (= Phou Hin Poun) National Biodiversity Conservation Area, Ban Chocksavang Village, eaten by residents of Ban Chocksavang, 22 May 1994, R. J. Timmins. FMNH 258882, carapace only, CL = 23.2, CW = 19.1, Savannakhet Province, Thaphangthong District, Xe Bang Nouan National Biodiversity Conservation Area. Ban Houay Meun Village, eaten by residents of Ban Houay, 20 June 1994, R. J. Timmins. FMNH 258883. carapace (broken) and plastron only, PL = 36.6. collect- ing information same as FMNH 258882 except collect- ed 19 June 1994. FMNH 258877, carapace only, CL = 34.1, CW = 24.2, Salavan Province, Toumlan District. Xe Bang Nouan National Biodiversity Conservation Area, Ban Nalan Village, eaten by residents of Ban Nalan, 15 June 1994, R. .1. Timmins. FMNH 258878. intact shell only, CL = 36.8. CW = 25.8, PL = 35.3, BD = 14.7, Salavan Province, Xe Bang Nouan National Biodiversity Conservation Area, Ban Konglur Village, eaten by residents of Ban Konglur, 10 June 1994, R. J. Timmins. FMNH 258891, carapace only, CL = 17.3, CW = 15.1, Salavan Province, Xe Bang Nouan National Biodiversity Conservation Area, Ban Nasompeng Village, eaten by residents of Ban Nasompeng, 09 June 1994, R. J. Timmins. FMNH 258890, carapace (broken) only, CL = 19.4, Champasak Province, Pathoumphon District, Xe Pian National Biodiversity Conservation Area, Xe Pian River upstream from Ban Phonsaat Village, 100 m, discarded in camp along Xe Pian River 2004 1 ‘V * \: T9 «L M a**® v. ✓ 1 N. ., S . 35 * % • ■ 'i v 3 Figures 1-15. See text for locality details and circumstances of the record. 1. Platysternon megacephalum Huaphahn Province, Laos (photo B. L. Stuart); 2. Platysternon megacephalum Khammouan Province, Laos (photo B. L. Stuart) 3. Platysternon megacephalum Xe Kong Province, Laos (photo D. Showier); 4. Cuora amboinensis Battambang Province, Cambodia (photo B. L. Stuart); 5. Cuora amboinensis Kampong Thom Province, Cambodia (photo S G Platt); 6. Cuora amboinensis Koh Kong Province, Cambodia (photo S. G. Platt); 7. Cuora amboinensis Koh Konq Province, Cambodia (photo S. G. Platt). 8. Cuora amboinensis Kien Giang Province, Vietnam (photo B L Stuart)' 9 Cyclemys atripons Koh Kong Province, Cambodia (photo S. G. Platt); 10. Cyclemys tcheponensis Bolikhamxav Province, Laos (photo B. L. Stuart); 11. Heosemys grandis Koh Kong Province, Cambodia (photo S G Platt) 12 Hieremys annandalii Battambang Province, Cambodia (photo B. L. Stuart); 13. Hieremys annandalii Siem Reap Province, Cambodia (photo S. G. Platt); 14. Hieremys annandalii Kampong Thom Province, Cambodia (photo S G Platt); 15. Malayemys subtrijuga Battambang Province, Cambodia (photo B. L. Stuart). 2004 Asiatic Herpetological Research Vol. 10, p. 135 Figures 16-27. See text for locality details and circumstances of the record. 16. Malayemys subtrijuga Kampong Thom Province, Cambodia (photo S. G. Platt); 17. Malayemys subtrijuga Kandal Province, Cambodia (photo S. G. Platt); 18. Malayemys subtrijuga Vientiane, Laos (photo W. G. Robichaud). 19. Manouria impressa Koh Kong Province, Cambodia (photo S. G. Platt). 20. Manouria impressa Ratanakiri Province, Cambodia (photo Suon Phalla/TRAFFIC). 21. Manouria impressa Xe Kong Province, Laos (photo B. L. Stuart). 22. Chelonia mydas Kampong Speu Province, Cambodia (photo B. L. Stuart). 23. Eretmochelys imbricata Sihanoukville Province, Cambodia (photo S. G. Platt). 24. Dermochelys coriacea Gulf of Thailand, near Sihanoukville, Cambodia (photo Vanna Nhem). 25. Amyda cartilaginea Koh Kong Province, Cambodia (photo S. G. Platt). 26. Amyda cartilaginea Khammouan Province, Laos (photo B. L. Stuart). 27. Pelochelys cantorii Kratie Province, Cambodia (photo D. Gambade). after being eaten by hunters. May 1995, T. D. Evans. FMNH 255266, complete specimen, CL = 7.5, CW 6.4, PL = 6.2, BD = 2.5, Champasak Province, Mounlapamok District, Dong Khanthung Proposed National Biodiversity Conservation Area, 14° 07' N 105° 29' E, 60 m, grassland with dry dipterocaip and ever- green forest along Xe Lepou River, found in mud at bot- tom of flooded marsh, water depth about 15 cm, 1 1 July 1998, B. L. Stuart. FMNH 255272, carapace only, CL = 28.6, CW = 22.2, Champasak Province, Mounlapamok District, Dong Khanthung Proposed National Biodiversity Conservation Area, Ban Thahin Village, on Xe Lepou River, 14° 08' N 105° 35' E. 60 m, eaten by residents of Ban Thahin, 17 July 1998. B. L. Stuart. Vol. 10, p. Asiatic Herpetological Research 2004 CM O CM 110 E 2004 Asiatic Herpetological Research Vol. 10, p. 137 2 Z o m o CNJ T- T- o CNI m 110 E Vol. 10, p. 138 Asiatic Herpetological Research 2004 o CM in o T — 110 E 2004 Asiatic Herpetological Research Vol. 10, p. 139 z o CM CO CNI h- CN CO CM lO CM CM CO CM CM CN c Q E 'o CD CO N; CD T- CD CD o CO T- o CD CO CO q o CD cm 00 T- CO 00 co CO m CM CO N- CO D- co cb CO Is- 00 Is- CO N" CO 00 Is- Is- 00 Is- 00 CO Is- 00 CO CO 00 Is- LO q CN in r- m CO CO Is- T- in 00 co m CD co 00 T- N" T- q in < CO CO cd T — Is- - CD CD o cd d - CD o co CD x— CD o T — T — d - CD - - d d z Is- O in in < q < m in < < CD < < CO < < CO < o T- < < N; < < Is- CO ib CN CN m Is- z N- Z Is- N" N" z z CO N" z z cb N" z z CO N- z x — in 00 N" z z cd m z z ib co to Is- q q h- q N; T- q r- CD o 00 o T- q in N; m CN q O q co CO CO ib d cd N- d T— cd T— N" 00 r- CM cb in N1 T— cb cd cd r- cd in o N- N- CN N- CN N" CO CO N" CO N- N" CO co CN CO N" CO N" N" N" n- co N" N- co CO 35 + + + + + + + + + + + + + + + + + + + + + + + + + VN I I I I I I I I I I I I I + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + I I I I I I I I I I I I I I I I I I I I I I I I i + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + i i i i i i i • + + + + + + + + + + +2+ + + + + + +z + ' + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + ^SS^^cOCOCOCOCOCOCOCON-N-N-N-N-N-N'N-N-N-iOini'- SSSSSS5SSS5555S55oocooococococooooo Vol. 10, p. 156 Asiatic Herpetological Research 2004 assigned to a subgenus, but is regarded as a member of the "Tibeto-Himalayan" group. Our somewhat cursory examination of other members of this group, including C. baturensis (Khan, 1992), C. chitralensis (Smith, 1935) (synonymized with C. walli (Ingoldby, 1922) by Khan, [1992]), and C. mintoni (Golubev and Szczerbak, 1981), clearly shows that this group is highly artificial. Khan and Rosier (1999) did not consider Cyrtopodion stoliczkai as a member of the Pakistan gecko fauna, confining it to the upper Indus River valley in Kashmir. However, the morphological characters of our series from Skardu closely match those of Szczerbak and Golubev (1986, 1996) and Khan and Rosier (1999). Therefore, C. stoliczkai does indeed occur in Pakistan. Cyrtopodion stoliczkai (Steindachner, 1867) was described from a single specimen collected by Ferdinand Stoliczka in 1865 near Karoo, north of Dras, in northern Kashmir (Blanford, 1878). Some previous authors have cited Steindachner (1869) as the original description, but this actually refers to a reprint of the ear- lier work (K. Adler, pers. comm.). This single specimen was subsequently transferred to the Naturhistorisches Museum, Wien (Vienna, Austria), where Steindachner designated it as the holotype (NMW 16756) in honor of its collector. The holotype is well-illustrated by Szczerbak and Golubev (1986, 1996:Fig. 92). During the Second Yarkand Expedition (1873 - 1874), Stoliczka collected an additional 46 specimens from the type locality and a few localities eastward to Leh in the Indus River valley of central Ladakh, Kashmir. These speci- mens were subsequently deposited in the Indian Museum, Calcutta (Blanford, 1878). Annandale (1913) enumerated only 31 specimens in the Indian Museum collections in his treatment of Indian Gymnodactylus. Zugmayer (1909) and Brongersma (1935) reported this species from Lamayuru and Leh, respectively, both localities being in central Ladakh. Gruber (1981) col- lected 14 specimens from a few localities in the same general area as Stoliczka in 1865, which were deposited in the ZSM. Khan and Rosier (1999) presented a detailed redescription of C. stoliczkai based on this last series, but were unable to examine the holotype and the series in the Indian Museum. Khan and Rosier (1999) erroneously referred to a specimen of C. stoliczkai in the 2004 Asiatic Herpetological Research Vol. 10, p. 157 Museum of Comparative Zoology (MCZ 7132) as both a syntype and paratype. This specimen cannot be consid- ered either, as Steindachner mentioned only one speci- men in the original description, and Constable (1949:84) did not provide any type designation for this MCZ spec- imen. This specimen was received by the MCZ via exchange with the Indian Museum, Calcutta, in June 1908 (Constable, 1949:61; J. Rosado, pers. comm.). Stoliczka is given as the collector (Constable, 1949:61). Stoliczka died in 1874 during the Second Yarkand Expedition's return to India, indicating that his large series from Ladakh in the Indian Museum was collected in 1873 during the outbound portion of the expedition. We must assume that the MCZ specimen originates from this large series, however, it is possible that Stoliczka made additional collections in Ladakh between 1865 and 1873. Nevertheless, it is clear that Steindachner (1867) examined only one specimen at the time C. stoliczkai was described and the MCZ specimen cannot be a type. Additionally, Khan and Rosier (1999) referred to MCZ 7132 as a topotype. This may or may not be cor- rect, as Stoliczka's journal from the Second Yarkand Expedition indicated that only some of his specimens were collected at the type locality (Blanford, 1878) and others were collected elsewhere. However, since the only locality information available for the MCZ speci- men is "Ladakh", it cannot be ascertained that it is actu- ally one of the specimens collected at the type locality. Gymnodactylus walli Ingoldby, 1922 and G. yarkan- densis Anderson, 1872 were regarded as synonyms of Cyrtopodion stoliczkai by Smith (1935), a view fol- lowed by virtually all subsequent authors. Minton (1966) referred to a single specimen from Udigram, Swat District, Northwest Frontier Province, Pakistan as C. stoliczkai , which was later found to be a distinct species (Mertens, 1969:26; Khan, 1980:14; described as Gymnodactylus mintoni by Golubev and Szczerbak in 1981). Khan (1992) produced a compelling argument to consider Cyrtopodion walli distinct from C. stoliczkai based on an examination of the type specimens in the British Musuem. We are unable to resolve the synonymy of C. yarkandensis (Anderson, 1872). Blanford (1878) relegated C. yarkandensis to the synonymy of C. stoliczkai (Steindachner, 1 867), and subsequent authors followed this view (Annandale, 1913; Boulenger, 1890; Kluge, 1991, 1993, 2001; Mertens, 1969; Minton, 1966; Smith, 1935; Szczerbak and Golubev, 1986, 1996; Wermuth, 1965; Zhao and Adler, 1993). Khan (1994) resurrected C. yarkandensis based on an examination of a single specimen housed in the British Museum (BMNH 72.3.22.4). A comparison of color transparen- cies taken during a study of the same specimen by W. Auffenberg in the early 1990s, along with our series from Skardu, indicate that this specimen is probably best assigned to C. stoliczkai. However, important morpho- logical characters cannot be ascertained from the trans- parencies or Khan's (1994) description. Szczerbak and Golubev (1986, 1996) also assigned this specimen to C. stoliczkai. Whether C. yarkandensis is a distinct taxon or a synonym of C. stoliczkai can be determined only with a thorough examination of the types housed in the Indian Museum, Calcutta. We provide the following notes on C. yarkandensis at this point merely for a his- torical perspective. Anderson (1872) mentioned two specimens in his description of Cyrtodactylus yarkandensis. These were supposedly collected in Yarkand (= Shache, Xinjiang, China; Zhao and Adler, 1993) during the First Yarkand Expedition in 1870 (Blanford, 1878). This locality was doubted by Blanford (1878:12-13), maintaining that the types of C. yarkandensis were identical to the C. stoliczkai specimens collected by Stoliczka during the Second Yarkand Expedition in Ladakh, some of which were taken from the type locality of C. stoliczkai. Blanford (1878:13) stated "The specimens described by Dr. Anderson as Cyrtodactylus yarkandensis were brought, with others, by a collector, who accompanied Dr. Henderson on the mission which was sent to Yarkand in 1870; this mission traversed precisely the same route through Kashmir and Leh as the second in 1 873 - 74, and I do not think there can be any reasonable doubt that the real locality whence Cyrtodactylus yarkandensis was obtained must have been Ladak, and not Yarkand." Annandale (1913:316) incorrectly attrib- uted the collection of the types of C. yarkandensis to Stoliczka during the Second Yarkand Expedition. That mission embarked in 1873, about one year after Anderson's (1872) description of C. yarkandensis. Khan (1994) referred to this specimen (BMNH 72.3.22.4; "Yarkhand") as a syntype. The specimen catalogue at the British Museum indicates that this specimen was "Presented by [the] Indian Museum Calcutta through Dr. Anderson" and cataloged on March 22, 1872 (C. McCarthy, pers. comm.), the same year the species was described. Anderson (1872:381) mentioned only two specimens in the original description and Annandale (1913:316) referred to two specimens (ZSI 3792 - 93) as types of C. yarkandensis (as a synonym of Gymnodactylus stoliczkai Steindachner) in the Indian Museum, Calcutta. It can be assumed that these were the specimens on which Anderson based his description, thus the status of the British Museum specimen remains nebulous. Alsophylax ( Altiphylax ) boehmei Szczerbak, 1991 was described from two specimens collected by G. Osella from Skardu, Pakistan in July 1976. Although we did not examine the holotype, we have no doubt that the description of this species is based on subadult Vol. 10, p. 158 Asiatic Herpetological Research 2004 Cyrtopodion stoliczkai. This relationship was originally suggested by Golubev (in Szczerbak and Golubev, 1996:200, footnote). Morphological characters for A. boehmei provided by Szczerbak (1991) fall within the range of variation in those we recorded for C. stoliczkai (Table 1). The holotype (ZFMK 38773, see Fig. 3 in Szczerbak, 1991) matches the subadults in our complete growth series collected in Skardu in 1991 (Table 2). The whorls of the anterior third of the tail of C. stoliczkai do not become swollen and lobed until maturity, but Szczerbak (1991) lacked a sufficient series of specimens to make this determination. Golubev (in Szczerbak and Golubev, 1996:200, footnote) also suggested that Tenuidactylus baturensis Khan and Baig, 1992 may also be conspecific with Cyrtopodion stoliczkai. Our examination of one speci- men collected near the type locality of T. baturensis indi- cates that although it is similar in overall morphology, this species appears to be distinct. Khan (200 1 ) divided the Tibeto - Himalayan group of Cyrtopodion into three subgroups: Stoliczkai sub- group = C. baturensis (Khan and Baig, 1992), C. stoliczkai (Steindachner, 1867), and C. yarkandensis (Anderson, 1872); Tibetinus subgroup = C. battalensis (Khan, 1993), C. dattanensis (Khan, 1980), C. himalayanus (Duda and Sahi, 1978), C. mintoni (Golubev and Szczerbak, 1981), and C. tibetinus (Boulenger, 1905); and the Walli subgroup - C. walli (Ingoldby, 1922) (including C. chitralensis [Smith, 1935] as a synonym) and C. kirmanense (Nikolsky, 1900). Our preliminary examination of most of these taxa reveals that Khan's system has merit concerning overall morphological and ecological data. Further investigations into the Pakistan gecko fauna and that of adjacent areas will undoubtedly lead to further discover- ies of new species and more clearly define those already described. Acknowledgments We would like to thank the Division of International Conservation, United States Fish and Wildlife Service (Washington, D. C.), Deutscher Akademischer Austauschdienst, Bonn, Germany, and the Office of Sponsored Research, University of Florida for the fund- ing support awarded to W. Auffenberg that made possi- ble the fieldwork and museum visits by him. We also thank Kraig Adler (CU), Steve Anderson (University of the Pacific), M. S. Khan (Secane, Pennsylvania), Arnold Kluge (UMMZ), Colin McCarthy (BMNH), and Jose Rosado (MCZ) for pertinent literature, insight, and the examination of specimens under their care. We also rec- ognize Max Nickerson and Wayne King (Florida Museum of Natural History) for their encouragement and support. Tammy Johnson prepared Fig. 2. We thank the dedicated staff of the Zoological Survey Department of Pakistan, particularly Mohammad Farooq Ahmed, former Director, Hafizur Rehman, Shamim Fakhri, and Aleem Khan, for their support and assistance throughout the fieldwork portion of this project. Literature Cited Anderson, J. 1872. On some Persian, Himalayan, and other reptiles. Proceedings of the Zoological Society of London 1872(2):37 1-404. Anderson, S. C. 1999. The Lizards of Iran. Society for the Study of Amphibians and Reptiles, Ithaca, NY. pp. 442. Annandale, N. 1913. The Indian geckos of the genus Gymnodactylus. Records of the Indian Museum 9:309-326, pis. 16-17. Blanford, W. T. 1878. Scientific results of the Second Yarkand Mission; based on the collections and notes of the late Ferdinand Stoliczka, Ph.D. Reptilia and Amphibia, Calcutta, 26 pp. Boulenger, G. A. 1890. The Fauna of British India, including Ceylon and Burma. Reptilia and Batrachia. Taylor and Francis, London, xviii + 541 pp. Boulenger, G. A. 1905. On some batrachians and reptiles from Tibet. Annals and Magazine of Natural History (7th series) 11:379-380. Brongersma, L. D. 1935. Amphibien und Reptilien. Pp. 446-451. In Visser (ed.), Karakorum I. Wissensch- aftliche Ergebnisse der Niederlandischer Expeditionen in den Karakorum und die angrennzenden Gebiete 1922, 1925 und 1929/30. Brockhaus, Leipzig. Constable, J. D. 1949. Reptiles from the Indian penins- ula in the Museum of Comparative Zoology. Bulletin of the Museum of Comparative Zoology 1 03(2):59- 1 60. Duda, P. L. and D. N. Sahi. 1978. Cyrtodactylus himalayanus: A new gekkonid species from Jammu, India. Journal of Herpetology 12(3):35 1-354. Fitzinger, L. J. 1843. Systema reptilium. Fasciculus Primus: Amblyglossae (Conspectus Geographicus). Braumuller und Seidel, Vienna, iv + 106 pp. 2004 Asiatic Herpetological Research Vol. 10, p. 159 (Reprint 1973 Society tor the Study of Amphibians and Reptiles, Ithaca, New York). Golubev, M. L. and N. N. Szczerbak. 1981. [A new species of the genus Gymnodactylus Spix, 1823 (Reptilia, Sauria, Gekkonidae)]. Vestnik Zoologii 198 1(3):40-45. (In Russian). Gruber, U. 1981. Notes on the herpetofauna of Kashmir and Ladakh. British Journal of Herpetology 6:145- 150. Ingoldby, C. M. 1922. A new stone gecko from the Himalaya. Journal of the Bombay Natural History Society 28:1051. Khan, M. S. 1980. A new species of gecko from north- ern Pakistan. Pakistan Journal of Zoology 12(1): 11-16. Khan, M. S. 1992. Validity of the mountain gecko Gymnodactylus walli Ingoldby, 1922. Herpetological Journal 2:106-109. Khan, M. S. 1993. A new angular-toed gecko from Pakistan, with remarks on the taxonomy and a key to the species belonging to genus Cyrtodactylus (Reptilia: Sauria: Geckkonidae). Pakistan Journal of Zoology 25(l):67-73. Khan, M. S. 1994. Validity and redescription of Tenuidactylus yarkandensis (J. Anderson, 1872). Pakistan Journal of Zoology 26(2): 139-143. Khan, M. S. 2001. Taxonomic notes on angular-toed Gekkota of Pakistan, with description of a new species of genus Cyrtopodion. Pakistan Journal of Zoology 33(1): 13-24. Khan, M. S. and K. J. Baig. 1992. A new Tenuidactylus gecko from northeastern Gilgit Agency, North Pakistan. Pakistan Journal of Zoology 24(4):273- 277. Khan, M. S. and H. Rosier. 1999. Redescription and generic redesignation of the Ladakhian gecko Gymnodactylus stoliczkai Steindachner, 1969 [sic], Asiatic Herpetological Research 8:60-68. Kluge, A. G. 1991. Checklist of gekkonid lizards. Smithsonian Herpetological Information Service 85:1-35. Kluge, A. G. 1993. Gekkonoid Lizard Taxonomy. International Gecko Society, San Diego, 245 pp. Kluge, A. G. 2001. Gekkotan Lizard Taxonomy. Hamadryad, Special Publication 26(1): 1-209. Leviton, A. E., R. H. Gibbs, Jr., E. Heal, and C. E. Dawson. 1985. Standards in herpetology and ichthyology: Part I. Standard symbolic codes for institutional resource collections in herpetology and ichthyology. Copeia 1985:802-832. Mertens, R. 1969. Die Amphibien und Reptilien West- Pakistans. Stuttgarter Beitrage zur Naturkunde 197:1-96. Minton, S. A., Jr. 1966. A contribution to the herpeto- logy of West Pakistan. Bulletin of the American Museum of Natural History 1 34(2):29- 1 84. Nikolsky, A. M. 1899 [1900]. Reptiles, amphibiens et poissons, recueillis pendant la voyage de Mr. N. A. Zarudny en 1 898 dans la Perse. Annuaire du Musee Zoologique de l'Academie Imperiale des Sciences de St. Petersbourg, 4:375-417. (In Russian). Smith, M. A. 1935. The Fauna of British India, Including Ceylon and Burma. Reptilia and Amphibia, vol. 2, Sauria. Taylor and Francis, London, xiii + 440 pp. Steindachner, F. 1867. Reptilia. In: F. Steindachner (ed.) Reise der Osterreichischen Fregatte Novara um die Erde in den Jahren 1857, 1858, 1859 unter den Befehlen des Commodore B. von Wullerstorf- Urbair, Zoology, vol. 1, part 3. Kaiserlich- Koniglichen Hof-Staatsdruckerei, Vienna, 98 pp.. Steindachner, F. 1869. Reptilia. In: F. Steindachner (ed.) Reise der Osterreichischen Fregatte Novara um die Erde in den Jahren 1857, 1858, 1859 unter den Befehlen des Commodore B. von Wullerstorf- Urbair, Zoology, vol. 1, part 3. Kaiserlich- Koniglichen Hof-Staatsdruckerei, Vienna, 98 pp., 3 pis. (Reprint of 1867 Edition). Szczerbak, N. N. 1991. Eine neue Gecko-Art aus Pakistan: Alsophylax ( Altiphylax ) boehmei sp. nov. Salamandra 27(l):53-57. Szczerbak, N. N. and M. L. Golubev. 1986. [Gecko Fauna of the USSR and Adjacent Regions]. Nauka Dymka, Kiev, 232 pp. (In Russian). Szczerbak, N. N. and M. L. Golubev. 1996. Gecko Fauna of the USSR and Adjacent Regions. [English edition translated from the Russian by M. L. Golubev and S. A. Malinsky; A. E. Leviton and G. Vol. 10, p. 160 Asiatic Herpetological Research 2004 R. Zug, eds.]. Society for the Study of Amphibians and Reptiles, Ithaca, New York, 232 pp. Wermuth, H. 1965. Liste der rezenten Amphibien und Reptilien: Gekkonidae, Pygopodidae, Xantusidae. Das Tierreich 80:xxiv + 246 pp. Zhao, E. and K. K. Adler. 1993. Herpetology of China. Society for the Study of Amphibians and Reptiles, Oxford, Ohio, 522 pp. Zugmayer, E. 1909. Beitrage zur Herpetologie von Zentral-Asien. Zoologischen Jarhbuchem 27(5):481-508. 2004 Asiatic Herpetological Research Vol.10, pp. 161-163 Antimicrobial Activity in the Skin Secretion of Bufo viridis (Laurenti, 1768) Ba§aran Dulger1’*, Ismail Hakki Ugurta§2, and Murat Sevinc2 Canakkale Onsekiz Mart University, Faculty of Science & Arts, Department of Biology, C an akkal e-Turkey 2 Uludag University, Faculty of Science & Arts, Department of Biology, Bursa-Turkey * Correspondence author, E-mail: dbasaran@comu.edu.tr; Fax number: +90.286.2 180533 Abstract. - In this study, antimicrobial activity of various extracts prepared from Bufo viridis skin secretion were tested against the microorganisms by disk diffusion method. Escherichia coli ATTC 10536, Listeria monocytogenes ATCC 19117, Klebsiella pneumoniae UC57, Salmonella typhi ATCC 19430, Staphylococcus aureus ATCC 6538P, Mycobacterium smegmatis CCM 2067, Rhodotorula rubra and Saccharomyces cerevisae ATCC 9763 were used. According to our results, the extracts prepared from Bufo viridis skin secretion have high antimicrobial activity against the tested microorganisms. Keywords. - Bufo viridis, Amphibia, antimicrobial activity, skin secretion. Introduction Amphibians have skin glands producing mucous and poison. Amphibians have been studied and have attracted special attention from a toxicological point of view. Various substances with antimicrobial activity have been isolated from skin secretions of amphibian species (Dapson, 1970; Croce et al., 1973; Dapson et al., 1973; Preusser et al., 1975; Cevikbas, 1978). Several toxins in amphibian poisons have been used as experi- mental tools and contributed to significant progress in physiology. Some toxins (Batrachotoxins) specifically block the inactivation of the voltage regulated Na+ chan- nels in nerve and muscle cells, which causes a massive inflow of Na+. The cells become irreversibly depolar- ized, which, among other things, produces heart arrhyth- mia and respiratory failure and finally cardiac insufficiency. In humans, some amphibian toxins (Bufo- tenin) produce symptoms similar to those of LSD (Lutz, 1971; Edstrom, 1992). In previous studies, some skin secretions showed remarkable cytotoxic activity against eukaryotic cells (Kolbe et al., 1993; Sanna et al. 1993). The aim of the this study is to test the antimicrobial activity of Bufo viridis skin secretions against Gram- positive, Gram-negative bacteria and yeast cultures for future possible use in providing pharmacological tools for the study of new drugs and aid in benefitting human health. Materials and Methods Specimens of Bufo viridis were collected from different regions in Bursa, Turkey in March 1998. Collected frogs were brought to the laboratory and kept in an aquarium. Before experimentation, the frogs were washed first with tap water and then with distilled water. They were placed for 3-5 minutes in a glass jar containing a piece of cotton soaked with ether to stimulate skin secretions. The secretion accumulated on the skin was obtained by scraping the body of the animals with a spatula. The foamy secretion thus obtained was placed in a tube, left in an 80°C water bath for 30 min and centrifuged at 5,500 rev/min for 30 min. After centrifugation, the pre- cipitate was used in the experiments. Before using in the experiments, the precipitate was diluted with distilled water 0.1 M HC1, 0.1 M NH4OH, and 1 M phosphate buffers (pH: 4 and pH: 7). In this study, Escherichia coli ATTC 10536, Liste- ria monocytogenes ATCC 19117, Klebsiella pneumo- niae UC57, Salmonella typhi ATCC 19430, Staphylococcus aureus ATCC 653 8P, Mycobacterium smegmatis CCM 2067 bacteria cultures and Rhodot- orula rubra and Saccharomyces cerevisae ATCC 9763 yeast cultures were used. In vitro antimicrobial activity studies were carried out by the Agar-Disc Diffusion Method. Mueller Hinton Agar (Oxoid) was preferred as the most suitable medium for antimicrobial activity studies. Each extract was implemented into a sterile disc in varying concen- trations starting from 20 pi. Each disc was 6 mm in diameter. Bacteria and yeast cultures were suspended in 4-5 ml Brain Heart Infusion Broth (Oxoid) and Malt Extract Broth (Difco). Bacteria were incubated in 37°C for 2-5 hours. Yeast cultures were incubated in 30°C for 5-7 hours. A visible turbidity was obtained at the end of this time. The turbidity of bacterial suspension was adjusted © 2004 by Asiatic Herpetological Research Vol.10, p. 162 Asiatic Herpetological Research 2004 Table 1. Antimicrobial activity of various extracts of Bufo viridis skin secretions on microorganisms. Microorganisms / solvents 0.1 N HCI 0.1 N NhUOH Phosphate Buffer pH: 4 pH: 7 Distilled water Escherichia coli ATCC 10356 ++ +++ (+) (+) ++ Listeria monocytogenes ATCC 19117 +++ +++ ++ ++ ++ Staphylococcus aureus ATCC 6538P +++ ++ ++ ++ +++ Klebsiella pneumoniae UC57 +++ ++ +++ ++ +++ Salmonella thyphi ATCC 19430 +++ ++ +++ ++ ++ Mycobacterium smegmatis CCM 2067 +++ +++ ++ ++ +++ Rhodotorula rubra +++ +++ +++ +++ ++ Saccharomyces cerevisiae ATCC 9730 +++ +++ +++ +++ +++ (+) : Inhibition zone less than 1 mm surrounding the 6 mm paper disk. + : Inhibition less than ++ : Inhibition comparable to +++: Inhibition more than 10 pg penicillin or sulconazole / disk; Inhibition zones of references : 12-16 mm diameter. according to Macfarland Standard Tube [0,5] with phys- iologic serum and inoculation performed. Prepared bac- terial suspension was mixed with a sterile applicator and excess fluid of applicator was removed by rotating the applicator to one side of the tube. We streaked the entire Mueller Hinton Agar surface in three different direc- tions by rotating the plate 60° angles after each streak- ing. Yeast cultures were inoculated into Muller Hinton Agar (102 cfu/ml). All petri dishes after inoculation were allowed to dry for 15-20 min at room temperature (bac- teria at 35°C and yeast at 30°C). Inhibition zone diame- ters were measured after 24-48 hours (Collins et al. 1987, NCCLS 1993). In addition, continued only sol- vent was used as negative control disc and antibiotic penicillin and sulcanozole discs were used as references. Experiments were repeated three times and results were expressed as average values. Results and Discussion Antimicrobial activity effects of five different extracts, which were prepared by using distilled water, 0.1 N HC1, 0.1 N NH4OH, 1 M phosphate buffers (pH: 4 and pH: 7), were obtained from the skin secretions of Bufo viridis against bacteria and yeast cultures, results are given in Table 1. According to our findings, all the extracts of skin secretion against the yeast cultures exhibit higher anti- microbial activity than that of a compared antibiotic. The 0.1 M HC1 extract shows more effect than that of the other extracts against bacteria. 1 M phosphate buffer (pH: 4 and 7) extracts exhibited minor effects against Escherichia coli. However, phosphate buffer (pH: 4 and 7) extracts exhibited strong effects against the other bac- teria. It can be said that the active substance obtained from Bufo viridis skin secretion dissolves easily in the 0.1 M HC1 and has high antimicrobial activity as a con- sequence. It has been reported that sensitivity of the microorganisms to the chemotherapeutic agents changes from strain to strain (Cetin et al., 1989). Our results are in agreement with the other authors’ results. Inhibition zone diameters around the control disc were measured as 0-1 mm. In this study, antimicrobial effects of the prepared extracts on the tested microor- ganisms were determined by using different solvents. Croce et al. (1973) investigated antimicrobial activ- ity of skin secretions from Bombina variegata pachy- pus. They homogenized skin secretion with phosphate buffers (pH: 4 and 7) 1 M HC1, 1 M NH4OH and dis- tilled water. These homogenates show high antimicro- bial activity against Staphylococcus aureus but they do not show any antimicrobial effect against Aspergillus niger, Trichophyton mentagrophytes ATCC 8757 and Candida albicans. Cevikbas (1978) examined antibacterial activity in the skin secretions of Rana ridibunda. The author reported that skin secretion of Rana ridibunda shows antibacterial activity at different levels. However, in our 2004 Asiatic Herpetological Research Vol.10, p. 163 piesent study, skin secretions of Bufo viridis against the yeast cultures shows more antimicrobial activity than that of the bacterial cultures. Our findings parallel those reported in the above studies. In Amphibia, antimicro- bial activity of skin secretions differ at both the generic and specific levels. Although the antimicrobial activity of skin secre- tions from Bufo viridis , Bufo vulgaris , Salamandra mac- ulosa and Salamandra atra were determined (Pavan, 1962; Pavan and Nascimbene, 1948), antimicrobial activity ot skin secretion from Bufo marinus, Triturus, and Xenopus were not observed (Preusser et al., 1975; Kolbe et al., 1993; Ozeti and Yilmaz, 1994). Antiyeast activity observed in our study was not observed in Croce’s et al. study (1973). Our results show that skin secretion components from Bufo viridis may be different from Bombina variegata pachymus. Acknowledgments The authors are grateful to Aegean University, Science Faculty, Department of Biology, and Basic and Indus- trial Microbiology for supplying the strain of microor- ganism used in the study. References Cetin, T. E. and N. Gurler. 1989. Bakterilerin Antibiyo- tiklere Duyarlilik Deneyinin Yapilmasi. Kukem 12:2-5. Cevikbas, A. 1978. Antibacterial activity in the skin secretion of the frog Rana ridibunda. Toxicon 16(2): 195-197. Collins, C. M., P. M. Lyne, and J. M. Grange. 1989. Microbiological Methods, Six Edition. Butter- worths & Co. Ltd., London. 410 pp. Croce, R., N. Giglioli, and L. Bolognani. 1973. Antimi- crobial activity in the skin secretions of Bombina variegata pachymus. Toxicon 1 1 :99-100. Edstrom, A. 1992. Venomous and Poisonous Animals. Kriger Publishing Company. Malabar, Florida. USA. 226 pp. Kolbe, FI. V. L, A. Fluber, P. Cordier, U. B. Rasmussen, B. Bouchon, M. Jaqunod, R. Vlasak, E. Delot, and G. Kreil. 1993. Xenoxins: A Family Of Peptides From Dorsal Gland Secretion Of Xenopus laevis Related To Snake- Venom Cytotoxins And Neuro- toxins. Journal of Biological Chemistry 268(22): 16458-16464. Lutz, B. 1971. Venomous Animals and Their Venom. Academic Press, London. 423-427 pp. NCCLS. 1993. Performance Standards for Antimicro- bial Disk susceptibility Tests. Approved Standard NCCLS publication M2- A5, Villanova, PA, USA. Ozeti, N. and F Yilmaz. 1994. [Amphibians of Turkey]. Aegean University Press, Izmir. 151 pp. Pavan, M., 1962. Die Antibiotica tierischer Herkunft. Zeitschrift Fur Hygiene und Infektionskrankheiten 34:136-138. Pavan, M., and A. Nascimbene. 1948. Studi sugli antibi- otici di origine animale. Bolletino Societa Medico- chirurgica, Pavia 1-2:229. Preusser, H. J., G. Habermehl, M. Sablofski, and H. D. Schmall. 1975. Antimicrobial activity of alkoloid from amphibian venoms and effects on the ultras- tucture of yeast cells. Toxicon 12: 285. Sanna, A. P., A. Bamabei, and G. Delfino. 1993. The Cutaneous Venom of Bombina orientals : Cytotoxic Effects on the human HL 60 Cell Line and A Com- parison with Bombina variegata. Journal of Natural Toxins 2(2): 16 1-173. Analysis of the Stomach Contents of the Lycian Salamander Mertensiella luschani (Steindachner, 1891) (Urodela: Salamandridae), Collected from Southwest Turkey Serdar Du§en, Mehmet Oz, and M. Rizvan Tunc Akdeniz University, Faculty of Arts and Sciences, Department of Biology, 07058, Antalya, Turkey Abstract. - In this paper, the stomach contents of 116 specimens (39 males, 47 females, and 30 juveniles) from the Southwest Turkey Mertensiella luschani populations are analyzed. A total of 342 prey items were identified and their frequency of occurence and percent of diet were tabulated. The majority of the diet consisted of Insecta (50.58%), and within Insecta, Coleoptera (65.32%) was the major order represented. In addition to insects, M. luschani feeds on Gastropoda (19.59%), Arachnida (16.08%), Myriapoda (8.57%), Clitelliata (3.50%) and Crustacea (1.75%). Key words. - Mertensiella luschani , stomach contents, prey, southwest Turkey. Baran and Atatiir, 1980, M. 1. billae Franzen and Kle- wen, 1987, and M. 1. flavimembris Mutz and Steinfartz, 1995. Mertensiella luschani is not dependent on water, it inhabits humid soils and crevices under the Pinus bru- tia forests, Mediterranean maquis, and open rocky areas. Its vertical distribution ranges between 15-1300 m. Various studies have been done on M. luschani in terms of its taxonomy (Franzen et all, 2001), ecology (Klewen, 1991; Steinfartz and Mutz, 1998), and repro- ductive biology (Ozeti, 1973; Ozeti, 1980). The aim of Figure 1. Collecting localities of Mertensiella luschani in Southwest Turkey. 1-Kocagol, 2-Dodurga, 3- Letoon, 4-Nadar lar, 5-Finike, 6-Buyukgaltlcak, 7-Hurma, 8-Fersin Introduction Nine subspecies of the Lycian Salamander, Mertensiella luschani, are distibuted along the coast of Southwestern Turkey and on some islands (e.g., Kastellorizon, Meis, Kekova, and Karpathos) (Baran and Atatiir, 1997; Ba^- oglu et al., 1994; Veith et ah, 2001). These are M. 1. lus- chani Steindachner, 1891, M. 1. helverseni Pieper, 1963, M. 1. atifi Ba?oglu, 1967, M. l.fazilae Ba^oglu and Atat- ur, 1974, M. l.finikensis Ba^oglu and Atatiir, 1975, M. 1. antalyana Ba^oglu and Baran, 1976, M. 1. basoglui © 2004 by Asiatic Herpetological Research Vol.10, p. 165 Asiatic Herpetological Research 2004 Table 1. Composition of the stomach contents of Mertensiella luschani (39 males, 47 females, 30 juve- niles) collected from the Southwest Turkey. N: The numbers of every prey found in all stomachs, n: The number of stomachs every prey type was found in. Taxon N (%) n (%) GASTROPODA Pulmonata 67 19.59 36 31.03 CLITELLIATA Neogliochaeta (=Prospora) Lumbricidae 12 3.50 9 7.75 CRUSTACEAE Isopoda 6 1.75 6 5.17 MYRIAPODA Diplopoda 1 0.29 1 0.86 Julidae 17 4.97 14 12.07 Chilopoda 8 2.34 6 5.17 Geophilidae 3 0.88 3 2.58 ARACHNIDA Aranae 37 10.82 27 23.27 Pseudoscorpionida 18 5.26 15 12.93 (=Chelonethi) INSECTA Collembola 4 1.17 3 2.58 (=Podura) Dermaptera Forficulidae 8 2.34 8 6.89 Isoptera 1 0.29 1 0.86 Heteroptera 4 1.17 5 4.31 Lygaidae 3 0.88 1 0.86 Homoptera 1 0.29 1 0.86 Coleoptera 113 33.04 51 43.96 Hymenoptera Formicidae 31 9.06 19 16.37 Diptera 3 0.88 3 2.58 Lepidoptera 5 1.46 5 4.31 2004 Vol.10, p. 166 Hvmenoptera 1 7 . 92% Dermaptera 4.62% Heteroptera 4.05% .Lepidoptera 2.89% ■ Collembola 2.31 % - Diptera 1.73% Others 1.16% - Coleoptera 65.32% Figure 2. Distribution of the insect groups in numerical percentages. the present preliminary investigation on seven M. lus- chani subspecies (except M /. flavimembris and M. I helverseni ) from eight localities in Southwest Turkey is an analysis of stomach contents. Materials and Methods Specimens for this study were collected from the eight localities in Southwest Turkey during the known activ- ity period of M. luschani (December-February, 1999). A total 116 (39 males, 47 females, and 30 juveniles) M luschani specimens were collected by hand under stones. Collection sites are shown in Figure 1. Once collected, the salamanders were taken to the laboratory to undergo stomach-flushing. A thin pipe wash bottle is inserted in salamander’s esophagus and stomach. Gentle pressure on the wash bottle forces dis- tilled water in to the stomach and forces the food out throught mouth (modified from Gittins, 1987). The prey items obtained from each specimen were labeled and stored in 10 cc. bottles containing 70% ethanol. Dried pieces from both undigested and partially digested prey were placed on microscope slides and held in place with cellophane tape (Du§en and Oz, 2001). These pieces consisted of whole body, wings, thorax with abdomen, head, and mouth parts. Through this approach, identifi- cation to the lowest taxonomic categories was attempted, samples were examined using a stereomicro- scope with 10-25x magnification. Prey items were iden- tified and grouped utilizing methods described elsewhere (Demirsoy, 1998a,b; Grzimek, 1979a,b, Lodos, 1986; Riehm, 1984) Results We did not observe any significant differences in the stomach contents of seven subspecies males, females and juveniles; they were thus evaluated together. Of the 116 specimens (39 males, 47 females, and 30 juveniles), only two females had empty stomachs. Small unrecog- nizable insect remains (parts of heads and larvae, anten- nae, wings, etc.) of the stomach contents are not included to the numerical analysis. Other non-food materials such as small pebbles, sand grains, plant parti- cles, and pieces of feather, possibly ingested during prey capture were not included either. A total of 342 prey items were counted from the investigated stomach contents (Fig. 2); Insecta 173 (50.58%), Gastropoda 67 (19.59%), Arachnida 55 (16.08%), Myriapoda29 (8.57%), Clitelliata 12 (3.50%) and Crustacea 6 (1.75%). Table 1 presents the stomach contents with respect to prey groups (their taxonomic grouping, number of prey items, and percentages of preyers). Insects were identified to the ordinal level. The total number of prey and their percentages are as follow: coleopters 113 (65.32%), hymenopterans 31 (17.92%), dermapterans 8 (4.62%), heteropterans 7 (4.05%), lepi- dopterans 5 (2.89%), dipterans (1.73%), collembolan 4 (2.31%), homopterans 1 (0.57%), and isopterans 1 (0.57%) (Fig. 2). The same insect orders can be ranked from the viewpoint of the number of prey eaten; their percentages are as follows: coleopterans 51 (43.96%), co i JD Q. O © O O co © o_ o c © E >. x CO t_ 0) Q_ 03 E © Q co © Q. O T3 Q. © CO i_ © Q. 0 1 © © X JO o -O E © o O ■ Prey Eater □ Relative Percentage CO L_ 0) Q_ b CO 2 1 (-2)? 1 1 1? 1? Balancer rudimentary rudimentary? well devel- oped No? No? No No? No? Vol. 10, p. 173 Asiatic Herpetological Research 2004 dis- tance min. max snout number eye dist. hind distance hight to ant. of rec- total head body body tail head diame- eye to foreleg leg between dorsal point ognized mass d.a.h, length length lengthl Iength2 length width ter snout length length feet fine dorsal fin costal grooves (9) 1 MEAN 19.28 2.40 11.05 11.63 7.49 2.54 0.73 0.89 0.65 1.04 4.01 0.05 1 MIN 19.05 2.25 10.80 11.40 7.06 2.00 0.67 0.77 0.50 1.00 3.50 0.05 1 MAX 19.61 2.50 11.40 11.80 7.80 2.95 0.80 0.97 0.80 1.05 5.00 0.05 1 SD 0.28 0.11 0.25 0.17 0.37 0.40 0.06 0.09 0.15 0.03 0.67 0.00 N = 4 (1) 4 MEAN 19.30 2.67 11.00 11.73 7.37 2.45 0.77 0.90 0.65 1.12 3.50 0.05 4 MIN 18.50 2.40 10.50 11.20 7.00 2.30 0.70 0.90 0.55 1.10 3.00 0.04 4 MAX 19.79 2.90 11.40 12.10 7.60 2.65 0.85 0.90 0.80 1.15 4.00 0.06 4 SD 0.70 0.25 0.46 0.47 0.32 0.18 0.08 0.00 0.13 0.03 0.50 0.01 N = 3 (1) 8 MEAN 21.76 2.79 12.11 12.73 8.86 3.19 0.96 1.06 1.34 1.33 5.09 0.07 8 MIN 21.27 2.25 11.80 12.20 8.40 2.75 0.90 0.90 1.00 1.10 4.75 0.06 8 MAX 22.20 3.60 12.50 13.00 9.40 3.50 1.02 1.22 1.70 1.50 5.50 0.08 8 SD 0.30 0.53 0.27 0.29 0.37 0.26 0.04 0.11 0.23 0.12 0.23 0.01 N = 8 (1) 14 MEAN 24.61 3.28 13.68 14.44 10.15 3.61 1.03 1.22 2.14 0.70 7.97 1.45 5.90 0.10 14 MIN 23.13 2.65 12.76 13.50 9.50 2.90 0.95 1.00 1.50 0.55 7.45 1.35 4.00 11 0.09 14 MAX 25.53 3.70 14.00 14.90 10.70 4.10 1.10 1.45 2.60 0.90 8.25 1.60 8.00 14 0.12 14 SD 0.92 0 39 0.41 0.53 0.44 0.47 0.06 0.14 0.35 0.14 0.26 0.09 1.29 0.01 N = 7(1) 16 MEAN 27.06 4.03 14.65 15.54 11.47 3.86 1.19 1.36 2.72 1.39 8.37 1.49 6.56 0.12 16 MIN 26.45 3.25 14.10 15.10 10.36 3.50 0.95 1.00 2.00 1.00 8.15 1.40 5.05 12 0.11 16 MAX 27.93 4.40 15.54 16.65 12.00 4.00 1.30 1.55 3.10 1.55 8.60 1.55 8.15 14 0.15 16 SD m — n 0.43 0.35 0.41 0.48 0.53 0.17 0.10 0.16 0.32 0.19 0.16 0.05 0.86 0.01 19 In — y MEAN 24.87 3.28 13.83 14.56 10.09 2.83 1.08 1.08 1.86 1.41 8.56 1.22 5.29 0.06 19 MIN 20.90 2.35 11.80 12.50 8.00 2.00 0.75 0.75 0.75 0.25 7.25 0.95 3.00 13 0.04 19 MAX 36.90 5.40 19.79 20.53 16.28 5.30 1.80 1.70 5.20 4.00 9.70 2.15 8.00 14 0.21 19 SD 4.28 0.80 2.04 2.10 2.32 0.88 0.29 0.28 1.21 1.22 0.88 0.30 1.69 0.04 N =15 (3) 24 MEAN 26.67 3.70 14.75 15.63 10.77 3.11 1.09 1.18 1.78 0.60 8.93 1.30 4.94 0.08 24 MIN 22.94 3.00 13.30 14.10 8.90 2.65 0.90 0.85 1.25 0.50 8.00 1.10 3.50 0.07 24 MAX 30.15 4.80 16.40 17.20 13.00 3.75 1.30 1.65 2.50 0.75 9.70 1.50 7.75 0.11 24 SD 2.58 0.76 1.26 1.22 1.49 0.43 0.17 0.27 0.44 0.13 0.86 0.14 1.53 0.02 N = 6 (2) 34 MEAN 30.46 4.68 16.33 17.12 13.10 4.15 1.43 1.67 3.15 1.91 8.95 1.58 6.92 0.13 34 MIN 29.04 4.20 15.20 16.10 12.10 3.80 1.25 1.50 2.30 0.80 8.40 1.50 6.00 12 0.11 34 MAX 32.74 4.90 16.90 17.60 14.20 4.50 1.65 1.80 3.80 2.85 9.50 1.70 8.50 12 0.16 34 SD 1.71 0.25 0.60 0.53 0.79 0.26 0.16 0.13 0.62 1.00 0.42 0.08 0.97 0.02 N = 6 (2) 46 MEAN 31.69 4.42 16.58 17.56 13.95 4.18 1.40 1.59 3.53 2.19 8.92 1.45 7.83 0.10 46 MIN 28.12 4.00 16.00 16.80 12.30 3.30 1.15 1.20 2.10 1.00 8.50 1.25 7.00 0.07 46 MAX 33.80 4.75 17.57 18.68 15.30 5.00 1.60 1.90 4.55 3.70 9.50 1.65 9.00 0.13 46 SD 2.03 0.25 0.56 0.68 1.12 0.72 0.17 0.27 1.03 1.24 0.48 0.15 0.67 0.02 N = 6 (3) 54 MEAN 31.78 4.71 17.24 18.13 14.08 4.23 1.35 1.53 3.32 1.72 8.66 1.61 7.23 0.13 54 MIN 25.90 3.35 14.90 15.70 10.50 3.00 1.05 1.25 1.95 0.50 7.60 1.40 5.00 12 0.08 54 MAX 36.20 5.50 19.24 20.35 16.66 5.50 1.65 1.90 4.75 4.10 9.40 2.05 9.50 13 0.22 54 SD 3.69 0.73 1.65 1.78 2.26 0.94 0.22 0.26 1.03 1.14 0.51 0.22 1.57 0.05 N = 8 (3) 62 MEAN 34.65 5.50 18.52 19.43 15.60 4.90 1.38 1.53 4.12 2.40 9.40 1.92 7.90 0.24 62 MIN 31.82 4.60 16.70 17.60 13.50 4.30 1.15 1.25 3.35 1.60 8.70 1.55 7.50 12 0.16 62 MAX 37.20 6.00 20.35 21.27 16.65 5.40 1.50 1.75 4.60 3.20 10.00 2.25 8.60 12 0.32 SD 2.70 0.78 1.83 1.84 1.82 0.56 0.20 0.26 0.67 0.80 0.66 0.35 0.61 0.08 2004 Asiatic Herpetological Research Vo). 