Asian Herpetological Research 2017, 8(2): 131–136 ORIGINAL ARTICLE DOI: 10.16373/j.cnki.ahr.160066

First Record of the Blue-banded Sea Krait (Laticauda laticaudata, Reptilia: Squamata: Elapidae: Laticaudinae) on Jeju Island, South Korea

Jaejin PARK1#, Kyo-Soung KOO1#, Il-Hun KIM1,2, Woo-Jin CHOI1 and Daesik PARK3*

1 Department of Biology, Kangwon National University, Chuncheon, Kangwon 24341, South Korea 2 National Marine Biodiversity Institute of Korea, Seocheon, Chungnam 33662, South Korea 3 Division of Science Education, Kangwon National University, Chuncheon, Kangwon 24341, South Korea

Abstract We report the first recorded capture of a blue-banded sea snake (Laticauda laticaudata Linnaeus, 1758, Jobeuntti KunBadabam in Korean) in South Korea based on one male specimen collected from Marado-ri, Seogwipo- si, Jeju-do on 20 October 2016. The morphological features of the lateral nostrils, the much wider ventrals than adjacent dorsals, the horizontally undivided rostral, the two prefrontals, and the uniform black bands on the body indicate that the specimen is L. laticaudata. An analysis of the partial mitochondrial cytochrome b gene sequence indicated that the specimen fits well into the known L. laticaudata phylogenetic group, which confirms that the sea krait is L. laticaudata. Including this report, five species of sea snakes L.( laticaudata, L. semifasciata, Hydrophis platurus, H. cyanocinctus, and H. melanocephalus) have now been reported in Korean waters.

Keywords sea snake, Hydrophiinae, Laticaudinae, blue-banded sea snake, Laticauda laticaudata

1. Introduction approaches, studies to determine the species distributions and biological and ecological characteristics are necessary Marine (Hamann et al., 2007; Rasmussen et al., 2011a; (Hamann et al., 2007; GBRMPA, 2012). In particular, Wallace et al., 2016) and terrestrial reptiles (Reading new records of sea snake species beyond their typical sea et al., 2010) are declining globally, and the major factors snake distribution ranges must be reported. underlying the decline in marine reptiles include climate Globally, 70 sea snakes (aquatic elapids) are distributed change, coastal development, water pollution, and in the two subfamilies Hydrophiinae (true sea snakes) overexploitation (GBRMPA, 2012; Cao et al., 2014). and Laticaudinae (sea kraits), and these snakes primarily Sea snakes are a representative marine reptile that is inhabit the tropical and subtropical waters of the Indian vital to marine ecosystems because of their roles as and Pacific oceans (hppt://www.reptile-database.org). top predators and their use of relatively wide habitats In East Asia, which includes China, Japan, and Korea, (Brischoux et al., 2007; Ineich et al., 2007). Similar to a total of 16 sea snake species have been reported, with marine turtles (GBRMPA, 2012; Wallace et al., 2016), 13 in the Hydrophiinae and 3 in the Laticaudinae (Kang sea snakes are also declining (Hamann et al., 2007; and Yoon, 1975; Mao and Chen, 1980; Zhao and Adler, Goiran and Shine, 2013; Lukoschek et al., 2013). To 1993; Toriba, 1994; Kim and Han, 2009). Recently, understand the declining trends and develop mitigation several sea snake have been captured outside of typical sea snake distribution ranges. For example, H. platurus # Both authors contributed equally to this paper. was found in Ventura County, California, USA (Gordon, * Corresponding author: Prof. Daesik Park, from Division of Science Education, Kangwon National University, Chuncheon, South Korea, 2015), L. semifasciata and L. laticaudata were found on with his research focusing on basic and conservation ecology of the east and west coasts of mainland Japan (Tandavanitj amphibians and reptiles. E-mail: [email protected] et al., 2013a), and three L. semifasciata specimens Received: 20 November 2016 Accepted: 14 April 2017 were reported at Jeju Island, Korea in 2016 (Park et al., 132 Asian Herpetological Research Vol. 8

