Seven specimens of the sharpbelly Hemiculter leucisculus (Basilewsky, 1855) were collected by angling from a stream connected to the Hyakken River emptying into Kojima Bay, Okayama city, Okayama Prefecture, western Honshu, Japan, on 19 July 2016. This collection represents a new Japanese record for the species as an alien fish. The specimens are identified based on their morphology and genetic analysis of CO1 of mitochondrial DNA. One of the sequences of CO1 from six specimens is accorded with a previously reported haplotype H2, and that from the remaining one specimen has a newly obtained haplotype (H5) close to H2. Although the species has not yet been confirmed for its establishment in the stream, some individuals were found on the sampling date to jump out of the water, and this behavior resembles one associated with the spawning of the species observed in China. It is necessary to monitor the species in the stream and adjacent waters.This study was partially supported by JSPS KAKENHI grands (no. 15J05777 to M. N. and no. 15K07527 to K. N.

Kawai, Koichiro

Nagasawa, Kazuya

Nitta, Masato

Name not available

広島大学学術情報リポジトリHiroshima University Institutional RepositoryTitle First Japanese record of the sharpbelly Hemiculterleucisculus (Basilewsky, 1855) (Cypriniformes: Cyprinidae)from Okayama Prefecture, western HonshuAuther(s) Nitta, Masato; Kawai, Koichiro; Nagasawa, KazuyaCitation Biogeography , 19 : 17 - 20Issue Date 2017-09-20DOISelf DOIURL http://ir.lib.hiroshima-u.ac.jp/00044323Right (c) 日本生物地理学会論文ファイルの利用は著作権の範囲内に限り認められます。RelationBiogeography 19.First Japanese record of the sharpbelly Hemiculter leucisculus (Basilewsky, 1855) (Cypriniformes: Cyprinidae) from Okayama Prefecture, western HonshuMasato Nitta*, Koichiro Kawai and Kazuya NagasawaGraduate School of Biosphere Science, Hiroshima University,Kagamiyama 1-4-4, Higashi-Hiroshima, Hiroshima 739-8528, JapanDuring a sampling of a domestic alien cyprinid, Ischikauia steenackeri (Sauvage, 1883), to examine its parasite fauna in a stream connected to the Hyak-ken River flowing into Kojima Bay, Okayama city, Okayama Prefecture, seven cyprinid specimens simi-lar to I. steenackeri were collected but differed from the species. They were later identified as the sharp-belly Hemiculter leucisculus (Basilewsky, 1855) based on their morphology and molecular analysis. We report here this collection as the first record of the species from Japan.The specimens were collected from the stream (34°37ʹ19.1ʺN, 133°58ʹ48.8ʺE) by angling and brought alive or on ice to the laboratory of Hiroshi-ma University on 19 July 2016. Five dead fish were stored frozen until dissection. All fish were examined for their parasites, fixed in 10% formalin, and later preserved in 70% ethanol. Before fixation, their right pectral fins were removed from the fish and stored in 99% ethanol for molecular analysis. Voucher speci-mens are deposited in the fish collection of the Na-tional Museum of Nature and Science, Tsukuba city, Ibaraki Prefecture, Japan (NSMT-P131564). Counts and measurements followed Hubbs & Lagler (2004). Gill rakers were counted from the left gills removed for the parasitological survey. DNA was extracted from the right pectral fins using the DNeasy blood and tissue kit (Qiagen) in ac-cordance with the manufacturer’s instructions. The DNA was amplified by polymerase chain reaction (PCR) using the primer pair FishF1 (5'-TCAAC-CAACCACAAAGACATTGGCAC-3') and FsihR1 (5'-TAGACTTCTGGGTGGCCAAAGAATCA-3') to amplify first subunit of cytochrome oxidase (CO1) (Ward et al. 2005). A total of 25 μL PCR reaction consisted of 1 μL of DNA template, 1× ExTaq Buf-———————————————————————*Corresponding author:licht.bsn.mono@gmail.comAbstract.　Seven specimens of the sharpbelly Hemiculter leucisculus (Basilewsky, 1855) were collected by angling from a stream connected to the Hyakken River emptying into Kojima Bay, Okayama city, Okayama Prefecture, western Honshu, Japan, on 19 July 2016. This collection represents a new Japanese record for the species as an alien fish. The specimens are identified based on their morphology and genetic analysis of CO1 of mitochondrial DNA. One of the sequences of CO1 from six specimens is accorded with a previously reported haplotype H2, and that from the remaining one specimen has a newly obtained haplotype (H5) close to H2. Although the species has not yet been confirmed for its establishment in the