10, p. 174 Table 5 (previous page). Morphometric data on early larvae of Batrachuperus gorganensis. d.a.h. = days after hatching; SD = standard deviation; Min = minimum; Max = maximum; N = number of larvae measured (the value in parentheses shows from how many clutches the measured larvae orig inated) According to these studies, larvae from the first season may reach a total length of 30 to 62 mm (persicus, between June and August) and 41 to 50 mm {gorganen- sis, mid of June), respectively, and must overwinter before they form “large larvae”, which can enter meta- morphosis in the following year. Conclusions and Future Research Our initial studies support the relationship of the three taxa Ranodon sibiricus, Batrachuperus mustersi and the Iranian hynobiid(s), i.e. B. persicus and B. gorganensis. These taxa have apparently more features in common than with the Batrachuperus taxa East of Tibet. In addi- tion to the essential need for molecular data, it would be valuable and interesting to study egg deposition in the natural habitats in more detail and to collect data throughout the range of the Iranian hynobiid salaman- ders, especially from the central and western portions. During proof reading, one of us (HGK) provided infor- mation that MMTT 452, 453, and 454 are eggs of B. per- sicus, found near the road from Asalem to Khalkhal, col- lected May, 20, 1975, by M. Thireau, R. Khazaie and R. G. Tuck. Acknowledgments M. E. and H. G .K. wish to thank G. Yolmeh for provid- ing material and initial information on the spawning pond, as well as G. Soltanpour for help during the field work. M. S. thanks David Wake for references and advice, Tate Tunstall for translation from Chinese, S. L. Kuzmin for data on R. sibiricus, G. Wogan and J. Vindum for references, Robert Hijmans for cartographic support, J. 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Tehran University Publication, Tehran, 177 pp. (In Farsi). Brame, A. H. 1985. Family Hynobiidae. Pp. 562-568. In D. R. Frost (ed.), Amphibian species of the world. A taxonomic and geographical reference. Allen Press and Association of Systematics Collections Lawrence, Kansas. Brushko, Z. K., and S. P. Narbaeva. 1988. [Reproduction of Ranodon sibiricus in the Borokhudzir River val- ley (South Eastern Kazakhstan)]. Ekoloyia (Sverdlovsk) 2:45-49 (In Russian). Clergue-Gazeau, M., and J. P. Farcy. 1978. Un Batrachuperus adulte dans une grotte d’lran. Espece nouvelle? International Journal of Speleology 10:185-193. Clergue-Gazeau, M., and R. Thom. 1979. Une nouvelle espece de salamandre du genre Batrachuperus en provenence de l’lran septentrional (Amphibia, Caudata, Hynobiidae). Bulletin de la Societe d’Histoire naturelle, Toulouse 114(3/4):455-460. Crawford, A.J., and D. Wake. 1998. Phylogenetic and evolutionary perspectives on an enigmatic organ: the balancer of larval caudate amphibians. 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Phylogenetic systematics of salamanders (Amphibia:Urodela), a review, p. 31-108 In Sever, D., ed., Reproductive Biology and Phylogeny of the Urodela, Enfield, New Hampshire, USA, Science Publishers, Inc. Litvinchuk, S. N., L. J. Borkin, and J. M. Rozanov. 2004. Intraspecific and interspecific genome size variation in hynobiid salamanders of Russia and Kazakhstan: determination by flow cytometry. Asiatic Herpetological Research 10:279-291. Liu, C.C. (1950): Amphibians of western China. Fieldiana: Zool. Mem. (Chicago) 2:80-102. Nawabi, S. 1965. A rare representative of the amphibi- ans in Afghanistan Batrachuperius mustersi. Science, Quarterly Journal of the Institute of Zoology and Parasitology published by the Faculty of Science, Kabul-University: 21-25. (In Farsi). Regel, E.D. 1968. [The development of the cartilaginous neurocranium and its connection with the upper part of mandibular arch in Sibirian salamander Ranodon sibiricus (Hynobiidae, Amphibia)] pp. 5-168 In Morfologiya nizshikh pozvonochnykh zhivotnikh [Morphology of lower vertebrates], Trudy Akademiya Nauk SSSR [Works of the Academy of Sciences of the USSR], IsdateFstvo Nauka, Leningrad (In Russian). Reilly, S.M. 1983. The biology of the high altitude sala- mander Batrachuperus mustersi from Afghanistan. Journal of Herpetology 17(1): 1-9. Reilly, S.M. 1987. Paradactylodon : a junior synonym for Batrachuperus. Amphibia-Reptilia 8:283-284. Schmidtler, J.F., and J.J. Schmidtler. 1971. Eine Salamander-Novitat aus Persien. Batrachuperus persicus. Aquarien-Magazin 11:443-445. Sparreboom, M. 1979. [Eggs of Batrachuperus muster- si ]. Lacerta 37(5):83-88 (In Dutch). Steiner, H.M. 1973. Beitrage zur Kenntnis von Verbreitung, Okologie und Binomie von Batrachuperus persicus (Caudata, Hynobiidae). Salamandra (Frankfurt/M.) 9(1): 1-6. Stock, M. 1999. On the biology and the taxonomic sta- tus of Batrachuperus gorganensis Clergue-Gazeau et Thorn, 1979 based on topotypic specimens (Amphibia: Caudata: Hynobiidae). Zoologische Abhandlungen des Staatlichen Museums fur Tierkunde Dresden 50(1 4):2 17-241. Thom R., and J. Raffaelli. 2001. Les Salamandres de FAncien Monde. Boubee, France, 449 pp. [partly revised version of the original publication 1968]. Zhao, Er-mi, and Qixiong Hu. 1988. Studies on Chinese tailed amphibians. Pp. 1-43. In Er-Mi Zhao (ed.) Studies on Chinese salamanders. Contributions to Herpetology, no. 4. Society for the Study of Amphibians and Reptiles, Oxford, Ohio. Histochemical Characterization of the Lingual Salivary Glands of the House Gecko, Ptyodactylus hasselquistii (Squamata: Gekkonidae) Bashir M. Jarrar and Noory T. Taib Zoology Department, College of Science, King Sand University, P.O.Box 2455, Riyadh 11451 Saudi Arabia. Abstract. - Histochemical investigations of the lingual salivary glands of the house gecko, Ptyodactylus hasselquistii have been conducted. The glands are comprised of mucous and mucoserous cells. Mucous cells secrete or elaborate neutral mucosubstances, neuraminidase sensitive carboxylated mucins, hyaluronidase resistant sulfomucins, but are devoid of proteins. The mucoserous cells secrete and elaborate neutral mucosubstances and glycoproteins but are devoid of sialomucins and sulfomucins. The results are discussed in the context of the feeding habits and phylogeny of reptiles. Key words. - Histochemistry, lingual, salivary glands, house gecko, Ptyodactylus hasselquistii, Gekkonidae. Introduction Histochemical studies on the lingual salivary glands of vertebrates have mainly been concerned with mammals, whereas little attention has been paid to the lingual sali- vary glands of non-mammalian vertebrates. Most stud- ies on the lingual salivary glands of reptiles have focused with morphological and histological aspects while few histochemical studies have been carried on these glands (Raynaud, 1961; Gabe and Saint-Girons, 1969; Lopes et al., 1982; Taib and Jarrar, 1985a; 1985b; 1985c; 1986; Taib, 1986, Asgah et al., 1990). Nevertheles, the literature on the lingual secretions of lizards is rather scanty and their consitituents have yet to be determined. The present study is a detailed histochemical char- acterization of the lingual salivary glands of the house gecko, Ptyodactylus hasselquistii. Materials and Methods Twenty adults of each male and female house gecko Ptyodactylus hasselquistii were trapped from different houses in Riyadh city, Saudi Arabia. They were killed by etherization and the whole tongue was removed from each animal and quickly immersed for 24 hrs in one of the following fixatives: neutral buffered formalin, Bouin's fluid and Gendre's fluid. They were then thor- oughly washed in running water, processed for serial sectioning at 4-5 pm thickness and the sections were stained with haematoxylin-eosin or with Mallory trichrome for histological examination, whereas the sec- retary cells of the glands were characterized by the cri- terion of Gabe and Saint-Girons (1969). Other sections were used for the following histochemical reactions: Neutral mucosubstances. - Periodic acid-Schiff (PAS) technique (Gurr, 1962), PAS after diastase digestion (McManus and Mowry, 1964), PAS after alpha-amylase digestion (Luna, 1968), PAS after acetylation blockade (McManus and Cason, 1950), PAS after acetylation- saponification (Oxello et al., 1 958), PAS after phenylhy- drazine treatment (Spicer et al., 1967) and PAS after treatment with chloroform and methanol. Acid mucosubstances. - Alcian blue (AB) at pH 2.5, 1.0, and 0.4 (Mowry, 1956; Luna, 1968). Distinction between acidic and neurtal mucosub- stances. - AB (pH 2.5)-PAS (Mowry and Winkler, 1956) and AB (pH 1.0)-PAS (Spicer et al., 1967). Distinction between sulfomucins and sialomucins. - Aldehyde fuchsion (AF) and AF-AB, pH 2.5 (Spicer and Meyer, 1960); weak (25°C, 16 hr), mild (37°C, 4hr) and strong (60°C h hr) methylation-saponification- AB (PH 2.5) (Spicer, et al., 1967); toluidine blue (TB) buffered at pH 1.7 and 3.4 (Landsmeer, 1951), critical electrolyte concentration (CEC) technique for extinction of alcianophilia at pH 5.6 in the presence of gradual con- centation of Mg2+ (Scott and Dorling, 1965). Enzyme digestion tests. - Diastase-PAS technique (McManus and Mowry, 1964); neuraminidase (Sialidase, Vibrio chlolerae, type V)-AB (pH 2.5) (Spicer and Warren, 1960); hyaluronidase (testicular)- AB (pH 2.5) (Spicer et al., 1967). Ribonuclease diges- tion (Love and Rabotli 1963); neuraminidase-TB (pH 3.7), hyaluronidase-TB (pH 2.0) were employed. In each case control sections were incubated for the same length of time at the same temperature in buffer solu- tions without the enzyme. © 2004 by Asiatic Herpetological Research Vol. 10, p. 177 Asiatic Hcrpetological Research 2004 Figure 1. Lingual glands of P hasselquistii after staining with haematoxylin-eosin. Note that the mucous cells of the lingual glands are located in the papillar space of the papillae. x950. Figure 2. Lingual glands of P. hasselquistii after staining with PAS. The reactivity of the glands confirms the pres- ence of the neutral mucosubstances. x950. Figure 3. Lingual glands of P hasselquistii after staining with AB (2.5), confirming the presence of sialomucins and sulfomucins. x950. Figure 4. Lingual glands of P. hasselquistii after staining with AB (I.O)-PAS. The bluish purple color indicates the presence of neutral and sulfated mucosubstances simul- taneously. x950. Figure 5. Lingual glands of P. hasselquistii after staining with CEC at 0.3M Mg++, confirming that the mucosub- stances produced by the glands contain carboxyl and sulfated groups. x950. 7- • Figure 6. Lingual glands of P. hasselquistii after staining with MBPB, indicating the protein contents of the glands secretion. x700. 2004 Asiatic Herpetological Research Vo). 10, p. 178 Proteins. - Mercuric bromophenol blue method (Mazia et al., 1953); ninhydrin-Schiff (Yasuma and Itchikawa, 1953), mercuric-bromophenol blue (MBPB) and PAS after trypsin digestion (Pearse, 1972). Photographs. - Photographs were taken with a 35mm Zeiss Ikon camera on Kodacolor NR 100 film. Results The lingual salivary glands of the house gecko, Ptyodactylus hasselquistii occupy the papi liar invagina- tion of the posterior two-thirds of the dorsal surface together with the lateral sides of the tongue. The anteri- or part of the tongue is devoid of any glandular structure and covered by keratinized squamous epithelium. These glands are made of mucous cells located in the inner papillar space of the filiform papillae (Fig. 1) together with simple tubular structures made of mucoserous cells seen at the most posterior part of the dorsum. The mucous cells have an alveolar cytoplasm and flattened, basally located nuclei with clear apical ends resting on a delicate basement membrane. As summarized in Table 1, the mucous cells of the lingual glands of P. hasselquistii exhibited strong PAS reactivity (Fig. 2) which was neither labile to alpha- amylase nor to saliva digestion but completely lost by phenylhydrazane treatment. Flowever, this reactivity was completely blocked by acetylation and was partly restored by deacetylation-PAS sequential techniques. They showed marked alcianophilia at both pH 2.5 (Fig. 3) and 1 .0 but to lesser extent at pH 0.4. They also react- ed with both PAS and AB and stained bluish purple with AB (2.5)- PAS and AB (1.0) PAS (Fig. 4). These glands also reacted with AF as well as with AF-AB (2.5) and AF-AB ( 1 .0). The alcianophilia of the glands was part- ly lost at pH 2.5 with acid hydrolysis and weak methy- lation and there after restored by saponificiation tech- niques. They demonstrated alcianophilia with the CEC techniques at 0.1M, 0.2M, and to some extent at 0.3M Mg2+ (Fig. 5) and showed metachromasia at pH 3.4 and 1.7 but reacted negatively to all protein detection tests. The mucoserous cells of the glands showed PAS reaction, exhibited no alcianophilia at pH 2.5 and 1.0 and were orthochromatic at pH 3.4 and 1.7 but reacted positively to all protein detection tests (Fig. 6). No sex- ual dimorphism was observed in the lingual secretion of the species under study. Discussion The lingual salivary glands present great diversity in morphology amongst the various groups of reptiles. These glands are entirely absent from Varan idae. Amphisbaenia, Ophidia and some species of Cheloma such as Chelonia mydas (Kochva, 1978). On the other hand, these glands are simple consisting of three differ- ent types of goblet cells in most species of Testudinidae (Nalvade and Varute, 1976; Taib and Jarrar, 1984). Some lizard possess mainly goblet cells together with simple tubular glandular structures in their tongues (Nalvade and Varute, 1976; Shevliuk, 1976; Taib and Jarrar 1985 b; 1985c and 1986), while others have more developed lingual salivary glands as seen in some Agamidae, Iguanidae, Gekkonidae, Anguidae and Chamaleonidae (Gabe and Saint-Girons, 1969; Kochva, 1978; Asgah et al., 1990). On the bases of the results of the present study and in view of the criterion of Gab and Saint-Girons (1969), the lingual salivary glands of Ptyodactylus hasselquistii are made ot unicellular mucous goblet cells lining the dorsal epithelium of the tongue with mucoserous simple tubular glandular appa- ratus at the base of the tongue. The structure of the lin- gual salivary glands of P. hasselquistii is different from those of Tupinambis teguixin , Agama blandfordi, Uromastyx microlepis, Acanthodactylus schmidti and Scincus mitranus , which have only mucous cells in their lingual glands (Lopes et al, 1974; Taib and Jarrar, 1985b; 1985c; 1986; Taib, 1986). According to Gabe and Saint-Girons (1969), the lingual glands are mucous in Gekkonidae, mucoserous on Sphenodontidae, Anguidae and Pygopodidae, but seromucous in Chamaleonidae and serous in some speices of Iguanidae and Agamidae. The grading from non-glandular tongues through unicellular with or without simple tubular glan- dular structure to only simple tubular and to then tubu- lo-alveolar ones may reflect developmental stages towards the definitive lingual glands of higher verte- brates (Shevliuk, 1976; Kochva, 1978). A tentative interpretation of the types of mucosub- stances in the lingual glands of P. hasselquistii can be made from the results of the different histochemical reactions used in the present investigation and from the classification of mucosubstances proposed by Mowry and Winkler, 1956; Spicer and Meyer, 1960; Scott and Dorlling, 1965; Pearse, 1972). Neutral mucosubstances are PAS positive, diastase resistant, as well as unstain- able by cationic dyes. Acetylation produces derivatives of primary and secondary amines which prevent 1, 2 glycol groups, from reacting with PAS indicating the presence of neutral mucosubstances or sialic acid, sepa- rately or simultaneously. Alcian blue is generally con- sideied as being specific for identifying acid mucosub- stances where alcianophilia at pH 2.5 and 1 .0 is specific tor sialomucins and sulformucins respectively (Mowrv and Winkler, 1956). In the combined aldehyde fuchsin- alcian blue sequential techniques, sulfomucins stain pur- ple blue and sialomucins blue (Spicer and Meyer. 1960) Vol. 10, p. 179 Asiatic Herpetological Research 2004 Table 1. The histochemical reactions in the lingual salivary glands of Ptyodactylus hasselquistii. Histochemical reaction Results MC MSC PAS ++,P ++,P Diastase digestion -PAS Nb Nb Acetylation-deacetylation-PAS ++,P ++.P Phenylhdrazine-PAS Cb Cb AB (pH 0.4) +B - AB (pH 1.0) +B - AB (pH 2,5) ++> B - AB (pH 1.0) -PAS +,Bp - AB (pH 2. 5)- PAS ++,Bp - AF +,P - AF- (AB pH 1.0) +,Bp - AF- (AB pH 2.5) ++,Bp - Acid hydrolysis- AB (pH 2.5). Pb - W. methylation - AB (2.5) Pb - W. methylation-saponification - AB (pH 2.5) ++,Bp - M. methylation-AB (pH 2.5) Cb - M. methylation-saponification -AB (pH 2.5) +.B - S. methylation-AB (pH 2.5) Cb - S. methylation-saponification- AB (pH 2.5) +,B - TB (pH 1.7) + - TB (pH 3.4) + - CEC (AB, 0.1 M) + - CEC (AB, 0.2 M) + - CEC (AB, 0.3 M) ± - CEC (AB, 0.5 M) - - Neuraminidase-AB (pH 2.5) +B,pb - Hyaluronidase-AB (pH 2.5) ++B,Nb - Ninhydrin-Schiff - + Hg- bromophenol blue - + Chloramine T-Schiff - + Trypsin digestion-PAS Nb - (Chloroform + methanol)-PAS Nb - Reactions: - negative; ± weak; +, moderately positive; ++, intensely positive; Cb, complete blockade; M, mild; Pb, partial blockade; Nb, no blockade; S, strong; TB, toluidine blue; W, weak. Colors: B, blue; Bp, bluish purple; P, pink. Glands: MC, mucous cells; MSC, mucoserous cells. Sialomucins can be identified by alcianophilia at pH 2.5 which is partially lost following acid hydrolysis and completely removed after neuraminidase digestion, but neuroaminidase did not affect the staining of sulfated mucosubstances. A loss of alcianophilia after hyaluronidase digestion is due to the removal of hyaluronic acid and chondroitin sulfates. Methylation blocks subsequential stainig of simple mucosubstances by esterification of carboxyl groups and complex sulfat- ed mucosubstances desulphation. Subsequent treatment with potassium hydroxide (saponification) after methy- lation will restore the staining of carboxyl groups (Drury et al., 1967). The mucosubstances that are stained at 0. 1 M MgCl2 in the CEC reaction, but not at 0.2M MgCl2 are believed to contain carboxyl group and no sulfate groups. Sulfated mucosubstances, on the other hand, stain strongly and selectively at 0.2M Mg2" but lose their alcianophilia at different levels with increasing MgCl2 concentration (Spicer and Lillie, 1960). The lin- gual glands of the species under study resisted trypsin digestion and the action of (chloroform-methanol) which excludes the possibility of lipids and proteins. Accordingly, the lingual salivary glands of of the house gecko, Ptyodactylus hasselquistii contain neutral muco- substances, sialadase labile carboxylated mucosub- stances and hyaluronidase resistant sulfomucins and gly- coproteins. The lingual secretions of P. hasselquistii are differ- ent from those of some lizards such as Tupinambis teguixin and Agama blandfordii which contain neutral 2004 Asiatic Herpetological Research Vol. 10, p. 180 mucosbstances and sialomucins but no sulfomucius (Lopes et al., 1974; Taib and Jarrar, 1985c). They also differ from the secretions of the lingual glands of Uromastyx microlepis, Acanthodactylus schmiditi, Scincus mitranus and Stenodactylus slevini which con- tain neutral mucosubstances, sialomucins and sulfor- mucins but no glycoprotein (Taib and Jarrar, 1985b, 1986; Taib, 1986; Asgah et al., 1990). Neutral mucosub- stances have been demonstrated in the secretions of all studied reptiles that possess salivary glands while phylo- genetically, the absence of sulfomucins in the lingual glands would favour the concept that sialomucins secre- tive cells are more primitive than sulfated mucosub- stances secretive ones. In addition, the heterogenous his- tochemical recactivity of the lingual glands might have appeared in the evolutionary lines of reptiles to meet the different changes in the feeding habits of various species. Neutral mucins were present in the lingual glands of almost all studied reptile species while sialo- mucins together with neutral mucosubastances were identified in the lingual glands of all insectivorous rep- tiles so far studied (Nalvade and Varute, 1972; Taib and Jarrar, 1985c, 1986; Taib, 1986). The lingual glands of all insectivorous and carnivorous reptiles studied thusfar exhibited sulfomucins. More work is needed to elucidate whether the lingual secretion diversity of reptiles imply phylogenetic relationships or different feeding habits. Literature Cited Asgah, N. A., N. T. Taib, and B. M. Jarrar 1990. Morphology, histology and histochemistry of the cephatic glands of Slevin's ground gecko, Stenodactylus slevini Hass, 1957. Tropical Zoology 3:209-217. Bums, B. and V. G. Pickwell. 1972. Cephalic glands in sea snakes ( Pelamis , Hydrolis, and Laticauda ). Copeia 1972 (3):547-559. Carmignani, M. A. and G. Zaccone. 1975. 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Journal of Histochemistry and Cytochemistry 8:135-137. Spicer, S. S., R. G. Horn, and T. J. Leppi. 1967. Histochemistry of connective tissue mucopolysac- charides. pp 251-303. In: The connective tissues. International Academy of Pathology Monograph No.7. Williams and Wikins, Balthimore. Taib, N. T. 1986. Histochemical observations on the lin- gul salivary glands of the skink Scincus mitranus (Anderson, 1871) (Scindidae, Reptilla): Bulletin of the Chicago Herpetological Society 2 1(1 -2): 14-22. Taib, N. T. and B. M. Jarrar. 1985a. Histochemical char- acterization of musousbstances in the tongue of the terrapin, Mauremys caspica (Gmelin) (Reptilia, Testudins, Emydidae). Jorurnal of Biological Sciences Research, Iraq 16(2):239-249. Taib, N. T. and B. M. Jarrar 1985b. Histochemical stud- ies on the lingual salivary glands of the spiny-tailed lizard Uromastyx microlepis (Blandford). Bulletin of the Institute of Zoology, Academia Sinica 24(2):203-212. Taib, N. T. and B. M. Jarrar. 1985c. Histochemical analysis of mucosubstances in the lingual salivary glands of the lizard A gam a blandfordi (Agamidae, Reptilia). Sudan Journal of Science 1:97-101. Taib, N. T. and B. M. Jarrar. 1986. The histochemistr of the lingual salivary glands of the lizard Acanthodactylus schmidti (Wiegmann) (Reptilia, Lacertilia, Lacertidae). Bulletin of the Maryland Herpetological Society 22(2):27-36. Yasuma, A. and T. Itchikawa. 1953. Nihydrin-Schiff and alloxan Schiff staining, A new histochemical method for protein. Journal of the Laboratory Clinical Medicine 41:296-299. 2004 Asiatic Herpetological Research Vol. 10, pp. 182-190 The Biology of the Persian Mountain Salamander, Batrachuperus persicus (Amphibia, Caudata, Hynobiidae) in Golestan Province, Iran Haji Gholi Kami Department of Biology, Faculty of Sciences, University of Agricultural Sciences and Natural Resources, P O. Box 49165, Gorgan, Iran. Abstract. - The Persian Mountain Salamander, Batrachuperus persicus, is a hynobiid endemic to Iran and is distrib- uted in specific localities of Hyrcanian forests in four northern provinces of Iran. The biology of this salamander was studied at four localities in Golestan Province of Iran, especially in Shirabad Cave, between 1996 and 1999. Information is presented about the cave and other localities. This salamander has four fingers and toes. The larval stages of the salamander are found at all times of year and probably don't transform during the first year. The head form of small larvae is wider posteriorly while the head form of large larvae, juveniles and adult specimens is more or less rectangular. Juveniles have more yellow spots than adults. Juvenile and adult specimens are found inside and outside of water in the cave but in other localities they can be found in burrows around springs and are not active dur- ing the day. They feed on larval and adult forms of insects and other arthropods. Adults also feed on small specimens of bats ( Myotis blythyii) inside of the cave. Some large specimens are cannibalistic and feed on larvae and juveniles of B. persicus in natural habitats and in the laboratory. This species does not hibernate inside the cave and is active all times of the year. The total length of the longest specimen was 268.5 mm. Key words. - Amphibia, Hynobiidae, Batrachuperus persicus, Iran. Introduction The Persian Mountain Salamander, Batrachuperus per- sicus Eiselt and Steiner 1970, was described based on five salamander larvae collected near Asalem in the Talesh Mountains in Gilan Province of Iran (Eiselt and Steiner, 1970) (See editorial note and Ebrahimi et al., 2004). Subsequently J. J and J. F Schmidtler collected some larvae in Weyser, southeast of Chalus, in Mazandaran Province, Iran, in 1970. These transformed in captivity and a brief description of juvenile specimens was presented (Schmidtler and Schmidtler, 1971). Primary information was presented on adult specimens in Ardabil Province of Iran (Baloutch and Kami, 1995). New distribution records were published some years ago (Kami and Vakilpoure, 1996). Adult specimens of this species were described for the first time together with their habitats in Gilan and Ardabil provinces of Iran (Kami, 1999). The biology of this salamander has been studied in the laboratory and in natural habitats especially in Shirabad Cave of Khanbebain. Students of Gorgan University and I have visited Shirabad Cave twelve times between 1996 and 1999. Detailed information on the other localities is sparse and these localities must be studied more in the future. Study areas. - Batrachuperus persicus is distributed in the four northern provinces of Iran (Fig. 1). Figure 2 shows new localities of Batrachuperus persicus in Golestan Province. Most of the research was done at locality 1 (Shirabad Cave), other localities were visited only one time. Climatic information for five cities of Golestan Province is summarized in Table 1 . Locality 1. Shirabad Cave- Shirabad Cave (36° 57' N, 55° 03' E) is situated 70 km East of Gorgan, southeast of Khanbebain and Shirabad Village at about 420 m above sea level (Fig. 3, 4). The cave and waterfalls were desig- nated as a National Park by the Department of © 2004 by Asiatic Herpetological Research Vol. 10, p. 183 Asiatic Herpetological Research 2004 Tab 1. Summary of climatic information (15 years) in five cities of Golestan province of Iran. station (city) elevation Annual (m) precipitation (mm) Mean of Air temperature (°C) Mean of minimum temperature (°C) Mean of maximum temperature (°C) Mean of relative moisture (%) Annual evaporation (mm) Azadshahr 129 847.6 17.1 0.7 31.9 71.6 — Ramian 200 883.1 16.2 -1.0 35.0 65.0 497.0 Minoudasht 155-180 765.3 17.3 -1 36.0 82.1 482.5 Gonbad-e- 30-45 388.3 18.2 -3.5 36.5 68.6 510.0 kavous Gorgan 160 642.7 17.7 2.8 36.4 — — Figure 2. Localities of of Batrachuperus persicus in Golestan Province, Iran: 1- Shirabad Cave, 2 - Vantakhteh; 3- Near Shirabad Cave; 4- Spring of Khouklou; 5- Barankouh (36° 45' N, 54° 25' E), about 10 km south west of Gorgan, almost 1100 m elevation; 6- Spring of Khonakou, situated south southwest of Gorgan, Jahannama Protected Area, Valley of Sorkhcheshmeh, below elevations of Pir-e-zan (elevations of Pahlavan Ghaleh), 1800 m elevation; 7- elevations of Yakhkesh (36° 42' N, 54° 23' E) about 15 km south of Gorgan, 2300 m elevation; 8- Water falls of Shirabad; 9- Valley of Loushan, inside of valley, (36° 42' N, 54° 41' E) about 30 km southeast of Gorgan; 10- Region of Aram-e- Sorkhcheshmeh opposite side of Siah Marzkouh, Village of Aliabad. Circles are cities: 1- Gorgan; 2- Aliabad; 3- Ramian; 4- Azadshahr; 5- Gonbad-e-kavous; 6- Minoudasht; 7- Kalaleh. 2004 Asiatic Herpetological Research Vol. 10, p. 184 Table 2. Dates of study, air and water temperature of four localities in Golestan province of Iran. 37°00 Locality Dates of study Air tem- Water perature temperature (°C) (°C) Time Shirabad 1996/11/1 — 12 Cave 1997/2/28 — 11 1997/4/11 — 10-10.5 1997/4/17 12.5-13 11 1997/7/10 19.5-21 13 1045- 1100 1997/10/2-3 — 12 998/4/17 — 11 998/5/8 18 10 1200 1998/5/15 18 13 1430 1998/5/19 15 12 1998/7/10 — 13 1998/9/2 17 11 Vantakhteh 1996/11/25 2-6 6.5-7 1330- 1530 Near 1997/7/10 20-23.5 19 1000 Shirabad Khouklou 1999/5/26 26-27 12 1230 Spring Environment of Gorgan and Gonbad-e-Kavous in 1998. There are seven waterfalls below the cave. The entrance of the cave is about 15 m high and is at least 3-4 m high in other parts of the cave. It is almost 240 m long and is completely dark. Water emerges from the mouth of the cave and flows to the river and waterfalls at all times of the year. No plant species live inside the cave, but there is Lycopodium sp on the floor of cave from the entrance to about 10 m inside of it on large flat stones. Some plant species that were identified outside the entrance of the cave on 28 February 1997 and 17 April 1997 are: Pteridum aquilinum, Adianthum capillus-veneris, Athyrium flix-mas, Phyllitis scolopendrium, Funaria sp, Celtis australis, Evonymus latifolia, Convulvulus (=Calistegia) sepium, Hedera pastochowii, Lamium album, Carex pendula, Rubus hyrcana, Ficus carica, Danae recemosa, Acer insigne, Parrotia persica, Carpinus betulus Cyclamen elegans, Marcantia sp. The water temperature of cave is 10-13°C (Mean 1 1 .6°C) and is constant from the entrance to end of cave. Air temperature of cave varied between 1 2.5-2 1°C over six visits. The inside of the cave is a little warmer than the entrance of the cave (Table 2). Locality 2. Vantakliteli. - The Spring of Vantakhteh (36° 40' N, 54° 25' E ) is about 1 8 km south of Gorgan City and 5 km southwest of Ziarat Village at about 1200-1300 55°00' + 55°05' + 37°00' kouh-e-NargesI 0 331 m Daland water resource To Khanebain \ \__ Shirabad village Figure 3. That National park that includes the cave and waterfalls of Shirabad (1650 Hectares). m elevation. Salamanders were observed in this locality in 1979 and later in Yakhkesh (2300 m elevation) on 25 November, 1999. There are two springs in this locality that are formed from soil and limestone and situated to east. Water flows from springs to the Souteh River. The sides of the river were frozen and snowy. The springs are 3-4 m above the river. Water temperature of one spring was 6.5 and other 7°C. Air temperature was 6°C at 13:30 and 2°C at 15:30. Snow was melted in east side of the springs. Some plant species found around the springs are as follows: Rosa albicans, Berberis vulgaris, Juniperus communis, Carpinus orientalis, Juncus effusus, Circium nekarmanicum, St achy s bizantica. Five salamanders, all metamorphosed, were found inside of burrows near the springs. Locality 3. Near Shirabad Cave. - Locality 3 is a small pond (about 2m x 2m) in southwest of Shirabad Village and 20 m above Shirabad Cave. Water depth was almost 50 cm. Around this shady pond were stones, lichens and trees {Danae racemosa, Quercus sp). A Grass Snake (Natrix natrix), Marsh Frogs (Rana ridibunda), crabs {Potamon sp.; and larvae of Batrachuperus persicus with total length of 3-4 cm were observed. No adult or metamorphosed salamanders were seen. Air temperature was 20-23. 5°C and water temperature was 19°C at 1000 on 10 July 1997. Some larvae such as Gerridae (Heteroptera), Chironomidae (Diptera), Ephemeroptera, and Amphipoda (Gamaridae), earthworms (Lumbricidae), and Gastropoda were collected inside and around the pond under decaying logs. Vol. 10, p. 185 Asiatic Herpetological Research 2004 Table. 3. Measurments of living Batrachuperus persicus inside of Shirabad cave of Golestan province of Iran. All spec- imens were released after measuring. All measurements are in mm. Dateand number Form Total length Head length Trunk length Tail length 1996/11 /I n=6 larva 80 10 28.4 41.8 larva 104.4 11.8 36.1 54.1 larva 88.2 9.8 33.1 45.2 larva 79.2 9.7 29.6 40.0 larva 84.8 10.6 30.9 43.2 adult 236.5 24.5 85 127 1997/2/28n=4 larva 70 - - - larva 83 - - - larva 79 - - - larva 74 - - - 1 997/4/1 7n=4 larva 50 - - - larva 80 - - - adult 240 - - - adult 245 - - - 1 997/7/1 0n=1 9 adult 229.2 26.6 78.6 124 adult 227.7 24.8 81.9 121 adult - 29.3 84.2 - juvenile 97.3 11.9 37.5 48.6 larva 45 - - - larva 57 - - - juvenile 90 - - - larva 80 - - - larva 86 - - - larva 62 - - - larva 52 - - - larva 50 - - - larva 67 - - - larva 56 - - - larva 103 - - - larva 108 - - - larva 94 - - - larva 55 - - - adult 217 - - - 1 998/5/1 9n=1 3 larva 105 - - - adult 225 - - - adult 225 - - - adult 230 - - - adult 235 - - - larva 100 - - - larva 75 - - - larva 40 - - - larva 41 - - - larva 42 - - - larva 42 - - - larva 44 - - - larva 55 - - - 1999/4/26n=5 larva 37.9 - - “ larva 36.8 - - - larva 36.2 - - - larva 36.9 - - - larva 75.5 - - - 2004 Asiatic Herpetological Research Vol. 10, p. 186 co CD 0 CM x — CO X > ^ x — X“ ( n o 1 1 1 1 1 | 1 | 1 X” | 1 o o 6 CNJ x — CM 1 CO X — CD i CD c N- LD CO CO O o 00 o ID y o l 1 O F i F ib 1 K o cb o d CM < CD T~ T_ CM T— co CM ID -Q x — x — E (f) A A A A A A A A A A A X X X M" M- M- M- M" M- M" CM "D cr ' 1 II A 1 A A 1 A A A A A A A C CM CM CM CM CM CM CM CM CM CM X A A A A A A A A A A A X CO CO CO CO CO CO CO CO CO CO CO -Q E CD CD X h- CO CD ID CM CO CD CM T3 C 1 CM lb 1 CM F 1 d CM x — CD x — cb CD 00 CM X -Q T— M" x— X— T- T— T— x— x— X — x — X— E CO II A II ii A A II A A A A A X X — X M" M- M- M" M" M" M" M" X CD CD CD 1 1 A A A 1 1 A A 1 A A A A A A A L— CO CO CO CO CO CO CO CM CM CO CM CM O o A II A A A A II A A A A A '*+— o LL CM CM CM CM CM CM CM CO CO CM CO CO 0 O c _Q E Is- CM X X CO o h- CO ID q 03 00 > CD 1 1 1 CD CD CD 1 ib CO 00 1 d x — x— cb CM cb o o Q. u_ c CD ro O CD CD M- T- ID (/) CD O T- o CM cb o O CD co 3 1 1 1 , CM F O CO CO d , CO d M" CM 25 , , CO o , X T_ CM ID o CM CM CM a> ib CD CM X LD CO ib Q. co 1- T_ T_ CM CM CM co CM M" CO CO h- 2 d) a c CD CD JA ’d CM CM O CD r^. 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CD 0 L_ CD 0 L_ CD 0 i_ CD 0 CD 0 O 0 O 0 03 03 O 0 CO 0 0 i— i_ CD 0 CD 1 CD T3 CD JC X CD CD ID CO CO ID q CO 03 h- 0 X o $ 1 1 CO F F CD F d d CM X — CM CO o ib _C a y (/) ■o CD -C m CD CD 00 CD CO o CD co co ID o CO \- CD 0 x c 0 CD 1 CD CD CD CD F fr 00 o o c\i X — CM fr x — CNJ E ~o c .c ID CM X O co ID CO 00 X — LD CD o o C" CXD co CD o o CM CM LD ib ib 1 1 NT CM fr CD T — o 'u F 0 CO co CO X X M" X id r- 00 CO CD C= 0 0 0 F CD CD CD CD CD CD CD CD CD CD CD CD CD CD — • — • -C u. 2 2 2 2 2 2 2 2 2 2 2 2 2 2 c (D c CD c (D a. L— o LL. CD CD CD CD CD CD CD CD CD CD CD CD CD CD > D > D > 13 Z3 n CD CO CD CM CD h- CO CO M" CO ID CO CD CO O 00 CD CD C\J CM CO CO 00 CM h- 00 CO CO 00 M- M- co D- K- f N X X CM CM CM CM CM CM CO CM CO CM CM M" CM CM Locality 4. Spring of Khouklou. - The Spring of Khouklou (36° 44' N, 54° 53' E) is situated almost 23 km south of Aliabad at about 1500 m elevation. This spring is 200 m west of Chenarbin and along side of the Khouklou River and situated to north beneath a large rock. This locality was studied on 26 May 1999. Six specimens were seen and three of them were collected. All specimens had external gills. Materials and Methods Shirabad Cave was studied 12 times and the other local- ities only one time between 1996 and 1999. Air and water temperature of the cave and two other localities measured on some dates. Important plant species of localities were identified. Measurements (total, head, trunk, and tail lengths) were done on living specimens of salamanders inside the cave. Some specimens (larval and transformed) collected and brought to aquaria at the zoology laboratory of Gorgan University and kept with ice. Some specimens (30) were fixed in alcohol or for- malin. Almost all fixed transformed salamanders were dissected and stomach contents and sexes were noted. Morphometric and meristic characters of specimens were taken. The behavior of salamanders was studied inside the cave and in the laboratory. On each visit the total number of salamanders was counted from the entrance to the end of the cave. Preserved specimens of Batrachuperus persicus studied for this research are as follows: ZMGU 67, 273, Shirabad Cave collected by H. Naghghash and M. Rahmani in 1994; ZMGU 246, 281, 282, 283, 284, near Shirabad Cave collected by H. Kami, A. Maghsoudlou, M. Rahmani, M. Azma on 10 July 1997; One specimen without number collected in Shirabad Cave by A. Maghsoudlou on 8 May 1998; ZMGU 267, Shirabad Cave, collected by A. Maghsoudlou on 21 May; ZMGU 266, 268, 269, 270, 272, 285, 286, 427, 428, 429, col- lected by H. Kami, M. Fatemi, N. Okhli, N. Moghaddam, J. Ghasemi, M. Mahmoudi, R. Zakeri, on 19 May 1998; ZMGU 275, 430, Shirabad Cave, proba- bly collected by H. Kami, S. Afzali, R. Ghaemi on 28 February 1997; ZMGU 276, Jahannama Protected Area, south of Gorgan and Ziarat Spring of Khonakou, collect- ed by N. Torbatinejad on 10 June 1997; ZMGU: 277, 278, Vantakhteh, collected by H. Kami, S. Afzali, M. Firouznia, H. Rezaee, Y. Shakoumahalli, Y. Nariman, on 25 November 1996; ZMGU 279, 280 without correct information (probably Vantakhteh or Shirabad); ZMGU 333, 335, Shirabad Cave, collected by M. Goli, S. Afzali and eight other students of Gorgan University on 27 April 1999; ZMGU 343, Spring of Khouklou, collected by H. Absalan, S. Hosseini, on 26 May 1999. Vol. 10, p. 187 Asiatic Herpetological Research 2004 o Is- CN Z> o N Cl) o c > o X2 0 T3 < E in h" =) o N c 03 O — - 0-g O 03 o I' Q-S 0 Jl> 00 0 O o c E 0 oo o _0 CO =3 0 03 ^ TO o CO 0 0 *7. m ^ 2 5 0 o <-> 0 O C To c6 >- 0 0 <- c II e™ 0 c g ro i- 0 0 2 E c o 0 f! o 0 *> ^ in -o _ i_ -Q T3 lE 0 X 0 if) LL 1 LL. M F LL LL LL 0-* LL LL LL (/> 3 0 CM CO T— T— CM •5— CM CO CO CM CO T— V) O x — T~ T— 1 T— ■*— M" t— T— t— "t- o V-/ O 4- CO T_ CM A- cb CM cb CM CM 1 M- CM cb o k. 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O CM M- CM 00 CM cb 0 i_ 1- CO Is" Is- CO 00 03 CO 00 1^ 03 CO 03 C CO 0 0 $ c 4-1 Is- LO T- CO in 03 00 CO Is- in CO V) lO Is" cb Is" r- 00 Is- rz Is" M" d cb 0 0 CQ c C 0 0 c/> ■V in CO CO 03 03 co 1" in 00 O CM 1 0 in in 1 10 LO in cb in in in d d d 0 m 0 L_ C/3 0 a "O T~ in in ( 00 CO CM CM M; 1 cq Is- CO Q. "0 cb cb M’ in M- d d 3 > 0 L_ L— L— 1 1 U_ L "O C33 1— 03 03 u 03 03 03 i ~o E 0 0 0 > C 0 0 c c 0 c c c 0 re c c c 0 c c 0 0 C 0 0 0 c 0 X L- O <*- 0 0 1 0 0 L— 0 0 i 0 0 -* — 1 O 0 0 0 i 0 0 0 0 1 -4—' O 0 i_ 0 0 0 0 L— * 0 0 1 0 0 1 L— 0 0 i L_ \ L. -0 -C T_ CD CM cq M" co CM c M; CO M- Is" 0 CO re 0 X ■O in 00 in 06 d r— 03 T- T— d 00 00 T— d $ T_ T_ CM CM T“ CM CM CM T— T— CM T— -0 x: •*-> CO CO CD 03 03 CM Is- Is- M" O T- 03 O 00 re 0 X 03 v_ in 00 Is- CO 00 in CO in cb CO 00 L. _0 CM CM CM CM CM CM CM CM CM CM CM CM CM CM 3 .c ■*-> CO CO CM CO cq CM Is- T- co 00 CO in 03 1 cb O cb cb 06 03 d CM ib d Is" CO 00 O H c 0 03 CM CM CM CM co CO CO co CO co 3 O m O in 03 CO 03 O co CM Is- r" S co Is" Is- Is- Is- CO 00 co in Is- Is" co N CO CO CM 1 CM CM CM CM CM CM Is- CM CM CM Table 6. Summary of information on the feeding of B. per- sicus in Golestan province of Iran. ZMGU 75 is from Ardabil province. ZMGU Form Total length Stomach and intestine contents 267 Two hairless Bats (Myotis blythyii), one digested (~3 cm) and another undigested (~6 cm). Soft material is mixed with undigested bones in large intestine. 276 adult 268.5 One butterfly larva (chrysalid) 28 mm; ~45 ver- miform larvae of Diptera (probably Tabanidae), each ~1 cm; two species of black beetle, (Coleoptera), ~45 mm; stomach of salamander was full. 277 adult >164.5 Soft unidentifiable material. 75 adult 246.6 One larva of B. persicus with snout - vent length >32 mm, mostly digested. adult 236.6 Two mayfly larvae 333 adult 75.5 (Ephemeroptera). Specimen was fed on larvae of Iranian wood frog in laboratory before fixation. 335 adult Some algae and a semidi- gested Earwig (Dermoptera) found in feces. 