2016). To evaluate the effects of recent climate change were sequenced at the Macrogen Sequencing facility on sea snake population dynamics and potential range (Macrogen, Seoul, South Korea). expansions, records of sea snake captures beyond the To construct the phylogenetic tree, we additionally limits of their distribution range are critical. downloaded 14 sea snake mt-Cyt b sequences from In this study, we report for the first time the capture GenBank (Slowinski and Keogh, 2000; Lukoschek and of the blue banded sea krait Laticauda laticaudata Keogh, 2006; Sanders et al., 2008, Sanders et al., 2013a, (Linnaeus, 1758) in Korea based on a specimen collected b; Tandavanitj et al., 2013b) and aligned the all sequences from Jeju Island on 20 October 2016. In addition, we using Clustal X2 (Thompson et al., 2002). Uniform update the identification keys for species of Laticaudinae sequences were edited using the Bioedit software and Hydrophiinae in Korea and discuss the meaning of package (Hall, 2011) and then analyzed by the maximum recent sea krait captures in Korea. likelihood (ML) method using PAUP 4.0b10 (Swofford, 2002) and ModelTest (Posada and Crandall, 1998) and 2. Materials and Methods the Bayesian method using MrBayes 3.1 (Ronquist and Huelsenbeck, 2003). In addition, we determined the 2.1 Collecting the specimen and the morphological haplotype of the mt-Cyt b sequence of the specimen study The sea snake reported in this study was collected by comparing the known mt-Cyt b haplotypes of L. approximately 30 km south of Marado-ri, Seogwipo-si, laticaudata collected by Tandavanitj et al. (2013b) on the Jeju-do, South Korea (N 32° 51′ 0.00″, E 126° 20′ 60.00″) Taiwan-Ryukyu archipelagos. by a fisherman (Kang Y. N.) on 20 October 2016. When caught by a hand net, the sea snake was swimming on the 3. Results and Discussion water surface near a cutlassfish ship. The dead snake due to keeping it in a freezing storage on the ship was donated The male snake specimen had a prominent, vertically to the Herpetology Laboratory of the Kangwon National flattened, paddle-like tail, which is a representative feature University (KNU). On arrival at the lab, we determined of typical sea snakes (Figure 1 A, B). The ground body its sex by gonadal observation and measured its snout color was blue, the ventral color was yellowish, the upper vent and tail lengths to 0.1 cm using a tape ruler and its lip was pale blue, the dorsal bands were black, and the body weight to 0.1 g using a digital balance (ELT 4001, ventral bands were yellowish black (Figure 1 A, B). The Sartorius-Korea, Seoul, South Korea). Subsequently, we width of the uniform black cylindrical bands on the trunk investigated the key morphological and classification and tail was between three and four dorsal scales (Figure features, such as the number of scale rows and bands on 1 A). The specimen had 49 bands on the trunk and five on the head and body, photographed the snake, collected 2-3 the tail (Figure 1 A). The rostral visible from above was mm tail tissues for later molecular study, preserved it in not divided into two (Figure 2 A, B). The snake had two 95% ETOH, and finally deposited it in the herpetological prefrontals, two internasals, one frontal, and two parietals collection of the KNU (G535LL). We identified the (Figure 2B). In addition, we identified seven supralabials specimen on the basis of previous morphological studies (the 3rd and 4th touched the eye), one supraocular, one (Mao and Chen, 1980; Toriba, 1994; Rasmussen et al., preocular, two postoculars, and 1+2 temporals on each 2011b). dorsal side of the head (Figure 2 C) and one mental, four 2.2 DNA extraction, PCR, and phylogenetic analysis inframaxillaries and seven infralabials on each ventral of the mitochondrial cytochrome b gene We extracted side of the head (Figure 2 C, D). The snake had 239 whole genomic DNA from the tail muscle tissues using ventrals, 42 subcaudals and 2 anals (Figure 1 B). The the QIAGEN DNeasy Blood and Tissue kit (QIAGEN, number of scale rows around the neck, on the mid-body, Hilden, Germany) following the manufacturer’s and near the anus was 19, 19, and 18, respectively. The instruction and then amplified the partial mitochondrial snout-vent length was 108.6 cm, the tail length was 12.4 cytochrome b (mt-Cyt b) gene using the forward primer cm, and the body weight was 485.1 g. L14910 and the reverse primer H16064 (Burbrink et In the phylogenetic analysis of the partial mt-Cyt b al., 2000). We conducted the PCR using a SimpliAmp gene sequences, the specimen fit well into a previously Thermal Cycler (Life Technologies, Carlsbad, CA, USA) known L. laticaudata group (Figure 3). The specimen under the following conditions: 94ºC for 4 min; 35 cycles had a unique mt-Cyt b haplotype (GenBank accession of 94ºC for 30 s, 57ºC for 30 s, and 72ºC for 1 min; and No. KY114891) that had not been previously reported; a final extension of 72ºC for 7 min. The PCR products however, the haplotype sequence differed by only one No. 2 Jaejin PARK et al. First Record of Laticauda laticaudata in Korea 133

Figure 1 Male Laticauda laticaudata caught at Marado-ri, Seogwipo-si, Jeju-do, South Korea on 20 October 2016. Dorsal (A) and ventral (B) surfaces.

