Herpetology Notes, volume 14: 613-615 (2021) (published online on 02 April 2021)

First record of Amblyomma testudinarium Koch, 1844 (Acari: Ixodidae) parasitising Reeves’ Pond Turtle, Mauremys reevesii (Gray, 1831) (Testudines: Geoemydidae), in Japan

Chiaki Kambayashi1,*, Keitaro Hemmi2, and Atsushi Kurabayashi1,3

Ticks are obligate hematophagous ectoparasites and preserved in 99% ethanol for later identification. It of various terrestrial vertebrates, including turtles. was identified as an adult femaleA. testudinarium on the Although tick bites rarely cause serious illness in basis of its morphological characters. Specifically, the host turtles (Rundquist, 1995), they can be carriers of female A. testudinarium is easily distinguishable from blood-borne diseases that sometimes afflict humans two other Amblyomma species in Japan, A. geoemydae and other animals. It is thus important to document and A. nitidum Hirst & Hirst, 1910, because the denticles ticks and their associated hosts to understand pathogen of its inner file are much smaller than those of the other transmission routes. Amblyomma geoemydae (Cantor, files (Fig. 2A). Additionally, the A. testudinarium female 1847) is a reptile-associated tick species, and four has coxa I with two ventral spurs, the external of which turtle species of the family Geoemydidae Theobald, is slightly longer (Fig. 2B), unlike in the other two 1868, Cuora flavomarginata (Gray, 1863), Geoemyda Amblyomma species (Yamaguti et al., 1971). japonica Fan, 1931, Mauremys japonica (Temminck & Schlegel, 1835), and M. mutica kami Yasukawa et al., 1996, have been recognised as its hosts in Japan (Keegan and Toshioka, 1957; Yamaguti et al., 1971; Yabe and Hayashi, 1998; Takano et al., 2011). Although the congeneric A. testudinarium Koch, 1844, also found in Japan, has a broad host range including reptiles, its infestations of turtles are scarcely documented. In this study, we report the first example of parasitism of A. testudinarium on Reeves’ Pond Turtle, Mauremys reevesii (Gray, 1831). On 2 October 2018, we captured an adult female M. reevesii (straight carapace length 230 mm; Fig. 1A) in Hiroshima City Forestry Park, Hiroshima Prefecture, Honshu Island, Japan (34.4414°N, 132.5538°E; elevation 404 m). During inspection of this turtle, we found a tick of the genus Amblyomma Koch, 1844 attached on the skin at the base of the tail near the upper shell margin (Fig. 1B). The tick was removed from the host’s body using forceps

1 Department of Bio-Science, Nagahama Institute of Bio- Science and Technology, Nagahama, Shiga 526-0829, Japan. 2 Hiroshima City Insectarium, Higashi-ku, Hiroshima 732-0036, Japan. 3 Unit for Environmental Sciences and Management, North- Figure 1. (A) Dorsal view of Mauremys reevesii parasitised West University, Potchefstroom 2520, South Africa. by Amblyomma testudinarium. (B) Adult female of A. * Corresponding author. E-mail: [email protected] testudinarium attaching to the skin at the base of the tail near © 2021 by Herpetology Notes. Open Access by CC BY-NC-ND 4.0. the upper shell margin. Photos by Chiaki Kambayashi. 614 Chiaki Kambayashi et al.

Hasegawa, 1986; Takahashi et al., 2017), Rhabdophis tigrinus (Boie, 1826) (Suzuki et al., 2005; Takahashi et al., 2011; Suh et al., 2016), Gloydius blomhoffii (Boie, 1826) (Suzuki et al., 2005), G. brevicaudus (Stejneger, 1907) (Kim et al., 2018), G. ussuriensis (Emelianov, 1929) (Suh et al., 2016; Kim et al., 2018), and Protobothrops flavoviridis (Hallowell, 1861) (Kitaoka, 1977). However, there were no other recognised reptile hosts. Additionally, in a field survey conducted in South Korea, A. testudinarium were found in several snake species, but not in any lizards, which indicated that A. testudinarium prefers ophidian hosts among reptiles (Suh et al., 2016; Kim et al., 2018). The present study demonstrated that M. reevesii is one of the possible blood feeding hosts of A. testudinarium, which revealed that this tick species can also parasitise reptiles other than snakes in East Asia. Therefore, in future studies, other novel A. testudinarium hosts may be found when additional reptile taxa are examined.