343 adult 84.8 Two undigested larvae of B. persicus with a total lenght of 30 mm emerged one day after collecting. Results Measurements of specimens from the localities are pre- sented in Table 3. Morphometric and ineristic characters of preserved salamanders are summarized in Table 4 and Table 5. The maximum total length of this species expected 15-20 cm (Schmidtler and Schmidtler 1971). The total length of the longest specimen was 268.5 mm. Description of larvae. - In small larvae (Total length less than 80-100 mm), the head is large, depressed, more or less triangular with rounded end anteriorly, wider poste- riorly, with small eyes and poorly developed eyelids; black horny margin present in lower jaw; gills large; vomerine tooth-bundle arc-shaped, situated anterolater- ally, and extending in front of the choanae, short, in the middle hardly discernibly separated from one another. Trunk with 11-14 costal grooves, vertebral groove often present, forelimbs are longer than hind limbs, tips of fin- 2004 Asiatic Herpetological Research Vol. 10, p. 188 Figure 4. A- One of seven waterfalls of Shirabad Cave (Golestand Province, Iran). Batrachuperus persicus are found in the water near the edges of the pond (Photo by R. Ghaemi 2/28/97); B- The entrance of Shirabad Cave (Photo by R. Ghaemi 2/28/97); C- Myotis blythyi, an abundant bat species of Shirabad Cave and one of the food items of Batrachuperus persicus ; D- Larval specimen of Batrachuperus persicus from Shirabad Cave collected 2/28/97 (Photo by A. Sanee 3/11/1997); E- Juvenile specimen of Batrachuperus persicus about 10 days after metamorphosis. Collected from Shirabad Cave 2/28/97 (Photo by A. Sanaee 4/8/97). gers with black homy pads, their arrangements are 3>2>4>or2>4>1; adpressed limbs not overlapping; tail highly compressed from laterally, upper caudal fin very distinct, reaching to occiput in some specimens, lower caudal fin reaching to posterior of cloaca, cloacal aperture is oval or elliptical. In larger larvae the head is more or less rectangular, adpressed limbs overlapping, upper and lower caudal fins not clearly distinct. The 12 larvae specimens exam- ined (fixed or living) in this study (see Tables 3, 4) have a tail length which is smaller (Table 4) or longer (Table 3) than the head plus body length. Description of juveniles. - Head more or less elliptical, decreasing toward the rear, adpressed limbs overlap- ping, upper and lower caudal fins are not distinct, tail more or less rounded especially at base of it. Four juve- nile specimens examined in this study (see Tables 3, 4), have a tail length which is often smaller than the head plus body length. Description of adults. - Head form is more or less rec- tangular, or wider anteriorly. Eyelids well developed and movable, width of eyelid is less than distance of inter- eyelids (interoculars), distance of external nostrils are longer than distance of nostril to anterior of eye, nostrils are semi-circle; vomerine tooth-bundle is different from that of larvae, inner portion of it is longer than outer one. Trunk with 11-14 costal grooves, adpressed limbs over- lapping. Tail compressed laterally in some specimens and with thin upper and lower caudal fins, and in some specimens more or less rounded especially at base. Cloacal aperture is longitudinal and in some specimens cross-shaped, and longitudinal protuberance is present inside of cloaca in others. The 16 adult specimens exam- ined in this study (see Tables 3, 5) have a tail length which is longer than head plus body length except two specimens (ZMGU 273, 335) which have tail lengths smaller than head plus body length. Coloration - Small larvae (Total length 40 mm) are in general light yellow without any distinct spots; dark eyes very distinct in small larvae, larger larvae have irregular dark gray spots, ventral portion of larvae light and without spots; dorsum of ZMGU 285 is dark gray; Iris yellowish, pupil dark, bases of all limbs yellow^e!- low color of forelimbs not reaching to knee but in hindlimbs reaching to knee. Yellow spots of larvae are Vol. 10, p. 189 Asiatic Herpetological Research 2004 more than in adults. Juveniles are darker than larvae. Yellow spots are less in adult, and ZMGU 269 is deep violet and have only one yellow spot beside of vertebral groove. Yellow spots of adults are often in vertebral groove. Feeding. - Batrachuperus persicus feeds on larvae and adult forms of some orders of insects and probably other arthropods. They also feed on bats ( Myotis blythyi) in Shirabad Cave (Fig. 4). Some specimens are cannibalis- tic and feed on smaller specimens of B. persicus espe- cially in captivity. Algae, that may be eaten with other insects, was found in one larva. Stomach is white with a thin wall. Total length of digestive system of ZMGU 267 was 337 mm from anterior of stomach to posterior of cloaca. Contents of stomachs of some dissected B. per- sicus are shown in Table 6. Behavior. - Small larvae and adult large salamanders are usually almost motionless inside of cave. They have no reaction to light. Adults escape to water. Adults swim more slowly than larvae under water. Adults are active in Shirabad Cave in all times of year but in other localities are not active during the daytime. Parasites. - Many nematodes and mastigophorans were found inside the cloaca of one salamander from locality 2. Some nematodes moved freely and some were inside of a cyst. Metamorphosis. - Larvae are found at all times of the year in Shirabad Cave and probably don’t transform dur- ing the first year. A newly transformed juvenile was found on February 25 1997. Larvae transform rapidly in captivity probably as a result of starvation and higher temperature. Habitat. - Batrachuperus persicus was studied in four localities and observed in some other localities in Golestan Province of Iran. Larvae were found inside of small shady ponds. Juveniles and adults were found inside and outside of water in Shirabad Cave but in other localities live in borrows about 20 cm long. Some spec- imens found above stones, and some in grooves of stones near water in Shirabad Cave. One specimen was 1 meter away of water and moved 0.5 meter on the stone which was in an almost vertical position. Measurements. - Measurements are summarized in Tables 3, 4, and 5. Total length of the smallest larvae was 36.2 mm and of the longest one was 105 mm. Juveniles are smaller than the largest larvae. Total length of the longest adult was 268.5 mm. Distribution. - Batrachuperus persicus collected or observed by students of Gorgan University, staffs of Department of Environment of Gorgan and Gonbad-e- Kavous, and by me in many localities in Golestan Province of Iran. These localities are listed on figure 2. Discussion These salamanders are active at all times of year in Shirabad Cave, but in other localities found in borrows of near of springs during daytimes and are probably active at night. Larval salamanders have morphological adaptations that correlate with the environments that the larvae inhabit (Noble 1927 in Petranka 1988). One dichotomy is that between species that typically breed in running versus standing water habitats. In this division larvae of Batrachuperus persicus are “stream-type” lar- vae. A third group of aquatic larvae (“mountain brook” larvae) was recognized by Valentine and Dennis (1964) that is perhaps better viewed as an extreme form of the stream-type morphology (Petranka, 1998). It is better recognized larvae of B. persicus to this type of larvae. Feeding on bats by adults of B. persicus may be unique in the world of salamanders. Acknowledgments I am much indebted to N. Okhli for various assistance in the field and in laboratory and typing the manuscript. I would like to thank the students of Gorgan University (M. Sabbaghpour, R. Ramezani, S. Afzali, S.Ashrafi, R. Zakeri, M. Azma, M. Rahmani, H. Naghghash, G. Daryanabard, H. Zohouri, A. Sanaee, A. Maghsoudlou, M. Mahmoudi, J. Ghasemi, M. Goli) for their coopera- tion during field work in Golestan Province and for tak- ing the photographs. Thanks to staffs of Department of Environment of Gorgan and Gonbad-e-Kavous especial- ly R. Ghaemi, M. Firoznia Y. Shahkoumahalli and H. Abslan for their cooperations. I am grateful 1 to N. Abbasy and M. Fatemi, N. Moghaddam for identifica- tion of plants and A. Abdoli for identification of water insects. I am also thankful to T. Papenfuss, Museum of Vertebrate Zoology, University of California, Berkeley, for all his recommendations, encouragements, and read- ing the manuscript before publication. Editor’s Note The name Batrachuperus persicus Eiselt and Steiner, 1970. Ann. Naturhist. Mus. Wien, 74:77 (Holotype: NHMW 19435:4 [larval specimen], type locality: Talysch Mountains near Assalem, Gilan Province, Iran, small creek about 800 m) has priority over 2004 Asiatic Herpetological Research Vol. 10, p. 190 Batrachuperus gorganensis Clerque-Gazeau and Thom, 1979. Bull. Soc. Hist. Nat. Toulouse 114:455 (Holotype: MNHMP 1978-1982, type locality: At the edge of a cav- ernous stream on a clay bank 200 m inside the entrance of a cave, situated between the village of Gorgan and Ali-Abad, Elborz Mountain Range of north-central Iran, near the southeast shore of the Caspian Sea and with an elevation of 400 m above sea level). The type localities of the two nominal taxa lie at the extreme western and eastern ends respectively of the known Iranian range of Batrachuperus. Most recent authors have considered the two specimens conspecific and a small number of popu- lations distributed between the two type localities have been discovered. However, the taxonomic status of B. gorganensis as well as that of intermediate populations remains unsettled. Unsettled; see also discussion of the problem in Ebrahimi et al. (2004). Indeed, Risch (1984, Alytes, Paris 3:44) made B. gorganensis the type of his monotypic genus, Paradactylodon. SCA Literature Cited Baloutch, M and H. G. Kami. 1995. Amphibians of Iran. Tehran University Publication, Tehran. 177 pp. (In Farsi). Clergue-Gazeau, M. and R. Thom. 1979. Une nouvelle espece de salamander du genere Batrachuperus in provence de l’lran septentrional (Amphibia, Caudata, Hynobiidae). Bulletin Societe d’Histoire Naturellell4 (3/4):455-460, Toulouse. Ebrahimi, M., H. G. Kami, and M. Stock. 2004. First description of egg sacs and early larval develop- ment in Hynobiid Salamanders (Urodela, Hynobiidae, Batrachuperus) from north-eastern Iran. Asiatic Herpetological Research 10:168-175. Eiselt, J. and H. M. Steiner. 1970. Erstfund eines hyno- biiden Molches in Iran. Annalen des Naturhistorischen Museum Wien 74:77-90. Kami, H. G. and E. Vakilpoure. 1996. Geographic distri- bution of Batrachuperus persicus. Herpetological Review 27 (3): 147. Kami, H. G. 1999. Additional specimens of the Persian Mountain Salamander, Batrachuperus persicus from Iran (Amphibia: Hynobiidae). Zoology in the Middle East 19:37-42. Petranka, J. W. 1998. Salamanders of the United States and Canada. Smithsonian Institution Press, Washington and London. 587 pp. Schmidtler, J. J. and J. F. Schmidtler. 1971. Eine Salamander- Novitat aus Persien, Batrachuperus persicus. Aquarien Magazin 5(1 1). 443-445, Stuttgart. Steiner, H. M. 1973. Beitrage zur Kenntnis der Verbreitung, Okologie und Bionomie von Batrachuperus persicus. Salamandra 9:1-6. Valentine, B. D. and D. M. Dennis. 1964. A comparison of the gill-arch system and fins of three genera of larval salamanders, Rhyacotriton, Gryinophilus, and Ambystom a. Copeia 1964:196-201. | 2004 Asiatic Herpetological Research Vol. 10, pp. 191-201 Annotated Checklist of Amphibians and Reptiles of Pakistan Muhammad Sharif Khan Apt H A 17, 151-S. Bishop Ave., Secane, PA 19018, USA E-mail: typhlops99@hotmail.com Abstract. - From recent herpetological collections several new amphibian and reptilian taxa have been added to the herpetofauna of Pakistan. Thus raising the number of species from Minton's 144 and Mertens' 178 to 225. Key words. - Checklist, Pakistan, herpetofauna. Introduction Following is the checklist of amphibian and reptile species that, so far, have been recorded and described in the major works on the herpetology of the subcontinent from the areas now included in Pakistan (Gunther, 1864; Murray, 1884, 1886, 1892; Boulenger, 1890, 1896; Smith, 1931, 1935, 1943). Recently described and recorded species from Pakistan are also included in it (Anderson and Leviton, 1966, 1967, 1969; Anderson and Minton, 1963; Cherlin, 1981, 1983; Szczerbak, 1991; Golubev and Szczerbak, 1981; Ingoldby, 1922; Mertens 1969a,b, 1970, 1971, 1972, 1974; Baig, 1988, 1989, 1998, 1999; Baig and Bohme, 1996; Baig and Gvozdik, 1998; Dubois and Khan, 1979; Minton, 1966; Minton and Anderson, J. 1965; Minton et al., 1970; Khan, A. Q. and Khan, M.S. 1996; Khan, 1972, 1974, 1980, 1984a, b, 1988, 1989, 1991, 1992, 1993a, b, 1994, 1997a, b,c, 1998, 1999a, b, 2000, 2001a, b, 2003a, b; Khan and Baig, 1992; Khan and Khan, R.Z., 1997; Khan and Tasnim, 1989, 1990; Khan and Rosier, 1999; Rastegar- Pouyani, 1999. Frogs and Toads Bufo stomaticus Liitkin, 1 862 Bufo surdus Boulenger, 1891 Bufo viridis zugmayeri Eiselt and Schmidtler, 1973 Family: Microhylidae Microhyla Tschudi, 1828 Microhyla ornata (Dumeril and Bibron, 1841) Uperodon Dumeril and Bibron, 1841 Uperodon systoma (Schneider, 1799) Family: Ranidae Euphlyctis Fitzinger, 1843 Euphlyctis cyanophlyctis cyanophlyctis (Schneider, 1799) Euphlyctis cyanophlyctis microspinulata Khan, 1997 Euphlyctis cyanophlyctis seistanica (Nikolsky, 1900) Family: Bufonidae Bufo Laurenti, 1768 Bufo himalayanus Gunther, 1 864 Bufo latastii Boulenger, 1882 Bufo melanostictus hazarensis Khan, 200 1 Bufo olivaceus Blanford, 1874 Bufo pseudoraddei pseudoraddei Mertens, 1971 Bufo pseudoraddei baturae Stock, Schmid, Steinlein, and Grosse, 1999 Bufo siacheninsis Khan, 1997 Hoplobatrachus Peters, 1 863 Hoplobatrachus tigerinus (Daudin, 1802) Fejervarya Bolkay, 1915 Fejervarya limnocharis (Boie, 1834) Fejervarya syhadrensis ( Annandale, 1919) Nanorana Gunther, 1836 Nanorana pleskei (Gunther, 1896) © 2004 by Asiatic Herpetological Research Vol. 10, p. 192 Asiatic Herpetological Research 2004 Paa Dubois, 1975 Paa barmoachensis (Khan and Tasnim, 1989) Paa hazarensis Dubois and Khan, 1979 Paa sternosignata (Murray, 1885) Paa vicina (Stoliczka, 1872) Sphaerotheca Dumeril and Bibron, 1841. Sphaerotheca breviceps (Schneider, 1799) Turtles and Tortoises Family: Cheloniidae Caretta Rafiuesque, 1814 Caretta caretta (Linnaeus, 1758) Chelonia Brongniart, 1800 Chelonia my das (Linnaeus, 1758) Eretmochelys Fitzinger, 1843 Eretmochelys imbricata (Linnaeus, 1766) Lepidoc/ielys Fitzinger, 1843 Lepidochelys olivacea (Eschscholtz, 1824) Family: Dermochelyidae Dermochelys Blainville, 1816 Dermochelys coriacea (Vandelli, 1761) Family: Emydidae Geoclemys (Gray, 1821) Geoclemys hamiltonii (Gray, 1821) Har della Gray, 1870 Hardella thurjii Gray, 1 870 Kachuga Gray, 1856 Kachuga smithii (Gray, 1 863) Kachuga tecta (Gray, 1831) Family: Testudinidae Agrionemys Khozatsky and Mlynarsky, 1966 Agrionemys horsfieldii (Gray, 1844) Geochelone Fitzinger, 1835 Geochelone elegans (SchopfF, 1792) Family: Trionychidae Aspideretes Hay, 1835 Aspideretes gangeticus (Cuvier, 1 825) Aspideretes hurum (Gray, 1831) Chitra Gray, 1 844 Chitra indica (Gray, 1831) Lissemys Smith, 1 93 1 Lissemys punctata andersoni Webb, 1980 Crocodiles and Gavials Family: Crocodylidae Crocodylus Laurenti, 1768 Crocodylus palustris Lesson, 1831 Family: Gavialidae Gavialis Oppel, 1811 Gavialis gangeticus (Gmelin, 1789) Lizards Family: Agamidae Brachysaura Blyth, 1856 Brachysaura minor (Hardwicke and Gray, 1827) Cables Cuvier, 1817 Calotes versicolor versicolor (Daudin, 1 802) 2004 Asiatic Herpetological Research Vol. 10, p. 193 Calotes versicolor farooqi Auffenberg and Rehman, 1995 Japalura Gray, 1853 Japalura kumaonensis (Annandale, 1907) Laudakia Gray, 1845 Laudakia agrorensis (Stoliczka, 1872) Laudakia badakhshana (Anderson and Leviton, 1969) Laudakia caucasia (Eichwald, 1831) Laudakia fusca (Blanford, 1876) Laudakia himalayana (Steindachner, 1869) Laudakia lirata (Blanford, 1874) Laudakia melanura nasiri Baig, 1999 Laudakia melanura melanura Blyth, 1854 Laudakia microlepis (Blanford, 1874) Laudakia nupta (de Filippi, 1843) Laudakia nuristanica (Anderson and Leviton, 1969) Laudakia pakistanica (Baig, 1989) Laudakia pakistanica auffenbergi Baig and Bohme, 1996 Laudakia pakistanica khani Baig and Bohme, 1996 Laudakia tuberculata (Hardwicke and Gray, 1827) Phrynocephalus Kaup, 1 825 Phrynocephalus clarkorum (Anderson and Leviton, 1967) Phrynocephalus euptilopus Alcock and Finn, 1896 Phrynocephalus luteoguttatus Boulenger, 1887 Phrynocephalus maculatus Anderson, 1 872 Phrynocephalus ornatus Boulenger, 1 887 Phrynocephalus scutellatus Olivier, 1807 Trapelus Cuvier, 1816 Trapelus agilis Olivier, 1804 Trapelus agilis agilis (Olivier, 1804) Trapelus agilis pakistanensis Rastegar-Pouyani, 1999 Trapelus megalonyx Gunther, 1 864 Trapelus rubrigular is Blanford, 1876 Trapelus ruderatus baluchianus (Smith, 1935) Family: Chameleonidae Chamaeleo Laurenti, 1768 Chamaeleo zeylanicus Laurenti, 1768 Family: Eublepharidae Eublepliaris Gray, 1 827 Eublepharis macularius (Blyth, 1854) Family: Gekkonidae Agamura Blanford, 1 874 Agamura persica (Dumeril, 1856) Altigekko M.S. Khan, 2003 Altigekko baturensis (Khan and Baig, 1992) Altigekko boehmei (Szczerbak, 1991) Altigekko stoliczkai (Steindachner, 1 869) Bunopus Blanford, 1874 Bunopus tuberculatus Blanford, 1874 Crossobamon Boettger, 1888 Crossobamon lumsdeni (Boulenger, 1887) Crossobamon maynardi (Smith, 1933) Crossobamon orientalis (Blanford, 1876) Cyrtopodion Fitzinger, 1843 Cyrtopodion agamuroides (Nikolsky, 1900) Cyrtopodion kachhense kachhense (Stoliczka, 1872) Cyrtopodion kachhense ingoldbyi Khan, 1997 Cyrtopodion kohsulaimanai (Khan, 199 Id) Cyrtopodion montiumsalsorum { Annandale, 1913) Cyrtopodion potoharensis Khan, 2001 Vol. 10, p. 194 Asiatic Herpetological Research 2004 Cyrtopodion scabrum (Heyden, 1 827) Cyrtopodion watsoni (Murray, 1 892) Hemidactylus Oken, 1817 Hemidactylus brookii Gray, 1845 Hemidactylus flaviviridis RUppell, 1835 Hemidactylus frenatus Schlegel, 1836 Hemidactylus leschenaultii Dumeril and Bibron, 1836 Hemidactylus persicus J. Anderson, 1872 Hemidactylus triedrus (Daudin, 1802) Hemidactylus turcicus (Linnaeus, 1758) Indogekko M.S. Khan, 2003 Indogekko fortmunroi (Khan, 1993) Indogekko indusoani (Khan, 1980) Indogekko rhodocaudus (Baig, 1998) Indogekko rohtasfortai (Khan and Tasnim, 1990) Mediodactylus Szczerbak and Golubev, 1977 Indogekko walli (Ingoldby, 1 922) Ptyodactylus Goldfuss, 1820 Ptyodactylus homolepis Blanford, 1876 Rhinogecko de Witte, 1973 Rhinogecko femoralis (Smith, 1933) Rhinogecko misonnei de Witte, 1973 Siwaligekko M. S. Khan, 2003 Siwaligekko battalensis (Khan, 1993) Siwaligekko dattanensis (Khan, 1980) Siwaligekko mintoni (Golubev and Szczerbak, 1981) Teratolepis Gunther, 1 870 Teratolepis fasciata { Blyth, 1853) Teratoscincus Strauch, 1863 Teratoscincus microlepis Nikolsky, 1899 Teratoscincus scincus (Schlegel, 1858) Teratoscincus scincus keyserlingi Strauch, 1 863 Tropiocolotes Peters, 1880 Tropiocolotes depressus Minton and J. A. Anderson, 1965 Tropiocolotes persicus persicus (Nikolsky, 1903) Tropiocolotes persicus euphorbiacola Minton, S. Anderson, and J. A. Anderson, 1970 Family: Lacertidae Acanthodactylus Wiegmann, 1834 Acanthodactylus blanfordii Boulenger, 1918 Acanthodactylus cantoris Gunther, 1 864 Acanthodactylus micropholis Blanford, 1874 Eremias Wiegmann, 1 834 Eremias acutirostris (Boulenger, 1887) Eremias aporosceles (Alcock and Finn, 1896) Eremias fasciata Blanford, 1874 Eremias persica Blanford, 1874 Eremias scripta (Strauch, 1867) Mesalina G ray, 1838 Mesalina brevirostris Blanford, 1874 Mesalina watsonana (Stoliczka, 1872) Ophisops Menetries, 1832 Ophisops elegans Menetries, 1832 Ophisops jerdonii Blyth, 1853 Family: Scincidae Ablepharus Fitzinger, 1823 Ablepharus gray anus (Stoliczka, 1872) Ablepharus pannonicus (Fitzinger, 1823) 2004 Asiatic Herpetological Research Vol. 10, p. 195 Chalcides Laurenti, 1768 Chalcides ocellatus (Forskal, 1775) Eurylepis Blyth, 1854 Eurylepis taeniolatus taeniolatus (Blyth, 1854) Lygosoma Hardwick and Gray, 1827 Lygosoma punctata (Linnaeus, 1766) Mabuya Fitzinger, 1826 Mabuya dissimilis (Hallowed, 1860) Mabuya macular i a (Blyth, 1853) Novoeumeces Griffith, Ngo, and Murphy, 2000 Novoeumeces blythianus (J. Anderson, 1871) Novoeumeces indothalensis (M.S. Khan and M.R.Z. Khan, 1997) Novoeumeces schneider ii zarudnyi (Nikolsky, 1900) Ophiomorus Dumeril and Bibron, 1839 Ophiomorus blanfordi Boulenger, 1887 Ophiomorus brevipes (Blanford, 1874) Ophiomorus raithmai S. Anderson and Leviton, 1966 Ophiomorus tridactylus (Blyth, 1853) Scincella Mittleman, 1950 Scincella himalayana (Gunther, 1 864) Scincella ladacensis (Gunther, 1864) Family: Uromastycidae Uromastyx Merrem, 1 820 Uromastyx asmussi (Strauch, 1 863) Uromastyx hardwickii Gray, 1 827 Family: Varanidae Varanus Merrem, 1820 Varanus bengalensis (Daudin, 1 802) Varanus flavescens (Hardwicke and Gray, 1 827) Varanus griseus (Daudin, 1803) Varanus griseus caspius (Eichwald, 1831) Varanus griseus koniecznyi Mertens, 1954 Ophidia: Snakes Family: Leptotyphlopidae Leptotyphlops Fitzinger, 1843 Leptotyphlops blanfordii (Boulenger, 1890) Leptotyphlops macrorhynchus (Jan, 1 862) Family: Typhlopidae Ramphotyphlops Fitzinger, 1843 Ramphotyphlops braminus (Daudin, 1803) Typhlops Oppel, 1811 Typhlops ahsanai M.S. Khan, 1999 Typhlops diardii Schlegel, 1839 Typhlops diardii platyventris M.S. Khan, 1998 Typhlops ductuliformes M.S. Khan, 1999 Typhlops madgemintonai madgemintonai M.S. Khan, 1999 Typhlops madgemintonai shermanai M.S. Khan, 1999 Family: Boidae Eryx Daudin, 1803 Eryx conicus (Schneider, 1801) Eryx johnii (Russell, 1801) Eryx tataricus speciosus Zarevsky, 1915 Python Daudin, 1803 Python molurus (Linnaeus, 1758) Family: Colubridae Amphiesma Dumeril, Bibron and Dumeril, 1854 Vol. 10, p. 196 Asiatic Herpetological Research 2004 Amphiesma platyceps (Blyth, 1854) Amphiesma sieboldii (Gunther, 1860) Amphiesma stolatum (Linnaeus, 1758) Oligodon Boie, 1827 Oligodon arnensis (Shaw, 1 802) Oligodon taeniolatus (Jerdon, 1853) Argyrogena Werner, 1924 Argyrogena fasciolata (Shaw, 1 802) Boiga Fitzinger, 1826 Boiga melanocephala (Annandale, 1904) Boiga trigonata (Schneider, 1802) Platyceps Blyth, 1860 Platyceps rhodorachis kashmirensis (M. S. Khan and A. Q.Khan, 2000) Platyceps rhodorachis ladacensis (J. Anderson, 1871) Platyceps rhodorachis rhodorachis (Jan, 1 865) Platyceps ventromaculatus bengalensis (M. S. Khan and Coluber Linnaeus, 1758 A. Q.Khan, 2000) Platyceps ventromaculatus indusai (M. S. Khan and A. Coluber karelini karelini Brandt, 1838 Coluber karelini mintonorum Mertens, 1969 Q. Khan, 2000) Platyceps ventromaculatus ventromaculatus (Gray and Enhydris Sonnini and Latreille, 1802 Hardwicke, 1834) Enhydris pakistanica Mertens, 1959 Psammophis Fitzinger, 1826 Psammophis condanarus (Merrem, 1820) Hemorrhois Boie, 1826 Psammophis leithii leithii Gunther, 1869 Hemorrhois ravergieri (Menetries, 1832) Psammophis lineolatus lineolatus (Brandt, 1838) Lycodon Boie, 1826 Psammophis schokari schokari (Forskal, 1775) Lycodon aulicus aulicus (Linnaeus, 1758) Pseudo cyclop It is Boettger, 1888 Lycodon striatus Shaw, 1 802 Lycodon striatus bicolor (Nikolsky, 1903) Pseudocyclophis persicus (J. Anderson, 1 872) Lycodon striatus striatus (Shaw, 1 802) Ptyas Fitzinger, 1843 Lycodon travancoricus (Beddome, 1870) Ptyas mucosus (Linnaeus, 1758) Lytorhynchus Peters, 1862 Sibynophis Fitzinger, 1843 Lytorhynchus maynardi Alcock and Finn, 1896 Lytorhynchus paradoxus (Gunther, 1875) Sibynophis Sagittarius (Cantor, 1839) Lytorhynchus ridgewayi Boulenger, 1887 Spalerosopliis Jan, 1865 Matrix Laurenti, 1 768 Natrix tessellata (Laurenti, 1 768) Spalerosophis arenarius (Boulenger, 1890) Spalerosophis diadema diadema (Schlegel, 1837) Spalerosophis diadema atriceps (Fisher, 1885) Spalerosophis schirazianus (Jan, 1 865) 2004 Asiatic Herpetological Research Vol. 10, p. 197 Telescopus Wagner, 1830 Telescopus rhinopoma (Blanford, 1874) Xenochrophis Gunther, 1864 Xenochrophis cerasogaster cerasogaster (Cantor, 1839) Xenochrophis piscator piscator ( Schneider, 1799) Xenochrophis sanctijohannis (Boulenger, 1 890) Family: Elapidae Bungarus Daudin, 1803 Bungarus caerulens caeruleus (Schneider, 1801) Bungarus sindanus razai M. S. Khan, 1985 Bungarus sindanus sindanus Boulenger, 1847 Naja Laurenti, 1768 Najanaja (Linnaeus, 1758) Naja oxiana (Eichwald, 1831) Family: Hydrophiidae Astrotia Fisher, 1856 Astrotia stokesii (Gray, 1 846) Enhydrina Gray, 1849 Enhydrina schistosa (Daudin, 1803) Hydrophis Latreille, 1802 Hydrophis caerulescens (Shaw, 1 802) Hydrophis cyanocinctus Daudin, 1 803 Hydrophis fasciatus (Schneider, 1799) Hydrophis lapemoides (Gray, 1 849) Hydrophis mamillaris (Daudin, 1 803) Hydrophis ornatus (Gray, 1 842) Hydrophis spiralis (Shaw, 1 802) Lapemis Gray, 1835 Lapemis curtus (Shaw, 1 802) Microcephalophis Lesson, 1834 Microcephalophis cantoris (Gunther, 1 864) Microcephalophis gracilis (Shaw, 1802) Pelamis Daudin, 1803 Pelamis platurus (Linnaeus, 1766) Pr aes cut at a Wall, 1921 Praescutata viperina (Ph. Schmidt, 1852) Family: Viperidae Daboia Gray, 1842 Daboia russelii russelii (Shaw and Nodder, 1797) Echis Merrem, 1 820 Echis carinatus (Schneider, 1 820) Echis carinatus astolae Mertens, 1969 Echis carinatus multisquamatus Cherlin, 1981 Echis carinatus sochureki Stemmier, 1964 Eristicophis Alcock and Finn, 1896 Eristicophis macmahonii Alcock and Finn, 1897 Macrovipera Reuss, 1 927 Macroviper a lebetina obtusa (Dwigubsky, 1832 Pseudocerastes Boulenger, 1 896 Pseudocerastes bicornis Wall, 1913 Pseudocerastes persicus (Dumeril, Bibron, and Dumeril, 1854) Family: Crotalidae Gloydius Hoge and Romano-Hoge, 1981 Gloydius himalayanus (Gunther, 1864) Vol. 10, p. 198 Asiatic Herpetological Research 2004 Literature Cited Anderson, S. C. and A. E. Leviton. 1966. A review of the genus Ophiomorus (Sauria: Scincidae), with descriptions of three new forms. Proceedings California Academy of Sciences. 4th Ser. 33:499- 534. Anderson, S. C. and A. E. Leviton, 1967. 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The identity of Testudo punctata Lacepede, 1788 (Testudines, Trionychidae). Bulletin du Museum National d’Flistoire Naturelle. Paris. 4(2):547-557. 2004 Asiatic Herpetological Research Vol.10, pp. 202-207 A Morphological and Taxonomic Study on Lacerta parva Boulenger, 1887 (Sauria: Lacertidae) from West Taurus, Turkey Y. KUMLUTA^1’*, S. H. Durmu^1, Y. Kaska2 , M. Oz3 AND M. R. TUNg3 ' Dokuz Eyliil Universitesi, Buca Egitim Fakiiltesi, Biyoloji A.B.D. Buca- / zmir, Turkey. 2 Pamukkale Universitesi, Fen-Edebiyat Fakiiltesi, Biyoloji Boliimu Denizli-Turkey. J A kdeniz Universitesi, Fen-Edebiyat Fakiiltesi, Biyoloji Boliimii Antalya-Turkey. jjj To whom correspondence should be addressed E-mail: yusuf.kumlutas@deu.edu.tr Abstract. - The morphometric measurements of taxonomically important characters, coloration, and pholidosis fea- tures of 74 Lacerta parva specimens collected from West Taurus, Turkey were investigated. Statistical analyses were done and these results were compared with those from relevant literature. Some of the characters were found to be different on the specimens from different localities. New localities from southwest Turkey were also discovered dur- ing this study. Key words. - Lacerta parva, West Taurus, Turkey. Introduction Lacerta parva was first identified as a new species based on a female specimen collected from Kayseri, Turkey (Boulenger, 1887). In later studies, the distribu- tion of this species was extended to include all of Anato- lia and the Caucasian region (Werner, 1902; Nesterow, 1912; Nikolsky, 1915; Bird, 1936; Bodenheimer, 1944; Mertens, 1952; Ba§oglu and Baran, 1977; Baran et al., 1992; Baran and Atatiir, 1998). The distribution of this species was extended to Europe by giving the locality from Tekirdag, Turkey (Venchi-Bologna, 1996). Peters (1962) compared the variations and similari- ties between Caucasian and Anatolian populations by examining the 131 and 74 specimens, respectively. Atagiin (1984) also did a comparative study on the 208 specimens collected from six different sub-populations (Fethiye, Denizli, Konya, Ankara, Kayseri, Erzurum) from Anatolia. Recently, Mtilayim et al. (2001) studied 46 specimens collected from Golka§i Village, Bey^ehir- Konya and found much more similarities between Konya and Fethiye populations as previously mentioned by Atagiin (1984). This work investigates the distribution of this spe- cies in the west of Turkey and also provides morpho- metrical comparison of these specimens with previously collected specimens and relevant literature. We try to resolve the taxonomical situation of the sub-populations of Lacerta parva in Anatolia. Material and Methods Most of the specimens were obtained from Taurus, southwest of Anatolia. A sum of 22 male, 38 female and 14 juvenile specimens were collected. These specimens, collected during the years of 1995-1997, were kept at ZDEU (Zoology Department of Ege University). The locations where the samples were collected are given in Figure 1. The list of material is given below as the Departmental Identification Code, sex, number of speci- mens, locality, date, initials, and surname of the collec- tors respectively. List of Material 1) 140 / 1995, 1 male, Bey§ehir, 19.09.1995, Leg. M. Oz. 2) 238 / 1996 1-9 males, 10-22 females, Bozhoyiik Ovacik-Elmali, 18.06.1996, Leg. Y. Kumluta§ , R. Tun9, S. H. Durmu§ . 3) 239 / 1996, 1-5, 6-13, 14-27 Juv., gayryakas - Gazipa^a, 23.08.1996, Leg. Y. Kumluta§, M. Oz, R. Tun?. 4) 163 / 1997, 1-7, 8-24 , Beyobasi, 25.06.1997, Leg. Y. Kumluta§, M. Oz, R. Tun?. Coloration of living specimens was determined by eye, slides were taken, and then the specimens fixed with the traditional processes and kept in 70% alcohol. The morphometric measurements were done with a dig- ital calliper with an accuracy of 0.02 mm. The body Measurements taken and their descriptions and indexes of the characters are as follows. Pileus Width (PW): © 2004 by Asiatic Herpetological Research Vol.10, p. 203 Asiatic Herpetological Research 2004 Figure 1. The distribution of Lacerta parva in Turkey. Collection localities in this study. Refer to the materials list for details. Collection localities from literature (Peters, 1962; Atagiin, 1984; Ba§oglu-Baran, 1977; Baran et al. , 1992; Venchi-Bologna, 1996; and Miilayim et al. , 2001). 1- Bey§ehir-Konya (30 km) 15- Qubuk-Ankara 29- Yozgat 2- Bozhoyuk-Ovacik 16- Ankara 30- Sorgun-Yozgat 3- Qayiryakasi-Gazipa§a 17- Kulu-Konya 31- Between Mecitozu and Qorum 4- Beyobasi 18- inevi-Konya 32- Suluova-Amasya 5- Gokka§ikoyu-Bey§ehir 19- Konya 33- Tekneli Village-Tokat 6- Antalya 20- Karaman 34- Akdag-Sivas 7- Fethiye 21- Diimbelek Mount-Mersin 35- Erzurum 8- Akdag-Qivril 22- Akgol 36- Akdag (Northeast Anatolia) 9- Isparta 23- Uluki§la 37- Horosan-Erzurum 1 0- Afyon 24- Nigde 38- Sarikami§ 11- Kutahya 25- Nigde-Kayseri arasi 39- Kagizman 12- Eski§ehir-Alayurt arasi 26- Erciyes Dagi 40- Between Culfa and Lake Van 13- Gerede 27- Kayseri 41- Tekirdag 14- Kastomonu 28- Bunyan-Kayseri The widest distance between the parietal plates. Pileus Length (PL): The distance from the posterior point of parietal plates to the tip of rostrum. Head and Body Length (HBL): The distance between the front tip of rostrum and front edge of anus. Body Length (BL): The total length of body from tip of rostrum to the end of tail. Tail Length (TL): The length of tail from anus to the tip of tail. Forelimb Length (FL): The length of forelimb from the body connection to the tip of longest finger. Hind-limb Length(HL): The length of hind limb from the body connection to the tip of fourth fin- ger. Pileus Index (PI)=PW/PLxl00, Tail Index (TI)=TL/BLxl00 and Forelimb Index (FI)=FL/ BLxlOO were also calculated. The ANOVA statistical test were used in comparison of the measurements and the ratios (Minitab, 1991). The values of “Coefficient of difference (CD)” were used in comparison of some characteristics among the population (Mayr, 1969). Results Pholidosis and Morphometric Measurements 2004 Table 1 : The results of descriptive statistics on some of the characteristics of L. parva specimens (These measurem are given as miiimeter). Parameter N Mean (Min -Max) SD SE Pileus Width (PW) 60 5.28 (4.38-6.90) 0.48 0.06 Pileus Length (PL) 60 10.31 (8.72-11.76) 0.74 0.09 Head and Body Length (HBL) 60 49.84 (42.12-58.81) 3.41 0.44 Tail Length (TL) 32 74.34 (53-96) 10.50 1.87 Pileus Index (PI) 60 51.17 (44.78-60) 2.17 0.28 Tail Index (Tl) 29 60.32 (51.36-65.14) 3.25 0.60 Forelimb Index (FI) 59 29.70 (24.81-34.58) 2.34 0.30 Supra-cilliar Granule 74 6.24 (1-10) 2.03 0.24 Supra-cilliar Plate 74 5.36 (4-9) 0.76 0.08 Median Gularia 74 17.61 (15-19) 1.16 0.13 No. of Lateral Lines in Ventralia male 22 28.27 (27-30) 0.94 0.20 female 38 31.90 (28-34) 1.13 0.18 male+female+Juv 74 30.47 (27-34) 2.08 0.24 Dorsalia 74 37.13 (33-43) 1.94 0.22 Femoral Opening male 22 17.23 (14-26) 2.47 0.53 female 38 15.95(14-18) 1.31 0.21 male+female+Juv 74 16.16 (14-20) 1.36 0.16 No. of 4. Sub-digital Lamellae 74 20.52 (17-23) 1.11 0.13 Rostrale were connected to the nostril and the numbers of postnasal plate were occasionally two (96%), only one (3%) in two specimens and one specimen had one on the left and two on the right. The number of occipital plates were also commonly one (97%) but divided into two in two (3%) specimens. The numbers of supercili- ary plates was five in 44 specimens (59.4%), six in 23 specimens (31.1%), four in four specimens (5.4%), seven in two specimens (2.7%) and nine in one speci- men (1.4%). The numbers of supralabial plates, in front of the subocular plate, was usually four (74.3%); three in two specimens (2.7%); four (one small and three big) in three specimens (4.05%); five in two specimens as two small and three big ones; four big and one small in six (8.1%) other specimens; six (four big and two small) in two specimens, seven (four big and three small) in two specimens. The distal end of the collaria was jagged in shape and the numbers varied between five and eight with a mean of 6.7. The results from statistical analyses of the above mentioned measurements are presented in Table 1; comparative results with other literature are presented in Table 2. There were no statistical differ- ences (F-test, P>0.05) between the specimens collected from different locations in this study. Although the T-test does not show any statistical differences (P>0.05) between males and females, males have relatively higher values than females. For example, the mean length of the pileus was 11.15 (Min.= 9.64 - Max =16.60) in males and 9.99 (8.72 - 11.50) in females. The coefficient of differences (CD) was 0.59. The width of the pileus was 5.53 (4.74 - 6.23) in males and 5.14 (4.38 - 6.90) in females and the CD was 0.44. Vol.10, p. 205 Asiatic Herpetological Research 2004 x 0 >s 0 CQ ■o c CO E 3 L_ D LU 0 c/3 >, CO TO i— CO c < 0' >, c o N 'c 0 o cu > x cu LL E o vt CO £ CO Q. o CO c o 0 Cl 0 Cl E .£ o >s -^5 >s o Ul aS _ co Z co - 8 E c CO c ^ cu M- § CD 00 ^ UJ m O 2 05 5 &< T- CO CO C 2 8 .§ - S ™ E 'o)3s c 'o iS O) , c 0 E c :3 0 03' CD' CO v- CM 03' c 03 CO o o 0 03 0 Cl < z M" M" C/3 CM CM CM CM c CD 03 CO C/3 0 03 h- 03_ "n c 0 r— C :3 0 h-' ID CO t— CM 'c E 03 00 0 o 0 ■+— > 03 Q 0 CL C/3 < z 00 CO CM CM 28 c 00 M- ID O C/3 0 lO ID r- ID 0 c 0 E C 0 CD' ID' o' r--' > X :3 03 M- S 00 CO t- CM T — 0 LL o 0 CL 0 < 03 CM t- CM CM CO T — T — r— T— 0 0 E 0 i_ 0 CL CO CO CM t— h''- M" T_. LO ^ CO I ^ 00 I CD O ,,-T h-' o' 05 CO CM CD CO M" M- M" M" M" -^T O h- 1-- h- h» r-» N- CD O CO CM <•(-. I 00 Tf CD_ LT) CD 03 O CD" CO O trC co' 03 00 CO T— CM ^ T- CM M- CO CD CD CD CD CD CO M~ M" M" M“ M- M' M“ CD CO CO COt-N co' co' 03' CO X— T — CO 03 co' m in in io m M- oo co co co' co' t-~ CO T- CM M" M- M- h- r^- r^. in o d oo oo o h-' in' t-' CO T- CM in CM r-' in m m- m m m m in 0 •*-> 0 a o = ® 0 3 2>E c .E 0 2 g* 0 D)= o o -£ 2 .5 - ^ E cn o.~o c -1 o 0 ^ 3 ® 8 OQ Q IL Tt « 2 > I ro L_ a> +-» J5 .{2 o _0 o 3 Z 0 C:0 The tail was longer in males (76.59) than females (70.62) and the CD of this measurements was 0.28. Color and Pattern. - Dorsal coloration is more ground, grayish-brown in the Elmali population and lighter brown in the other three populations studied. A slim dark line was present from posterior of occipital scute towards the posterior. This line does not stretch to the base of forelimbs in juveniles. There were few dark spots on the vertebrals of some specimens (11%). Dark blotches extending dorso ventral ly from vertebal line formed bands, particularly on Beyobasi specimens. The supra-temporal line was usually continuous until the middle part of body; broken lines continued posteriorly to the base of the tail or sometimes until the tip of tail in some specimens. Small spots are present, their central parts are greenish-blue and surrounding areas are dark starting at the base of the forelimb and usually continu- ing posteriorly. The subocular line is dirty white in color and continues to the hindlimbs. Dark spots were present under this line in some specimens. Ventral scales are yellow in males, especially dur- ing the breeding season (which change to white later in the season), and lighter in females. However, the color under the hind limb was sometimes yellowish; the other parts were pinkish-white for females. Dark stains on ventralia were absent on the samples, except for the last ventral plate. Ecological Observations. - The three specimen collec- tion localities were new localities for L. parva , except for the Bey§ehir population. These habitats were nearly 2000 m in altitude (i.e., Bozhoyiik-Ovacik 1800 m., (fayiryakasi-Gazipa§a 1850 m., Beyobasi 1900 m.). The 22 specimens, from Bozhoyiik-Ovacik population, were caught between bushes and small vegetation. The weather was a bit cloudy and the temperature was approximately 24°C. Laudakia stellio and Ablepharus kitaibelii species were also observed in the same habitat. The specimens from Cayiryakasi and Beyobasi were caught while active or under stones at the temperature around 29°C. The col- lection habitats of the specimens were covered mainly small bushy vegetation not big trees. Lacerta danfordi , Mabuya vittata, Cyrtopodion kotschyi and Natrix natrix species were also observed in the same area. Evaluation and Discussion There were no statistical differences between the differ- ent populations in this study, but tail length, pileus length, fore-limb and hind-limb lengths, and the number of femoral openings were higher in males than females. Vol.10, p. 206 The head and body length and the number of lateral plate lines were higher in females than males. Atagiin (1984) reported that only one plate is present behind the postnasal plate in 26% of the specimens from the Erzu- rum population, but not present in the remaining five populations in his work. We did not record any such character from our specimens. Atagiin (1984) also reported that the division of the occipital plate was also different in the Erzurum population by having a higher number of divisions. This population was also studied by Peters (1962), but he did not mention such differ- ences. Only a small percent (3%) of our specimens showed a division in the occipital plate. Peters (1962), in his study comparing L. parva pop- ulations between Caucasia and Anatolia, found that the mean number of dorsalia were different (males = 35.97; females = 34.76) in Caucasian population than Anato- lian population (males= 38.52; females= 37.53). Our values from West Taurus (males= 37.68; females= 36.42) were very similar to Peters (1962) values from Anatolia. This value, along with other parameters, are presented in Table 2. As it can be seen from this table, the results of this work are very close to the results of the Bey§ehir population reported by Mulayim et al. (2001). Peters (1962) also reported the mean number of lamellae under the fourth finger to be very similar between the Caucasian (males= 21.6; females= 21.2) and Anatolian (males= 22.6; females= 22.1) specimens. Our results for this character were slightly lower than Peters’ results but very close to the results of Mulayim et al. (2001) (Table 2). The number of femoral openings reported by Peters (1962) were slightly higher (males= 17.56; females= 16.46) for Caucasian than for Anatolian (males= 17; females= 15.91) specimens. The number of femoral openings in this study were found to be very close to the most eastern Anatolian population of Erzu- rum (Table 2). The HBL, the mean numbers of supra- cilliar plate, lateral plates in ventralia were very similar with the results of Mulayim et al. (2001), but the mean number of median gularia in this study was very close to the Fethiye population reported by Atagiin (1984). There were no remarkable differences in color and pat- tern reported by others (Peters, 1962; Atagiin, 1984; Baran et al., 1992; Miilayim et al., 2001) and our results. As it can be seen from our present and other previ- ous studies, phenotypic variation among the reptile pop- ulations from Turkey have been quantified extensively using morphological characters. Comparison of mor- phometric measurements may yield a new subspecies, but the different populations of L. parva from Turkey may vary even genetically. Unfortunately, genetic diver- sity at the intra-specific level is not available for any species in Turkey. Sequencing DNA, in particular mtDNA, may help to solve the taxonomic problems present in the herpetofauna of Turkey, as it was done for other amphibian species (i.e. Garcia-Paris et al., 1998) and for sea turtles (i.e., Bowen et al., 1994; Kaska, 2000). Acknowledgments This work is some part of the project [Project No: TBAG-1475 (196T021)] supported by TUBITAK (The Scientific and Technical Research Council of Turkey). Literature Cited Atagun, F. 1984. Turkiye’de Lacerta parva (Reptilia, Lacertidae)’nm Taksonomik Arastirilmasi. (Yuk- sek Lisans Tezi) Bomova- IZMIR, 1-18. Baran, I., I.Yilmaz, R. Kete, Y. Kumluta? and S. H. Durmu§. 1992. Bati ve Orta Karadeniz Bolgesi’nin Herpetofaunasi. Doga Tropical Journal of Zoology 16(3):275-288. Baran, i. and M. K. Atatiir. 1998. Turkiye Herpeto faunas (Kurbaga ve Suriingenler). Qevre Bakanligi- Ankara: 1-214. Ba§oglu, M. and I. Baran. 1977. Turkiye Siiriingenleri. Kisim I. Kaplumbaga ve Kertenkeleler. Ege Univ. Fen Fak. Kitaplar Serisi, Izmir, No. 76: 1-272. Bird, C. G. 1936. The Distribution of Reptiles and Amphibians in Asiatic Turkey, with notes on a Col- lection from Vilayets of Adana, Gaziantep and Malatya. The Annual Magazine of Natural History, London 10(18):257-281. Bodenheimer, F. S. 1944. Introduction into the Knowl- edge of the Amphibia and Reptilia of Turkey. Revue de la Faculte des sciences de l’Universite d’lstanbul 9 (1): 1-78. Boulenger, G. A. 1887. Catalogue of The Lizards In The British Museum (Natural History). Second Edition, Vol. Ill (Lacertidae, Gerrhosauridae, Scincidae, Anelytropidae, Dibamidae, Chamaeleontidae), London, 1-575. Bowen, B. W., N. Kamezaki, C. L. Limpus, G. H. Hughes, A. B. Meylan and J. C. Avise. 1994. Glo- bal phylogeography of the loggerhead turtle {Caretta caretta ) as indicated by mitochondrial DNA haplotypes. Evolution 48:1820-1828. Garcia-Paris, M., M. Alcobendos, and P. Alberch. 1998. Influence of the Guadalquvir river basin on mito- Vol. 10, p. 207 Asiatic Herpetological Research 2004 chondrial DNA evolution of Salamandra salaman- dra (Caudata: salamandridae) from southern Spain. Copeia 1998:173-176. Kaska, Y. 2000. Genetic structure of Mediterranean Sea Turtle Populations. Turkish Journal of Zoology 24:191-197. Mayr, E. 1969. Principles of Systematics Zoology. Mac- Graw Hill Book Co., Inc. New York, 428 pp. Mertens, R. 1952. Amphibien und Reptilien aus der Ttirkei. Istanbul Universitesi Fen Fakiiltesi mec- muasi. ser. B 17:41-75. Minitab Reference Manual PC Version. 1991. Release 8.2 Qickest Inc., Rosemont, Pennsylvania. Mtilayim, A., C. V. Tok and D. Ayaz. 2001. Bey§ehir (Konya) Civarindan Toplanan Lacerta parva Bou- lenger, 1887 (Sauria: Lacertidae) Omekleri Ozer- inde Morfolojik Bir £ali§ma. Anadolu University Journal of Science and Technology, Vol. 2, No. 2:345-349. Nesterow, P. W. 1912. K gerpetologii judo-sapadnowo sakawkasja I pogranitschnoj tschasti Maloi Asii (Zur Herpetologi des Siidwestlichen Tran- skaukasiens und der Agrenzenden Teile von Klein- asien). Annuaire du Musee Zoologique de l’Academie des Sciences, St. Petersbourg 17:74-75. Nikolsky, A. M. 1915. Fauna Rossii i Sopredelnych stran.I. Presmykajuschtschijesja. Fauna of Russia and adjacent Countries. Reptiles. Vol. I. Chelonia and Sauria. 