Figure 2 Male Laticauda laticaudata caught at Marado-ri, Seogwipo-si, Jeju-do, South Korea on 20 October 2016. Frontal (A), dorsal (B), lateral (C), and ventral surfaces of head (D). bp from the Lati-1 haplotype, which has been identified identified the sea krait collected in Korea as a blue-banded throughout Taiwan and the Ryukyu Islands (Tandavanitj sea krait (L. laticaudata). In northeast Asia, three sea krait et al., 2013b). species (L. colubrina, L. laticaudata, and L. semifasciata) Based on the above morphological features, we have been reported (Kang and Yoon, 1975; Mao and 134 Asian Herpetological Research Vol. 8

Figure 3 Maximum likelihood (ML) tree based on the partial mitochondrial cytochrome b gene of Laticauda laticaudata collected at Jeju Island, South Korea. Three Hydrophis species, which previously known in Korea, and 11 Laticauda sequences were downloaded from GenBank (the accession number is listed after each scientific name) and used in the analysis. The phylogenetic tree was constructed with PAUP* v4.0b10 (Sowfford, 2002). On each branch, the Bayesian posterior probabilities are denoted.

Chen, 1980; Zhao and Adler, 1993; Toriba, 1994; Kim gene sequences, the specimen was well matched to the and Han, 2009). Among them, only L. semifasciata has a known L. laticaudata group. This finding clearly shows horizontally divided rostral and evident V-shaped bands that the specimen is L. laticaudata. Unfortunately, we on the body, while L. colubrina has a unique yellow to could not determine the origin of the specimen from white upper lip, which is the source of its English name the haplotype of the partial mt-Cyt b sequence. The ‘yellow-lipped sea krait’, and three prefrontals (Mao and haplotypes of the mt-Cyt b gene of L. laticaudata Chen, 1980; Rasmussen et al., 2011b; Park et al., 2016). are not highly differentiated throughout Taiwan- Compared with L. colubrina and L. semifasciata, L. Ryukyu archipelagos. Throughout the regions, only laticaudata has two prefrontals and the rostral is visible four haplotypes have been identified from 136 samples from above (Mao and Chen, 1980; Rasmussen et al., (Tandavanitj et al., 2013b). Although the haplotype 2011b). The features of the horizontally undivided rostral, of the mt-Cyt b gene of the L. laticaudata subject in two prefrontals, and uniform black bands on the body this study was unique, it differed by only one bp from of the specimen are all consistent with L. laticaudata. In previously reported Lati-1 haplotype from specimens in addition, the number of bands, ventrals, subcaudals, and Taiwan and the southern and northern Ryukyu Islands. scale rows on the body are all within the known range The morphological and molecular data obtained here of L. laticaudata (Mao and Chen, 1980; Toriba, 1994; consistently indicate that the sea krait collected at Jeju Rasmussen et al., 2011b). These features all indicate that Island is L. laticaudata; therefore, this paper represents the specimen collected at Jeju Island is L. laticaudata. the first official report of L. laticaudata in Korea. To In the phylogenetic analysis of the partial mt-Cyt b date, five species of sea snakes (L. laticaudata, L. No. 2 Jaejin PARK et al. First Record of Laticauda laticaudata in Korea 135 semifasciata, Hydrophis platurus, H. cyanocinctus, and Acknowledgements This research was supported by the H. melanocephalus have been reported in Korean waters. Basic Science Research Program through the National Our first capture of L. laticaudata at Jeju Island is Research Foundation of Korea (NRF) funded by the particularly meaningful because another sea krait species, Ministry of Education (2014R1A1A4A01005302). L. semifasciata, was newly reported in Korea within the This research was conducted within the guidelines last year (Park et al., 2016). Despite the capture of two and approval of the Institutional Animal Care and Use different sea krait species at Jeju Island, the L. laticaudata Committee of Kangwon National University (KW- specimen in this study might represent a simple accidental 161108-1). drifter via the Taiwan Warm or Kuroshio currents (Su, 2001; Xu et al., 2009; Zhou et al., 2015) because it is References unlikely that a sea krait population has been established at Jeju Island. Nevertheless, this specimen indicates Brischoux F., Bonnet X., Shine R. 2007. Foraging ecology of sea that sea kraits have newly or recently entered Korean kraits Laticauda spp. in the Neo-Caledonian lagoon. Mar Ecol Prog Ser, 350: 145–151 waters (Lee et al., 2013; Kim et al., 2016; Park et al., Burbrink F. T., Lawson R., Slowinski J. B. 2000. Mitochondrial 2016). 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