Acknowledgement. We are grateful to Takeo Yamauchi (Obihiro University of Agriculture and Veterinary Medicine) for providing the pre-peer review and helpful comments on this note. We also thank Mallory Eckstut (Edanz Group) for editing a draft of this manuscript. This study was financially supported by JSPS Figure 2. Microscope images of Amblyomma testudinarium. KAKENHI Grant number JP18H02497. (A) Ventral view of the hypostome. The denticles of the inner file are much smaller than those of the other files. (B) Coxa I References with two ventral spurs, the external of which is slightly longer. Guglielmone, A.A., Robbins, R.G., Apanaskevich, D.A., Petney, Scale bars: 500 μm. Photos by Chiaki Kambayashi. T.N., Estrada-Peña, A., Horak, I.G. (2014): The Hard Ticks of the World (Acari: Ixodida: Ixodidae). London, United Kingdom, Springer. Amblyomma testudinarium is a common tick species Hasegawa, H. (1986): The tick Amblyomma sp. parasitic on the distributed in the Australasian, Oriental, and Palearctic snake Entechinus semicarinatus collected at Mt. Yonaha, regions (Guglielmone et al., 2014). Parasitism by Okinawajima Island, Japan. Akamata 3: 29–31. (in Japanese) Hoogstraal, H., Lim, B.L., Nadchatram, M., Anastos, G. (1972): immature stages of this tick has been reported in small- The Gunong Benom Expedition 1967. 8. Ticks (Ixodidae) of and medium-sized mammals, birds, and reptiles, but Gunong Benom and their altitudinal distribution, hosts and adults mainly feed on large mammals and occasionally medical relationships. Bulletin of the British Museum (Natural reptiles (Petney et al., 2019). The reptile hosts of History). Zoology 23: 167–186. A. testudinarium largely include snakes, although Keegan, H.L., Toshioka, S. (1957): Ixodid Ticks of Japan, Korea, parasitism of two other species, Common Water and the Ryukyu Islands. Camp Zama, Japan, Department of Monitor, Varanus salvator (Laurenti, 1768) and Asian Entomology, 406th Medical General Laboratory and U. S. Army Medical Research Detachment, Far East. Brown Tortoise, Manouria emys (Schlegel & Müller, Kim, H.C., Chong, S.T., Suh, J.H., Yun, S.M., Lee, W.J., Kim, 1844), has also been recorded in the Malay Peninsula J.H., et al. (2018): Ixodes nipponensis Kitaoka and Saito and (Hoogstraal et al., 1972). Amblyomma testudinarium Koch (Acari: Ixodida: Ixodidae) In East Asia, a variety of snake species have been collected from reptiles (lizards, skinks, and snakes) in the recorded as hosts of A. testudinarium, such as Elaphe Republic of Korea, 2016. Systematic and Applied Acarology climacophora (Boie, 1826) (Suzuki et al., 2005), E. 23: 757–767. dione (Pallas, 1773) (Suh et al., 2016; Kim et al., 2018), Kitaoka, S. (1997): Ticks (lxodoidea). In: Animals of Medical Importance in the Nansei Islands in Japan, p. 239–250. Sasa, Lycodon semicarinatus (Cope, 1860) (Kitaoka, 1977), M., Takahasi, H., Kano, R., Tanaka, H., Eds., Tokyo, Japan, Ptyas semicarinatus (Hallowell, 1861) (Kitaoka, 1977; Shinjuku-shobo. First Record of Amblyomma testudinarium Parasitising Mauremys reevesii 615

Petney, T.N., Boulanger, N., Saijuntha, W., Chitimia-Dobler, L., Pfeffer, M., Eamudomkarn, C., et al. (2019): Ticks (Argasidae, Ixodidae) and tick-borne diseases of continental Southeast Asia. Zootaxa 4558: 1–89. Rundquist, E.M. (1995): Reptile and Amphibian Parasites. Neptune, New Jersey, USA, TFH Publications. Suh, J.H., Kim, H.C., Yun, S.M., Lim, J.W., Kim, J.H., Chong, S.T., et al. (2016): Detection of SFTS virus in Ixodes nipponensis and Amblyomma testudinarium (Ixodida: Ixodidae) collected from reptiles in the Republic of Korea. Journal of Medical Entomology 53: 584–590. Suzuki, S., Yamamoto, S., Shioyama, Y., Fujita, H. (2005): Survey of ixodid ticks infested on the snakes in Miyazaki Prefecture. Medical Entomology and Zoology 56: 59. (in Japanese) Takahashi, M., Misumi, H., Tamura, H. (2017): First record in Japan of tortoise tick Amblyomma geoemydae (Cantor 1847) (Acari: Ixodidae) parasitizing Pryer’s keelback snake Hebius pryeri (Boulenger, 1887) (Reptile: Colubridae). International Journal of Acarology 43: 314–319. Takahashi, M., Misumi, H., Kaito, T., Kuriyama, T., Fujita, H. (2011): First record of Amblyomma geoemydae tick on the snake Dinodon semicarinatum, with records of ticks on snakes in Japan. Annual Report of Ohara General Hospital 51: 35–38. Takano, A., Fujita, H., Kadosaka, T., Konnai, S., Tajima, T., Watanabe, H., et al. (2011): Characterization of reptile-associated Borrelia sp. in the vector tick, Amblyomma geoemydae, and its association with Lyme disease and relapsing fever Borrelia spp. Environmental Microbiology Reports 3: 632–637. Yabe, T., Hayashi, F. (1998): Amblyomma geoemydae (Acarina: lxodidae) parasitic on the turtles, Mauremys japonica, in the Atsumi Peninsula, Aichi Prefecture, Japan. Journal of the Acarological Society of Japan 7: 47–50. (in Japanese) Yamaguti, N., Tipton, V.J., Keegan, H.L., Toshioka, S. (1971): Ticks of Japan, Korea and the Ryukyu Islands. Brigham Young University Science Bulletin Biological Series 15: 1–226.

Accepted by Kanto Nishikawa