352pp., Petrograd (English Edition by the Israel Program for Scientific Translations, Jerusalem 1963). Peters, G. 1962. Die Zwergeidechse (Lacerta parva Boulenger) und ihre Verwandtschafts-beziehungen zu anderen Lacertiden, insbesondere zur Libanon- Eeidechse (L. fraasii Lehrs). Zoologisches Jahr- buch 89: 407-478. Venchi, A. and M. A. Bologna. 1996. Lacerta parva Boulenger, a new lizard species for the European fauna. Amphibia-Reptilia 17:89-90. Werner, F. 1902. Die Reptilien-und Amphibienfauna von Kleinasien. Sitz. -ber. Adademie der Wissen- schaften Mathematische - Naturwisssenschaften. Abt. I, 111:1057-1121. 2004 Asiatic Herpetological Research Vol. 10, pp. 208-214 Genetic Variation Among Agamid Lizards of the Trapelus agilis Complex in the Caspian-Aral Basin J. Robert Macey1’2’* and Natalia B. Ananjeva3 1 Department of Evolutionary Genomics, Joint Genome Institute, Lawrence Berkeley National Laboratory, 2800 Mitchell Drive, Walnut Creek, CA 94598-1631, USA 2 Museum of Vertebrate Zoology, University of California, Berkeley, CA 94720, USA 3 Zoological Institute, Russian Academy of Sciences, St. Petersburg, Russia *To whom correspondence should be addressed: E-mail: jrmacey@lbl.gov Abstract. - Allozyme variation is examined in eight populations of Trapelus from the Caspian-Aral Basin of the for- mer USSR. Thirty one loci (15 variable) exhibit remarkably low levels of genetic variation with only a Nei's genet- ic distance of 0.1 17 across 2500 km. An isolated population on the European side of the Caspian Sea is found to phe- netically cluster inside the Asian populations examined, suggesting that it should not be considered taxonomically dis- tinct. Key words. - Reptilia, Squamata, Agamidae, Trapelus , Central Asia, biogeography, allozyme electrophoresis. Introduction The Trapelus agilis complex is distributed on the Iranian Plateau and adjacent regions of southwestern Asia, as well as in the Caspian-Aral Basin to the north in the inte- rior of Asia. Two separate populations of the Trapelus agilis complex are separated by the Caspian Sea in the Caspian-Aral Basin (including regions draining to Lake Balkhash) of the former USSR. One population on the eastern side of the Caspian Sea ranges from western China, Kazakhstan, and Kirgizistan in the north, to Turkmenistan (Fig. 1), Uzbekistan, and Tadjikstan in the south. This Central Asian population is continuous with Iranian, Afghan, and other southwest Asian populations referred to Trapelus agilis. On the western side of the Caspian Sea in Europe a small population occurs in Chechenia and Dagestan, Russia. The two populations occurring in the Caspian-Aral Basin are placed either in a separate species, T. sanguinolentus, or subspecies, T. agilis sanguinolentus. Taxonomic controversy also exists as to the status of the isolated European popula- tion of Trapelus. Some authors consider the European population to be a distinct subspecies of T. sanguinolen- tus ( T. s. sanguinolentus ) with the Central Asian popula- tions being referred to T. s. aralensis (Ananjeva and Tsaruk, 1987). Others consider the European and Asian populations in the Caspian-Aral Basin to be a single taxon, either T. sanguinolentus (Bannikov et al., 1977) or T. agilis sanguinolentus (Wermuth, 1967). The focus of this study is on the relative position of the European and Asian populations in the Caspian-Aral Basin. The Caspian-Aral Basin populations are always grouped together either as a species or as one or two distinct sub- species relative to the southwest Asian populations referred to as T. agilis. Trapelus is an old genus of Agamid lizards with an Afro-Arabian origin (Macey et al., 2000b). Sequence divergence between Trapelus species in Africa ( Trapelus savignii ), Arabia (T. persicus ), the Iranian Plateau ( T. agilis ), and the Caspian-Aral Basin ( Trapelus sanguino- lentus population 6 of this study), which form a clade, is 10.7-13.9% for the mitochondrial DNA segment span- ning from nadl to coxl (Macey et al., 2000b). Applying the rate of 1.3% change per million years for pairwise comparisons as calculated for this segment of mitochon- drial DNA in agamid lizards of the genus Laudakia (Macey et al., 1998), divergence times among these species of Trapelus are estimated to be 8.3 to 10.7 mil- lion years before present (MYBP). These data suggest that the genus has been in Asia since the Miocene. Allozyme data are used to distinguish hypotheses of early divergence of the European and Asian trans- Caspian populations into discrete entities, verses colo- nization of the European side of the Caspian Sea by western Asian populations. High mountains in the Caucasus and Elburz ranges prevent colonization of the European population from the south, where a continuous land connection does exist to Trapelus populations in Iran. The Caspian-Aral Basin corresponds to much of the Paratethys Sea, which during the Miocene almost completely dried up 5-6 MYBP and then returned briefly in the Pliocene, 3.0-3.5 MYBP (Steininger and Rogl, 1984). Divergence following the early period (5-6 MYBP) when much of the Caspian-Aral Basin was © 2004 by Asiatic Herpetological Research Vol. 10, p. 209 Asiatic Herpetological Research 2004 Figure 1. A Trapelus from Repetek Desert Reserve Station, Repetek (38° 34' N, 63° 11' E), Chardjou Region, Turkmenistan. The photo was taken in May, 1989. This is a representative of population 5 of this study. available for colonization, should halt gene flow in the late Miocene or Pliocene and two discrete populations are expected to be detected, one in Europe and one in Asia. Alternatively, a more recent colonization from a founder event, when the Caspian Sea level fluctuated in the Pleistocene, should have a much later restriction in gene flow and therefore the European population may be expected to be nested within the Asian population. Material and Methods Laboratory Protocols. - Tissues were taken in the field, immediately frozen in liquid nitrogen, and later trans- ferred to an ultracold freezer and maintained at -80° C. For analysis of allozymic variation, liver and muscle tis- sues were homogenized separately. Horizontal starch- gel electrophoresis was employed to differentiate varia- tion in 3 1 presumptive loci. The 3 1 loci and eight buffer conditions utilized to resolve them are displayed in table 1. Allozymes were stained using standard methods (Harris and Hopkinson, 1976; Murphy et al., 1990; Richardson et al., 1986; Selander et al., 1971). Carboxylic ester hydrolase (Dimeric Esterase) was resolved using 4-methylumbelliferyl acetate as the sub- strate, Alcohol dehydrogenase (ADH) was resolved using 7h7ra-2-Hexen-l-ol as the substrate, an unidenti- fied peptidase (PEP-1) was resolved using L-leucyl-L- alanine as the substrate, and Peptidase D (PEP-D) and an unidentified peptidase (PEP-2) with the use of L-pheny- lalanyl-L-proline as the substrate. The isozymes, and loci if more than one, were labeled according to their migration from anode to cathode. Specimen Information. - Museum numbers and locali- ties for voucher specimens are presented below. Acronyms are CAS for California Academy of Sciences, San Francisco and MVZ for Museum of Vertebrate Zoology, University of California at Berkeley. Russia: (population 1) Tersko-Kumskaya nizmennast (the low- land between Terek and Kuma Rivers), 15 km WNW (airline) of Voskresenskaya, which is approx. 25 km NNW of Gudermes (43° 21' N 46° 06' E), Schelkovskaya District, Chechen-lngush Autonomous Republic (CAS 182952, 183032-183038). Kazakhstan; (population 2) Almaty (43° 15' N 76° 57' E), Almaty Region (MVZ 216014-216016, CAS 183047-183051). Uzbekistan: (population 3) sand dunes on the west side of the Surkhan Darya (River), on the Kumkurgan (37° 48' N, 67° 37' E) to Denau (38° 16' N, 67° 54’ E) Rd., Surkhan Darjinskaya Region (CAS 183004-183006). 2004 Asiatic Herpetological Research Vol. 10, p. 210 Table 1. The 31 protein loci scored and the electrophoretic conditions within which they were resolved. Enzyme or blood protein Electrophoretic abbreviation E. C. No. No. of Loci Tissue9 Conditions*5 Serum albumin AB - 1 L 1 Aconitase hydratase ACON 4.2. 1.3 1 L 2 Adenylate kinase AK 2.74.3 1 M 2 Alcohol dehydrogenase ADH 1.1. 1.1 2 L 3 Aspartate aminotransferase AAT 2.6.1. 1 1 L 4 Carboxylic ester hydrolase EST-DC 3.1.1.- 2 L 1 Creatine kinase CK 2.7.32 1 M 4 Fructose-bisphosphate aldolase FBA 4.1.2.13 2 L 5 Glucose-6-phosphate isomerase GPI 5.3.1. 9 2 L 4 Glycerol-3-phosphate dehydrogenase G3PDH 1.1. 1.8 1 L 1 D-2-Hydroxy-acid dehydrogenase HADH 1.1.99.6 1 L 5 L-lditol dehydrogenase IDDH 1.1.1.14 1 L 5 Isocitrate dehydrogenase IDH 1.1.1.42 2 L 2 L-Lactate dehydrogenase LDH 1.1.1.27 2 L 3 Malate dehydrogenase MDH 1.1.1.37 2 L 6 Mannose-6-phosphate isomerase MPI 5.3.1. 8 1 L 6 Peptidase (unidentified 1) PEP-1 3.4.-.- 1 L 1 Peptidase D PEP-D 3.4.13.9 1 L 7 Peptidase (unidentified 2) PEP-2 3.4.-.- 1 L 7 Phosphoglucomutase PGM 5.4. 2.2 1 L 8 Phosphogluconate dehydrogenase PGDH 1.1.1.44 1 L 6 Purine-nucleoside phosphorylase PNP 2.4.2. 1 1 L 7 Pyruvate kinase PK 2.7.1.40 1 L 8 Superoxide dismutase SOD 1.15.1.1 1 L 4 ^Tissue abbreviations are: L = liver; M = skeletal muscle. ^Electrophoretic conditions: (1) Lithium-borate/Tris-citrate pH 8.2, 250 v for 6 h (Selander et al., 1971); (2) Amine-cit- rate (Morpholine) pH 6.0, 250 v for 6 h (Clayton and Tretiak, 1972); (3) Tris-citrate/borate pH 8.7, 250 v for 5 h (Selander et al., 1971); (4) Histidine-citrate pH 7.8, 150 V for 8 hours (Harris and Hopkinson, 1976); (5) Phosphate-cit- rate pH 7.0, 120 v for 7 h (Selander et al., 1971); (6) Tris-citrate II pH 8.0, 130 v for 8 h (Selander et al., 1971); (7) Tris- HCL pH 8.5, 250 v for 4 1/2 h (Selander et al., 1971); (8) Tris-maleate-EDTA pH 7.4, 100 v for 10 h (Selander et al., 1971). CEST-D = Dimeric Esterase Turkmenistan: (population 4) SW bank of the Amur Darya (River), approx. 2 km NE of Nephtezavodsk which is 30 km WNW of Deynau (39° 15' N, 63° 1 T E), Chardjou Region (CAS 179552-179559); (population 5) 1 km north of Repetek Desert Reserve Station, Repetek (38° 34’ N, 63° 1 1 ' E), Chardjou Region (CAS 1 79 1 99- 179203, 179416-179420), and Repetek Desert Reserve Station, Repetek (38° 34' N, 63° 11' E), Chardjou Region (CAS 179331); (population 6) 55 km north of Ashgabat (37° 57' N, 58° 23' E) on the Ashgabat - Bakhardok (38° 46' N, 58° 30' E) Rd. then 21 km WNW on dirt Rd., Ashgabat Region (CAS 179758-179767); (population 7) Ashgabat (37° 57' N, 58° 23' E), Ashgabat Region (MVZ 216087-216092); (population 8) near Iolotan’ [YolotanJ (37° 18' N, 62° 21' E), Mary Region (MVZ 216013). Data Analysis, - Nei’s (1978) unbiased genetic distance and Rogers (1972) genetic similarity were calculated using BIOSYS-I (Swofford and Selander, 1981). Phenetic clustering was constructed using the neighbor- joining algorithm (Saitou and Nei, 1987), which does not require rate uniformity, using PAUP* 4.0 (Swofford, 1999) and Nei’s (1978) unbiased genetic distance. Results Variable Loci. - Fifteen of the 31 loci screened show variation among the sampled populations (Table 2). Up to five different allelic states are recognized per loci among populations with no more than four allelic states being present within a population. Genetic Distances. - Allozymic variation among sam- pled populations of Trapelus is surprisingly low (Table 3). The two geographically most distant samples, the European side of the Caspian-Aral Basin (West Caspian) and Kazakhstan (Almaty), have a Nei’s (1978) unbiased genetic distance of only 0.117 across 2500 km. The highest Nei’s (1978) unbiased genetic distances recov- Vol. 10, p.211 Asiatic Herpetological Research 2004 p*®c^romorPh frequencies for the 15 polymorphic loci from eight populations of Trapelus sampled. Localities are West Caspian (WCA), Almaty (ALM), Uzbekistan (UZB), Nephtezavodsk (NEP), Repetek (REP), 70 km NW Ashgabat (NWA), Ashgabat (ASH), lolotan' (IOL). See text for complete localities of all populations used. Locus Electromorph 1-WCA 2-ALM 3-UZB 4-NEP 5-REP 6-NWA 7-ASH 8-IOL AK a 0.125 b 0.875 0.063 0.167 0.938 0.909 1.000 0.929 1.000 c 0.938 0.667 0.063 0.091 0.071 d 0.167 EST-D-2 a 1.000 0.313 1.000 1.000 1.000 0.950 1.000 1.000 b 0.688 0.050 CK a 1.000 1.000 1.000 1.000 0.909 1.000 1.000 1.000 b 0.091 FBA-1 a 0.500 b 0.063 c 0.125 0.188 d 1.000 0.875 1.000 0.750 1.000 1.000 0.929 0.500 e 0.071 FBA-2 a 1.000 b 1.000 0.875 0.818 0.600 0.929 1.000 c 1.000 0.125 0.182 0.400 0.071 GPI-2 a 0.313 0.045 0.150 b 0.688 1.000 1.000 1.000 0.864 0.850 1.000 1.000 c 0.091 HADH a 1.000 1.000 0.667 1.000 1.000 1.000 0.929 1.000 b 0.071 c 0.333 IDDH a 0.182 0.050 0.071 b 1.000 1.000 1.000 1.000 0.818 0.950 0.929 1.000 IDH-1 a 1.000 1.000 0.833 1.000 1.000 1.000 1.000 1.000 b 0.167 LDH-2 a 0.375 0.091 b 1.000 1.000 1.000 0.625 0.909 1.000 1.000 1.000 MPI a 0.063 0.071 b 0.938 1.000 1.000 1.000 0.955 1.000 0.929 1.000 c 0.045 PEP-1 a 0.045 0.100 0.500 b 1.000 1.000 0.500 1.000 0.727 0.800 0.857 0.500 c 0.500 0.227 0.100 0.143 PGM a 0.063 0.091 b 0.125 0.125 0.150 c 0.875 1.000 1.000 0.813 0.909 0.850 1.000 1.000 PGDH a 0.091 b 1.000 0.063 0.091 c 0.333 0.125 0.045 d 1.000 0.667 0.813 0.773 1.000 1.000 1.000 PNP a 1.000 1.000 1.000 1.000 1.000 0.950 1.000 0.500 b 0.050 0.500 Table 3. Matrix of genetic distance and identity coefficients from the eight populations of Trapelus sampled. Nei's unbi- ased genetic distance (Nei, 1978) is above the diagonal, Rogers genetic similarity (Rogers, 1972) is below the diago- nal and sample sizes are on the diagonal. See text for specimen deposition and complete localities of all populations used. 1-WCA 2-ALM 3-UZB 4-NEP 5-REP 6-NWA 7 -ASH 8-IOL 1 . West Caspian (WCA) 8 0.117 0.066 0.039 0.034 0.026 0.035 0.064 2. Almaty (ALM) 0.865 8 0.088 0.102 0.094 0.094 0.108 0.145 3. Uzbekistan (UZB) 0.886 0.873 3 0.031 0.019 0.032 0.021 0.048 4 Nephtezavodsk (NEP) 0.927 0.872 0.910 8 0.006 0.009 0.006 0.029 5. Repetek (REP) 6 70 km NW Ashgabat (NWA) 0.928 0.865 0.920 0.952 11 0.002 0.001 0.026 0.949 0.875 0.909 0.947 0.958 10 0.003 0.026 7. Ashgabat (ASH) 8. lolotan' (IOL) 0.939 0.873 0.931 0.957 0.963 0.964 7 0.022 0.899 0.836 0.900 0.925 0.920 0.932 0.943 1 2004 Asiatic Herpetological Research Vol. 10, p. 212 Genetic Distances among USSR Populations of Trapelus agilis Complex Figure 2. Map of the Caspian-Aral Basin and southwest Asia showing the distribution of the Trapelus agilis complex. Dots depict populations sampled. Lines connect populations in major areas, West Caspian, Kazakhstan, Uzbekistan and Turkmenistan. Nei's unbiased genetic distance (Nei 1978) is plotted between areas. The Turkmen populations are averaged. The western most sample on the west side of the Caspian Sea is population 1 (table 3). The eastern most sample is Almaty in Kazakhstan (population 2). To the southwest of this sample is the Uzbekistan population (popula- tion 3). Four of the Turkmen samples are connected by lines. The most northeastern is population 4 from Nephtezavodsk and the most southeastern is population 5 from Repetek. The most northwestern is population 6 from 70 km NW Ashgabat and the most southwestern is population 7 from Ashgabat. Population 8 from lolotan' is not includ- ed in the average of Turkmen populations because of the low sample size of one and it is distributed between dodu- lations 5 and 7. H H ered are 0.088-0.145 (note that the highest value is with a sample size of one) between the northwestern popula- tion in Kazakhstan (Almaty) and all other populations sampled in the Caspian-Aral Basin. The European pop- ulation on the western side of the Caspian Sea is separat- ed from all other populations except the Kazakhstan (Almaty) population by Nei’s (1978) unbiased genetic distances of 0.026-0.066. The population in Uzbekistan is distinct from those in Turkmenistan by Nei’s (1978) unbiased genetic distances of 0.019-0.048. Mapping these genetic distances on geography reveals a pattern of isolation by distance in which all dis- tances appear relatively additive (Fig. 2). Clustering of these data in a neighbor-joining phenogram and rooting the tree on the longest path places the Kazakhstan (Almaty) population and Uzbekistan sample in a basal Vol. 10, p. 213 Asiatic Herpetological Research 2004 West Caspian (1) *— 70 km NW Ashgabat (6) •“ Ashgabat (7) lolotan’ (8) Nephtezavodsk (4) — Repetek (5) Uzbekistan (3) Airaa-Aty (2) 0.01 changes Figure 3. Neighbor-joining phenogram rooted on the longest path. Note that the European population on the western side of the Caspian Sea is nested inside the Asian populations sampled and appears as the sister population to its nearest Asian population (70 km NW Ashgabat). Population numbers corresponding to figure 1, and tables 2 and 3 are given adjacent to locality names. position. The European (West Caspian) population appears nested within the Turkmen populations and as the sister population to its nearest Asian population (70 km NW Ashgabat), (Fig. 3). This pattern is consistent with the observed genetic distances. Discussion The Age of Trapelus in the Caspian-Aral Basin. - The European population of Trapelus is found to cluster phe- netically within the Asian populations with little genetic differentiation, suggesting that these taxa do not repre- sent distinct forms. The low genetic diversity in Trapelus of the Caspian-Aral Basin probably indicates a recent dispersion of Trapelus throughout the Caspian- Aral Basin. Nei’s (1978) unbiased genetic distances do not exceed 0.117 (with the exception of Kazakhstan to lolotan’ Turkmenistan with a sample size of one). A very approximate estimate of divergence time and genetic distance is 14 million years for a Nei’s D of 1.0 (Maxson and Maxson, 1979). Given this rate these data suggest a divergence of Trapelus populations in the Caspian-Aral Basin at around 1.6 MYBP. This result suggests that Trapelus did not diverge in the Caspian-Aral Basin until the Pleistocene, well after the last drying of the Paratethys Sea 3.5 MYBP (Steininger and Rogl, 1984). Comparison to Other Taxa. - One additional genus of lizard has been sampled from the Caspian-Aral Basin for allozyme variation. The northern populations of the gekkonid genus Mediodactylus from Almaty and the Junggar Depression of China show a minimum of two fixed differences when compared to a southern popula- tion in the Kara Kum Desert (Macey et ah, 2000a). This divergence is greater than those observed among Trapelus where no fixed differences are detected between Almaty (population 2) and the Kara Kum Desert (populations 4-8). Because the Caspian-Aral Basin has had periods of inundation followed by drying over the last 6 million years, and the surrounding moun- tains of the Pamir-Tien Shan are older providing a land refuge (10 million years old; Abdrakhmatov et al., 1996), taxa in the Caspian-Aral Basin may show differ- ent levels of divergence. Taxonomic Recommendations. - Because Trapelus sanguinolentus in the Caspian-Aral Basin is distin- guished from Trapelus agilis of the Iranian Plateau by 10.9% sequence divergence for the mitochondrial DNA segment spanning from nadl to coxl (Macey et al., 2000b), we consider them separate species. No more than a single fixed difference is observed between pop- ulations of Trapelus in the Caspian-Aral Basin. Therefore, we interpret these populations to be a single taxon, T. sanguinolentus. Further work comparing pop- ulations in Southwest Asia is needed in order to deter- mine the specific status of these populations. Acknowledgments This work is LBNL-54654 and was performed under the auspices of the U.S. Department of Energy, Office of Biological and Environmental Research, under contract No. DE-AC03-76SF00098 with the University of California, Lawrence Berkeley National Laboratory. This work was also supported by grants from the National Geographic Society (4110-89 and 4872-93 to Theodore J. Papenfuss and J.R.M.), Russian Foundation of Basic Research (N 02-04-48720 to N.B.A.), Scientist School (NS 1647.2003.4 to N.B.A.), the California Academy of Sciences and the Museum of Vertebrate Zoology. We thank Tatjana N. Duysebayeva for tissue specimens. Nikolai Orlov, Theodore J. Papenfuss, Sakhat M. Shammakov, and Boris S. Tuniyev aided with field work. Kraig Adler and Allan Larson provided valu- able comments on an earlier draft of the manuscript. The first author thanks David B. Wake and Margaret F. Smith 2004 Asiatic Herpetological Research Vol. 10, p. 214 for the opportunity to collect allozymic data at the Museum of Vertebrate Zoology. Literature Cited Abdrakhmatov, K. Ye., S. A. Aldazhanov, B. H. Hager, M. W. Hamburger, T. A. Herring, K. B. Kalabaev, V. I. Makarov, P. Molnar, S. V. Panasyuk, M. T. Prilepin, R. E. Reilinger, I. S. Sadybakasov, B. J. Souter, Yu. A. Trapeznikov, V. Ye. Tsurkov, and A. V. Zubovich. 1996. Relatively recent construction of the Tien Shan inferred from GPS measurements of present-day crustal deformation rates. Nature 384:450-453. Ananjeva and Tsaruk. 1987. The taxonomic status of the steppe agama, Trapelus sanguinolentus in the Precaucasus. In Herpetological Investigations in the Caucasus. Proc. Zool. Institute, Leningrad 158:39- 46. (In Russian). Bannikov, A. G., I. S. Darevsky, V. G. Ishchenko, A. K. Rustamov, and N. N. Shcherbak. 1977. [Guide to amphibians and reptiles of the USSR fauna]. Prosveshchenie, Moskva. 415 pp. (In Russian). Clayton, J. W., and D. N. Tretiak. 1972. Amine-citrate buffers for pH control in starch gel electrophoresis. Journal of Fish. Res. Board Canada 29:1169-1172. Harris, H., and D. A. Hopkinson. 1976 et seq. Handbook of Enzyme Electrophoresis in Human Genetics, Oxford, North Holland Publishing Co., Amsterdam, (loose leaf with supplements in 1977 and 1978). Macey, J. R., Schulte II, J. A., Ananjeva, N. B., Larson, A., Rastegar-Pouyani, N., Shammakov, S. M., and Papenfuss, T. J. 1998. Phylogenetic relationships among agamid lizards of the Laudakia caucasia species group: Testing hypotheses of biogeograph- ic fragmentation and an area cladogram for the Iranian Plateau. Molecular Phylogenetics and Evolution 10:118-131. Macey, J. R., N. B. Ananjeva, Y. Wang, and T. J. Papenfuss. 2000a. Phylogenetic relationships among Asian gekkonid lizards formerly of the genus Cyrtodactylus based on cladistic analyses of allozymic data: Monophyly of Cyrtopodion and Mediodactylus. Journal of Herpetology 34:258-265. Macey, J. R., J. A. Schulte II, A. Larson, N. B. Ananjeva, Y. Wang, R. Pethiyagoda, N. Rastegar-Pouyani, and T. J. Papenfuss. 2000b. Evaluating trans-Tethys migration: An example using acrodont lizard phy- logenetics. Systematic Biology 49:233-256. Maxson, L. R., and R. Maxson. 1979. Comparative albumin and biochemical evolution in plethodontid salamanders. Evolution 33:1057-1062. Murphy, R. W. , J. W. Sites, Jr., D. G. Buth, and C. H. Haufler. 1990. Proteins I: Isozyme electrophoresis. In D. M. Hillis and C. Moritz (eds.), Molecular Systematics, pp. 45-126. Sinauer Associates, Sunderland, Mass. Nei, M. 1978. Estimation of average heterozygosity and genetic distance from a small number of individu- als. Genetics 89:583-590. Richardson, B. J., P. R. Baverstock, and M. Adams. 1986. Allozyme Electrophoresis. A Handbook for Animal Systematics and Population Studies. Academic Press, Sydney. Rogers, J. S. 1972. Measures of genetic similarity and genetic distance. Stud. Genet. VII, Univ. Texas Publ. 7213:145-153. Saitou, N., and M. Nei. 1987. The neighbor-joining method: A new method for reconstructing phyloge- netic tree. Molecular Biology and Evolution 4:406- 425. Selander, R. K., M. H. Smith, S. Y. Yang, W. E. Johnson, and J. R. Gentry. 1971. Biochemical polymorphism and systematics in the genus Peromyscus. I. Variation in the old-field mouse ( Peromyscus polionotus ). Studies in Genetics VI. Univeristy of Texas Publications 7103:49-90. Steininger, F. F., and F. Rogl. 1984. Paleogeography and palinspastic reconstruction of the Neogene of the Mediterranean and Paratethys. In J. E. Dixon and A. H. F. Robertson (eds.), The Geological Evolution of the Eastern Mediterranean, pp. 659-668. Blackwell Scientific Publications, Oxford. Swofford, D. L. 1999. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods), 4.0, Sinauer, Sunderland, Mass. Swofford, D. L., and R. K. Selander. 1981. BIOSYS-1: A FORTRAN program for the comprehensive analysis of electrophoretic data in population genet- ics and systematics. Journal of Heredity 72-282- 283. 2004 Asiatic Herpetological Research Vol.10, pp. 215-216 The Feeding Biology of Rana macrocnemis Boulenger, 1885 (Anura: Ranidae), Collected in llludag, Bursa, Turkey iSMAlL H. UGurta^, Hikmet S. Yildirimhan, and Mehmet Kalkan Uludag University’ Science-Art Faculty ; Department of Biology’ Bursa-Turkey Abstract. - In this study gut contents of 64 mature specimens (34 male, 30 female) from Rana macrocnemis popula- tion collected from Uludag(Bursa) are analyzed. The results indicate that the majority (68.05%) of the food items is composed of insects. Key words. - Rana macrocnemis , feeding, stomach content. Introduction Rana macrocnemis is a very common species in the northern and eastern Caucasus and has dispersed into Western and Northern Anatolia in Turkey. Also known as a “mountain frog”, its altitudinal distribution ranges from 1,000 to 2,300 m. It generally lives in open areas or in small brooks in or near the woods. It is also seen in areas w ith muddy bottoms or close to water. Many stud- ies have been conducted to find out the feeding biology of amphibia (Bohme 1975; Boulenger, 1897; Schreiber, 1912; Beschkov, 1970; Lamb, 1984; Yilmaz, 1984; Sampetro, 1986; Gittins, 1987; AtatUr, 1993; Ugurtas 1995), but no detailed study exists on the feeding biol- ogy of Rana macrocnemis. The aim of this study is to establish various animal groups that are taken as prey by this species. Materials and Methods The specimens of Rana macrocnemis used in this study were collected in three localities between June and July 1997 (Fig. 1). These localities are: Kirazlyayla (1 6 males, 30 females) Hotels Area (10 males) (^obankaya (8 females) The specimens were found between the hours 730 and 1930 hours in daylight, but were observed to appear more often between 1030 and 1500 hours. We used Parker (1982), Lodos (1983, 1986), and Caglar and Demirsoy (1992, 1999) to identify prey items. Results We did not observe any significant discrepancies in the stomach contents of males and females. Thus, they were evaluated together. Of the 64 specimens collected dur- Figure 1 . Localities where Rana macrocnemis speci- mens were collected. ing the feeding period, two had empty stomachs. Among stomach contents which were investigated, 626 prey items were counted. Of these prey items, 426 (68.05%) belonged to Insecta, 36 (5.75%) to Arach- nida, 44 (7.02%) to Gastropoda, 4 (0.63%) to Myri- apoda, 112 (17.89%) to lsopoda and 2 (0.31%) to Acarina groups. Two (0.31%) juveniles of Rana mac- rocnemis were also found as stomach content. The number of prey items found in stomachs and their taxonomy are listed below. It was found that the majority of food taken by Rana macrocnemis was com- posed of insects (68.05%). 144 (36.15%) were Coleoptera, 82 (19.24%) Plecoptera, 94 (22.06%) Diptera, 40 (9.38%) Hymenoptera. 36 (8.45%) Odonata, 6 ( 1 .40%) Orthoptera, 6 ( 1 .40%) Lepidoptera, 4 (0.93%) Homoptera and four (0.93%) in Hemiptera (Fig. 2). As a result of this study on the stomach contents, we con- © 2004 by Asiatic Herpetological Research □ Cole op be ra (36 15 %) ■ Plecoptera (1 9 24 %) □ Diptera (22 06 %) □ Hymenoptera (9 38 %) ■ Odonata (8 4 5 %) □ Orthcptera (1 40 %) □ Lep ideptera (140%) ■ Homoptera (0 93 %) M Hemiptera (0 93 %) Figure 2. The precentages of insect groups taken as prey. elude that Rana macrocnemis is an opportunity feeder that utilizes any prey in its environment that it has the ability to consume. Acknowledgments Demirsoy, A. 1999. Ya amin 7'emel Kurallar, Omurgas- zlar -Bocekler Di§inda, Cilt II, Kisim I, Meteksan A. Ankara, S, 724-939. Gittins, S. P. 1987. The Diet of the Common Toad ( Bufo bufo) Around A Pong in Mid-Wales. Amphibia- Reptilia 8: 13-17. Lamp, T. 1984. The Influence of Sex and Breeding Con- dition on Microhabitat Selection and Diet in the Pig Frog Rana gryllio. 1 he American Midland Natural- ist 1 1 1(2):31 1-318. Lodos, N. 1983. Tiirkiye Entomolojisi (Genel Uygula- mal ve Faunistik), Cilt I Ege Universitesi Matbaas Ziraat Fak. Yayinlari 282:134-336 Lodos, N. 1986. Tiirkiye Entomolojisi (Genel Uygula- mal ve Faunistik), Cilt II Ege Universitesi Matbaas Ziraat Fak. Yaynlar 429:57-498 This work was supported by grants from Uludag Uni- versity Scientific Research Fund (2001/60) Literature Cited Atatiir, M. K. 1993. A Preliminary Study on the Feeding Biology of Rana ridibunda (Anura, Ranidae) popu- lation from Bey^ehir Lake. Turkish Journal of Zool- ogy 17(2): 127- 131. Beschkov, V. 1970. Biologie und Verbreitung des Griechischen Froschers {Rana graeca BLGR.) (In Bulgarian) Academie Bulgare des Sciences. Bulle- tin de L'institut de Zoologie et Musee. Tome 31:5- 17. Boulenger, G. A. 1897. The Tailless Batrachians of Europe. Part. I. Addlard and Son. Hanover Square, London Bohme, W. 1975. Zum Vorkommen von Pelobates syri- aens Boettger, 1889 Griechland. Senck. Biol. 56:199-202. £aglar, M. 1974. Omurgasiz Hayvanlar, Anatomi- Sistematik. II. Kisim. U. Yayinilar, Fen Fak, Sayi 123, Fen Fakultesi Basimevi, Istanbul, S, 157-206. Demirsoy, A. 1992. Yaamn Temel Kurallari, Omurgasizlar - Entomoloji. Cilt II, Kisim II, Meteksan A. Ankara, S, 338-814. Parker, P. S. 1982. Editor Synopsis and Classifications of Living Organism. Vol. 1 and 2. McGraw-Hill Book Company. Sampedro, M. A., and L. Montanez. 1986. Food of Rana catesbeiana in Two Different Areas of Cuba. P. 413-416. In Z. Rocek (ed.). Studies in Herpetol- ogy. Charles University, Prague. Schreiber E. 1913. Herpetologia Europea. Gustav Fis- cher, Jena. Ugurta§, . H., and M. Oz . 1995. Bursa ve Sakarya li Pelobates syriacus (Anura, Pelobatidae) Populasyonlarmin Beslenme Biyolojisi Uzerine Bir On Cali§ma. Turkish Journal of Zoology 19:273- 275. Yilmazj. 1984. Trakya Kuyruksuz Kurbagalari Uzerine Morfolojik ve Taksonomik Bir Ara§tirma. Do§a Biy oloji, S. A2, 8(2): 244-264. [2004 Asiatic Herpetological Research Vol. 10, pp. 217-223 Morphological Observations on the Erythrocyte and Erythrocyte Size of Some Gecko Species, Turkey Murat Sevinc*, Ismail Hakki UGurta§, Hikmet Sami Yildirimhan Uludag University, Science and Art Faculty, Department of Biology, Bursa, Turkey * To whom correspondence should be addressed; E-mail: smurat@uludag.edu.tr Abstract. - In this study, erythrocyte size and morphology of the four gecko species [Asaccus elisae, Hemidactylus turcicus, Cyrtopodion scaber and C. heterocercus mardinensis (Gekkonidae)] from Turkey were examined. Forty-two specimens were used in this study, of which twelve were A. elisae, eight were H. turcicus , twelve were C. scaber, and ten were C. h. mardinensis. Erythrocyte morphology of these examined species was described using Wright’s tech- nique. The sizes of erythrocytes and their nuclei were measured using an ocular micrometer at a magnification of 1600x. The results of this study were compared with previous works on the other reptile species. The longest erythro- cytes were found in H. turcicus and the shortest in A. elisae. In terms of the studied species, the nucleus and erythro- cyte sizes were found to be correlated (Gekkonidae: r = 0.39; P < 0.001). Key words. - Gekkota, Turkey, erythrocyte. Introduction Initial studies on the blood of reptiles described the structures, often comparing them with those of the other vertebrates. Literature on the haematology of reptilian blood are based on a few studies where most were con- cerned with especially European species (Saint Girons, 1970). Various authors have focused on the different circu- lating blood cell types of different reptiles (Taylor and Kaplan, 1961; Heady and Rogers, 1963; Hartman and Lessler, 1964; Szarski and Czopek, 1966; Duguy, 1970; Saint Girons, 1970; Mateo et al., 1984; Canfield and Shea, 1988; Cannon et al., 1996; Alleman et al., 1999; Sevin? et al., 2000; Atatiir et al., 2001; Sevinc and Ugu- rta§, 2001; Ugurta§ et al., 2003). Some authors have studied seasonal (Hutton, 1960; Cline and Waldman, 1962; Haggag et al., 1966) or sexual (Altland and Thompson, 1958) variations in the number of blood cells of different reptile species. In addition, researchers have studied the number of blood cells of different rep- tiles (Baker and Kline, 1932; Charipper and Davis, 1932; Altland and Thompson, 1958; Hutton, 1961; Hutchinson and Szarski, 1965; Engbretson and Hutchinson, 1976; Mateo et al., 1984). Furthermore, authors have also studied haemoglobin and hematocrit content of blood and hematopoiesis of different reptiles (Altland and Thompson, 1958; Hutton, 1961; Goin and Jackson, 1965; Engbretson and Hutchinson, 1976; Newlin and Ballinger, 1976; Mateo etal., 1984; Alleman et al., 1999). In Turkey, hematological studies have generally been conducted on humans and some economically important animals. However, there are few hematologi- cal studies of the reptiles living in this country (Sevin? et al., 2000; Atatiir et al., 2001; Sevin? and Ugurta§, 2001; Ugurta§et al., 2003). In the current study, our aim was to describe and measure erythrocytes of Asaccus elisae (Werner, 1895), Hemidactylus turcicus (Linnaeus, 1758), Cyrtopodion scaber (Heyden, 1827) and C. heterocercus mardinensis (Mertens, 1924) which live in Turkey. This study is the first of its kind on Turkish species. Materials and Methods In this study, twelve (6 males, 6 females) individuals of Asaccus elisae, eight (4 males, 4 females) of Hemidactylus turcicus, twelve (8 males, 4 females) of Cyrtopodion scaber and ten (4 males, 6 females) of C. heterocercus mardinensis (Gekkonidae) were examined. Twenty-two specimens examined were male and twenty were female. The study was performed on 01-05 June 2000. H. turcicus species were collected from Hatay (36° 34' N, 36° 09' E) and the other specimens were from §anliurfa (36° 53' N, 39° 02' E) (Fig. 1; Table 1). Blood was obtained by cutting the tail (Duguy, 1974). Immediately after blood was obtained in heparinized capillary tubes, blood smears were prepared. Three or five blood smears were prepared per individual. The smears were air-dried and stained with Wright’s stain (Hartman and Lessler, 1964). Twelve drops of Wright’s stain were dropped on the slides and allowed to remain on the slide one and half minutes before rinsing with phosphate buffer (pH 6.5). The slides were allowed to stand for ten minutes at room temperature, were washed with distilled water, and allowed to dry. © 2004 by Asiatic Herpetological Research Vol. 10, p. 218 Asiatic Herpetological Research 2004 On each slide, fifty mature erythrocytes and their nuclei were measured by means of an ocular micrometer at a magnification of 1600x. In this way fifty erythrocyte sizes were calculated. Erythrocyte and nucleus measure- ments of examined species are given in tables 2-5. Erythrocyte and nucleus sizes were, respectively, calcu- lated according to the formulas [(EL x EW x pi) / 4] and [(NL x NW x pi) / 4]; where EL is the erythrocyte length, EW is the erythrocyte width, NL is the nucleus length and NW is the nucleus width. Results Erythrocytes, or red blood cells, of geckos are nucleat- ed, oval cells. Their nuclei are also oval and centrally located, like those of the other reptiles. The cytoplasm of mature erythrocytes appeared both light, dark pink, and homogeneous under Wright’s stain. Nuclei of mature erythrocytes are chromophilic (Figs. 2-5). Because there were no significant differences between the erythrocyte sizes of female and male geck- os, data from the females and males of individual species were combined. The longest erythrocytes were found in Hemi- dactylus turcicus. The mean length of mature erythro- cyte of H. turcicus was 16.98 mm (± 1.26 standard devi- ations, with a range of 14.64-19.52 mm) (Table 2; Fig. 6) and also erythrocyte size and length/width ratios of H. turcicus are given in table 2. The shortest erythrocytes were found in Asaccus elisae. The mean length of mature erythrocytes of A. elisae was 14.96 mm (± 0.79 standard deviations, with a range of 13.42-17.08 mm) (Table 3; Fig. 6). Erythrocyte size and length/width ratios of A. elisae are given in table 3. The widest erythrocytes were found in Cyrtopodion scaber. The mean width of mature erythrocytes of C. scaber was 10.26 mm (± 0.78 standard deviations, with a range of 8.54-12.20 mm) (Table 4; Fig. 7). Erythrocyte size and length/width ratios of C. scaber are given in table 4. The narrowest erythrocytes were found in Cyrtopodion heterocercus mardinensis. The mean width of mature erythrocyte of C. h. mardinensis was 9. 1 8 mm (± 0.70 standard deviations, with a range of 6.71-10.98 mm) (Table 5; Fig. 7) and also erythrocyte size and length/width ratios of C. h. mardinensis are given in table 5. The longest nuclei were found in Cyrtopodion scaber. The mean length of mature nuclei of C. scaber Table 1. Materials list. NM: number of males; NF: number of females; CD: collection date; CL: collection locality. All specimens are from the Zoology Museum in Uludag UniversityScience and Art Faculty, Department of Biology. Species NM NF CD CL Asaccus elisae 6 6 1-3 June Sanliurfa Hemidactylus turcicus 4 4 4-5 June Hatay Cyrtopodion scaber 8 4 1-3 June Sanliurfa C. heterocercus mardinensis 4 6 1-3 June Sanliurfa 2004 Asiatic Herpetological Research Vol. 10, p. 219 Figure 2. Erythrocyte and nucleus sizes of Hemidactylus turcicus. Figure 4. Erythrocyte and nucleus sizes of Cyrtopodion scaber. was 6.81 mm (± 0.60 standard deviations, with a range of 5.49-8.54 mm) (Table 4; Fig. 6). Nucleus size and length/width ratios of C. scaber are given in table 4. The shortest nuclei were found in Asaccus elisae. The mean length of mature nuclei of A. elisae was 6.06 mm (± 0.58 standard deviations, with a range of 4.88- 7.32 mm) (Table 3; Fig. 6). Nucleus size and length/width ratios of A. elisae are given in table 3. The widest nuclei were found in Cyrtopodion hete- rocercus mardinensis. The mean width of mature nuclei of C. h. mardinensis was 3.78 mm (± 0.44 standard devi- ations, with a range of 3.05-4.88 mm) (Table 5; Fig. 7). Nucleus size and length/width ratios of C. h. mardinen- sis are given in table 5. The narrowest nuclei were found in Hemidactylus turcicus. The mean width of mature nuclei of H. turcicus was 3.53 mm (± 0.42 standard deviations, with a range of 3.05-4.27 mm) (Table 2; Fig. 7). Nucleus size and length/width ratios of H. turcicus are given in table 2. Discussion Investigations carried out by various authors (Hartman Figure 3. Erythrocyte and nucleus sizes of Asaccus elisae. Figure 5. Erythrocyte and nucleus of Cyrtopodion hetero- cercus mardinensis. and Lessler, 1964; Szarski and Czopek, 1966; Saint Girons, 1970; Seving et al., 2000; Seving and Ugurta§, 2001; Atatiir et al, 2001; Ugurta§et al, 2003) reported that the sizes of erythrocytes vary in members of the four orders of reptiles. Within the class Reptilia, the largest erythrocytes are seen in Sphenodon punctatus , turtles, and crocodil- ians (Hartman and Lessler, 1964; Saint Girons, 1970; Alleman et al., 1984). Cryptodiran turtles have the largest erythrocytes from all previously studied reptiles (Saint Girons, 1970). The shortest erythrocytes are found in the Lacertidae family (Hartman and Lessler, 1964; Saint Girons, 1970; Seving et al., 2000; Seving and Ugurta§, 2001). Saint Girons (1970) reported erythrocytes and nuclei measurements of some gecko species. In Coleonyx variegatus, erythrocyte length is 18.9 pm and width is 9.6 pm; nucleus length is 7.3 pm and width is 3.7 pm. In Gehyra variegata, erythrocyte length is 17.2 pm and width is 1 1.5 pm; nucleus length is 6.3 pm and width is 3.8 pm. In Heteronota binoei, erythrocyte length is 21.4 pm and width is 10.7 pm; nucleus length is 8.1 pm and width is 3.4 pm. Vol. 10, p. 220 Asiatic Herpetological Research 2004 Table 2. Erythrocyte dimensions of Hemidactylus turcicus with standard deviations. EL: erythrocyte length, EW. ery throcyte width; ES: erythrocyte size; NL: nucleus length; NW: nucleus width; NS: nucleus size. EL (pm) EW (pm) EL/EW (pm) ES (pm) NS/ES (pm) Maximum Minimum Mean 19.52 ± 1.26 14.64 ± 1.26 16.98 ± 1.26 11.59 ±0.67 7.93 ± 0.67 9.69 ± 0.67 2.07 ± 0.14 1.42 ±0.14 1.76 ±0.14 166.50 ± 15.45 94.93 ± 15.45 129.50 ± 15.45 0.20 ± 0.02 0.09 ± 0.02 0.14 ± 0.02 NL (pm) NW (pm) NL/NW (pm) NS (pm) Maximum Minimum Mean 7.93 ±0.59 4.88 ± 0.59 6.41 ± 0.59 4.27 ± 0.42 3.05 ± 0.42 3.53 ± 0.42 2.40 ± 0.22 1.43 ± 0.22 1.83 ±0.22 26.58 ± 3.17 11 ,68± 3.17 17.84± 3.17 Table 3. Erythrocyte dimensions of Asaccus elisae with standard deviations. EL: erythrocyte length; EW: erythrocyte width; ES: erythrocyte size; NL: nucleus length; NW: nucleus width; NS: nucleus size. EL (pm) EW (pm) EL/EW (pm) ES (pm) NS/ES (pm) Maximum Minimum Mean 17.08 ±0.79 13.42 ± 0.79 14.96 ± 0.79 10.98 ± 0.66 7.32 ± 0.66 9.26 ± 0.66 2.08 ± 0.14 1.29 ± 0.14 1.62 ± 0.14 142.00 ± 10.38 83.54 ± 10.38 108.80 ± 10.38 0.27 ± 0.03 0.11 ± 0.03 0.16 ±0.03 NL (pm) NW (pm) NL/NW (pm) NS (pm) Maximum Minimum Mean 7.32 ± 0.58 4.88 ± 0.58 6.06 ± 0.58 4.88 ± 0.41 3.05 ± 0.41 3.62 ± 0.41 2.40 ± 0.25 1.14 ± 0.25 1.69 ± 0.25 23.37 ± 2.47 11.68 ±2.47 17.24 ± 2.47 Table 4. Erythrocyte dimensions of Cyrtopodion scaber with standard deviations. EL: erythrocyte length; EW: ery- throcyte width; ES: erythrocyte size; NL: nucleus length; NW: nucleus width; NS: nucleus size. EL (pm) EW (pm) EL/EW (pm) ES (pm) NS/ES (pm) Maximum Minimum Mean 18.30 ±0.88 14.64 ± 0.88 16.20 ± 0.88 12.20 ±0.78 8.54 ± 0.78 10.26 ±0.78 1.93 ±0.13 1.33 ±0.78 1.59 ± 0.78 175.30 ± 13.93 102.20 ± 13.93 130.60 ± 13.93 0.23 ± 0.02 0.10 ± 0.02 0.15 ±0.02 NL (pm) NW (pm) NL/NW (pm) NS (pm) Maximum Minimum Mean 8.54 ± 0.60 5.49 ± 0.60 6.81 ±0.60 4.88 ± 0.47 2.44 ± 0.47 3.65 ± 0.47 2.75 ± 0.26 1.38 ± 0.26 1.89 ± 0.26 30.38 ± 3.42 12.85 ±3.42 19.53 ± 3.42 Table 5. Erythrocyte dimensions of Cyrtopodion heterocercus mardinensis with standard deviations. EL: erythrocyte length; EW: erythrocyte width; ES: erythrocyte size; NL: nucleus length; NW: nucleus width; NS: nucleus size. EL (pm) EW (pm) EL/EW (pm) ES (pm) NS/ES (pm) Maximum Minimum Mean 17.69 ±0.85 14.03 ±0.85 15.65 ± 0.85 10.98 ±0.70 6.71 ± 0.70 9.18 ± 0.70 2.27 ± 0.16 1.39 ± 0.16 1.71 ±0.16 147.20 ± 11.00 80.33 ± 11.00 112.80 ± 11.00 0.28 ± 0.03 0.12 ± 0.03 0.17 ± 0.03 NL (pm) NW (pm) NL/NW (pm) NS (pm) Maximum Minimum Mean 7.93 ± 0.62 5.49 ± 0.62 6.56 ± 0.62 4.88 ± 0.44 3.05 ± 0.44 3.78 ± 0.44 2.40 ± 0.25 1.13 ± 0.25 1.76 ±0.25 30.38 ± 3.10 13.14 ± 3.10 19.49 ± 3.10 2004 Asiatic Herpetological Research Vol. 10, p. 221 12 Asaccus elisae Hemidactylus Cyrtopodion C. heterocercus turcicus scaber mardinensis Dl Erythrocyte @ Nucleus Examined species Figure 6. Erythrocyte and nucleus lengths of examined specimens. Cannon et al. (1996) reported the leukocyte mor- phology and size of the roughtail gecko Cyrtopodion scabrnm. However, they did not report any information on the erythrocyte of this species. In reptiles, the numbers of erythrocytes are smaller than in mammals or birds. Lizards have more erythro- cytes than snakes, and turtles have the fewest. Since lizards have the smallest erythrocytes of all reptiles, and turtles the largest, there may be an inverse correlation between the number of erythrocytes and their size; this hypothesis was advanced by Ryerson (1949) (Duguy, 1970). In this study, the longest erythrocytes were found in H. turcicus, the shortest in A. elisae, the largest in C. scaber and the narrowest in C. heterocercus mardinen- sis. The longest nuclei were found in C. scaber, the shortest A. elisae , the largest in C. heterocercus mardi- nensis and the narrowest in H. turcicus (Tables 2-5; Figs. 6,7). In the present study, erythrocyte morphology and the results of erythrocytes and nuclei sizes (Tables 2-5; Figs. 6,7) are agreement with the other results carried out by (Saint Girons, 1970). Acknowledgments The authors would like to thank to MSc. student Abdulmiittalip Akkaya for helping during studies. Literature Cited Alleman, A. R., E. R. Jacopson, and E. R. Raskin. 1992. Morphologic, cytochemical staining and ultrastruc- tural characteristics of blood cells from eastern dia- mondback rattlesnake ( Crotalus adamanteus ). American Journal of Veterinary Research 60:507- 514. Asaccus elisae Hemidactylus Cyrtopodion C. heterocercus turcicus scaber mardinensis ■ Erythrocyte S Nucleus Examined species Figure 7. Erythrocyte end nucleus widths of examined species. Vol. 10, p. 222 Asiatic Herpetological Research 2004 Altland, P. D. and E. C. Thompson. 1958. Some factors affecting blood formation in turtles. Proceedings of the Society of Experimental Biology and Medicine 99:456-459. Atatiir, M. K., H. Arkan, E. Qevik, and A. Mermer. 2001 . Erythrocyte measurements of some scincids from Turkey. Turkish Journal of Zoology 25:149-152. Baker, E. G. S. and L. E. Kline. 1932. Comparative ery- throcyte count of representative vertebrates. Proceedings of the Indian Academy of Science 41:417-418. Canfield, P. J. and G. M. Shea. 1988. Morphological observations on the erythrocytes, leukocytes and thrombocytes of blue tongue lizards (Lacertilia: Scincidae, Tiliqua ). Anatomia, Histologia, Embryologia 17:328-342. Cannon, M. S., D. A. Freed, and P. S. Freed. 1996. The leukocytes of the roughtail gecko Cyrtopodion scabrum : a bright-field and phase-contrast study. Anat. Histol. Embryol. 25:11-14. Charipper, H. A., and D. Davis. 1932. Studies on the arneth count. A study of the blood cells of Pseudemys elegans with special reference to the polymorphonuclear leukocytes. Quarterly Journal of Experimental Physiology 21:371-382. Cline, M. J. and T. A. Waldmann. 1962. Effect of tem- perature on red cells in the alligator. Proceedings fo the Society of Experimental Biology and Medicine 111:716-718. Duguy, R. 1970. Numbers of blood cells and their vari- ation. Pp. 93-104 In Gans (ed.), Biology of the Reptilia, Vol. 3, Morphology C. Academic Press, New York Engbretson, G. A. and V. H. Hutchinson. 1976. Erythrocyte count, hematocrit and haemoglobin content in the lizard Liolaemus multiformis. Copeia 1:186. Goin, C. J. and C. G. Jackson. 1965. Hemoglobin val- ues of some amphibians and reptiles from Florida. Herpetologica 21:145-146. Haggag, G., K. A. Raheem, and F. Khalil. 1966. Hibernation in reptiles II changes in blood glucose, haemoglobin, red blood cells count, protein and nonprotein nitrogen. Comparative Biochemistry and Physiology 17:335-339. Hartman, F. A. and M. A. Lessler. 1964. Erythrocyte measurements in fishes, amphibians and reptiles. Biological Bulletin 126:83-88. Heady, J. M. and T. E. Rogers. 1963. Turtle blood cell morphology. Proceedings of the Iowa Academy of Sciences 69:587-590. Hutchinson, V. H. and H. Szarski. 1965. Number of ery- throcytes in some amphibians and reptiles. Copeia 3:373-375. Hutton, K. E. 1960. Seasonal physiological changes in the red-eared turtle Pseudemys script a elegans. Copeia 4:360-362. Hutton, K. E. 1961. Blood volume, corpuscular con- stants and shell weight in turtles. American Journal of Physiology 200:1004-1006. Mateo, M. R., E. D. Roberts, and F. M. Enright. 1984. Morphologic, cytochemical and functional studies of peripheral blood cells from young healthy American alligators {Alligator mississippiensis). American Journal of Veterinary Research 45:1046^ 1053. Newlin, M. E. and R. E. Ballinger. 1976. Blood haemo- globin concentration in four species of lizards. Copeia 2:392-394. Saint Girons, M. C. 1970. Morphology of the circulating blood cells. Pp. 73-91 In Gans (ed.), Biology of the Reptilia, Vol. 3, Morphology C. Academic Press, New York Sevin9, M. and i. H. Ugurta§. 2001. The morphology and size of blood cells of Lacerta rudis bithynica (Squamata, Reptilia) Turkey. Asiatic Herpetological Research 9:122-129. Sevin?, M., i. H. Ugurta§, and H. S. Yidirimhan. 2000. Erythrocyte measurements in Lacerta rudis (Reptilia, Lacertidae). Turkish Journal of Zoology 24:207-209. 2004 Asiatic Herpetological Research Vol. 10, p. 223 Szarski, H. and G. Czopek. 1966. Erythrocyte diameter in some amphibians and reptiles. Bulletin de PAcademie Polonaise des Science. Classe 2. Serie des Sciences Biologiques 14(6):437-443. Taylor, K. and H. M. Kaplan. 1961. Light microscopy of the blood cells of pseudemyd turtles. Herpetologica 17:186-196. Ugurta§, I. H, M. Sevin9, and H. S. Yildirimhan. 2003. Erythrocyte size and morphology of some tortoises and turtles from Turkey. Zoological Studies 42(1): 173-178. 2004 Asiatic Herpetological Research Vol. 10, pp. 224-229 Distribution and Conservation Status of Neurergus microspilotus (Caudata: Salamandridae) in Western Iran Mozafar Sharifi* and Somayeh Assadian Department of Biology, Faculty of Science, Razi University, Kermanshah, Iran. E-mail: sharifimozafar@hotmail. com Abstract. - Field and laboratory observations of the Yellow Spotted Newt, Neurergus microspilotus (Nestrov, 1917), in western Iran have yielded preliminary data on conservation biology and distribution of this species. New distribu- tional ranges have been determined for Neurergus microspilotus in its Iranian range in the mid-Zagros Mountains. In streams occupied by Neurergus microspilotus, dissolved oxygen, temperature, discharge, N03, and P04 were meas- ured. Land use practices, adjacent riparian habitats and channel substrate were also determined. Four new stream habitats were identified. On the basis of interviews with local inhabitants, three other streams were identified as like- ly habitat for Neurergus microspilotus. Measurements of relative abundance of N. microsphilotus indicate that this animal is likely to occur in higher numbers in cold and first order streams located at high altitudes in the western edge of the Iranian plateau on the mid-Zagros Range. The limiting factor for the yellow spotted newts in western Iran appears to be human disturbance. In the last four years, one of the five known streams with N. microspilotus, in the area of Ghorighala, has virtually lost its entire population due to pollution by a tourist facility and local sewage efflu- ence. Key words. - Neurergus microspilotus , salamander, first order stream, distribution, conservation. Introduction Available information on the conservation biology of the western Iranian salamanders is scarce. Investigations made in the 1970s (Schmidtler and Schmidtler, 1975) indicated that three of four species of salamanders belonging to the genus Neurergus {N. crocatus, N. microspilotus and N. kaiseri ) occur in Iran. There is no recent information on distribution and abundance of these species for assessment of conservation. However, a world-wide concern over declines in amphibian popu- lations (Wake, 1991; Gardner, 2001) is equally pertinent in remote areas of western Iran. Amphibians are sensi- tive to land-use alteration (Wilkins and Peterson, 2000) and there is widespread concern that environmental pol- lution and land deterioration are responsible for their decline (Richardson, et al 2000). Several factors are known to have contributed to the declines, including habitat destruction (Sala et al, 2000), fragmentation of habitat (Sjogren, 1991, Marsh and Trenham, 2000), and alteration of species composition of communities through the introduction of exotic predators and pathogens (Beebee, 1977). In addition, acidification and other chemical pollution, alteration of climate (Pounds and Crump, 1994), disease and road kill (Carey, 1993, 2000) are candidates for the amphibian decline. Relatively few caudate species occur in Iran. These include seven species of the genera Triturus, Batrachuperus, Neurergus, and Salamandra (Balutch and Kami, 1995). Newts of the genus Neurergus have a relatively wide geographic distribution, ranging from western Iran (Zagros Mountains) and extending into Iraq and southern Turkey (Balutch and Kami, 1995). There is no sufficient information regarding the geographic dis- tribution of the three species of newts that occur in west- ern Iran. Previous investigations indicate that the pri- mary distribution range of Neurergus microspilotus is in the mid-Zagros range at the border of Iran and Iraq (Nesterov, 1917; Schmidtler and Schmidtler, 1975). This information also indicates that Neurergus kaiseri and Neurergus crocatus are expected to occur in southern and northern parts of the Zagros Range, respectively. Recent investigations on N. microspilotus confirms that this newt occurs in highland streams in the mid-Zagros region (Assadian and Sharifi, 2002; Rastegar Pouyani and Assadian, 2002). The aims of the present study are to determine the geographic distribution and conservation biology of Neurergus microspilotus. To improve conservation efforts related to this species, information is needed on physico-chemical characters of the habitat. For this rea- son, we measured some variables in the aquatic environ- ment and adjacent terrestrial habitats. © 2004 by Asiatic Herpetological Research Vol. 10, p. 225 Asiatic Herpetological Research 2004 Figure 1. Geographic distribution of Neurergus microspilotus in streams of the mid-Zagros Range in western Iran. Those streams with the species have been shown by (1). Those streams that expected to have the animal are shown by (2). Study Areas The Iranian basin is a large triangular depression flanked by Elbourz Mountains in the north and the Zagros Mountains in the west. The Zagros Mountains extend diagonally from eastern Turkey to the north of the Persian Gulf and Pakistan border. This range is part of a greater geographic unit arising from the east of the Anatolian Plateau of Turkey and expanding southward to include Iran, Afghanistan, Pakistan and further east to the western edge of the Tibetan Plateau. The Zagros Mountains act as barriers to the incoming air parcels from the west and receive precipitation according to their height and longitude. In general, the northern and western portions of the range receive considerably more rainfall than those in the south and east. The average annual precipitation in the northern Zagros ranges from 400 to 800 mm. per year. Most of the central and south- ern Zagros receive between 300 and 500 mm (Ghobadian, 1990). The western Zagros Range meets the northern Mesopotamian Plain, a low land with a hot and dry cli- mate. In some parts of the Zagros Range, where this meeting takes place over a relatively short distance a steep environmental gradient is encountered where high altitude and cold climate from the Iranian Plateau dif- fuse into the low altitude and warm Mesopotamian Plain in just few kilometers. The weather condition in the western edge of the Iranian Plateau in the mid-Zagros Range is characterized by a pronounced seasonal varia- tion including a long freezing period in winter and a mild summer. Although the average annual precipitation in this area is around 500 mm, most of this comes as snow. As a result, many seasonal and permanent streams at the western side of the Zagros Range are nourished by heavy snow accumulated on the high mountains. In the lowlands of the northern Mesopotamian Plain, which in some parts lie only 20 or 30 km from cold uplands, sum- mers are hot and dry and winters are free of frost. Precipitation in this area approaches 400 mm per annum, rarely appearing as snow. Information obtained from Ravansar Synoptic Station (20 km from Kavat Stream in the highlands) and Sarepolezahab on the northern Mesopotamian Plain (40 km away from Kavat Stream) summarizes the annual climatological data for these two contrasting environments. Materials and Methods Streams, ponds, and springs were searched for adults and larvae of Neurergus microspilotus in the mid-Zagros Range in Kermanshah and Kurdistan provinces in west- ern Iran in the spring and summer 2001 and 2002. In streams where the salamander was found, channel sub- strate, channel width, adjacent riparian plant community type, and land use practice were determined. Where pos- sible, relative abundance of N. microspilotus was deter- 2004 Asiatic Herpetological Research Vol. 10, p. 226 Table 1. Altitude at head stream, approximate length of the streams and amount of discharge (I/s) in streams where Neurergus microspilotus was sighted. Stream Altitude (m) Approximate Length (km) Position (l/s) Discharge Kavat 1500 4 34° 53’ N, 46° 3T E 625.7* Dorisan 1600 3 35° 21’ N, 46° 24’ E 35** Dareh Najar 1400 2 35° 06’ N, 46° 19’ E - Ghorighaleh 1600 0.1 34° 54’ N, 46° 30’ E 333.7* Paveh rood 1100 2.3 35° 06' N, 46° 17' E - Darian 1000 2 35° 08' N, 46° 19' E - Shamshir 1800 1.5 34° 59' N, 46° 25' E - Marakhil 1600 2.5 35° 02' N, 46° 11' E “ * Based on measurement made by department of water resource. ** - Discharge measured in field by determining the velocity of water and the extent of cross-section. mined and expressed as individuals per every ten paces. In small streams where there was no routine hydrologi- cal measurement of water discharge, the water discharge was estimated by measuring velocity of water and extent of cross-section width of the channel. In Kavat Stream, where highest relative abundance of N. microspilotus was found, visual estimates were made of percent chan- nel substrate composition (Wilkins and Peterson, 2000) by bedrock, boulder (>256mm diameter), cobble (64- 256 mm diameter), gravel (16-64 mm diameter), pebble (2-16 mm diameter), fine sediment and coarse woody debris. In these streams several water characteristics were measured. These include dissolved oxygen (Winkler method), temperature (glass thermometer), electrical conductivity (conductivity meter), N03 and P04 (spectrophotometer). Results The geographic distribution of Neurergus microspilotus in its Iranian range is shown in Figured. These are Kavat Stream (34° 53' N, 46° 31' E), Dorisan Stream (35° 21 ' N, 46° 24' E), Ghorighaleh Stream (34° 54' N, 46° 30' E), Najar Stream (35° 06' N, 46° 19' E), and Paveh Rood Stream (35° 06' N, 46° 17' E). Apart from streams in which the newt has already been observed, there are three other streams where, on the basis of inter- views with local inhabitants, the presence of this animal is likely. These streams are upstream of Marakhil River (35° 02’ N, 46° 1 V E), Shamshir Stream (34° 59’ N, 46° 25’ E) and Hajij Stream (35° 08’ N, 46° 19’ E). Altitude, approximate length of the streams in which N. microspi- lotus is expected to occur, geographic position, and water discharge are shown in Table 1 . Physico-chemical characteristics in streams with N. microspilotus are shown in Table 1. Water analysis has been carried out in upper and lower reaches of Ghorighala Stream in order to demonstrate the human impact on the water quality. Occurrence of the yellow spotted newt in different aquatic microhabitats has been evaluated using the Wilkins and Peterson (2000) classification of channel substrate including bedrock, boulder, cobble, gravel, pebble and fine sand sediment. The yellow spotted newt occupies an assortment of aquatic microhabitats during the breeding season. Visual determination of substrate texture in Kavat Stream indicated that this newt tended to occupy substrates that are gravel or pebble (60%). Figure 3 demonstrate the frequency distribution of sub- strate classes used by this newt. Table 2. Physico-chemical characteristics of water where Neurergus microspilotus was found. Streams DO no3-n (mg/I) po4-p (mg/I) EC (micm /cm) Temperature (°C) Head stream in Ghorighala 8.15 0.38 N.D 323 11 Lower reach of Ghorigala 6.65 0.35 N.D 356 11 Dareh nagar 7.8 1.14 N.D 549 15 Kavat - - - - 11.5 Dorissan - - - - 11 Vol. 10, p. 227 Asiatic Herpetological Research 2004 50 35 25 15 5 -5 Sarpolzahab Climograph Figure 2. Climographs representing the pattern of precip- itation and temperature in Ravansar at the western edge of the Iranian Plateau and Sarepolezahab in the northern Mesopotamian Plain. Data are 20 years mean monthly temperature and rainfall collected at the synoptic stations in these two cities. Discussion The presence of Neurergus microspilotus in Ghorighaleh Stream has also been reported in previous studies (Nesterov, 1917; Schmidtler and Schmidtler, 1975). Assadian and Sharifi (2002) and Rastegar Pouyani and Assadian (2002) have reported Neurergus microspilo- tus in this stream. Papenfuss and Sharifi also collected several salamanders from this stream in Spring 2000. No information is available regarding the occurrence of Neurergus microspilotus in other streams, therefore, the other four streams are new records for N. microspilotus in its Iranian range. All Neurergus streams reported in this study with originated from the western edge of the Iranian Plateau (Figure 2) and join to the Dez-Karkheh watershed sys- tem in the northern Mesopotamian Plain and finally enter into the Persian Gulf. All these stream are first order streams located at relatively high altitude (1100- 1600 m) and join to the main rivers in the lowland (300- 600 m) of the catchments (Table 1). Neurergus microspilotus is a medium size salaman- 40 Bedrock Boulder Cobble Gravel Pebble Sand Figure 3. Percent occurrence of Neurergus microspilo- tus in various substrate size group in Kavat Stream (n=42). der with a slender body. Adults reach a length (snout to vent) of 60-70 mm (mean=65.6, sd=4.63, n=20). Adults are black dorsally and laterally, with greenish yellow blotches. The spots are distributed on the salamander’s body without an obvious pattern. Neurergus microspilo- tus characteristically possess broad heads with blunt, rounded snout. N. microspilotus is also structured for swimming using their laterally compressed tails for propulsion and steering during swimming. The prevailing climatic conditions are distinctly different between and within streams. Since these streams are located at the western edge of the Iranian Plateau, the climatic conditions may vary considerably at the upper reaches compared with lower reaches of the same streams. At the same time it appears that streams closer to the Mesopotamian Plain are experiencing cli- matic conditions that are different with those that are located at the western edge of the Iranian Plateau. Because of the steep environmental gradient the streams occupied by Neurergus microspilotus can be convenient- ly divided into two groups. Those located in the high altitude and cold weather regions on the western Iranian Plateau (Kavat, Ghorighaleh, Dorisan and Shamshir streams) and those in the north and north eastern part of the range which because of lower elevation experience warmer climate (Marakhil River, Dareh Najar and Darian streams). In Dareh Najar Stream where very few Neurergus microspilotus were located, and also in Marakhil and Darian streams where the animal is report- edly seen, it is possible that the animal drifted by the action of water currents. It is also possible that the lower relative abundance in N. microspilotus in Dareh Najar and possibly in the other two streams is due to the lower altitude and the vicinity to the northern Mesopotamian Plain. Terrestrial habitats occupied by N. microspilotus include diverse community types including oak-pista- chio open woodlands dominated by Quersus branti and Pistachio spp. This woodland grows on various soil 2004 Asiatic Herpetological Research Vol. 10, p. 228 types, including deep sandy loam soils at the bottom of valleys or gravelly soils on the slopes of steep valleys. In warmer parts of its range, riparian vegetation may also contain willow ( Salix spp.) or shrubs such as Cerrasns and Amygdale (Amygdalus spp.). In colder parts of the range the riparian vegetation may be charac- terized by more hydrophobic plants such as sedges 0 Carex spp.) and sphagnum moss (, Sphagnum spp.). Neurergus microspilotus moves from its wintering site to the breeding streams as soon as the spring melt occurs, from late January through early March. Within its range, in high altitude-cold weather regions, egg-lay- ing was observed in early May. However, it appears that the reproductive pattern of N. microspilotus is not tightly synchronized because unhatched eggs have been observed as late as mid-June. No breeding activity, eggs, or juveniles of N. microspilotus have been observed in the low altitude warm climate part of the range. Eggs of N. microspilotus are laid singly or in small clumps on vegetation or on rocks. The number of oocytes in a female dissected in laboratory was 108. Laboratory observations of larval growth and development indicate that larvae complete the metamorphosis in the first year. In early autumn they still possess their gills. Larvae with large heads, well developed dorsal fins, and bushy gills have the ability to react suddenly with a whole body reaction to external stimulus. Although no information is available regarding wintering activity of Neurergus microspilotus in its Iranian range, the appearance of the animal in early spring and disappearance in summer implies that this newt requires both upland and wetland habitat that con- tain suitable aquatic environment during the breeding season and subterranean burrows appropriate for winter- ing. These normally include an aquatic environment for breeding and a terrestrial habitat where juveniles and adults spend most of their time. Habitat loss through divergence of streams for irri- gation of cultivated lands is probably the single most important factor that threatens Neurergus microspilotus in its Iranian range. Traditionally, due to the lack of land in steep valleys in the mid-Zagros Range, extensive attempts have been made to construct a complex of rein- forced terraces of land, which is cultivated for walnut and other orchard trees. Water is diverted from its natu- ral channel to irrigate these lands. Although no harm is directed toward N. microspilotus in these orchards, the impact of land use alteration especially in dry periods causes many of these creatures to be deprived of a healthy aquatic environment. Although human settlement in the mid-Zagros area is characteristically less developed compared with other localities in western Iran, Neurergus microspilotus is experiencing an environmental impact similar to that found in more urbanized areas in the country. For exam- ple, Ghorighaleh Stream originates from a cave that has been developed by a reclamation project for visitors. Since the construction of this unit the stream is suffering from gross pollution caused by thousands of visitors. Changes in water characteristics in the upper and lower reaches of the stream are shown in Table 2. Although in 2000 and 2001 numerous Yellow Spotted Newts were reported (Assadian and Sharifi, 2002; Rastegar Pouyani and Assadian, 2002) no newts were seen in 2002. The absence of Neurergus microspilotus is presumably due to the human impacts resulting from an ecotourism cen- ter developed in 1999 at the Ghorighaleh Cave where the source of the stream is located. Massive solid waste disposed by thousands of visitors together with raw sewage released to the stream by residents of Ghorighaleh Village can be observed in the upper reach- es of this stream although only changes in dissolved oxygen are evident in physico-chemical characteristics measured in this study. Conclusions Although Neurergus microspilotus has been virtually extirpated from one of five known breeding streams in its Iranian range, it does not appear to be in immediate danger of extinction because one is likely to find this newt occur in other streams in the area. However, the sit- uation for N. microspilotus is not promising as the major threatening factors such as habitat destruction and water pollution are operating. Robust populations occur in at least in one of its habitats (Kavat Stream). However, the lack of information essential to estimate population size and population trends makes it difficult to assess conser- vation status of this salamander. Future work should examine the long-term effects of anthropomorphic impacts associated with land use alteration and pollu- tion. Acknowledgments The authors wish to express their gratitude to Professor D. B. Wake for his great help in reviewing the manu- script. This study was supported by Razi University grant. Literature Cited Assadian, S. and Z. Sharifi. 2002. Distribution and con- servation status of Neurergus microspilotus in west em Iran. 1st Iranian conference on Animal Science and Biodiversity. Vol. 10, p. 229 Asiatic Herpetological Research 2004 Beebee, T. J. C. 1977. Enviromental change as a cause ot Natterjack Toad (Bufo calamita) declines in Britain. Biological Conservation 11:87-102. Baloutch, M. and H. G. Kami. 1995. Amphibians of Iran. Tehran University Publications. [In Farsi] Carey, C. 1993. Hypotheses concerning the causes of the disappearance of boreal toads from the mountains of Colorado. Conservation Biology 7:355-362. Carey, C. 2000. Infectious disease and worldwide declines of amphibian populations, with comments on emerging diseases in coral reef organisms and in humans. Enviromental Health Perspectives, 1 08(Suppl): 143-150. Gardner, T. 2001. Declining amphibian populations: a global phenomenon in conservation biology. Animal biodiversity and conservation 24.2:25-44. Ghobadian, A. 1990. Natural features of the Iranian Plateau. Kerman University Publication Centre. Kerman, Iran. Marsh, D. M. and P. C. Trenham. 2000. Metapopulation dynamics and amphibian conservation. Conservation Biology 15:40-49. Nesterov, P. V. 1917. Tri novych chvostatych amfibii is kurdistana. Annuaire du Musee Zoologique de L’Academie des Sciences, Petrograd 21:1-30. Pounds, J. A. and M. L. Crump. 1994. Amphibian declines and climate disturbance: the case the Golden Toad and the Harlequin Frog. Conservation Biology 8:72-85. Rastegar Pouyani, N. and S. Assadian. 2002. Sexual dimorphism in Neurergus microspilotus (Caudata:Salamandridae). 1st Iranian conference of Animal Science and Biodiversity. Sala, O. E., F. S. I. Chapin, J. J. Armesto, E. Berlow, J. Bloomfield, R. Dirzo, E. Huber-Sanwald, L. F. Huenneke, R. B. Jackson, A. Kinzig, R. Leemans, M. Lodge, H. A. Mooney, M. Oesterheld, N. L. Pofif, M. T. Sykes, B. H. Walker, M. Walker, and D. H. Wall. 2000. Global biodiversity scenarios for the year 2100. Science 287: 1770-1774. Sjogren, P. 1991. Extinction and isolation gradients in metapopulations: the case of the pool frog {Rana lessonae). Biological Journal of the Linnean Society 42:135-147. Schmidtler, J. J. and J. F. Schmidtler. 1975. Untersuchujngen an westpersischen Bergbach- molchen der Gattung Neurergus Caudata, Salamandridae). Salamandra 11:84-98. Wake, D. B. 1991. Declining amphibian populations. Science 253:860 2004 Asiatic Herpetological Research Vol.IO, pp. 230-234 An Investigation on the Blood Cells of the Leopard Gecko, Eublepharis angramainyu (Reptilia: Sauria: Eublepharidae) Murat Tosunoglu1, Dinner Ayaz2, Cemal Varol Tok1, and Ba^aran Dulger1 1 Qanakkale Onsekiz Mart University, Faculty of Science-Literature, Department of Biology, Terzioglu Campus, Qanakkale, Turkey 2Ege University, Faculty of Science, Department of Biology, 35100 Bornova, Izmir , Turkey Abstract. - In this study, blood cell counts and sizes in three adult Eublepharis angramainyu specimens (one male, two female) collected from SE Anatolia (Sanlurfa - Birecik). The number of erythrocytes in 1 mm3 ranged between 870,000 and 950,000 (average 910,000). The mean total length of erythrocytes was calculated as 20.35 pm, the width as 10.59, the size as 169.68 pm2; the mean nucleus length as 7.50 pm, the width as 4.15 and the size as 24.47 pm2. Small lymphocytes had a mean diameter of 9.78 pm, big lymphocytes 13.71 pm, monocytes 15.37 pm, neutrophiles 16.56 pm, eosinophiles 17.59 pm, and basophiles 12.87 pm. The mean length of thrombocytes was measured at 8.82 pm, and the width at 5.93 pm. Key words. - Eublepharis angramainyu , Sauria, blood cell count, blood smears, erythrocytes, leucocytes, thromb- ocytes. Introduction Eublepharis angramainyu Anderson and Leviton, 1966, also known as the “leopard gecko”, was first found between Masjid Soleyman and Batsvand in the Khuzes- dan province of Iran. The species was reported to range in the western foothills of the Zagros Mountains and Mesopotamia in Iraq and Iran, and NE Syria with a ver- tical distribution of 300 to 1000 meters (Anderson and Leviton, 1966; Leviton et al., 1992; Disi and Bohme, 1996; Anderson, 1999). Studies conducted in recent years (Go9men et al., 2002) established that the species also inhabited SE Anatolia (Sanliurfa-Birecik). Although there are a number of studies on the distribu- tion, morphology and ecology of the species (Anderson and Leviton, 1966; Leviton et al., 1992; Disi and Bohme, 1996; Anderson, 1999; Go9men et al., 2002), a literature review has not revealed any detailed haemato- logical studies. Most studies on the haematology of different spe- cies are related to blood cell counts (Alder and Huber, 1923; Hutchison and Szarski, 1965; Duguy, 1970; Arikan, 1989) and blood cell sizes (Szarski and Czopek, 1966; Hartman and Lessler, 1964; Atatiir et al., 1998, 1999). The number haematological studies related to amphibian and reptile species living in Anatolia has been increasing in recent years (Arkan, 1989; Atatiir et al., 1998, 1999, 2001; Sevin9 et al., 2000). In this study, the number and sizes of blood cells of Eublepharis angramainyu were determined and photographs of their blood cells presented. Materials and Methods Three adult specimens (one male, two female) examined in this study were collected near £i9ekalan Village between 2200-2400 hours at an altitude of 400 m during the species breeding season (01 July, 2002). Blood sam- ples were taken within the first three days after the spec- imens were collected live in the wild and brought to the laboratory. Blood cell counts were carried out by means of Neubauer hemocytometer. Hayem solution was used to dilute the erythrocytes. Wright- Stained blood smears were made use of in the measurement (erythrocytes, leu- cocytes and thrombocytes) and computation of blood cells. The necessary blood samples were obtained by cardiac (ventriculus) puncture, via heparinized hemat- ocrit capillaries. Blood cell measurements were taken by means of a MOB-l-15x ocular micrometer. On each blood smear, measurements related to 40 randomly cho- sen erythrocytes (total erythrocyte length, total erythro- cyte width, nucleus length and nucleus width) were made, and the nucleus size was calculated) according to the formula EL.EW./4 and the nucleus size according to the formula NL.NW./4 (Duguy, 1970; Atatiir et&al., 2001). Moreover, micrometric measurements were made on leucoytes and thrombocytes. Photographs of © 2004 by Asiatic Herpetological Research Vol.10, p. 231 Asiatic Herpetological Research 2004 A r ' » » 9 * 9 ' * 9 * • 0 B + * % * % ^ 1 » # w m ^ • ^ f | % 9 - * , . % % t % D * # 1 # r i * . 1 i d E „ ’ * * % *|L m % § % fc $ ~"4 # % * . # 0 * gT ** G » H % ^ 0 0 * ~ ' % $ t % , ' - #0~ * 0 % # # W 0 9 4^ ft * Figure 1 : The blood cells of Eublepharis angramainyu. A- erythrocytes, B- small lymphocyte, C- large lymphocyte, D- monocyte, E- neutrophil, F- eosinophil, G- basophil, H- a cluster of thrombocytes. blood cells were taken using a Carl Zeiss Jena micro- scope at 40x magnification. Results Male and female specimens were assessed together as there were no significant differences between them with respect to the number and size of blood cells. As in other lizrds, the erythrocytes belonging to Eublepharis angra- mainyu are also ellipsoidal cells with nuclei. The nuclei are also ellipsoidal, somewhat regular and centrally located (Figure 1A). The mean total length of the eryth- rocytes (L) was calculated as 20.35 pm the width (W) as 10.59 pm, the size (S) as 169.68 pm2; the mean nucleus length (NL) as 7.50 pm, the width (NW) as 4.15 pm, and the size (NL) as 24.47 pm2. The number of erythro- cytes in 1 mm3 of blood ranged between 870,000 and 950,000 (average 910,000) (Table 1). Lymphocytes have a spherical shape. Both small and large lymphocytes were examined in the blood smears prepared (Figure IB, C). Large lymphocytes were 13.71 pm in diameter and had a large cytoplasmic zone and a centrally-located, large, round nucleus. The cytoplasm is stained pale blue and the nucleus purplish blue using the Wright Stain. No granule formation was observed in the cytoplasm. Small lymphocytes had a mean diameter of 9.78 pm (Table 1). The large nucleus covers the majority of the cell’s area; the cytoplasm is in the shape of a thin ring. Lymphocytes are the most com- monly seen leucocytes in the preparates. Although resembling the large lymphocytes in size, monocytes are easily distinguished from the shape of the nucleus (Figure ID). They have a mean diameter of 15.37 pm Granule formation was observed in the cyto- plasm (Table 1). The nucleus is not oval, but depressed on one side and occupies at least half of the cell. The cytoplasm is stained light purple and the nucleus dark blue. They are the second most common leucocytes after lymphocytes and neutrophiles. Neutrophiles are spherical cells with a mean diame- ter of 16.56 pm (Table 1). Using the Wright Stain, the cytoplasm is stained light blue and the nucleus dark blue. There are very fine granules in the cytoplasm (Fig- ure IE). The nucleus is a structure with lobes and seg- ments. They are the most common leucocytes second to lymphocytes. Eosinophiles have a diameter of 1 7.59 pm (Table 1 ). The cytoplasm is stained light blue and the nucleus dark blue. Large, round, bright red granules within the cyto- plasm strike eye as the most distinctive characteristic of these cells (Figure IF). The nucleus was seen to have two lobes. These cells take the fourth place in the prepa- ration of smears after lymphocytes, monocytes and neu- trophiles. Basophiles are oval-shaped with a mean diameter of 12.87 pm (Figure 1G and Table 1). When stained by means of the Wright Stain, dark bluish purple granules within the light blue cytoplasm are in a position marking the dark blue nucleus. These cells are rarely seen in the preparates. Thrombocytes are spindle-shaped cells with a mean length of 8.82 pm, a width of 5.93 pm (Figure 1H and Table 1). In the Wright-Stained preparates, dark stained cells with large oval nuclei and small irregular cytopla- sic zones form groups of two or more. 2004 0, p. 232 Table 1: The established counts, measurements and sizes concerning the blood cells of Eublepharis angr amainyu (in m and m2). N: Number of specimens; n: Number of measurements/computings in each speci men; Ext: extreme values; SD and SE: standard deviations and the standard errors of the means, respectively. Blod Cells N n Ext Mean SD SE Number of Erythrocytes 2 3 870,000-950,000 910,000 - Total Erythrocyte Length 2 40 15.00-22.50 20.35 2.15 0.24 Total Erythrocyte Width 2 40 7.50-12.50 10.59 1.00 0.11 Total Erythrocyte Size 2 40 91.26-220.78 169.68 26.87 3.00 Nucleus Length 2 40 6.25-8.75 7.50 0.41 0.05 Nucleus Width 2 40 3.25-5.25 4.15 0.52 0.06 Nucleus Size 2 40 17.86-31.40 24.47 3.44 0.38 Lymphocyte (big) Diameter 2 20 12.50-16.25 13.71 1.21 0.19 Lymphocyte (small) Diameter 2 20 7.50-11.25 9.78 1.11 0.17 Monocyte Diameter 2 40 11.25-20.00 15.37 2.00 0.31 Neutrophile Diameter 2 40 15.00-20.00 16.56 1.61 0.36 Eosinophile Diameter 2 20 15.00-20.00 17.59 1.56 0.24 Basophile Diameter 2 5 10.00-15.20 12.87 1.81 0.40 Thrombocyte Length 2 40 7.25-11.25 8.82 1.16 0.18 Thrombocyte Width 2 40 5.00-8.25 5.93 0.77 0.12 Table 2: The number of erythrocytes in 1 mm3 of blood in different lizard species. Researchers Species Number of Erythrocytes Present study Eublepharis angramainyu 870,000-950,000 Duguy (1970) Hemidactylus turcicus 866,000 Chalcides ocellatus 806,000 Agama atra 1,250,000 Lacerta agilis 945,000-1.420.000 Lacerta viridis 840,000-1,600,000 Anguis fragilis 466,000-1,615,000 Vol.10, p. 233 Asiatic Herpetological Research 2004 Table 3: Sizes of erythrocytes and nuclei in different lizard species according to various researchers (EL/ EW: Erythrocyte Length/Erythrocyte Width Ratio, ES: Erythrocyte Size (pm2), NL/NW: Nucleus Length/ Nucleus Width Ratio, NS: Nucleus Size (pin2), N/C: Nuclear surface/Cell surface ratio). Researchers Species EL/EW ES NL/NW NS N/C Present study Eublepharis angramainyu 1.92 169.68 1.80 24.47 0.14 Atatiir, et al. (2001) Ablepharus chernovi 1.87 84.12 2.45 12.01 0.14 Chalcides ocellatus 1.86 91.33 1.98 10.70 0.12 Mabuya aurata 1.90 84.88 2.01 10.02 0.12 Ophiomorus punctatissimus 1.96 92.08 2.30 12.70 0.14 Eumeces schneideri 1.97 92.31 2.81 14.20 0.15 Seving, et al. (2000) Lacerta rudis 1.63 87.46 1.64 16.66 0.19 Lacerta viridis 1.86 125.00 1.94 14.6 0.11 Duguy (1970) Eumeces algeriensis 1.61 154.80 2.10 26.4 0.17 Anguis frag il is 1.88 143.90 1.62 22.5 0.14 Discussion Literature Cited As stated in the ‘Results’ section, E. angramainyu spec- imens were collected during the mating season and did not display sexual dimorphism with respect to the num- ber of erythrocytes and sizes. It has been found that there are significant differences among lizard families with respect to the number and size of erythrocytes, and the members of Gekkonidae have the highest number of erythrocytes among lizards (Alder and Huber, 1923; Hutchison and Szarski, 1965; Duguy, 1970). Values belonging to the number of blood cells determined in diffeent lizard species (Table 2), by various researchers were compared with those we obtained for E. angra- mainyu in the present study, and it was established that the number of erythrocytes in 1 mm3 of blood was very close to that of Hemidactylus turcicus (Gekkonidae) species, but different from that of other lizard species. Values we obtained in E. angramainyu with respect to the sizes of erythrocytes and nuclei were compared with those determined for some lizard species (Table 3), and it was found that values concerning the sizes of erythrocytes and nuclei were very close to those of spe- cies belonging to Gekkonidae family. When compared with the other lizard species, E. angramainyu can be said to have the largest erythrocytes with respect to the sizes of erythrocytes and nuclei. Alder, A. and E. Huber. 1923. Untersuchungen iiber Blutzellen und Zellbildung bei Amphibien und Reptilien. Folia Haematologica 29: 1-22. Anderson, S. C. and A. E. Leviton. 1966. A new species of Eublepharis from Southwestern Iran (Reptilia: Gekkonidae). Occasional Papers of the California Academy of Science 53:1-5. Anderson, S. C. 1999. The Lizards of Iran (Contribu- tions to Herpetology Vol. 15). Society for the Study of Amphibians and Reptiles, Missouri, USA. 422 pp. Arikan, H. 1989. Anadolu'daki Rana ridibunda (Anura: Ranidae) populasyonlarinin kan hticrelerinin sayisi bakirmndan incelenmesi. Turkish Zoology 13:54- 59. Atatiir, M. K., H. Arikan, and A. Mermer. 1998. Eryth- rocyte sizes of some Urodeles from Turkey. Turkish Journal of Zoology 22:89-91. Atatiir, M. K., H. Arikan, andi. E. Qevik. 1999. Erythro- cyte sizes of some Anurans from Turkey. Turkish Journal of Zoology 23:111-114. Atatiir, M. K., H. Arkan, i. E. £evik, and A Mermer. 2001. Erythrocyte measurements of some Scincids from Turkey. Turkish Journal of Zoology 25:149- 152. 2004 Vo) JO, p. 234 Disi, A. M., and W. Bohme. 1996. Zoogeography of the amphibians and reptiles of Syria, with additional new records. Herpetozoa 9(l/2):63-70. Duguy, R. 1970. Numbers of blood cells and their varia- tion. Pp. 93-109. In C. Gans and F. H. Pough (eds.), Biology of Reptilia, Volume 3. Academic Press, London and New York. Go9men, B., M. Tosunoglu, and D. Ayaz. 2002. First Record of the Leopard Gecko, Eublepharis angra- mainyn (Reptilia: Sauria: Eublepharidae) from Anatolia. Herpetological Journal 12(2):79-80. Hartman, F. A. and M. A. Lessler. 1964. Erythrocyte measurements in Fisches, Amphibia and Reptiles. Biological Bulletin 126:83-88. Hutchison, H. V. and H. Szarski. 1965. Number of erythrocytes in some Amphibians and Reptiles. Copeia 3:373-375. Leviton, A. E., S. C. Anderson, K. Adler, and S. A. Minton. 1992. Handbook to Middle East Amphibi- ans and Reptiles. In Contributions to Herpetology, Vol. 8., Society for the Study of Amphibians and Reptiles. Missouri, USA. Sevin9, M., i. H. Ugurta§, and H. S. Yildirimhan. 2000. Erythrocyte measurements in Lacerta rudis (Rep- tilia, Lacertidae). Turkish Journal of Zoology 24:207-209. Szarski, H. and G. Czopek. 1966. Erythrocyte diameter in some amphibians and reptiles. Bulletin of the Polish Academy of Sciences Biological Sciences 14(6):433-437. 2004 Asiatic Herpetological Research Vol. 10, pp. 235 A Record of Boiga ochracea walli (Stoliczka, 1870) from Bangladesh M. Farid Ahsan* and Shayla Parvin Department of Zoology, University of Chittagong, Chittagong 4331, Bangladesh. 5jc Corresponding author E-mail: mfahsan@ctgu.edu Abstract. - Two specimens of Boiga ochracea specimens from Bangladesh are referred to Boiga ochracea walli. The locality data for these specimens are lost, but they are probably form the University of Chittagong campus. These are the first records of this subspecies for Bengladesh. Key words. - Boiga ochracea , Bangladesh. While identifying snake species preserved in the Departmental Museum of Zoology, Chittagong University (CU), two specimens of Boiga ochracea walli (Stoliczka, 1870) were found. One was collected in 1975, but the localities where they were found are unknown. Both specimens were probably collected form the University of Chittagong campus. The occurrence of Boiga ochracea in Bangladesh was first reported by Khan (1982) based on a specimen collected from Chittagong Hill Tracts. Khan (1982) did not identify the specimen to subspecies. Smith (1943) reported the sub- species range as Burma (now Myanmar) south of Lat. 25°; Tenasserim; the Andaman and Nicobar Islands. Both localities of Boiga ochracea walli from Chittagong (this report) and B. ochracea from Chittagong Hill Tracts (Khan 1982) are close to Myanmar and south of the latitude mentioned by Smith (1943). This report extends the subspecies range up to Bangladesh and it may occur in other parts of the country like Greater Sylhet, Cox's Bazar, and the districts of Chittagong Hill Tracts (i.e., Rangamati, Khagracheri and Bandarbans) as they have similar habitats. The CU specimens have the following characters (although the natural colour may have changed due to the effects of preservation): faded greyish above, verte- bral series of scales paler than others, ventral side of body whitish. Smith (1943) described the subspecies as "greyish, reddish or yellowish brown above (coral red in life), some of the scales finely edged with black and forming more or less distinct transverse lines or bars, best marked in the young; the vertebral series of scales sometimes lighter than the others; paler below; lips and chin whitish". The CU specimens have eight supra labials, 4th, 5th and 6th below the eyes; one pre and two post-oculars present. Smith (1943) stated that there is normally one pre-ocular, not reaching the upper surface of the head; anterior genials about as long as the posterior, latter in contact with one another or separated by small scales; vertebrals strongly enlarged. The measurements of the CU specimens are compared below with those reported by Smith (1943). Literature Cited Khan, M. A. R. 1982. Wildlife of Bangladesh - a check- list. Dhaka University Press, Dhaka. 173 pp. Smith, M. A. 1943. The fauna of British India, including Ceylon and Burma. Reptilia and Amphibia, Vol. 3 Serpentes. Taylor and Francis Ltd., London. 583 pp. Table 1. - The table shows that the Smith's (1943) specimens and the present ones are similar. Cat. No. Date locality Total length (cm) Tail length (cm) Dorsal scales Ventral scales Anal scale Caudal scales Temporal scale Source 31CU 04.03.75 88 16 19:19:15 236 1 79 2+2 Present 37CU 50.3 9.3 17:19:15 229 1 97 2+2 work 105-110 23.5-21.5 19:19:15 221-246 1 89-107 2+2 Smith or (1943) 2+3 © 2004 by Asiatic Herpetological Research 2004 Asiatic Herpetological Research Vol. 10, pp. 236-240 Some Aspects of Breeding Biology of the Bengal Lizard (Varanus bengalensis) in Bangladesh M. Farid Ahsan1 and M. Abu Saeed2 ^ Department of Zoology, University of Chittagong, Chittagong 4331, Bangladesh; E-mail: mfahsan(fctgu.edu -AGROCARE, Golden Plaza (1st Floor), 58 Shaheed Taj uddin Ahmed Sharani, Mohakhali, Dhaka 1212, Bangladesh Abstract. - Some aspects of breeding biology of the Bengal, or Gray, Monitor Lizard ( Varanus bengalensis ) were studied in the farm area of Azra Produces Impex (a private enterprise) at Bhaluka, Mymensingh from 1995 to 1997. The feeding success, caring, egg-laying, clutch-size, incubation of neonate care were observed. The eggs were laid between August and October with a mean clutch-size of 21.1 (range 10-32, n=25). The mean incubation period was 192.7 days (range 189-216 days, n=678) with a hatching success of 3.3% which was very low due to many reasons. Some problems regarding farming of the species are discussed. Key words. - Varanus bengalensis , Bengal lizard, breeding, farming, Bangladesh. Introduction The Bengal, or Gray, Monitor Lizard ( Varanus ben- galensis) is one of the three varanid species found in Bangladesh. It is most widely distributed throughout the country, including many islands, in both forested and non-forested open wooded areas. It is economically important for its valuable skin and its role in the ecosys- tem, especially in controlling some pests. In Bangladesh, some tribes like the Shawtal, Kulee, Kukis, etc., also eat its meat. The few research works that have been done on varanids in Bangladesh mainly deal with their distribu- tion. However, Whitaker and Hikida (1981) and Akond et al. (1982) briefly worked on the ecology and stomach contents of varanids. There is no published report on the captive breeding of varanids of Bangladesh. Azra Produces Impex, a private enterprise, started a project in Bangladesh on the farming of V bengalensis. As advisor (MFA) and consultant (MAS) we looked into the bio- logical aspects of the project. This paper deals with the preliminary observations on feeding, caring, egg-laying, clutch-size, incubation of eggs, and caring of hatchlings. Some problems regarding farming of the species have also been discussed. Study Area and Study Animals The Varanus breeding farm of Azra Produces Impex is situated at Habirbari of Bhaluka Thana (Mymensign District, Bangladesh, 24° 21' N and 90° 21' E). It is 71 km north of Dhaka City and located adjacent to the Dhaka-Mymensign Highway. The farm was inaugurated in June, 1995 within a concrete boundary wall (about 3 Figure 1. Adult Varanus bengalensis eating supplied food. m high including wire rope) enclosure with an entrance (gate) on the western side along the said Highway. The total area of the farm is 37.74 acres (16.77 ha). There are 50 ponds inside the farm and a lake excavated around the periphery of the farm. One open wire-net enclosure (37 m x 61 m x 1 m) with a concrete base (of 25.4 cm) has also been made inside the farm area for some of the lizards (about 200 individuals). A total of 2, 112 Bengal lizards (685 [32.4%] males and 1427 [67.6%] females) were released inside the farm area. These lizards were captured by professional hunters from wild stock (with the permission of the con- cerned authority, Ministry of Environment and Forests, Government of the People's Republic of Bangladesh) of Greater Mymensign and Tangail districts between 4 June and 9 September, 1995. Before releasing, each lizard was physically checked and sex recorded. Injured and immature lizards were rejected. © 2004 by Asiatic Herpetological Research Vol. 10, p. 237 Asiatic Herpetological Research 2004 Figure 2. Eggs of Varanus bengalensis in a natural nest (left) and being prepared for artificial incubation (right). Methods Food. Cleaned pieces of stomach (omasum part only) of bovines, collected from different slaughter houses of Dhaka city, were supplied to the lizards as food. Food was supplied only inside the wire-net enclosure in trays (25.4 cm x 25.4 cm x 5 cm, made up of tin sheet). In other parts of the farm, food was thrown near the roost- ing places (close to bushes) of the lizards. Usually foods were supplied to the lizards every alternate day (except for Sunday) - half of the farm area was covered in one day and the rest half in the next day. About 100-150 such stomachs were offered to the lizards on every feeding day (Fig. 1). Eggs. Eggs (Fig. 2, Table 1) were collected from holes in the soil (dug obliquely) and holes of termite mounds (either natural or made by lizards) and placed inside arti- ficial holes made in the incubation cages. Special pre- cautions were taken during handling of eggs to avoid shaking or turing upside down, so that, embryos do not get loose or shift from the original position. Incubation. An incubation area was selected inside the farm where two cages were built (Fig. 3), each one hav- ing a small entrance. Cage-I was closed cart-shaped (12.5 m x 4.6 m x 2.6 m), made of wire-net (mesh size 5 mm x 5 mm) with a concrete base and supported by rods. Inside the cage three artificial incubation beds were built - western (1240 cm x 1 16 cm x 55 cm), mid- dle (1240 cm x 110 cm x 48 cm) and eastern bed (1240 cm x 95 cm x 32 cm). Spaces between western and mid- dle beds were 53 cm, and between middle and eastern were 43 cm. In each bed, two storied oblique holes were excavated - the western bed had holes only in the east- ern side, the eastern only in the western side, and the middle bed had holes on both sides. Sandy, granular, and dry soils were placed on the floor of each hole and dry sands on the hole mouth. The roof of each hole was made by moist sandy loam. The top of each bed was covered with grasses. Each hole (length 30 cm and diameter 10 cm) was marked by numbered stick plate. Altogether there were 190 holes in cage-I and 30 eggs were placed in each hole. When all holes of cage-I were filled with eggs (totalling 5,700), the second cage (cage- II) was built in the eastern periphery of the cage-I. Cage- II was closed rectangular-shaped (12.5 m x 7.9 m x 2.5 m) formed of wire-net (mesh size 5 mm x 5 mm) and supported by rods and poles. Here, only pits were dug, and 30 eggs were placed in each pit. There were 500 such pits (containing 14,499 eggs in total) in this cage and each one was marked with numbered stick plate. These pits were filled with loose soils in such a way that a 5-7 cm thick soil layer was on the eggs. The pits were covered with broad (palm) leaves during hot days to retain moisture in the soil. When topsoil of cage-II was too dry, water was added. Later on, some grasses grew naturally. Hatching and Hatchability. After two months of incu- bation, several nests were excavated weekly to check for hatching and 2-5 eggs were opened to see the develop- ment of embryos (Fig. 4). Those hatched (Fig. 5) were immediately transferred into baby nursery. When suffi- cient number of hatchlings were obtained in cage-I, all the remaining eggs which seemed to be still alive were transferred into 7 big trays (0.5 m x 2.5 m x 0.25 m) in the nursery, these trays were kept separate from the hatched lizards by a wall of 1 m within the nursery. The earlier mentioned precautions were taken during this transfer. Eggs were placed half-buried in the tray soil with the usual cover. Each tray was checked twice a day (morning and late afternoon) and the number of hatch- lings was recorded. Then hatchlings were released immediately in the nursery after careful noting of their measurements (Table 2). 2004 Asiatic Herpetological Research Vol. 10, p. 238 Figure 3. Incubation cages for Varanus bengalensis. Cage I (left), Cage II (right). See text for description. Table 1. Varanus bengalensis egg sizes (n=24). Size Mean ± SD Range Length (cm) 5.71 ± 0.41 4.92-6.18 Width (cm) 2.92 ± 0.06 2.80 - 3.04 Weight (gm) 26.93 ± 5.45 25.2-31.5 Baby Nursery. The concrete floor of the nursery (9.5 m x 3.6 m x 3 m) was covered with sandy loam soil. Tin sheets (40 cm high) were fitted against the nursery walls to prevent the escape of the baby lizards. Lumps of ter- mite mounds containing termites (adults, mostly eggs and larvae) were placed in the periphery of the nursery as food for the hatchlings. Other foods offered to the babies were crushed boiled poultry eggs, minced beef and minced clean stomach of bovine. Two artificial small water reservoirs (25.4 cm x 25.4 cm x 5 cm) were made in the nursery. All non-hatched eggs were piled and randomly 100 eggs from incubation cage-I and 150 from incubation cage-II were opened to determine the percentage of undeveloped eggs and dead embryos (Fig. 4). Observations, Results, and Discussion Food. Besides the supplied food, lizards were also seen to eat arthropods especially beetles and grubs from cow- dung, and small fish (mainly Tilapia , which were Table 2. Size of hatchling Varanus bengalensis (n=39). Size* Mean ± SD Range Total length (cm) 19.72 ± 1.31 17.2-22.2 Body length (cm) 9.24 ± 0.59 8.3-10.4 Tail length (cm) 10.52 ± 1.02 7.5- 12.5 Total weight (gm) 13.61 ±3.73 8.3-22.1 * Total length (snout to tail tip), body length (snout to anus), and tail length (anus to tail tip). released in the lake for propagation). Caring. Sick lizards were provided with food closer to them. Medical treatment was not given. Eggs. Eggs were white, oval, with soft leathery skin and contained a large yolk supply. The farm staff collected a total of 20,499 eggs from the holes of termite mounds during 11 September to 30 October, 1995. Although the first clutch of eggs (18) were found on 10 August, 1995 inside a termite mound on the embankment of a pond inside the farm, this was not recorded by the farm staff (so it is excluded from the total count). The average col- lection of eggs was 512.67 + 134.49 (range 160-1210, n=38 days) per day. Whitaker and Hikida (1981) and Akond et al. (1982) stated that the egg-laying period of Bengal lizard in Bangladesh is November and December. Daniel (1983), however, reported that eggs were collected from mid April to October in India. In Sri Lanka, the peak breeding period of Bengal lizard is January to April, but eggs also occur during June to December in ground logs or termite mounds (Deraniyagala, 1958). In the present study the egg-laying period was much earlier than that recorded by Whitaker and Hikida (1981) and Akond et al. (1982). The mean size of egg was 5.71 cm in length, 2.92 cm in width and 26.93 gm in weight (Table 1). From India, Daniel (1983) reported that the average egg size of gray lizard was 4.9 x 3.8 cm (range 4.7 x 3.6 to 5.5 x 4.4 cm, n=50) and weighed 1 1.4 gm (range 8.3-14.3 gm, n=25). The size of eggs in this study (Table 1) is close to that reported by Daniel (1983), but the weight data are very different. Clutch size, dutch size varies according to the size and age of the females, larger and older females lay more eggs than younger and smaller ones. The average clutch size was 21.1 + 7.4 (range 10-32, n=25). The clutch size of Bengal lizard was 8-32 (Whitaker and Hikida, Vol. 10, p. 239 Asiatic Herpetological Research 2004 Table 3. Minimum and maximum air temperatures and soil temperature (°C) during study period. Month Air Soil Minimum Maximum Morning Late afternoon November 1995 20.4 25.6 22.4 24.8 December 1995 18.0 22.4 18.0 22.1 January 1996 16.7 20.8 16.5 22.5 February 19.8 20.8 19.6 25.1 March 22.7 26.7 22.3 28.1 April NR NR NR NR May 27.3 35.0 29.8 30.4 June 25.4 32.8 27.7 29.6 NR- Not recorded. 1981; Akond et al., 1982) and 20-30 (Khan, 1987) in Bangladesh, while 8-30 in India (Daniel, 1983). The range of present observation is close to the mentioned works except for Khan (1987). Incubation period. On average, the incubation period of egg was 192.72 + 4.59 days (range 189-216 days, n=678 eggs). The first lizard hatched on 18 March and the last on 4 June, 1996. Most of the lizards hatched late at night or early in the morning; some also hatched dur- ing the day. Previously recorded incubation periods for the Bengal Monitor Lizards of Bangladesh were 7-8 months (i.e., 210-240 days) (Whitaker and Hikida, 1981); 6-8 months (i.e. 180-240 days) (Akond et al., 1982) and 7-8 months (i.e. 210-240 days) (Khan, 1987). Daniel (1983) mentioned that the incubation period of Bengal lizard in India was 8-9 months (i.e. 240-270 days). The present incubation period is closer to that recorded by Whitaker and Hikida (1981) and Akond et al. (1982), but smaller than that reported by Daniel (1983). The egg-hatching month has been mentioned as July (Whitaker and Hikida, 1981) and June-July (Akond et al., 1982) while in the present observation it spreads over mid March to early June. The variation in the incubation period of the present work and those of the above mentioned works could be due to the effect of some ecological factors like temper- ature, moisture, rainfall, etc. We recorded air tempera- ture in the farm and soil temperature of nest of the incu- bation cage-I (and later baby nursery) which give an indication of these conditions (Table 3). Hatchlings and hatching success. The average total length and weight of the hatchlings were 19.72 cm and 13.61 gm, respectively (Table 2). Out of 20,499 eggs, only 678 babies hatched. The hatching success, in this case, was 3.3%. (All the hatched eggs [678] were from cage-I only and the hatching success was 11.9%, but the eggs from cage-II resulted the poor hatching success i.e., 3.3%). After hatching out, a baby lizard did not eat for the next 2-3 days due to a continued absorbance of its yolk reserve. Neonate lizards ate termite eggs and larvae from the supplied lumps of termite mounds inside the Figure 4. Varanus bengalensis in two stages of development: embryo (left), newly hatched (right). 2004 Asiatic Herpetological Research Vol. 10, p. 240 Figure 5. Varan us bengalensis neonates. nursery and crushed boiled poultry eggs from the feed- ing trays. They showed less interest to eat minced beef and minced bovine stomach. Babies also drank water and preferred to roost in cold, damp areas inside grasses or water hyacinths, which were kept in a few places inside the nursery. The poor hatching success of eggs in this study was most probably due to: (1) soil in the incubation cage-II became compact due to rain and killed embryos; (2) mis-handling of eggs by the staff during egg transplan- tation; (3) unregulated temperature and moisture in the incubation cages. Of these reasons, the first one was most important because only 12 eggs hatched from the incubation cage-II (where 14,799 eggs were transplant- ed) and dead embryos or babies were found in 70% of the eggs (n=140). On the other hand, we were not sure whether all the unhatched eggs were fertilized or not. The additional reason for this huge damage of eggs was the negligence of the Managing Director of the project to implement our suggestions in constructing incubation cage-II. Problems regarding farming The following problems were faced during the study period: 1 . The set up of the project is not well designed and scientific. 2. Lack of electricity. 3. Instructions/suggestions given (jointly by the advisor and consultant) to the Managing Director (MD) of the project were not properly followed. 4. Research facilities provided by the farm are poor. Recommendations 1. Lizards should be caged rather than distributing them throughout the farm. A few small cages should be built for research. 2. Needs devoted staff. 3. Needs electricity. 4. Incubation cages should be constructed like cage-I. 5. Needs incubation chamber, or at least a place(s) where temperature and moisture fluctuaions are not drastic. 6. Needs separate “nursery” cages for neonates and juveniles. 7. Above all, instructions and suggestions proposed jointly by the advisor and consultant should be con- sidered in all activities. Acknowledgments Dr. M. A. G. Khan, Department of Zoology, University of Chittagong has kindly reviewed this manuscript. The proprietor and farm staff of Azra Produces Impex helped us in various ways. We thank them all. Literature Cited Akond, A. W., F. Ahsan, and M. Rahman. 1982. Monitor lizards of Bangladesh. Proceedings of the Second National Conference on Forestry held in 21-26 January 1982:540-545. Daniel, J. C. 1983. The book of Indian reptiles. Bombay Natural History Society, Bombay. 141 pp. Deraniyagala, P. E. P. 1958. Reproduction in the moni- tor lizard, Varanus bengalensis (Daudin). Spolia Zeylanica. 28:161-166. Khan, M. A. R. 1987. Bangladesh er banayaprani, part I (Wildlife of Bangladesh, vol. I). Bangla Academy Press, Dhaka. 175 pp. (In Bengali). Whitaker, R. and T. Hikida, 1981. Report of project for mation mission to Bangladesh (monitor lizards). FAO, Rome. 2004 Asiatic Herpetological Research Vol. 10, pp. 241-244 A New Locality for the Rare Bornean Skink, Lamprolepis vyneri (Shelford, 1905) (Sauria: Scincidae) Indraneil Das Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, 94300, Kota Samar ahan, Sarawak, Malaysia; E-mail: idas@ibec.unimas.my Abstract. - A specimen of the Bornean arboreal skink, Lamprolepis vyneri (Shelford, 1905), hitherto known from the holotype from Gunung Balingan, Sibu Division, Sarawak, and a second possible specimen from the upper reaches of Sungei Mahakam, Kalimantan, is reported from Bukit Balian, near the Kayan settlement of Kelep, at Sungei Asap, at the base of Gunung Dulit, Kapit Division, Sarawak. The species is illustrated for the first time. Key words. - Lamprolepis vyneri , redescription, Scincidae, Sarawak, Borneo. Introduction The genus Lamprolepis Fitzinger, 1843, which was revived from the synonymy of Dasia Gray (1829), by Greer (1970) contains four nominal species of arboreal skinks. Two of these are endemic to Borneo (L. nieuwen- huisii and L. vyneri ), a third ( L . leucosticta ) to Java (Manthey and Grossmann, 1997:263) and the fourth {L. smaragdina ) is widespread in the Philippines, Sulawesi, Lesser Sundas, the Republic of Belau, the Carolines, New Guinea, the Solomons and Santa Cruz Islands (Brown and Alcala, 1980:76-79; Greer, 1970). The first two species are arguably the least well known of all Bornean lizards. L. nieuwenhuisii (Lidth de Jeude, 1905) was described from "Long Bloe" (= Long Blu or Bloeoe, 00° 43' N; 114° 25' E), on the upper reaches of Sungei Mahakam, Kalimantan Tengah Province, Indonesia; RMNH 4455, holotype). It has subsequently been col- lected from isolated localities in northern Borneo, including Nanga Tekalit Camp on Sungei Mengiong, Kapit Division (reported as Dasia vyneri by Lloyd et al., 1968, based on FMNH 138542; 147562); and Pangkalan Lobang at Niah National Park, Miri Division (FMNH 131528), both in Sarawak State; and Kiau, Gunung Kinabalu National Park, Ranau District (MCZ 43494; BMNH 1929.12.22.96 and ZRC 2.1595); and Mahunbayon, Gunung Kinabalu National Park, Ranau District (MCZ 43495), both in Sabah State, East Malaysia. Lamprolepis vyneri (Shelford, 1905) is more poorly known. Named for Charles Vyner Brooke (1874-1963), the Rajah Muda of Sarawak at the time of description of the species, and subsequently, the Third Rajah of Sarawak between 1917-1946, it is only known from the holotype, BMNH 1946.8.15.56 (ex-BMNH 1909.8.18.2), from "Mount Balineau, Muka district, Sarawak" (= Gunung Balingan, 01° 25' N; 1110 28' E, Sibu Division, East Malaysia), according to the original description. However, in the records of the Sarawak Museum (Anon., 1903), the type locality is given as "Mt. Balingean" (in Muka District, Sibu Division, Sarawak). Lidth de Jeude (1905) questionably assigned to this species a specimen from the upper reaches of Sungei Mahakam (00° 30' S; 117° 15' E), Kalimantan Timur Province, which apparently differed from Shelford's (1905) species in some trivial details of squa- mation and body proportions. The location of this spec- imen is unknown, but was examined by De Rooij (1915), who allocated it to the present species. This species has never been illustrated. A second specimen (ZRC 2.5513; Figs. 1-2) of Lamprolepis vyneri is reported here from Bukit Belian (03° 08' 34.4" N; 113° 55' 45.5" E), near the Kayan set- tlement of Kelep, at Sungei Asap, situated at the base of Gunung Dulit, Kapit District, Sarawak. It was collected dead on 6 November 2001 from a logging road. Material and Methods The specimen was photographed upon collection, fixed in neutral buffered formalin and subsequently trans- ferred to 70% ethanol, within a week of collection. The following measurements were taken with Mitutoyo™ dial caliper (to the nearest 0.1 mm): snout-vent length (SVL; from tip of snout to vent), tail length (TL; from vent to end of unregenerated tail; tip missing), tail width (TW; measured at base of tail); head length (HL; dis- tance between posterior edge of last supralabial and snout-tip), head width (HW; measured at angle of jaws), head depth (HD; maximum height of head, from occiput to throat), ear length (EL; greater ear length); eye diam- eter (ED; greatest diameter of orbit), eye to nostril dis- tance (E-N; distance between anteriormost point of eyes and nostrils), eye to snout distance (E-S; distance © 2004 by Asiatic Herpetological Research Vol. 10, p. 242 Asiatic Herpetological Research 2004 Figure 1. The Sungei Asap specimen of Lamprolepis vyneri (ZRC 2.5513), showing (left) general view of body and (right) close-up of head and forebody. between anteriormost point of eyes and tip of snout), eye to ear distance (E-E; distance from anterior edge of ear opening to posterior comer of eyes), intemarial distance (IN; distance between nares), interorbital distance (10; shortest distance between orbits), tibia length (TBL; straight length of tibia, from knee to sole), in addition to measurements of digits, taken on the left limbs, from the base to tip. Scale counts and external observations of morphology were made using an Olympus SZX9 dis- secting microscope. Institutional abbreviations follow Leviton et al. (1985), except ZRC is retained for USDZ, following conventional usage. Description of Lamprolepis vyneri from Bukit Belian, Sungei Asap (ZRC 2.5513). - Habitus relatively slen- der, snout- vent length 55.2 mm; head elongate (HL/SVL ratio 0.20), narrow (HW/HL ratio 0.65), moderately depressed (HD/HL ratio 0.11), slightly distinct from neck; snout long (E-S/HW ratio 0.72), longer than the eye diameter (ED/E-S ratio 0.75), projecting slightly beyond mandible; interparietal distinct; parietal eye absent; supraoculars four; second and third largest; supraciliaries 8/8; first supraciliary contacts frontal; scales on snout and forehead smooth; rostral contact frontonasal posteriorly; rostral small, wider than deep (rostral width = 2.0 mm; rostral depth = 1.2 mm; width/depth ratio 1.67), contacted posteriorly by nasal and frontonasal; posteroventrally, rostral in contact with first supralabial; nares slit-like, situated on upper level of nasal, oriented laterally; nasal in broad contact with first supralabial; supranasals moderate in size, separat- ed; frontonasal trapezoid, wider than long, contacting frontal and prefrontals posteriorly; frontal longer than frontonasal, not constricted laterally; frontoparietals in contact with each other and with three supraoculars, and posteriorly, with interparietal and parietals; a single pair of parietals contacts interparietal; parietals separated behind by an azygous scale; loreals two, anteriormost longer than deep; a small dorsal presubocular, and a wider ventral one; eye large (ED/HL ratio 0.35); post- suboculars two; supralabials seven, with supralabials 4- 6 in suborbital position; supralabials three, fifth and sixth larger than the others; infralabials six; lower eyelid scaly; a single preocular between loreal and orbit; pos- toculars two; pretemporals two; two anterior and two posterior temporals; ear opening narrow, measuring 1.9 mm; situated laterally at a level slightly higher than jaws; a few lobules around ear opening present; tympa- num deeply sunk; eye-to-ear distance less than eye-to- nostril distance (E-E/E-N ratio 1 .26); a pair of enlarged nuchals, partially separated by a single cycloid scale; mental large, semicircular, wider than deep; postmental single, trapezoidal, larger than mental, its width 1.8 mm or 25.4 per cent head width; postmental contacts first infralabial only, bounded posteriorly by a pair of smooth, squarish, juxtaposed chin shields that are in contact; three pairs of enlarged chin shields, the first in contact with each other, the second separated by a single scale, the third separated by three scales; tongue narrow- ly elongate, narrowed distally, with a median cleft and scattered papillae on the dorsal surface; maxillary and mandibular teeth small, undifferentiated. Body slender, elongate (SVL/BW ratio 6.81); dor- sum and venter with smooth scales, with faint striae, scale size subequal dorsally as well as ventrally; anals six, smooth; outer overlapping inner; preanals three, not greatly enlarged, overlapped by last ventral, third pre- anal exceeding its posterior level, over vent; flank scales reduced in size. Limbs well developed, pentadactyle; adpressed limbs meeting at level of heels; lamellae under finger IV numbering 18; lamellae under toe IV numbering 20; rel- ative length of fingers (measurements in mm, in paren- theses): 4 (4.5) > 3 (4.4) > 2 (3.5) > 5 (2.7) > 1 (2.0); rel- ative length of toes: (measurements in mm, in parenthe- ses): 4 (7.8) > 3 (5.6) > 5 (5.5) > 2 (4.7) > 1 (3.0). Tail long, preserved tail length over 40.5 mm (tip missing), longer than snout-vent length; tail base slight- ly swollen; ventral surface of tail with smooth; subcau- dals very wide; scales on the postanal region and at the proximal part of the tail base smooth. Coloration. - Forehead olive-yellow, edged with black; dark smudges on forehead scales; scales on dorsum of 2004 Asiatic Herpetological Research Vol. 10, p. 243 Figure 2. Head of Lamprolepis vyneri (ZRC 2.5513) in dorsal (left) and ventral (right) views. Scale bars = 10 mm. body bright yellow, edged with black, appearing as four dark longitudinal lines that extend along the body to slightly beyond the base of tail; yellow dorsolateral stripe, 2-3 scale wide, runs from behind level of the axil- la, across the inguinal region, continuing along the side of the tail; venter, including the gular, pectoral and abdominal regions, undersurface of tail and of limbs yel- lowish-green, unpattemed; scales on flanks black-edged, reddish-orange, with scattered yellow scales; the same coloration is found on the upper surfaces of the fore and hind limbs; tail alternately banded yellowish-brown, each band one scale wide, and pale yellow; tongue and inner lining of mouth yellowish-pink; inner lining of body cavity yellowish-pink in preservative. Measurements (in mm). - BW 8. 1 ; ED 3.8; E-E 4.8; EL 1.2; E-N 3.8; E-S 5.1; IN 1.7; IO 4.5; HD 5.8; HL 10.9; HW 7.1; SVL 55.2; TBL 7.7; and TL 40.5 - original unregenerated, tip missing; TW 5.3. Scutellation. - Ventrals (between postmental and pre- anal) 49; midbody scale rows 22; subcaudal count unknown (tail-tip missing); supralabials seven (fourth, fifth and sixth in suborbital position) and infralabials six. Variation. - The Sungei Asap specimen differs from the holotype in the following particulars: SVL 55.2 vs 52.0 mm; supraoculars on left side four (vs five in the holo- Figure 3. Map of Borneo showing the known localities for Lamprolepis vyneri. 1 = Gunung Balingan, Sibu Division (type locality); 2 = Bukit Balian, near Sungei Asap, Kapit Division. type, as shown on the apparently anomalous right side of the head of the Bukit Belian specimen). The bright red and yellow coloration of the flanks of the Bukit Belian specimen turned to dark brown after three months of storage in preservative. Shelford (1905), who presum- ably examined a preserved specimen, reported the flanks as being olive-gray, van Lidth de Jeude's (1905) speci- men was 63 mm in SVL, and showed five black stripes, but only three entering the sacral region and tail-base. This poorly-preserved specimen was described as "putty grey", with several cephalic scales edged with black. Notes on Natural History. - The specimen being reported here was found freshly dead on a logging track at the base of Bukit Belian (03° 08' 34.4" N; 113° 55' 45.5" E), near Kelep, Sungei Asap, a Kay an resettlement colony in Kapit (Seventh) Division, central Sarawak. It may have fallen off a log that was being transported, because the members of the genus are highly arboreal, and the present specimen was otherwise physically intact, except for the missing tail-tip, and not run over. The area lies within a lowland dipterocarp forest with strands of the Bornean ironwood tree, Eusideroxylon zwageri (Iban name: Belian, which gives the hill its name) at 186 m elevation. Perhaps coincidentally, the holotype was taken at a similar-sounding locality, Gunung Balingan, for which no general habitat descrip- tion is available. One is therefore tempted to speculate Vol. 10, p. 244 Asiatic Herpetological Research 2004 that both localities derive their names for their strands of the Bornean ironwood, a dipterocarp much in demand from the timber industry for its durability, and hence threatened by logging. The new locality, at the base of Gunung Dulit, is ca. 190 km east of the type locality, across the Lumut Range (Fig. 3). Acknowledgments I thank the Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, for sup- porting my research on the herpetofauna of Borneo, and Esther Bala for field assistance. For loans of the holo- types of Lamprolepis vyneri and L. nieuwenhuisii, I am grateful to C. J. McCarthy, BMNH, and M. S. Hoogmoed, RMNH, respectively. I would like to thank R. F. Inger, A. Resetar and H. K. Voris, FMNH; J. E. Cadle, J. Rosado and the late E. E. Williams, MCZ, and K. K. P. Lim, P. K. L. Ng and C. M. Yang, ZRC, for per- mitting me to examine comparative material under their care and Allen Greer and an anonymous reviewer for comments on a draft manuscript. Finally, thanks are due to Gary Geller, Jet Propulsion Laboratory, National Aeronautics and Space Administration, for generating Fig. 3. Literature Cited Anonymous. 1903. Museum. The Sarawak Gazette 33:236. Brown, W. C., and A. C. Alcala. 1980. Philippine lizards of the family Scincidae. Silliman University. Natural Science Monograph Series 2. Silliman University Press, Dumaguete. 264 pp. De Rooij, N. 1915. The reptiles of the Indo- Australian Archipelago. Vol. I. Lacertilia, Chelonia, Emydosauria. E. J. Brill, Leiden. 384 pp. Greer, A. E. 1970. The relationships of the skinks referred to the genus Dasia. Breviora (348): 1-30. Leviton, A. E., S. C. Anderson, R. H. Gibbs, E. Heal, and C. E. Dawson. 1985. Standards in herpetology and ichthyology. Part I. Standard symbolic codes for institutional resource collections in herpetology and ichthyology. Copeia 1985(3):802-832. Lloyd, M., R. F. Inger, and F. W. King. 1968. On the diversity of reptile and amphibian species in a Bornean rain forest. American Naturalist 1 02(928):497-5 1 5. van Lidth de Jeude, T. W. V. 1905. Zoological results of the Dutch scientific expedition to central Borneo. The reptiles. Part I. Lizards. Notes from Leyden Museum 25(16): 187-202. Manthey, U., and W. Grossmann. 1997. Amphibien and Reptilien Siidostasiens. Natur und Tier Verlag, Munster. 512 pp. Shelford, R. 1905. A new lizard and a new frog from Borneo. Annals & Magazine of Natural Histoiy, Series 7, 15:208-210. Asiatic Herpetological Research Vol. 10, pp. 245-246 [ Leptobrachium smithi Matsui, Nabitabhata, and Panha, 1999 (Anura: Megophryidae), an Addition to the Fauna of Myanmar (Burma) Indraneil Das1 and Sh yamal Kumar Chanda2 1 Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, 94300, Kota Samar ahan, Sarawak, Malaysia; E-mail: idas@ibec.unimas.my “ Amphibia Section, Zoological Survey of India, Fire-Proof Spirit Building, 27, J. L. Nehru Road, Kolkata 700 019, India Abstract. - Three specimens of Leptobrachium from the collections of the Zoological Survey of India are identified as Leptobrachium smithi. These specimens were collected of Leptobrachium smithi for Myanmar. Key words. - Anura, Leptobrachium , Myanmar, Burma. Leptobrachium smithi Matsui et al. (1999) was described from peninsular Thailand, based on popula- tions that were formerly referred to L. hasseltii Tschudi, 1838 (see Frost, 1985). This species was recently report- ed from Chandubi in the Mayeng Hill Reserve Forest and Garbhanga Reserve Forest, Kamrup District, Assam State, north-eastern India by Sengupta et al. (2001). We here report specimens from Myanmar in the collection of the Zoological Survey of India (ZSI) that are allocat- ed to L. smithi. Three specimens of Leptobrachium smithii were examined: ZSI 10439-40, from "Ahsoon" (unlocated), in Tenasserim, Myanmar, altitude "2,000 feet", collected by the Swedish journalist, novelist, poet and ship cap- tain, Gustaf Arthur Ossian Limborg (1849-1908) in 1 877. Limborg's expedition to what was then Burma was sponsored by Lord Tweeddale (Kjellgren, 1983) and his collections are distributed in Sweden and the US). Also examined was ZSI 11841, from Lampi Island, Mergui, collected by John Anderson, in 1882 (referred to by Anderson, 1889, as from "Sullivan Island", an older name for Lampi, 10° 50' N; 98° 15' E). The material from Myanmar match the description of original description of Leptobrachium smithi , in addi- tion to additional specimens examined from Assam State (see Sengupta et al., 2001), in showing the following characteristics: moderate body size (snout-vent length 22.4-43.4 mm; head width 6.6-18.1 mm; n = 3); small inner metatarsal tubercle; dorsum smooth; and absence of rows of dermal ridges on dorsal surface of limbs. All specimens referred to here are discolored, hence other characters used in separating L. smithi from L. hasseltii , such as absence of white spots on sides of body and on thigh; absence of dark spots on ventrum; and absence of dark markings on dorsum, that differentiates the north- Limborg in 1877. These are the first confirmed records em L. smithi from the southern L. hasseltii , are indis- cemable. The known distribution of Leptobrachium smithi is thus north-eastern India, Myanmar (first country record on the basis of ZSI specimens reported here) and Thailand. Matsui et al. (1999) suspected the occurrence of the species in southern Myanmar, based of the larval description of L. hassseltii by Annandale (1917:153- 157, as Megalophrys hasseltii ), from the Dawna Hills of the Tenasserim. We have examined these specimens (ZSI 16735-43) that carry the following locality "Misty Hollow, w side of Dawna Hills, L. Burma". Surprisingly, Annandale, neither in his 1917 monograph, nor in any other works, have referred to the specimens from Burma mentioned earlier, although all of these were available to him (see Sclater, 1892). Acknowledgments We thank J. R. B. Alfred, Director, Zoological Survey of India, Kolkata and Colin J. McCarthy, The Natural History Museum, London, for permission and facilities at their respective institutions. Saibal Sengupta, Arya Vidyapeeth College, made comparative material avail- able to us, and Erik Ahlander, Swedish Museum of Natural History, Stockholm, provided details on the life of Ossian Limborg. Literature Cited Anderson, J. 1 889. Report on the mammals, reptiles, and batrachians, chiefly from the Mergui Archipelago, collected for the Trustees of the Indian Museum. Journal of the Linnean Society (Zoology) 21:331- 350. © 2004 by Asiatic Herpetological Research Vol. 10, pp. 246 2004 Annandale, N. 1917. Zoological results of a tour in the Far East. Batrachia. Memoirs of the Asiatic Society of Bengal 6:119-155; PI. V-VI. Frost, D. R. (Ed). 1985. Amphibian species of the world. A taxonomic and geographical reference. Allen Press, Inc., and Association of Systematics Collections, Lawrence, (iv) + v + 732 pp. Kjellgren, L. 1983. En prastson som alskade havets vag. Om kaptenen och litterataren Ossian Limborg. Sumlen (Svenskt Visarkiv) 1983:57-74. Matsui, M., J. Nabhitabhata, and S. Panha. 1999. On Leptobrachium from Thailand with a description of a new species (Anura: Pelobatidae). Japanese Journal of Herpetology 18(1 ): 1 9-29. Sclater, W. L. 1892. List of the Batrachia in the Indian Museum. Indian Museum, Calcutta, viii + 43 pp. Sengupta, S., N. K. Choudhury, and I. Das. 2001. Leptobrachium smithi Matsui, Nabhitabhata and Panha, 1999 (Anura: Megophryidae), a new record for India. Journal of the Bombay Natural History Society 98(2):289-291. Asiatic Herpetological Research Vol. 10, pp. 247-279 Species Diversity and Checklist of the Herpetofauna of Pulau Tioman, Peninsular Malaysia, With a Preliminary Overview of Habitat Utilization Jesse L. Grismer1, L. Lee Grismer1, Indraneie Das2, Norsham S. Yaakob3, Lim Boo Liat4, Tzi Ming Leong5, Timothy M. Youmans1, and Hinrich Kaiser1 * Department of Biolog)’, La Sierra University, Riverside, CA 925 1 5-8247, USA -Institute of Biodiversity and Environmental Conservation, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia ' Forest Research Institute Malaysia, Kepong, 52109 Kuala Lumpur, Malaysia 4 Taman Negara (PERHILITAN), Km 10 Jalan Cheras, 50664 Kuala Lumpur, Malaysia 3 Department of Biological Sciences, National University of Singapore, Singapore 119260 Abstract. - The environmental diversity of Pulau Tioman, a 48 km2 island off the eastern coast of Peninsular Malaysia, supports a remarkably diverse herpetofauna (97 species) with 22 frogs, one caecilian, one non-marine turtle, 34 lizards, and 39 snakes. The majority of this herpetofauna (74%) occurs in lowland dipterocarp forests. Fifteen new island records and eight newly described, or as yet undescribed, species are reported, bringing the number of endem- ic species to at least 11. Key words. - Pulau Tioman, Malaysia, herpetofauna, habitat diversity, checklist. Introduction Pulau Tioman (Tioman Island) is centrally located on the Sunda Shelf 38 km off the southeast coast of Peninsular Malaysia in the South China Sea (Fig. 1). Despite its small size of approximately 48 km2, it supports a diverse array of habitats. The island's coastline and low-lying periphery is dominated by mangrove and coastal vegeta- tive communities whereas inland areas support lowland dipterocarp forest on the alluvial foothills and hill dipte- rocarp forest at upper elevations (Latiff et al. 1999). Topographically, Pulau Tioman is characterized by steep mountainous terrain reaching 1,035 m in elevation. Exposed granitic outcroppings consisting of large boul- ders define much of the island's rugged interior and its slopes are cut by several fast-flowing, boulder-strewn streams. As discussed below, this environmental diver- sity contributes to the island's remarkable herpetological diversity with 23 amphibians, one non-marine turtle, 33 lizards, and 39 snakes now confirmed as present on the island (Table 1). This is in contrast to the relative depau- perate herpetofauna of the surrounding islands of Tulai (Grismer et al., 2001b), Aur (Escobar et al., 2002a; Grismer et al., 2001a), Dayang (Wood et al., 2003), Pemanggil (Youmans et al., 2002), Sembilan and Seribuat (Wood et al. in prep), Sibil and Besar (Wood et al., 2004a,b) and Tinggi (Escobar et al., 2002b). Prior to Elendrickson (1966a,b), no herpetofaunal survey had been undertaken on Pulau Tioman and only Figure 1. Location of Pulau Tioman, West Malaysia, in the South China Sea. limited accounts on particular taxa existed (i.e., Boulenger, 1912; Smith, 1930; de Haas, 1949). However, despite the thoroughness of Hendrickson (1966a,b) and subsequent efforts by Day (1990), Lim and Lim (1999), Hien et al. (2001), and Grismer et al. (2002a), the herpetofauna of this small island still © 2004 by Asiatic Herpetological Research Vol. 10, p. 248 Asiatic Herpetological Research 2004 Mangrove swamps (0 m) Coastal vegetation (0-80 m) Lowland dipterocarp forest (80-300 m) Hill dipterocarp forest (300-950 m) Ridge forest (950-1,035 m) Distribution of vegetation zones on Pulau Tioman. Modified from Latiff et al. (1999). remains incompletely known. This is evidenced by the 13 new island records since Grismer et al. (2002a) and Hien et al. (2001) and eight newly described and unde- scribed species reported herein. Additionally, there has been no attempt to establish the distribution or habitat use of each species on Pulau Tioman. Therefore, the intent of this paper is to report the results of the latest herpetofaunal surveys which not only list new additions to the island but new island localities of species known to be present. The latter will serve as the basis for a pre- liminary categorization of habitat use for each species based on its presence in different vegetation zones. Figure 3. Mangrove swamp at Kampung Paya. Vegetation Zones Vegetation zones generally serve to highlight broad cat- egorical differences between habitats across sizable geo- graphic areas. On Pulau Tioman, as elsewhere, these categorical differences lack well-defined geographic boundaries (Ashton, 1995) and with the exception of mangrove communities, each zone transitions smoothly and continuously into another along an altitudinal tran- sect. We use five different vegetation zones (Fig. 2), modified after Latiff et al. (1999), to characterize habi- tat differences on Pulau Tioman. Mangroves (0 m; Fig. 3). - Mangrove swamps are dis- junctly distributed along the island's coastline. Characteristic plant species include Rhizophora apicula- ta, Bruguiear gymnorhiza, Excoecaria agallocha , and Avicennia alba , which in some localities are unusually tall with large girth, attesting to the old age of the grove. Coastal vegetation (0-80 m; Fig. 4). - Coastal vegeta- tion forms a relatively narrow zone between the man- grove swamps (when present) and the lower reaches of the lowland dipterocarp forest. It is characterized by 2004 Asiatic Herpetological Research Vol. 10, p. 249 Fig. 6. Hill dipterocarp forest on Gunung Kajang. Fig. 7. Ridge forest at summit of Gunung Kajang. 2004 Figure 8. Collecting localities on Pulau Tioman. G. = Gunung (mountain); Kg. = Kampung (village); S. - Sungai (river); Tk. = Telok (bay); U. = Ulu (headwater). palms, such as Pandcmus dubius, and moderately-sized trees such as Scaevola taccada , Calophyllum inophyl- lum, and Vitex trifolia. Dipterocarp trees are noticeably absent. Lowland dipterocarp forest (80-300 m; Fig. 5). - Lowland dipterocarp forest occurs on the alluvial slopes between coastal vegetation and hill dipterocarp forest and is usually dominated by large non-dipterocarp trees such as Arenga pinnata , Caryota mitis, and Nenga macrocarpa. A few large dipterocarp species such as Anisoptera curtisii and N eobalanocarpus heimii exist as emergents. Hill dipterocarp forest (300-950 m; Fig. 6). - This zone is situated immediately above and adjacent to the lowland dipterocarp forest with which it is continuous. The transition from lowland dipterocarp forest to hill dipterocarp forest at approximately 300 m is essentially imperceptible and many plant species common to the lowland dipterocarp forest occur at lower elevations in the hill dipterocarp forest, a pattern paralleled by some species of amphibians and reptiles. To illustrate this we use the term low hill dipterocarp forest (300-500 m) and high hill dipterocarp forest (500-950 m). Hill diptero- carp forest is dominated throughout by large species of Shorea and Dipterocarpus. Ridge forest (hill top summits between 950-1035 m; Fig. 7). - Ridge forest occurs on summits where mosses, ferns, lichens, and bryophytes predominate. Due to increased exposure to sun and wind, trees are relatively short. At this altitude, species such as Garcinia penan- giana , Licuala tiomanensis , and Scleria sumatrensis dominate and presumably have adapted to live in the damp wind-blown environment typical of ridge forests. Materials and Methods Our data were collected from various localities (Fig. 8) on five trips; 12-24 March and 7-16 July, 2001 and 19- 27 March, 13-21 July, and 6-19 August, 2002 unless indicated otherwise. Individuals sighted but not collect- ed or photographed are listed but considered to be unconfirmed records. Collecting was done during the day by hand and blowpipes and at night by torch light. During July 2001 and 2002, three pitfall trap arrays with three 1 5-m drift fence sections at each array were sta- tioned in lowland dipterocarp forest at 142 m and 241 m along the Tekek-Juara trail. A third array was estab- Asiatic Herpetological Research Vol. 10, p. 251 X X X X X x x X X X XXX * a ***** a 53 CO a to a X a a s § -a X X X X X X X X X X X X X XX X XXX XX X XX X X X x XX XX XX X X X X X .5b • **** ■K* CO a a a a 3> a to a a **^ -a a L) X a Q •*»*» X. a -a a a “a "a -a a, a a a a a a '*'■** *"-**>* a a X X o i- o CJ a *s a & to a a a a 13 a a .£ ;a -a a ~S % a X' 3 ^ -a ~a a 03 &o ~ a a a ■*-» rS a a a* a a a a a ?* a a a ^ c & -I -I X ’"■3 X a a a a a -a a a a a a -a •** b a to a -a a a a a a a a a a X X X a a a .a X a a a X to a -*«» a * »** X js qj a S3 .& T3 a © £2 CL O u C3 a £ to a a a X to a a J3 • **** -a X I a a a a to a ■a ~a •*>* a a a ~a a a ~a XX aa a a -a X x a. 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CO Co 3 V 3 co 3 CO .3 3602 Stock, 1999 P. mustersi 43.3 Morescalchi et 1979 al., - - >3602 Reilly, 1983 Ranodon sibiricus 55.4 Present paper 3. 0-5.0 21-422 230-7202 Lebedkina, 1964; Brushko, Narbaeva, 1988; our data Salamandrella keyserlingii 66.4 Present paper 2. 1-3.5 144 78-1101 Ishchenko et al., 1995a, b; Berman, 1996 1 Laboratory conditions (temperature unknown); 2nature conditions; 3t = 10°; 4t = 16°. Table 4. Genome size (2C), diploid number of chromosomes (2n) and number of uniarmed macrochromosomes (UM) in some hynobiids. Taxon 2C Reference 2n UM Reference Hynobius dunni 33.8 Olmo, 1973 56 0 Morescalchi et al., 1979; Seto et al., 1986 H. nebulosus 38.4 Olmo, 1973 56 0-2 Seto et al., 1986; Kuro-o et al., 1987 H. tsuensis 33.0 Olmo, 1973 56 0 Seto et al., 1986 H. retardatus 38.3 Olmo, 1973 40 0 Azumi and Sasaki, 1971; Kuro- o et al., 1987 H. naevis 40.9 Olmo, 1973 58 0 Kohno et al., 1987 Onychodactylus fischeri 107.7 Present paper 78 0 lizuka and Yazawa, 1994 O. japonicus 104.0 Olmo, 1983 78 0 Morescalchi et al., 1979; Kohno et al., 1991 Paradactylodon gorganensis 34.8 Stock, 1999 62 0 Stock, 1999 P mustersi 43.3 Morescalchi et al., 1979 62 10 Morescalchi et al., 1979 Ranodon sibiricus 55.4 Present paper 66 18 Morescalchi et al., 1979 Salamandrella keyserlingii 66.4 Present paper 62 26 Kohno et al., 1991 2004 Vol. 10, p. 286 65 66 67 68 Genome Size (pg) Figure 2. Genome size distribution in Salamandrella key- serlingii. of samples proved to not be chaotic and the "Euro- Siberian" (8 samples) and "Primorsky" (2 samples) groups were recognized. The data ranges of the groups did not overlap, and the gap between both groups was equal to 0.3 pg (Fig. 2). The "Primorsky" samples were characterized by smaller genome size in comparison with the "Euro-Siberian" ones: 64.4-65.2 pg vs. 65.5- 68.1 pg; the means were 64.8 pg vs. 66.9 pg. The differ- ences (RD) between means were equal to 3.2%, and were observed in five separate comparisons. Therefore, these groups were shaped both by genome size and geo- graphically. The samples of R. sibiricus were taken from two semi-isolated populations from upper parts of Borokhudzir and Oy-Saz rivers. Both samples demon- strated different genome sizes (Table 1). The ranges of genome size values in these two samples slightly over- lapped. The Oy-Saz sample was characterized by small- er genome size in comparison with the Borokhudzir sample: 54.5-55.1 pg vs. 55.1-56.7 pg; the means are 54.8 pg vs. 56.0 pg. The differences (RD) between means were equal to 2.2%. The coefficient of variation (CV) ranged between 0.3% and 0.9% in S. keyserlingii , between 0.3% and 1.0% in R. sibiricus , and was equal to 0.7% in the sam- ple of O. fischeri (Table 1). The overall within-species genome size variation in S. keyserlingii and R. sibiricus were quite similar (1.5% and 1.3%, respectively). Among the "Euro-Siberian" samples of S. keyserlingii , the CVs ranged between 0.3% (Nizhny Novgorod Province) and 0.9% (Tomsk Province), whereas it was equal to 0.4% in the "Primorsky" samples of S. keyser- lingii. Discussion 1) Genome size values in hynobiids: literature data Presently, the nuclear DNA content has been determined for eleven hynobiid species. However, among them, only four species were examined through flow cytome- try (Table 2). The first data collected about genome size of nine hynobiid species was obtained by Italian researchers with an application of Feulgen densitometry (Olmo, 1973, 1983; Olmo and Morescalchi, 1975; Morescalchi et al., 1979). According to these data, genome size in hynobiids ranged from 32.96 pg in Hynobius tsuensis to 102-106 pg in Onychodactylus japonicus (Table 2). According to Mazin (1978), the nuclear DNA content in two Russian species, Onychodactylus fischeri and Salamandrella (" Hynobius") keyserlingii, measured by Feulgen densit- ometry, were similar to each other (45.5 and 42.5 pg, respectively). Grafodatsky and Grigoriev (1982) esti- mated genome size of Salamandrella keyserlingii (38 pg), perhaps, by means of Feulgen densitometry. Vladychenskaya et al. (1988) studied kinetics of DNA reassociation; they found that the nuclear DNA content of Salamandrella keyserlingii was equal to 33.2 pg. Using DNA flow cytometry, Vinogradov (1998) esti- mated genome size values as 95.08 pg for Onychodactylus fischeri, 45.59 pg for Ranodon sibiri- cus, and 55.48 pg for Salamandrella keyserlingii. Finally, Stock (1999) determined that genome size of Batrachuperus gorganensis is 34.77 pg, using DNA flow cytometry as well. The literature values of genome size for Onychodactylus fischeri, Ranodon sibiricus , and Salamandrella keyserlingii expressed in absolute units vary sometimes more than two-fold (ranges are 45.5- 95.1 pg, 45.6-50.7 pg, and 33.2-55.5 pg, respectively). Therefore, the comparison of data provided by various authors should be made cautiously. The contradictions may be explained by an application of different tech- niques (Feulgen densitometry, kinetics reassociation, and flow cytometry), dyes, and laboratory conditions (cell preparation methods, devices for measurements, types of reference cell standards, etc.). It has been shown that genome size measured with fluorochromes of dif- ferent nucleotide specificity may differ markedly (e.g., Johnston et al., 1987; Birshtein et al., 1993; Vinogradov and Borkin, 1993). For instance, the determination of genome size by means of flow cytometry for cell sam- ples of Salamandrella keyserlingii stained with olivomycin and Hoechst (GC- and AT-specific fluo- rochromes, respectively) provided 6.68 and 4.77 arbi- trary units (Rana temporaria was taken as an internal reference; Vinogradov, 1998). To exclude the influence Vol. 10, p. 287 Asiatic Herpetological Research 2004 of AT/GC-structure, it is necessary to use ethidium bro- mide or propidium iodide (Vinogradov and Borkin, 1993), which were used in our research. To convert genome size from the relative units to picograms, it is necessary to have data about genome size of reference cells. Such data should be obtained without using stains. Unfortunately, the data available today, mentioned by various authors, do not correspond to each other. For instance, Vinogradov (1998) reported a genome size of reference standard Mas musculus to be 6.5 pg. Our estimations of genome size of some mam- mals ( Homo sapiens , Mas mas cal as, Rattas norvegicas ) were the closest to that mentioned by Bianchi et al. (1983). In our work, we used the genome size of males of Mas musculus (C57B1) as a basic reference standard with value of 6.8 pg. Other authors preferred other ref- erence standards, which have different base-pair-speci- ficity of some stains widely used in flow cytometry. Vinogradov and Borkin (1993) listed the AT- and GC- pair specific DNA contents (CAT and CGC) for many species of amphibians. For instance, CAT/CGC was equal to 1 .42 in Xenopas laevis (Mazin's reference stan- dard), and 1.00 in Rana lessonae (= Rana " esculenta Olmo's and Morescalchi's standard). The estimations of Italian authors (Olmo, 1973, 1 983 ; Morescalchi et al., 1 979) and our genome size val- ues for Ranodon sibiricas (50.7 and 54.5-56.7 pg, respectively) and members of genus Onychodactylus (102.0-106.0 and 106.7-109.0 pg, respectively) are quite similar. However, data for Salamandrella keyserlingii (42.3 and 64.4-68.1 pg, respectively) are in obvious dis- cordance. The genome size of Onychodactylus fischeri , Ranodon sibiricas, and Salamandrella keyserlingii men- tioned by Vinogradov (1998), was lower than our val- ues, who used other stains and lower genome size esti- mation of basic reference standard. Unfortunately, some authors did not supply any information about sample sizes and localities. However, some differences in genome size might be influenced by intra-population and geographic variation as well. 2) Within-species variation Some authors discussed the levels of intraspecific varia- tion in genome size. We recognized two kinds of such a variation; namely, the "within-population" variation and "between-population" (or geographical) variation. A) Within Population Variation Among amphibians, the greatest intrapopulational vari- ation (CV = 7.5%, the data of Licht and Lowcock, 1991 were recalculated by us) was recorded for the Western Red-back Salamander {Plethodon vehicalam). However, the variation in other amphibian species was consider- ably lower (Licht and Lowcock, 1991; MacCulloch et al., 1996; Murphy et al., 1997; Lizana et al., 2000). The variation within populations of three hynobiid species (CVs were 0.3- 1.0%, mean was 0.67 ± 0.12%) was quite similar to that in salamandrids (range is 0.1 -1.7%, mean is 0.64 ± 0.03%, 99 populations studied), pelobatids, and other anurans studied in our laboratory at the same con- ditions (Litvinchuk et al., 1997, 1999, 200 1 a, b, 2003; Rosanov and Vinogradov, 1998; Borkin et al., 2001b, 2003; our data). Sexual dimorphism in genome size has been regis- tered in some amphibian species (Schmid et al., 2002; our data). Unfortunately, in the hynobiids examined by us, sexual differences are not expressed in external char- acters beyond the breeding time. Our study was based mostly on juvenile and non-breeding adult animals, and, therefore, we failed to reliably identify the sex without anatomical dissections. B) Geographical Variation The significant geographical variation in genome size was revealed for several amphibian species (Licht and Lowcock, 1991; Murphy et al., 1997; Litvinchuk et al., 1999, 2001b). In a few cases, differences (RD) exceed- ed 8%. However, in the majority of species studied, such differences were about 1% (Licht and Lowckock, 1991; Borkin et al., 1997, 2000, 2001, 2003; Litvinchuk et al., 1997, 1999, 2001b, 2003; our data). In Salamandrella keyserlingii, the maximum genome size difference (RD = 4.5%) was found between samples from the Khandyga (Yakutia Republic) and Kedrovaya Pad' Reserve (Primorsky Territory). The average differences (RD) between the "Euro-Siberian" and the "Primorsky" sam- ple groups of the species were equal to 3.2%. 3) Interspecies differences: developmental and kary- ological correlations Eleven species of the family Hynobiidae may be divid- ed into two groups by their genome size. Two species of the genus Onychodactylus form a group with the largest genomes (104-108 pg). They also have the longest embryonic and larval development periods (Table 3), as well as the greatest number of chromosomes (Table 4). Another group includes the remaining nine species (33-67 pg). Among them, Salamandrella keyserlingii has the largest genome size, and the seven species from the genera Hynobius and Batrachuperus have smaller sizes. Ranodon sibiricas has an intermediate genome size. Such a distribution of genome sizes in the second group does not seem to be associated with ovum diame- ter, and, perhaps, with time of embryonic and larval development (Table 3). The comparison of genome size values in the sec- ond group with karyological data evidenced no signifi- 2004 cant relations between the nuclear DNA content and diploid chromosome numbers (Table 4). Nevertheless, we found the positive correlation (r = 0.9998) between genome size and uniarmed macrochromosome numbers. 4) Intergeneric relationships Based on recent studies (Fei and Ye, 2000a; Fu et al., 2001), approximately eight or ten genera of the hynobi- ids could be recognized. The most speciose genus, Hynobins, consists of about 24 species, which may be arranged into three subgenera (Matsui et al., 1992; Mizuno et al., 1995; Borkin, 1999), namely Pseudosalamandra (7 species from Japan and Taiwan), Satobius (1 species from Hokkaido Island), and Hynobius (11 species from Japan, and, perhaps, 5 species from Korea and China). The family also includes the genera Liua (1 species), Onychodactylus (2 species), P achy hynobius (1 species), Protohynobius (1 species), Pseudohynobius (2 species), Ranodon (1 species), Salamandrella (1 species, which penetrates to eastern Europe), and Batrachuperus (sensu lato) (about 9 species). The taxonomic position of Ranodon , Liua, and Pseudohynobius was discussed (Fei and Ye, 2000b; Kuzmin and Thiesmeier, 2001). Based on mitochondrial DNA study, T. Papenfuss (Pers. comm, in Fu et al., 2001) showed that the genus Batrachuperus is para- phyletic, and consists of two groups with distinct geo- graphic distributions. The Eastern (Chinese) group belongs to Batrachuperus (sensu stricto) and consists of about six species (Fei and Ye, 2000b; Fu et al., 2001; Song et al., 2001). The western (Iran and Afganistan) group includes two or three species (Stock, 1999), and may be allocated to Paradactylodon. Various authors suggested several configurations of evolutionary relationships among hynobiids. For instance, based on reproductive biology characters, Thom (1969) proposed to recognize two families: Ranodontidae, with the genus Ranodon only, and Hynobiidae, with remaining genera. Using a set of mor- phological and biological data, Zhao and Hu (1984) sep- arated two "natural" groups among Chinese species: the Hynobius group with predominantly terrestrial species (Hynobius and Salamandrella ), and the Ranodon group with predominantly aquatic inhabitants ( Ranodon , Onychodactylus , Batrachuperus , and Liua). Later, Zhao and Zhang (1985) assigned the genus Pachyhynobius to a third group. Combining morphological characters and mitochondrial DNA data, Larson and Dimmick (1993) found the closest relationships between the genera Salamandrella and Hynobius , thus confirming the tradi- tional acceptance of the similarity of these taxa. However, the genus Onychodactylus was more closely related to that lineage, whereas Batrachuperus (the east- ern group) proved to be more distant. Recently, Fei and Ye (2000a) have erected a new subfamily for the newly discovered Protohynobius puxiongensis, whereas all other hynobiids were allocated to another subfamily. Genome size data are not in agreement with all these suggestions, which did not recognize separate position of the genus Onychodactylus. Apart from the largest genome size, distincton of the genus was also supported by morphological and karyological data (Morescalchi et al., 1979; Olmo, 1983; Kohno et al., 1991; Iizuka, Yazawa, 1994; Kuro-o et al., 2000; Litvinchuk and Borkin, 2003). 5) Taxonomic considerations Based on the intraspecific analysis, Bassarukin and Borkin (1984), and Borkin (1994) outlined the peculiar- ities of Salamandrella keyserlingii from the southern part of the Russian Far East. Moreover, these authors suggested that local populations would be considered as a geographic race of the species (the species type terri- tory is the Kultuk Village, Baikal Lake, Irkutsk Province, Russia; restricted by Borkin, 1994). Indeed, the "Euro-Siberian" and "Primorsky" groups of popula- tions are different in allozymes (Litvinchuk et al., 2001a), in some morphological characters (Ostashko, 1981; Bassarukin and Borkin, 1984; Borkin, 1994; Litvinchuk and Borkin, 2003), in breeding sites (Kuzmin, 1990), and in the shape of egg sacs and time of larval development (Korotkov, 1977; Bassarukin and Borkin, 1984; Sapozhnikov, 1990; Vorobyeva et al., 1999). Therefore, geographic differences in genome size revealed by us are in concert with differences in other characters. Based on a concordance of various charac- ters, including genome size, we would recognize a dis- tinct status of the "Primorsky" samples, at least, of a sub- specific rank. Formerly, Nikolsky (1906) and Dybowski (1928) have coined the names Salamandrella keyser- lingii var. tridactyla and Salamandrella keyserlingii var. kalinowskiana, respectively, for animals from the Russian Far East. Obviously, the first name has the pri- ority. Therefore, populations from Russian Far East (Primorsky Territory) should be named Salamandrella keyserlingii tridactyla Nikolsky, 1906. The range of Ranodon sibiricus is quite limited. Nevertheless, it consists of several semi-isolated areas (Brushko et al., 1988; Kuzmin et al., 1998; our data). The populations from the upper parts of Borokhudzir and Oy-Saz Rivers are separated from each other by a mountain range. Therefore, it is not surprising that the genome size difference (RD) between them is relatively large (2.2%). However, geographical variation in the species is poorly studied, and so the taxonomic value of that difference is still unclear. Vol. 10, p. 289 Asiatic Herpetological Research 2004 Acknowledgments We thank A. G. Borissovsky (Izhevsk), V. G. Ishchenko (Ekaterinburg), V. N. Kuranova (Tomsk), J. C. Lacroax (Paris), I. V. Maslova (Spassk-Dal'ny), N. Panfilova (Almaty), M. V. Pestov (Nizhniy Novgorod), G. P. Sapozhnikov (Vilnus), V. T. Sedalishchev (Yakutsk), E. I. Vorobyeva (Moscow), Y. Zhuravlyev (Almaty), and the staff of Almaty and Moscow Zoo terraria for help us in the field or providing animals. We also thank Theodore Papenfuss (Berkeley) for valuable comments and English corrections of the earlier version of the paper. The research was funded by the INTAS (grant no 97-11909), the Russian Foundation for Basic Research (the grant No. 02-04-49631), and the Federal State Program "Integration" (the grant No. E-0121). Literature Cited Azumi, J., and M. Sasaki. 1971. Karyotypes of Hynobius retardatus Dunn and Hynobius nigrescens Strineger. Chromosome Information Service 12:31-32. Bachmann, K. 1972. Nuclear DNA and developmental rate in frogs. Quarterly Journal of Florida Academy of Sciences 35:227-23 1 . Bachmann, K. and M. Nishioka. 1978. Genome size and nuclear size in Palaearctic frogs ( Rana ). Copeia 1978:225-229. Bassarukin, A. M., and L. J. Borkin. 1984. 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E-mail: schaedla@asu.edu Abstract. - I observed feeding by the Agamid lizard Calotes emma during the early part of the Thai monsoon season. During this period, one individual took advantage of swarming termite reproductives and fed noctumally. Nocturnal activity has not been reported for this genus. The lizard's behavior may have resulted from conditions created by arti- ficial lighting. Alternately, it may constitute a normal response to a rich annually-available food resource. Key words. - Calotes emma, feeding, termites, nocturnal, Introduction Agamid lizards belonging to the genus Calotes are widely distributed throughout South and Southeast Asia. They are characterized by semi-arboreal or arboreal behavior, strongly diurnal activity patterns, and insectiv- orous diets (Erdelen, 1988; Gunther, 1864; Subba Rao 1970, 1975; and Subba Rao and Rajabai, 1972). Calotes emma is a typical member of the genus. The species occurs from Assam through Yunnan in the north, and Peninsular Thailand in the south. It prefers moist forested habitat and is arboreal to semi-arboreal it its habits (Gunther, 1 864). Here I report observations of C. emma feeding at night in a manner apparently con- trary to norms for the genus. Methods This study took place at the Sakaerat Environmental Research Station. Sakaerat is a scientific and education- al facility located in Northeastern Thailand at 14° 30.46' N Latitude by 101° 55.92' E Longitude. The station grounds cover approximately 80 km2. Small and medium-sized wildlife is plentiful in the area. As of this writing, 70 species of mammals, 50 species of birds, and 25 species of amphibians have been recorded from Sakaerat. Reptile fauna is also abundant -82 species are known to occur (Lawanyawatna and Schaedla, 2000). Of the reptiles, C. emma is among the most common because of the station's abundant forest cover. Seasonal (monsoonal) and perennial dipterocarp forests comprise the bulk of Sakaerat's habitats. I worked specifically in an area of Dry Evergreen Forest, which is a Dipterocarp mosaic containing other floral components. It is a four storied forest. The upper story extends from 2 1 to 40 m in height and consists mostly of Hopea ferrea , H. odorata, Shorea sericeiflora , and atypical behavior. Irvingia malayana. The middle story ranges from 15 to 20 m in height and contains Hydrocarpus ilicifolius, Memocylon ovatum, and Walsura trichostemnon. The lower story is between 4 and 24 m in height and is char- acterized by Baccaurea sapida, Apodytes dimidiate, and Olea salicifolia. Undergrowth is less than 4 m from the ground, leafy, and composed mainly of Ardisia, Canthium, and Clausena. Average humidity at Sakaerat runs about 76% over the course of the year. Average annual precipitation is 1,222 millimeters, and average annual temperature is 26°C. March is the hottest month with a maximum recorded high of 37°C. January is the coolest with a minimum recorded low of 8°C. Sakaerat generally experiences a 3.5 month long rainy season that lasts from early June through mid September. Conversely, rainfall is rare from December through February (Tongyai, 1980). I made behavioral observations of Sakaerat's Calotes emma on the evenings of 7, 10, and 12 June, 2001. These dates followed the onset of the local mon- soon season, but were not monsoon days themselves. Weather conditions were overcast, but there was no heavy or sustained precipitation. Rainfall was light and intermittent, accompanied by occasional lightning. Ambient temperature was moderate, ranging from 27° to 30°C. My observations took place from approximately 6:00 PM (dusk) to 9:00 PM. I watched from an area in a semi-secluded part of the research station. My vantage was the front porch of a bungalow near Sakaerat's sta- tion headquarters, but offset in the forest and away from the main complex of office, visitor's center, cafeteria, and general housing. This area is dimly lit by two over- head fluorescent lights attached to the sides of buildings. These lights attract large numbers of insects, especially during seasonal monsoon periods. In particular, termite alates (winged reproductives) were present in high num- © 2004 by Asiatic Herpetological Research Vol. 10, p. 296 Asiatic Herpetological Research 2004 bers on the evenings I observed. Large swarms of Odontotermes sp. and Macrotermes sp. clouded the local area and eventually dropped to the ground. When termites fell to the ground, 1 observed a sin- gle C. emma feeding them. The lizard was present all three rainless nights, and seemed unperturbed by my presence. It was there from dusk, or before, to nearly 9:00 PM on all three evenings. It was active, and its behavior was restricted to the terrestrial environment. It did not climb nearby trees or the sides of wooden build- ings. All of its movements were directed; it displayed no signs of disorientation in the relative darkness of its surroundings. On the contrary, it focused on the termite alates and fed vigorously on them as they landed. On June 10, I captured the lizard to verily its iden- tity as C. emma. It had been correctly identified and was a mature female. Her stomach was distended from ter- mite consumption, but she was not visibly gravid with eggs. In addition to the lizard, other predators feeding on the termite alates included toads (Bufo melanostictus), geckoes ( Cyrtodactylus spp .), and centipedes (Scolopendr amorpha). Discussion Little is known about the exact feeding preferences of C. emma. However, some indication of its diet might be inferred from studies of a closely related arboreal species. Subba Rao (1972) found that C. nemoricola in India fed mostly on ants, while Sitana ponticeriana, an unrelated ground-dwelling lizard, fed on termites. In another study he found that C. nemoricola consumed a wide variety of invertebrates ranging from beetles, to gastropods, to earthworms. Analysis of gut contents showed a predominance of ants but no termites. He also noted a distinct absence of flying insects in the lizards' stomachs (Subba Rao, 1975). Hence, feeding on winged termites by C. emma may represent a departure from its normal dietary habits. Likewise, nocturnal feeding has not been reported in the literature surrounding Calotes. In fact, members of the genus are usually active only during the day (Erdelen, 1988; Gunther, 1864; Subba Rao 1970, 1975; and Subba Rao and Rajabai, 1972). C. emma at Sakaerat is decidedly diurnal in its habits. At night they tend to sleep on the ends of low-hanging tree branches. Spotlighting does not wake them and they can be cap- tured easily by anyone walking through the forest with a headlamp. I have worked at Sakaerat for three years, and, with the exception of the observations reported herein, I have never seen them active at night. Calotes is apparently physiologically predisposed towards diurnal activity. Light has a positive affect on both the pituitary and the hypothalmamo-neuro secreto- ry systems of C. versicolor (Banerjee, 1972). However, Kar (1987) found that day length (photoperiod) played a less important role than ambient temperature in scale regeneration by C. versicolor. He speculated this hap- pens because the lizards' thyroid activity is elevated by heat, rather than light. Evening temperatures were warm on the nights I made my observations. The lizard I observed may have been able to extend her activity because of this. It is also possible that the lack of sunlight was mitigated by the presence of overhead florescent lights (albeit dim). My subject may simply have been disrupted from its normal circadian routine by the local environment. Some sup- port for this possibility comes for another observational study. Subba Rao et al. (1984) noted that abrupt changes in light intensity, temperature, and relative humidity dur- ing a total solar eclipse actually stimulated activity in C. versicolor. Of course, the feeding I observed may have been part of C. emma' s normal behavioral regime. Termite alates constitute a rich, but seasonally discrete food source, induced by the onset of the monsoons. They are available in great numbers at specific times of the year, and they attract a wide variety of predators, including retiles. It is therefore possible that C. emma takes advan- tage of the bonanza via seasonal changes in behavior. Whatever the case, nocturnal feeding by C. emma is unusual and unreported. Even if not anomalous it deserves future attention. Literature Cited Banerjee, S. K. 1973. Effect of light and darkness on the hypothaamo-neurohypophyseal system of the gar- den lizard, Calotes versicolor. Experientia 29(6):7 13-714. Erdelen, W. 1988. Population dynamics and dispersal in three species of Agamid lizards in Sri lanka: Calotes calotes , C. versicolor , and C. nigrilabris. Journal of Herpetology 22(l):42-52. Gunther, A. C. L. 1864. The reptiles of British India. Robert Hardwick, London. Kar, A. 1987. Relative importance of temperature and photoperiod on the physiology of Indian garden lizard, Calotes versicolor. Current Science 56(10):497-499. Lawantayana K. and W. H. Schaedla. 2000. A Brief Overview of the Sakaerat Environmental Research Station in Thailand. Tigerpaper 27(2):29-32. 2004 Asiatic Herpetological Research Vol. 10, p. 297 Pomprasertchai, K. and D. Disbanjong 1998. Changes in the forest area at Sakaerat Environmental Research Station (In Thai). Thai government publication. Subba Rao, M. V. 1975. Studies on the food and feeding behaviour of the Agamid garden lizard Calotes ver- sicolor. British Journal of Herpetology 5(4):467- 470. Subba Rao, M. V. 1970. Studies on the biology of two selected lizards of Tirupati. British Journal of Herpetology 4:151-154. Subba Rao, M.V. and B. S. Rajabai, 1972. Ecological aspects of the Agamid lizards Sitana ponticeriana and Calotes nemoricola in India. Herpetologica 28:285-289. Subba Rao, M. V., A Eswar, and K. Kameswara Rao, 1984. Effect of pre, during, and post total solar eclipse on the activity pattern of an Agamid garden lizard, Calotes versicolor , Daudin. Journal of Environmental Biology 5(3): 197-201. Tongyai, M. L. P. 1980. The Sakaerat Environmental Research Station, Its role as a knowledge base for the determination of forest lands conservation poli- cies for establishing maximum sustained yields on forest resources. Thai government publication. Karyological Studies on Amphibians in China Yuhua Yang Molecular Biology Labortary, Department of Bioengineering, Sichuan University, Chengdu, 610064, R.P.China Abstract. - Since 1978, 79 species of anurans have been studied karyotypically using conventional Giemsa staining and various banding techniques. The chromosome numbers are 2n=22-64 and the karyotypes are variable. The homo- morphic and heteromorphic sex chromosomes of some species have been identified. The mitotic and meiotic chromo- somes of 14 species in two families of Urodela have also been investigated. The family Hynobiidae is karyotypical- ly more primitive than the family Salamandridae. Key words. - Karyology, amphibian, Anura, Urodela. Introduction Karyological studies of amphibians in China were pio- neeered by two scholars. In 1952, famous cytologist T. C. Hsu, an American of Chinese origin, developed the hypotonic technique for chromosome separation and observation. In 1956, cytogeneticist J. H. Tjio, a Swede of Chinese origin, reported that the number of human chromosomes is 46, not 48. In China, karyological stud- ies of amphibians began in 1978. Ninety-three species of amphibians have been studied so far, about 42% of the 220 living amphibian species in China (Table 1). Changes of chromosome numbers. - Kuramoto (1990) summarized the chromosome numbers of 983 species of anurans from 21 families (2n= 14-64). Chromosome number changes occurred in 12 of 21 families with poly- ploids in some species. In China, the chromosome num- bers of 79 species from seven families are 22-64 and variations of chromosome numbers were found in four genera, three families (Table 2). Only two species in the family Discoglossidae have been studied: Bombina orientals and B. maxima. Tian and Hu (1985) subdivided the genus Bombina into two subgenera, Bombina and Glandula. Bombina ( B .) orien- tals has 2n=28 chromosomes (Zhao, 1986), consistent with the chromosome number of Discoglossus pictus in Europe (Morescalchi, 1965; Schmid et al., 1987). Bombina (G.) maxima has 2n=24 chromosomes (Jiang et al., 1984), as reported by Okumoto (1974) and Schmid et al. (1987). The geography of the Hengduan Mountain Ranges greatly influence the evolution of pelobatid frogs, pro- viding refugia for some species as well as discontinuous population distributions that promote allopatric specia- tion (Yang et al., 1983; Hu et al., 1985). The pelobatids distributed in the Hengduan Mountain region have dis- tinct morphological differences adaptive to the unusual geographic conditions. These species belong to two sub- families, Megophryinae (Br achy tar sophrys and Atympanophrys ) and Oreolalaxinae ( Scutiger , Vibrissaphora, Leptobrachium, and Oreolalax). Available karyotypical information showed that species studied have 2n=26 chromosomes, but karyotypical dif- ferentiations are prominent in the family. The subgenus Vibrissaphora is the most specialized, all five species have 2n=26, consisting of six pairs of large chromo- somes and seven pairs of small ones, NF=52 and one stable secondary constriction is located in 6q (in the NoRs region; Zhao et al., 1983). The karyotypes of three species in genus Scutiger are similar to those of sub- genus Vibrissaphora , except the secondary constriction located in 2P not in 6q. Polymorphic chromosome num- ber occured in Oreolalax. Two of the three specimens of O. schmidti are 2n=28 and one is 2n=26 (Zheng andWu, 1989). A similar phenomenon was found in O. liang- beiensis and C-banding showed that the extra pair of small chromosomes is C-band negative and not B chro- mosome (Li et al., 1990). Polymorphic chromosome numbers were also found in three genera of Megophryinae. Wu (1987) observed one male triploid in Atympanophrys shapingensis. Tan et al. (1987) reported the karyotypes of Brachytar sophrys carinensis. Among 12 individuals examined, seven males and one female have 2n=26, three males have 2n=27, and the remaining male has 2n=28. The extra chromosomes are metacen- tric and between No. 10 and No. 11 in size. In addition, there are four pairs of small chromosomes which are telocentric in the2n=26 karyotype and two pairs of small telocentric chromosomes were observed in M. omei- montis (Wu, 1987; Zheng and Wu, 1989). The karotypes of pelobatids reported by foreign authors are 2n=26, NF=52 with no polymorphic chro- mosome numbers found, except Leptolalax pelody- toides with 2n=24 (Morescalchi, 1973; Morescalchi et al. 1977; Schmid, 1980, 1987). It is interesting that a special karyotype was observed in Rana phrynoides dis- tributed in Hengduan Mountains. In this species, 2n=64 consisting of all telocentric chromosomes. Only one homologous pair of NORs being found in interstitial © 2004 by Asiatic Herpetological Research Vol. 10, p. 299 Asiatic Herpetological Research 2004 Table 1. A list of amphibian species studied karyologically. Famlies Number of species Known Number of species studied Number of species Banded Anura: Discoglossidae 4 2 0 Pleobatidae 46 18 12 Bufonidae 13 4 4 Hylidae 6 4 1 Ranidae 77 35 24 Rhacophoridae 32 7 3 Microhylidae 14 9 4 Total anurans 192 79 48 Caudata: Hynobiidae 17 7 0 Salamandridae 17 7 2 Total caudates 34 14 2 segment of No. 20 chromosome, i.e, position of the sole secondary constriction, and No. 32 being sat-chromo- somes (Liu and Zan, 1984; Wu and Zhao, 1984). This karyotype is unique for anurans. Karyological Studies on wood frogs in China. - In China, wood frogs include five species ( R . altaica, R. amurensis, R. japonica, R. chaochiaoensis, and R. chensinensis', Tian and Jiang, 1986). Rana chensinensis had been called R. temproaria chensinensis (Pope and Boring, 1940; Liu and Hu, 1961). Wu (1981) reported the karyotype of R. chensinensis from Beijing, which has 2n=24 chromosomes, including six pairs of large chromosomes (relative length>7%) and six pairs of small ones (relative length<6%), while R. temporaria in Europe has 2n=26 (Guillemin, 1967), including five pairs of large chromosomes and eight pairs of small ones. Consequently, he suggested that R. chensinensis should be a good species, not a subspecies of R. tempo- raria. Wei et al. (1990) compared the C-bands and NORs between R. chensinensis from type locality and R. temporaria in Europe. As a result, in the former species, centric C-bands located in Nos. 9 and 10 chromosomes and telocentric C-bands in the terminations of a few chromosomes, 28 interstitial C-bands, one pair of stan- dard NORs in llq, and small additional NORs are found. However, in the latter species, both centric and telocentric C-bands are located in all chromosomes and only three interstitial C-bands and 1 pair of NORs in lOq were developed (Schmid, 1978). Evidently, this compar- ison supports Wu’s suggestion. Jiang et al. (1984), Luo and Li (1985) and Ma (1987) indicated that R. chensinensis from different localities have the same 2n=24 pattern, but the numbers of subtelocentric chromosomes and the positions of sec- ondaiy constrictionas are locality specific by comparing the karyotypes of R. chensinensis from Beijing, Qingdao, Lanzhou, Harbin, Hongyuan and Yanbei. Consequently, it was suggested that R. chensinensis might contain different subspecies . The five species of wood frogs have 2n=24 or 2n=26 chromosomes and may be divided into two groups: R. japonica , R. chaochiaoensis and R. amuren- sis belong to 2n=26 group and R. chensinensis and R. altaica to 2n=24 group. The karyotypic differences between two groups are listed in Table 3. Rana japonica from Hiroshima, Japan has 2n=26 chromosomes, Nos. 8 and 9 are subtelocentric and the secondary constriction is located in 9q; R. amurensis coreana from Korea has 2n=26, Nos. 10 and 13 subte- locentric and secondary constriction in 9q; while R. chensinensis from Hokkaido, Japan has 2n=24, No.l 1 is subtelocentric and the secondary constriction in lOq (Nishioka, et al., 1987). Matsui (1991) described R. chensinensis from Hokkaido as a new species, R. pirica, based on morphometric and electrophoretic studies. The No. 6 chromosome of the species having 2n=24 is nearly the same in relative length as the sum of chro- mosomes No. 11 and No. 12 or No. 13 of the species with 26 chromosomes, For instance, the average relative length of No. 6 chromosomes of R. chensinensis and R. altaica is 8.19, while the average relative lengths of Nos. 11, 12 and 13 chromosomes of the other three species are 4.52,4.31 and 3.82 respectively. In addition, it is clear from Table 3 that there are more subtelocen- tric chromosomes and secondary constrictions in the species having 26 chromosomes than those in the species having 24 chromosomes, and they are con- cerned with small chromosomes. Accordingly, it is speculated that all wood frog species would have a com- mon ancestor, from which the species having 26 chro- mosomes were derived and the species having 24 chro- mosomes evolved via fusion of two pairs of small chro- mosomes of the former. Then, the species possessing 2004 Vo). 10, p. 300 Table 2. A summary of chromosome numbers of 79 anuran species. * = Polymorphic chromosome number occurs in some species. Taxon Number of species 22 24 26 28 64 Total Discoglossidae Bombina 1 1 2 Pelobatidae Atympanophrys . 1 - - 1 1 Brachytarsophy - - 1 - - Megophrys* - - 2 - - 2 Oreolalax* - - 6 - - 6 Scutiger - - 3 - - 3 Vibrissaphora - - 5 - - 5 Bufonidae Bufo 4 _ _ - 4 Hylidae Hyla 4 2 _ - 6 Ranidae Rana 1 2 21 _ 1 26 Amolops - - 7 - - 7 Ooeidozyga - - 2 - - 2 Altirana - - 1 - - 1 Rhacophoridae Rhacophorus . 5 _ _ 5 Philautus - - 1 - - 1 Polypedates - - 1 - - 1 Microhylidae Microhyla _ 4 1 _ _ 5 Kaloula - - - 1 - 1 Kalophrynus - - 1 - - 1 Total 5 11 60 2 1 79 different subtelocentric chromosomes and secondary constriction positions were developed into two groups through inversions and translocations. The high resolu- tion R-bands of R. japonica were prepared and analyzed (Heng, 1984), providing a practical technique for study- ing karyotypic evolution of amphibians. Sex chromosomes and sex-determining mechanisms. - The first successful demonstration of sex-determining mechanism was made by making use of reversal and breeding experiments (Humphrey, 1942, 1945, 1957). The applications of cytogenetic techniques, such as C- banding, quinacrine mustard staining, Ag-NORs stain- ing and in situ hybridization of nucleic acids, have been helpful to the investigations on sex chromosomes and sex-determining mechanisms in amphibians. So far, eleven species with cytologically detected sex chromo- somes, including XY and ZW systems, even an OW/OO system of sex determination and multiple sex chromo- somes in one genome(Schmid et al., 1992) were discov- ered. Few sex-specific chromosome pairs in heteroga- metic individuals are heteromorphic and most of them are homomorphic. The homomorphic chromosome pair No.4 in Rana esculenta was identified as sex-specific chromosomes of XX/XY type by BrdU replication banding technique. All males have an extremely late-replication band in the long arm of Y, which is lacking in the X (Schempp and Schmid, 1981). The homomorphic chromosome pair No. 10 in Bufo gargarizans was demonstrated to be sex chromosomes of ZZ/ZW type. The Z chromosomes in all males replicated synchronously, while Z and W chro- mosomes of females revealed heteromorphic replication bands at the late replication stage. There was a replica- tion band on W chromosome’s long arm and Z chromo- some lacked the band (Wen et al., 1982;Shang and Deng, 1982). Similarly, the chromosome No. 6 pair in Bufo raddei was identified as XX/XY sex chromosomes (Deng and Shang, 1984) and the No. 9 chromosomes of Rana nigromaculata as XX/XY sex chromosomes (Wu and Zhang, 1985). The sex chromosomes of species mentioned above are homomorphic and could only be recognized by BrdU replication banding. So they are at the initial stage of sex chromosome differentiation. The Y or Z chromosomes of homomorphic sex chromosomes in some anurans heterochromatinized so Vol. 10, p. 301 Asiatic Herpetological Research 2004 Table 3. karyotypic comparison between wood frog groups. Species 2N Subtelocentric pair(s) Position of S.C. R. japonica 26 No. 7 or No. 9 2-5p,6-7q R. chaochiaoensis 26 No. 8 Nos. 9, 10, 12, 13 5-7p,8q,10p R. amurensis 26 8q R. chensinensis (from type locality) 24 No. 9 1 1 q R. altaica 24 — 1q highly that they could be recognized by C-banding or other specific staining of constitutive heterochromatin, for example, the XX/XY sex chromosomes in genus Triturus (Schmid, et ah, 1979). Chinese scholars Wu and Chen (unpublished data) determined the homomorphic chromosome pair No. 9 as XX/XY sex chromosomes in Rana margaratae using C-banding and quinacrine mus- tard staining. There is one interstitial C-band, i.e, the brightest fluorescense band, on both No. 9 chromosomes in females. The interstitial C-band is located only in one No. 9 chromosome, while one telocentric C-band in the other No. 9 chromosome shows no fluorescense differen- tiation. The first-discovered highly heteromorphic ZW type sex chromosomes occured in Pyxicephalus adspersus (Schmid, 1980). The W chromosome is much smaller than Z chromosome and its short arm is completely het- erochromatic. Wu and Zhao (1984) and Wu et al. (1987) demonstrated that Amolops mantzorum has well-differ- entiated XY type sex chromosomes, the Y chromosome is subtelocentric and mainly composed of euchromatin, but has strong C-band in the middle of long arm and X chromosome is metacentric by conventional Giemsa staining and C-banding methods. Karyological studied in urodeles. - Only two species were studied by C-banding and the others by conven- tional Giemsa staining. The karyotypic comparisons are listed in Table 4. The family Hynobiidae has a wide geo- graphical distribution. Twenty-six species in five genera out of more than 34 species in eight genera have been studied karyologically. The chromosome number vary from 40 to 80 . Twelve species in Hynobius and one in Salamandrella had been studied by C-banding, Ag- NORs staining, and R-banding. The relationships in the two genera were discussed by comparing banded kary- otypes and Southern hybridizations. It is suggested that the family is the most primitive living caudate (Morescalchi, 1973; Kohno et al., 1991).The same con- clusion is derived from morphological comparisons (Zhao and Hu, 1984). There are 17 species in seven genera in Hynobiidae known from China. The family can be divided into two groups: Hynobius group and Ranodon group. The Ranodon group evolved by adaptation towards two dif- ferent life-forms: aquatic and terrestrial. Liua and Batrachuperus are aquatic and are closely related. Table 4 shows that seven species have high chromosome num- bers: 2n=62-68. Their karyotypes are bimodal and sym- metrical, with more microchromosomes (Yang, 1992). Salamandrella keyserlingii has 2n=62 chromosomes, the karyotype formula being 4M + 2SM + 10ST + 10T + 36 m (Wang et al., 1983) in accordance with that reported by Morescalchi (1975), Morescalchi et al. (1979), Grafodatsky et al. (1978), Kuro-o(1986), and Ikebe et al. (1990). The bivalent number in cells of male Batrachuperus pinchonii in diakinesis is 3 1 . We expect the diploid chromosome number to be 2n=62 (Yang and Zhao, 1984), but this species also has 2n=66 chromo- somes (Kuro-o et al., unpublished data). There is little detailed information on the cause of the difference. The karyotypic differentiation of Hynobiidae is more complex. First, variations in diploid chromosome number occurred not only at the intergeneric level, but also at the intrageneric level. For instance, Salamandrella has 2n=62, Batrachuperus 2n=62-68, both Liua and Pachyhynobius 2n=64. Secondly, the numbers of microchromosomes vary from 36 to 46. Finally, the morphology of microchromosomes are vari- able. The numbers of M, SM, and ST are species-specif- ic. Telocentric macrochromosomes were found in S', key- serlingii, L. shihi and P. shangchengensis, but not in Batrachuperus. The L. shihi and P. shangchengensis studied were from the type localities (Zhao and Hu, 1983; Fei et al., 1983). Both two species have 2n=64 chromosomes, different from the known genera. However, they are different in morphology of macrochromosome and microchromosome number (Table 4) . Therefore, the cytogenetic data provide evi- dence supporting those genera and species. There are four species in Batrachuperus. Batrachuperus karlschmidti and B. yenyuanensis are only found higher than 3000M in the Hengduan Mountains, having 2n=68, no telocentric macromo- somes and 44 and 46 microchromosomes respectively. Batrachuperus pinchonii and B. tibetanus distributed higher than 1600M of Hengduan Mountains and adja- cent areas have 2n=62. Obviously, the karyotypic differ- 2004 Asiatic Herpetological Research Vol. 10, p. 302 Table 4. Karyological comparisons of urodela in China. M-metacentric macrochromosomes, SM-submetacentric macrochromosomes; ST-subtelocentric macrochromosomes; T-telocentric macrochromosomes, m-microchromo somes. Species 2N Number of Bivalent M SM ST T m Band Hynobiidae Salamandrella keyserlingii 62 4 2 10 10 36 Batrachuperus karlschmidti 68 6 - 18 - 44 — B. yenyuanensis 68 4 2 16 - 46 — B. pinchonii (62) 31 — B. tibetanus 62 — Liua shihi 64 6 2 4 10 42 — Pachyhynobius shangchengensis 64 4 - 2 18 40 — Salamandridae Tylototriton kweichowensis 24 12 16 6 2 - - — T. verrucosus (24) 12 — Cynops cyanurus yunnanensis (24) 12 — C. orientalis 24 12 16 8 - - - C Paramesotriton chinensis (24) 12 — Pachytriton brevips 24 16 8 - - - c P labiatum (24) — entiations in Batrachuperus are in conformity with the geographic distribution. Salamandridae is the advanced family in Caudata either from the viewpoint of karyotypic information or from that of morphological characteristics (Morescalchi, 1973, 1975, 1979; Zhao and Hu, 1984; Yang, 1992). There are five genera of Salamandridae found in China: Tylototriton, Echinotriton, Cynops, Paramesotriton, and Pachytriton. Of the five genera, Tylototriton is the most primitive and Pachytriton the most advanced . The seven species studied in five gen- era have the same chromosome number, 2n=24, without microchromosomes. The karyotypes are unimodal and symmetrical (Table 3). Interspecific differences were found in Tylototriton. The karyotypic formula of T. kwe- ichawensis is 16M+6SM+2ST, the same as that of T. verrucosus. However, Nos. 6, 8 and 11 are submetacen- tric and no secondary constriction was found in the for- mer (Yang, 1990), while Nos. 6, 7 and 11 are submeta- centric and secondary constriction were found at every chromosomes except No. 12 in the latter (Seto et al., 1982). Echinotriton andersoni has 2n=24 chromosomes, 14M + 8SM+ 2 ST, one more submetacentric chromo- some pair than both T. kweichowcensis and T.verrucosus (Seto et ah, 1982). In addition, the relative length of chromosome No. 1 is the longest and that of No. 12 the shortest in E. andersoni among the three species men- tioned above. The karyotypic formulas of Pachytriton brevipes and Cynops orientals are nearly identical and only the differences of C-band patterns are found (Zhu and Wei, 1981). The predominant mode of karyotypic evolution in Caudata is that the unimodal symmetrical karyotypes with fewer chromosome number are derived from the bimodal and asymmetrical karyotypes with more chro- mosome number, via Robertsonian centric fusions and pericentric inversions (Morescalchi, 1975). Robertsonian centric fusions, which could occur between telocentric macrochromosomes, between stable microchromosomes, and between telocentric macrochromosomes and stable microchromosomes, cause reduction of the diploid number and the microchromosome number and increase of the metacen- tric chromosomes. Consequently, the karyotypes tend toward stability. Pericentric inversions do not change the diploid number, but could increase the number of meta- centric chromosomes and the stability of karyotypes. There are some differences in the evolutionary trends of Hynobiidae and Salamandridae. The karyotyp- ic evolution in Hynobiidae involves Robertsonian cen- tric fusion as well as pericentric inversion. However, the phylogeny of the family could not be established on the available data. It is obvious that the karyotypes of Salamandridae are more stable than those of Hynobiidae. Morescalchi (1975) proposed that all species studied possess similar karyotypes that differ very little even at the intergeneric level. The differences between the karyotypes mainly lie in the absolute size of chromosomes and quantity of DNA. Accordingly, the karyotypic diversity among the species has chiefly resulted fiom pericentric inversions and reciprocal translocations that result in differences between individ- ual chromosomes by changing the telocentric chromo- somes into metacentric ones or changing the metacentric chromosomes into submetacentric, subtelocentric and telocentric chromosomes. Vol. 10, p. 303 Asiatic Herpetological Research 2004 Acknowledgments I want to express my thanks to three professors: Zhao Ermi, Chen Wenyuan and Wang Xizhong. Under their encouragement and guidence this paper was worked out. Sincere thanks are also extended to professor Zheng Xuejing, who read and made some corrections to this paper. Literature Cited Deng, C. and K. Shang. 1984. A cytogenetics demon- stration of XY sex determination in Bufo raddei. Acta Genetica Sinica. 11:395-399. (In Chinese). Fei, L. and C. Ye. 1983. A new genus and species of Flynobiidae from Henan, China. Amphibian Research 1:1. (In Chinese). Grafodatsky, A. S., O. V. Grigoriev, and A. A. Isaenko.1978. Differential staining of chromosomes in four species of amphibians. Zoological Zhumal 57:1279-1281. Heng, H.-Q. 1984. Studies on high resolution R-band of the chromosomes of Rana japonica japonica. Acta Herpetological Sinica. 3(2):55-59. (In Chinese) Hu, Q., Y. Jiang, and E. Zhao. 1985. Studies on the influ- ence of the Hengduan Mountains on the evolution of the amphibians. Acta Herpetological Sinica 4(3):225-233. (In Chinese). Ikebe, C., M. Kuro-o, T, Yamamato, and S. Kohno. 1990. Cytogenetic studies of Hynobiidae (Urodela). XI. Banding karyotype of Salamandrella keyser- lingii Dybowski and a comparison with those of Hynobius species. Cytogenetic Cell Genetics 54:169-171. Jiang, S., C. Shen and Y. Meng. 1984. Preliminary observations on the karyotype of Bombina orien- tals. Acta Herpetological Sinica 3(1): 19-23. (In Chinese) Kohno, S., M. Kuro-o, and C. Ikebe. 1991. Cytogenetics and evolution of Hynobiid salamanders. In D. M. Green and S. K. Session (eds). Amphibian Cytogenetics and Evolution. Academic Press., San Diego. Kuramoto, M. 1990. A list of chromosome numbers of Anuran amphibians. Bulletin of Fukuka University of Education 39:83-127. Kuro-o, M. 1986. Cytogenetic studies of genus Hynobius by means of DNA repliction pattern (R- banding). M. Sc. Thesis. Toho University, Funabashi, Japan. Li, S., L. Fei and C. Ye. 1990. Studies of karyotype, Ag- NORs and C-banding on five mountain pelobatoid toads from China. Acta Zoologica Sinica. 36:315- 323 (In Chinese). Liu, C. and S. Hu. 1961. Chinese Tailless Amphibians. Science Press, Beijing, China. 184 pp. (In Chinese). Liu, W. and R. Zan. 1984. A special karyotype in the genus Rana - an investigation of the karyotype, C- banding and Ag-stained NORs of Rana phry nodes Boulenger. Acta Genetica Sinica 11:52-60. (In Chinese). Luo, X. and J. Li. 1985. Comparative studies on kary- otypes of Rana temporaria chensinensis from Harbin, Lanzhou, and Hongyuan. Acta Herpetological Sinica 4 (1):5-11. (In Chinese). Ma, T. 1987. The karyotype of Rana chensisnensis found in Yanbei prefecture, Shan-xi Province. Acta Herpetological Sinica 6(l):70-73. (In Chinese) Matsui, M. 1991. Original description of the brown frog from Hokkaido, Japan (Genus Rand). Japanese Journal of Herpetology 14:63-78. Morescalchi, A. 1965. Osservazioni sulla carioilogia di Bombina. Bolletin Zoology 32:207-218. Morescalchi, A. 1973. Amphibia, pp. 233-348 In: A. B. Chiarelli and Capanna (eds). Cytotaxonomy and Vertebrate Evolution. Academic Press. New York. Morescalchi, A. 1975. Chromosome evolution in the caudate Amphibia. Evolutionary Biology 8:339- 387. Morescalchi, A. E., E. Olmo and V. Stingo. 1977. Trends of karyological evolution in pelobatid frogs. Experientia 33:1577-1578. Morescalchi, A., G. Odierna and E. Olmo. 1979. Karyology of the primitive salamanders, family Hynobiidae. Experientia 35:1434-1435. Nishioka, M., H. Okomoto, H. Ueda and M. Ryuzaki. Karyotypes of brown frog distributed in Japan, 2004 Asiatic Herpetological Research Vol. 10, p. 304 Korea, Europe and North America. Hiroshima University Bulletin 9:165-212. Okumoto, H. 1974. Chromosomes of amphibians. Zoological Magazine 76:39. Takeshi S., Y. Utsunomiya and T. Utsunomiya. 1982. Karyotypes and banding patterns of Tylototriton andersoni Boulenger, a newt endemic to the Ryukyu islands. Japanese Journal of Genetics 57:527-534. Schempp, W., and M. Schmid. 1981. Chromosome banding in Amphibia: Replication patterns in anura and demonstration of XX/XY sex chromosomes in Rana esculenta. Chromosoma 83:697-710. Schmid, M., L. Vitelli and R. Batistoni. 1978. Chromosome banding in Amphibia: Constitutive heterochromatin and nucleolus organizer regions in Ranidae, Microhylidae and Rhacophoridae. Chromosoma. 68:141-148. Schmid, M. 1980. Chromosome banding in Amphibia: Highly differentiated ZW/ZZ sex chromosomes and exceptional genome size in Pyxicephalus adspersus (Anura, Ranidae). Chromosoma 80:69-96. Schmid, M., J. Olert, and J. Klett. 1979. Chromosome banding in Amphibia: Sex chromosome in Triturus. Chromosoma 71:29-55. Schmid, M., C. Steinlein, and W. Feichtinger. 1992. Chromosome banding in Amphibia: First demonstration of multiple sex chromosomes in amphibians: Elentherodactylus massi (Anura, Leptodactylidae). Chromosoma 101: 284-292. Shang, K. and C. Deng. 1983. A cytogenetic demonstra- tion of ZW sex determination in Bufo bufo gar- garizans. Acta Genetica Sinica. 10:298-305. (In Chinese). Tan, A., X. Zeng, G. Wu and E. Zhao. 1987. Cytotaxonomical studies on Chinese pelo- batids: Preliminary study on the karyotype of Brachytarsophrys carinensis and the variation in their chromosome number. Acta Herpetological Sinica 6(2): 1-4. (In Chinese). Tian, W. and Q. Hu. 1985. Taxonomical studies on the primitive anurans of the Hengduan Mountains, with descriptions of a new subfamily and subdivision of Bombina. Acta Herpetologica Sinica 4(3):2 19-224. (In Chinese). Tian, W. and Y. Jiang.1986. Identification guide to the amphibians and reptiles of China. Science Press, Beijing. 120 pp. (In Chinese). Wang, X., J. Fang and X. Tang. 1983. Preliminary obser- vation on karyotype of Salamandrella keyserlingii. Acta Herpetological Sinica 2(2): 19-22. (In Chinese). Wei, G., F. Chen, and N. Xu. 1990. An investigation for the karyotypic, C- banding and Ag-NORs pattern on Rana chensinensis from type locality. Hereditas 12:24-26 (In Chinese). Wen, C, Q. Lu, and W. Xiang. 1983. Studies of chromo- some banding and sister chromatid exchange in Bufo bufo gargarizans. Acta Genetica Sinica. 10:291-297. (In Chinese). Wu, G. 1987. Cytotaxonomical studies on Chinese pelo- batids: The analysis of the karyotypes of Megophrys lateralis and Atymponophrys shapingensis . Acta Herpetological Sinica 6(3):45-48. (In Chinese). Wu, G. and E. Zhao. 1984. A rare karyotypes of anurans, the karyotype of Rana phrynoides. Acta Herpetological Sinica 3(l):29-32. (In Chinese). Wu, G. and E. Zhao. 1984. Two rare karyotypes of anu- rans, the karyotypes of Staurois mantzorum and S. liangshanensis. Acta Herpetological Sinica 3(4):5- 9. (In Chinese). Wu, H. and R. Zhang. 1985. A study of sex chromosome in Rana nigromaculata by BrdU-Hoechst 33258- Giemsa technique. Acta Genetica Sinica 12:462- 469. (In Chinese). Wu, W. and W. Chen. 1990. The studies on banded chro- mosomes of Rana margaratae Liu. M. Sc. thesis. Sichuan University.Chengdu, China. (In Chinese). Wu, Z. 1981. Karyotype of Rana chensinensis from Beijing. Acta Genetica Sinica. 8:138-144. (In Chinese). Wu, Z., A. Tan and E. Zhao. 1987. Cytogenetic studies on four species of Amolops in the Hengduan Range. Acta Genetica Sinica 14:63-68. (In Chinese). Yang, D., C. Su and S. Li. 1983. A study on amphibians and reptiles from the Hengduanshan Mountains of Yunnan. Acta Herpetological Sinica 2(3):37-49. (In Chinese). Vol. 10, p. 305 Asiatic Herpetological Research 2004 Yang, Y. 1992. Karyotypic studies on nine Chinese sala- manders. Asiatic Herpetological Research 4: ISO- 158. Yang, Y. and E. Zhao. 1984. Meiotic chromosomes and chromosome set in male Batrachuperus pinchonii and B. tibetanus. In E. Zhao and Q. Hu (eds). Studies on Chinese tailed amphibians. Sichuan Scientific and Technical Publishing House, Chengdu. (In Chinese). Zhao, E., G. Wu and W. Yang. 1983. Studies on Genus Vibrissaphora (Amphibia: Pleobatidae): A compar- ative study of the karyotypes of the genus Vibrissaphora. Acta Herpetological Sinica 2( 1 ): 1 5- 20. (In Chinese). Zhao,Y. 1986. Studies on the karyotype of Bombina maxima. Acta Herpetological Sinica 5(3):227-228. (In Chinese). Zhao, E. and Q. Hu. 1983. Taxonomy and evolution of Hynobiidae in Western China, with a description of a new genus. Acta Herpetological Sinica 2(2):29- 35. (In Chinese). Zhao, E., Q. Hu., Y. Jiang, Y. Yang. 1988. Studies on Chinese Salamanders. Contribution to Herpetology 4:1-67. Zheng, X. and G. Wu. 1989. Cytotaxonomocal studies on Chinese pelobatids: the karyotypes, C-bands and Ag-NORs of Megophrys omeimontis and Oreolalax schmidti. Chinese Herpetological Research 2:37-45. The History of the Journal Asiatic Herpetological Research Theodore J. Papenfuss Museum of Vertebrate Zoology, University of California, Berkeley, C A 94720, USA The origins of Asiatic Herpetological Research go back 34 years. Our present editor. Professor Ermi Zhao start- ed a scientific periodical. Materials for Herpetological Research, in 1972. Four issues were published, the last in 1978 (Zhao and Adler, 1993). The text of all articles was in Chinese only. The four issues were produced by the Sichuan Biological Research Institute (now the Chengdu Institute of Biology). Materials was followed by Acta Herpetologica Sinica, which was also edited by Zhao. There were two series between 1979 and 1987. The “old series” consisted of six volumes published from 1979-1982 and the “new series” from 1982-1987 (Zhao and Adler, 1993). The articles were in Chinese, but English titles and often, English abstracts were included. Volume 1 of a new journal, Chinese Herpetological Research, also edited by Zhao, was published in Chongqing for the Chinese Society for the Study of Amphibians and Reptiles, with Zhao continuing as edi- tor. In 1988 Zhao visited the Museum of Vertebrate Zoology, University of California at Berkeley as part of his collaboration with J. Robert Macey and E The MVZ’s single Mac Classic computer that could be used for desktop publishing impressed him. At Zhao’s request we agreed to transfer printing and distribution from China to Berkeley. Macey and I also agreed to serve as Associate Editors and to help assemble an inter- national editorial board. We changed the name of the journal to Asiatic Herpetological Research with volume 3 in 1990. With this volume 10, our journal will return to the Chengdu Institute of Biology for printing and dis- tribution. The quality of journals published in China is now of world standard and the Internet, unknown in 1988, allows for easy electronic transfer of manuscripts, and easy editing Although our society is not large in membership, it is very international. We have published articles by authors from 29 countries. Fourteen new species of amphibians and reptiles have been described since 1987. We organized the First Asian Herpetology Meeting, held in China in 1992, the Second Asian Herpetology Meeting held in Turkmenistan in 1995, the Third Asian Herpetology Meeting held in Kazakhstan in 1998, and a Fourth Asian Herpetology Meeting in 1991 again in China. Literature Cited Zhao, E. and K. Adler. 1993. Herpetology of China. Society for the Study of Amphibians and Reptiles. 522 pp. * IT 4- ACTA HEKPETOUX11CA SINICA - k * a ♦ -r n Materials of Acta Herpetologica Acta Herpetologica Chinese Herpetology Sinica "old series" Sinica "new series" Hemetolooical 1972-1978 1979-1982 1982-1987 Research 1987 © 2004 by Asiatic Herpetological Research Vol. 10, p. 307 Asiatic Herpetological Research 2004 2004 Asiatic Herpetological Research Vol. 10, pp. 308-3 1 2 Guidelines for Manuscript Preparation and Submission Summary Manuscripts must: 1) be written in English. 2) be of letter quality (laser printed or typewritten on bond paper). 3) include camera ready figures (if any). 4) include complete and accurate literature citations. 5) include complete and accurate localities with latitude and longitude. 6) include a camera ready map illustrating regions discussed (when applicable). Tips for electronic submission • Do not use multiple tabs or spaces to separate columns in tables. Either use the table feature of your word processor, a spreadsheet program (e.g. Excel), or separate columns with a single tab. Table 1 .Example of improper use of rrmltinlejahs to separate columns in a table. Column 1 Column 2 ^^V^CA^Column 2 Column 2 12.4 .5 9. 1 0.01 12.1 9 1020.4 0.6 0.02 • Do not type authors names in all capitals in literature cited. • Do not use two spaces following a period, or for any other purpose. • Do not attempt to recreate the format of the journal in your manuscript. Please use only simple formatting limited to italics, boldface, and underline. Manuscripts failing to meet these criteria will be returned without review for correction. Purpose and Content Asiatic Herpetological Research publishes articles concerning but not limited to Asian herpetology. The editors encourage publications from all countries in an attempt to create an open forum for the discussion of Asian her- petological research. Articles should be in standard scientific format and style. The following sections should be included: Title The title should reflect the general content of the article in as few words as possible. The editors encouraee titles that summarize the main findings of the article. Names and Addresses The names and addresses of all authors must be complete enough to allow postal correspondence. Please include email and World Wide Web addresses if applicable. Abstract The abstract should briefly summarize the nature of the research, its results, and the main conclusions. Abstracts should be less than 300 words. © 2004 by Asiatic Herpetological Research 2004 Asiatic Herpetological Research Vol. 10, p. 309 Key Words Key words provide an index for the filing of articles. Key words provide the following information (when appli- cable): 1) Taxonomy (e.g. Reptilia, Squamata, Gekkonidae, Gekko gecko). 2) Geography (e.g. China, Thailand). 3) Subject (e.g. taxonomic validity, ecology, biogeography). The order of taxonomy, geography, and subject should be observed. Text Manuscripts must be in English and spelling must be correct and consistent. Use Webster's New International Dictionary for reference. For clarity, use active voice whenever possible. For example, the following sentences in active voice are preferable to those in passive voice. Active voice: “Fizards were extremely common on the site.” and “I examined three female snakes.” Passive voice: “Fizards were observed to be extremely common on the site.” and “Three female snakes were examined.” Abbreviation Do not abbreviate unless the full phrase has already appeared. Scientific names may be abbreviated only if they have appeared fully in the same paragraph. Never begin a sentence with an abbreviation of a scientific name. Statistics Statistics must be accompanied by sample sizes, significance levels, and the names of any tests. Investigators should pay careful attention to independence and applicability of tests, and randomness of samples. One of the most frequent examples of nonindependence is the use of multiple, paired t-tests instead of analysis of variance (anova). In general, multiple tests on the same data set are not valid. Descriptive statistics are in many cases more appropriate than inferential statistics. Standard Format Manuscripts following standard format should include introduction, methods, results, and discussion sections. While other formats are acceptable, the editors encourage the use of standard format. Please do not type in all capital letters. Introduction The introduction typically states the significance of the topic and reviews prior research. Material and Methods This section should clearly state where, when, and how research was carried out. Include sample sizes. Protocols designed by other investigators must be properly cited. Research materials and their manufacturers should be listed. The reader must be able to replicate the methods of the author(s). Results This section states the results and their significance to the investigation. Figures and tables may be used to clarify, but not to replace, results statements in the text. Statistics should be used when applicable. Large amounts of data should be avoided, or included as an appendix at the end of the article. Discussion The discussion is a synthesis of the introduction and the results. No new information should be discussed unless it was presented in the results section. New findings should be discussed in relation to prior research. The author(s) should feel free to present several possible interpretations of the results. The editors particularly encourage suggestions of future research in Asian herpetology. Vol. 10, p. 310 2004 Manuscript Preparation Overview Please do not attempt to replicate the formatting style of AHR in your manuscript. All formatting except italics will be removed in the production process. Bold and underlined text should be used only to identify section head- ing levels (see below). Extraneous formatting is counterproductive and increases the production costs of the jour- nal. There are a few simple guidelines that authors must follow. Section Headings Articles will be published using three section heading styles. All heading levels must be on their own line, and left justified. For the purposes of manuscript submission, Level 1 heading is bold, and generally reserved for Introduction, Material and Methods, Results, and Discussion; Level 2 is italic , and Level 3 is underlined. Figures Figures must be referenced in order in the text. Each figure illustration (line art or photograph) submitted must be “camera ready” for publication with no modifications necessary other than reduction. AHR does not publish “plates”; please refer to these as figures numbered sequentially. Do not write on figure; do not mount more than one figure to a sheet. AHR cannot be responsible for redrawing, touching up, or otherwise modifying figure illus- trations for authors. In addition, figure illustrations submitted must: 1) be of publication quality with typeset text. 2) be mounted on a separate 21.5 x 28 cm (8.5 x 1 1 inch) sheet with figure number on back. 3) be on a separate sheet from figure legend. 4) not have poor type or handwriting on the face of the figure. 5) The TIFF file format is preferable for electronic versions of figures, but Photoshop, JPEG, or PICT formats are acceptable. Resolution of electronic versions of figures must be at least 600 dpi for line art, or 300 dpi for gray- scale and color images. 6) Figures will be reduced to either 1 column (3.25”) or two columns (6.5”). Times Roman typeface is preferred. In order to avoid wasted effort, please follow the above instructions care- fully. Please note: AHR will not alter or lay out figures for publication. Any figure requiring modification will be returned, and may cause significant delay in publication. Figure Legends. Figure legends should be typed on a separate sheet. Legends should explain the figure without reference to the text. A figure and legend should make sense if separated from the rest of the article. For example: Figure 2. Lateral view of live Psammodynastes pulverulentus holding a prey lizard (Anolis car- olinensis ). Note buccal tissue surrounding the enlarged anterior maxillary and dentary teeth of the snake. Color Figures. AHR may publish color figures at the discretion of the editors. AHR is now published both on paper and electronically. Printing costs of color figures may be required for the paper version of AHR. Color fig- ures will be published free of charge in the electronic version. If you submit color figures, please indicate if you wish them to be considered for publication in color in the paper version. Otherwise, they will be converted to black and white in the paper version. Tables Tables must be referenced in order in the text. Each table should be typewritten, double spaced on a separate sheet. For electronic submission, prepare tables as columns separated by one tab only. Do not use spaces to sepa- rate columns. End rows with a single carriage return. Typeface Twelve point type is preferred. Supply a detailed list of special characters (greek letters, male or female symbols, etc.) that are not part of a standard font. 2004 Asiatic Herpetological Research Vol. 10, p. 311 Literature Cited Accurate and standard references are a crucial part of any article. This is especially important when dealing with publications from many different countries. The reader must be able to precisely identify any literature cited. References in the text must be checked for consistency with references in the literature cited section. All refer- ences cited in the text must be in the literature cited section. The literature cited section may not contain any ref- erences not mentioned in the text. Articles containing inaccurate or inconsistent literature citations will be returned for correction. References in Text. 1) References to articles by one or two authors must include both surnames in the order they appear in the original publication. References to articles by more than two authors must include the first author's surname, followed by “et al.” 2) The year of article follows the authors, separated only by a space. 3) References with the same author and year are distinguished by the lower case characters “a, b, c, . . .” 4) References cited in text are listed in alphabetical order by first author. For example, “My results also incorporate literature records (Marx et al., 1982; Marx and Rabb, 1972; Mertens, 1930; Pope, 1929; Wall, 1909, 1910a, 1910b, 1910c).” References in Literature Cited. 1) References must include all authors, in the order that they appear in the orig- inal publication; “et al.” is never used in a literature cited section. 2) The first author is listed surname first, ini- tials) last. All other authors are listed initial(s) first, surname last. 3) References with the same author and year are distinguished by the lower case characters, “a, b, c, . . .” 4) References cited are listed in alphabetical order by first author. 5) Names of journals are not abbreviated. See below for examples: Journal article Dial, B. E. 1987. Energetics and performance during nest emergence and the hatchling frenzy in loggerhead sea turtles ( Caretta caretta). Herpetologica 43(3):307-315. Journal article from a journal that uses year instead of volume Gatten, R. E. Jr. 1974. Effect of nutritional state on the preferred body temperatures of turtles. Copeia 1 974(4):9 1 2-9 17. Journal article, title translated, article not in English Ananjeva, N. B. 1986. [On the validity of Megalochilus mystaceus (Pallas, 1776)]. Proceedings of the Zoological Institute, Leningrad 157:4-13. (In Russian). Note that for Acta Herpetologica Sinica, the year must precede the volume number. This is to distinguish between the old and new series, and between 1982-1987, Vols.1-6 (new series) and 1988 with no volume number, numbers 1 and 2 (new series). Cai, M., J. Zhang, and D. Lin. 1985. [Preliminary observation on the embryonic development of Hynobius chin- ensis Guenther]. Acta Herpetologica Sinica 1985, 4(2): 177-180. (In Chinese). Book Pratt, A. E. 1892. To the snows of Tibet through China. Longmans, Green, and Co., London. 268 pp. Article in book Huey, R. B. 1982. Temperature, physiology, and the ecology of reptiles. Pp. 25-91. In C. Gans and F. H. Pough (eds.), Biology of the Reptilia, Vol. 12, Physiological Ecology. Academic Press, New York. Government publication United States Environmental Data Service. 1968. Climatic Atlas of the United States. Environmental Data Ser- vice, Washington, D. C. Abstract of oral presentation Arnold, S. J. 1982. Are scale counts used in snake systematics heritable? SSAR/HL Annual Meeting. Raleigh, North Carolina. [Abstr]. Vol. 10, p. 312 Asiatic Herpetological Research 2004 Thesis or dissertation Moody, S. 1980. Phylogenetic and historical biogeographical relationships of the genera in the Agamidae (Rep- tilia: Lacertilia). Ph.D. Thesis. University of Michigan. 373 pp. Anonymous, undated Anonymous. Undated. Turpan brochure. Promotion Department of the National Tourism Administration of the People's Republic of China, China Travel and Tourism Press, Turpan, Xinjiang Uygur Autonomous Region, China. Copyright Asiatic Herpetological Research reserves the copyrights to all material published therein, except that excluded by permission of the editors. Any material under a prior copyright submitted to Asiatic Herpetological Research must be accompanied by the written consent of the copyright holder. Submission of Manuscripts Authors should submit letter quality, double spaced, single-sided manuscripts both in English and in the original language on 21.5 x 28 cm (8.5 x 11 inch) white bond paper. If possible, include a computer diskette containing the manuscript. Macintosh diskettes, Zip disks, or 3.5” magneto-optical (MO), containing Adobe FrameMaker, Word Perfect, Microsoft Word, Claris Works, Macwrite, Write Now, or text files, or 3.5" MS/PC DOS diskettes, Zip disks, or 3.5” MO with Adobe FrameMaker, Word Perfect, Microsoft Word, RTF, or ASCII files are prefera- ble. Please indicate author, computer, file format, and file name in writing on the disk. The TIFF file format is preferable for electronic versions of figures, but Photoshop, JPEG, or PICT formats are acceptable. Resolution of electronic versions of figures must be at least 600 dpi for line art, or 300 dpi for grayscale and color images. Fig- ures will be reduced to either 1 column (3.25”) or two columns (6.5”). Manuscripts will be reviewed. The editors will attempt to choose reviewers whose research knowledge most closely matches the content of the manuscript. Asiatic Herpetological Research requests $25 US per printed page from authors with funds available. Please indicate if funds are available. Send manuscripts by postal mail to Editors, Asiatic Herpetological Research, Museum of Vertebrate Zoology, 3101 Valley Life Sciences Building, University of California, Berkeley CA 94720-3160 USA. Send manuscripts by Internet email to asiaherp@berkeley.edu as a MIME attachment with binhex or uuencode encoding. If you use email to submit, an editor will acknowledge receipt of your manuscript. Please note that it is possible for your email message to disappear on the Internet without being delivered. If your mes- sage is returned, or not acknowledged, you may want to try again or to send your manuscript by postal mail. Kuinulta§, Y., S. H. Durma§, Y., Kaska, M. Oz, and M. R. Turn;. A Morphological and Taxonomic Study on Lacerta parva Boulenger, 1887 (Sauria: Lacertidae) from West Taurus, Turkey 202-207 Macey, J. R. and N. B. Ananjeva. Genetic Variation Among Agamid Lizards of the Trapelus agilis Complex in the Caspian-Aral Basin 208-214 Ugurta§, i. H., H. S. Yildirimhan, and M. Kalkan. The Feeding Biology of Ran a macrocnemis Boulenger, 1885 (Anura: Ranidae), Collected in ULUDAg, Bursa, Turkey 215-216 Sevin^, M., \. H. Ugurta§, and H. S. Yildirimhan. Morphological Observations on the Erythrocyte and Erythrocyte Size of Some Gecko Species, Turkey 217-223 Sharif i, M. and S. Assadian. Distribution and Conservation Status of Neurergus microspilotus (Caudata: Salamandridae) in Western Iran 224-229 Tosunoglu, M., D. Ayaz, C. V. Tok, and B. Dulger. An Investigation on the Blood Cells of the Leopard Gecko, Eublepharis angr.4mainyu (Reptilia: Sauria: Eublepharidae) 230-234 Ahsan, M. F. and S. Parvin. A Record of Boiga ochracea walli (Stoliczka, 1870) from Bangladesh 235 Ahsan, M. F. and M. A. Saeed. Some Aspects of Breeding Biology of the Bengal Lizard ( Varan us BENGALENSIS) IN BANGLADESH 236-240 Das, I. ANew Locality for the Rare Bornean Skink, Lamprolepis vyneri (Shelford, 1905) (Sauria: Scincidae) 241-244 Das, I. and S. K. Chanda. Leptobrachium smithi Matsui, Nabitabhata, and Panha, 1999 (Anura: Megophryidae), and Addition to the Fauna of Myanmar (Burma) 245-246 Grismer, J. L., L. L. Grismer, I. Das, N. S. Yaakob, L. B. Liat, T. M. Leong, T. M. Youmans, and H. Kaiser. Species Diversity and Checklist of the Herpetofauna of Pulau Tioman, Peninsular Malaysia, With a Preliminary Overview of Habitat Utilization 247-279 Guo, P. and E. Zhao. Pareas stanleyi - A Record New to Sichuan, China and a Key to the Chinese Species 280-281 Litvinchuk, S. N., L. J. Borkin, and J. M. Rosanov. Intraspecific and Interspecific Genome Size Variation In Hynobiid Salamanders of Russia and Kazakhstan: Determination by Flow Cytometry 282-294 Schaedla, W. H. Anomalous (?) Nocturnal Feeding by the Agamid Lizard Calotes emma in Northeastern Thailand 295-297 Yang, Y. Karyological Studies on Amphibians in China 298-305 Papenfuss, T. J. The History of the Journal Asiatic Herpetological Research 306-307 Editors. Guidelines for Manuscript Preparation and Submission 308-312 Asiatic Herpetological Research is created using QuarkXPress 6.0. Adobe Illustrator 10. Adobe Acrobat 6. and Adobe Photoshop 6 on both Macintosh OS 10 and Microsoft Windows XP operating system platforms. Body text is in Times New Roman and the Headings in Arial. Using digital technology, we consumed less than 400 sheets of paper in the prepress production of this issue. ISSN 1051-3825 Diaz, R. E., M. T. Leong, L. L. Grismer, and N. S. Yaakob. A New Species of Dibamus (Squamata: Dibamidae) from West Malaysia Grismer, L. L., j. L. Grismer, and T. M. Youmans. A New Species of Leptolalax (Anura: Megophryidae) FROM PlILAU TlOMAN, WEST MALAYSIA I I Leong, T. M. and L. L. Grismer. A New Species of Kukri Snake, Oligodon (Colubridae), from Pulau Tioman, West Malaysia 12-16 Stuart, B. L. and H. Heatwole. A New Philautus (Amphibia: Rhacophoridae) from Northern Laos 17-21 Diesmos, A. C., G. V. A. Gee, M. L. Diesmos, R. M. Brown, P. J. Widmann, and J. C. Dimalibot. Rediscovery of the Philippine Forest Turtle, Heosemys leytensis (Chelonia; Bataguridae), from Palawan Island, Philippines 22-27 Feldman, C. R. and J. F. Parham. Molecular Systematics of Old World Stripe-Necked Turtles (Testudines: Mauremys ) 28-37 Hutchison, J. H„ P. A. Holroyd, and R. L. Ciochon. A Preliminary Report on Southeast Asia’s Oldest Cenozoic Turtle Fauna from the Late Middle Eocene Pondaung Formation, Myanmar 38-52 Joyce, W. G. and C. J. Bell. A Review of the Comparative Morphology of Extant Testudinoid Turtles (Reptilies: Testudines) 53-109 Le, Minh, T. Hoang, and D. Le. Trade Data and Some Comments On the Distribution of Mauremys ANNAMENS1S (SlEBENROCK, 1 903) 110-113 Perala, J. and R. Bour. Neotype of Testudo terrestris ForsskAl, 1775 (Testudines, Testudinidae) 114-119 Schilde, M., D. Barth, and U. Fritz. An Ocadia sinensis x Cyclemys shanensis hybrid (Testudines: Geomydidae) 120-125 Shi, H., Z. Fan, F. Yin, and Z. Yuan. New Data on the Trade and Captive Breeding of Turtles in Guangxi Province, South China 126-128 Stuart, B. L. and S. G. Platt. Recent Records of Turtles and Tortoises from Laos, Cambodia, and Vietnam 129-150 Auffenberg, K., K. L. Krysko, and W. Auffenberg. Studies on Pakistan Lizards: Cyrtopodion stoliczkai (Steindachner, 1867) (Gekkonidae: Gekkoninae) 151-160 Diilger, B., j. H. Ugurta§, and M. Sevin^ Antimicrobial Activity in the Skin Secretion of Bufo viridis (Laurenti, 1768) 161-163 Du§en, S., M. Oz, and M. R. Tunq:. Analysis of the Stomach Contents of the Lycian Salamander Mertensiella luschani (Steindachner, 1891) (Urodela: Salamandridae), Collected from Southwest Turkey 164-167 Ebrahimi, M., H. G. Kami, and M. Stock. First Description of Egg Sacs and Early Larval Development in Hynobiid Salamanders (Urodela, Hynobiidae, Batrachuperus ) from North-Eastern Iran 168-175 Jarrar, B. M„ and N. T. Taib. Histochemical Characterization of the Lingual Salivary Glands of the House Gecko, Ptyodactylus hasselqu/stii ( Squamata: Gekkonidae) 176-181 Kami, H. G. The Biology of the Persian Mountain Salamander, Batrachuperus persicus (Amphibia, Caudata Hynobiidae) in Golestan Province, Iran 182-190 Khan, M. S. Annotated Checklist of Amphibians and Reptiles of Pakistan 191-201 (Continued on inside back cover) Herpetological Research Volume 11 • 2008 Chengdu Institute of Biology of the Chinese Academy of Sciences Asiatic Herpetological Research Society at the Museum of Vertebrate Zoology, University of California MCZ ERNST MAYR LIBRARY 3 2044 118 665 082