stream, some individuals were found on the sampling date to jump out of the water, and this behavior resembles one associated with the spawning of the species observed in China. It is necessary to monitor the species in the stream and adjacent waters. Key words: Hemiculter leucisculus, alien fish, new country record, Japan17–20. Sep. 20, 2017− 17−First records of Sphyraena iburiensis (Perciformes: Sphyraenidae) from the Amami Islands and southern Kyushu, Japanfer (TaKaRa), 0.2 mM of each dNTP, 1 μM of each primer, and 2.5 units of TaKaRa Ex Taq DNA Poly-merase (TaKaRa). PCR was carried out with the following protocol: 94°C for 60 sec followed by 35 cycles of 94°C for 30 sec, 54°C for 30 sec, and 72°C for 1 min, and 10 min of final hold at 72°C. PCR products were purified using NucleoSpin Gel and PCR Clean-up kit (Macherey-Nagel) and sequenced with a 3130X Genetic Analyzer (Applied Biosys-tems) with the same primers that were generated the PCR products and submitted to the DDBJ (DNA Data Bank of Japan) database (LC316767–316773). CO1 sequences of H. leucisculus were downloaded from GenBank (HQ536384, HQ536385, HQ536386, HQ536387, HQ536388, HQ536389, KF492988, KF492989, KF647872, KF956522), edited and aligned with newly obtained sequences (625bp) us-ing MEGA6 (Tamura et al. 2013).Hemiculter leucisculus (Basilewsky, 1855)Berg, 1909Japanese name: Kawa-iwashi(Fig. 1)Fig. 1.  Hemiculter leucisculus (Basilewsky, 1855) (NSMT-P 131564, 98.3 mm in standard length) collected from Okayama Prefecture, western Honshu, Japan.Specimen 1 2 3 4 5 6 7Standard length (mm) 98.3 86.6 113.6 101.7 107.7 102.7 98.0Percent of standard lengthHead length 22.6 24.0 21.1 22.3 21.3 18.6 21.4Body depth 23.8 22.3 22.3 19.8 22.0 22.8 22.3Body width 10.2 10.3 12.1 9.0 11.5 10.7 10.1Depth of caudal peduncle 9.2 9.9 9.3 8.2 9.7 9.8 9.1Length of caudal peduncle 15.7 15.9 15.6 13.3 15.1 14.7 14.8Preanal length 69.2 68.4 71.7 69.2 71.1 70.3 72.0Prepelvic length 47.3 49.1 50.4 48.7 50.5 50.8 50.3Height of dorsal fin 17.0 19.5 18.8 20.0 17.5 16.5 15.4Length of dorsal fin base 9.2 10.6 10.0 9.7 10.2 10.0 9.8Height of anal fin 11.4 10.4 11.3 10.3 11.8 9.5 11.8Length of anal fin base 12.1 12.5 11.3 12.5 11.7 12.0 12.2Pectoral fin length 20.2 20.4 21.1 20.0 19.5 21.8 19.2Percent of head lengthHead width 42.5 37.5 54.9 43.9 52.2 52.7 44.8Snout length 25.7 22.9 30.8 29.1 26.8 33.9 24.7Orbit diameter 25.2 22.1 29.3 26.0 25.9 33.9 23.3Interorbital width 28.3 24.6 33.1 30.0 32.9 39.3 28.9Table 1.  Measurements of seven specimens of Hemiculter leucisculus collected from Okayama Prefecture, western Honshu, Japan.− 18−New record of Hemiculter leucisculus in Japan Masato Nitta, Koichiro Kawai and Kazuya NagasawaSpecimens. NSMT-P 131564, seven specimens, 86.6–113.6mm in standard length, a stream connect-ed to the Hyakken River emptying into Kojima Bay, at Kuwano, Naka District, Okayama city, Okayama Prefecture, Japan, 19 July 2016.Description. Morphometric data are shown in Table 1. Body elongate. Mouth oblique, without bar-bels. Caudal fin strongly forked. Gill rakers 17–20. Belly with keel from throat to anus. Lateral line bending down under pectoral fin, rising behind end of anal fin obliquely in three scales, extending along middle of caudal peduncle. In life, dark dorsally, sil-very laterally, whitish ventrally.Dorsal fin with 3 simple and 6–7 branched rays. Anal fin with 3 simple and 11–12 branched rays. Pectoral fin with 1 simple and 12–13 branched rays. Pelvic fin with 1 simple and 7–8 branched rays. Pored lateral-line scales 51–56. Scale rows between lateral-line scales and dorsal-fin origin 8–9; between lateral-line scales and anal-fin origin 2.Remarks. This fish was originally described by Basilewsky (1855) as Culter leucisculus from Pe-kin, China. The Japanese name, “Kawa-iwashi” was proposed by Oshima (1923) for Cultriculus kneri (Warpachowski, 1888) collected in Taiwan, but the species has been regarded as Hemiculter leucisculus (Berg 1909; Wu 1964; Vasil’eva & Kozlova 1988). The specimens examined in this study approximately conform to the descriptions of H. leucisculus collect-ed in China (Wu 1964) and Russia (Vasil’eva & Ko-zlova 1988). Also, the sequences of CO1 determined from our seven specimens are almost identical to the reported sequences of H. leucisculus: one of those from six specimens is accorded with a haplotype H2 determined from Azerbaijan (Mustafayev et al., 2015), while that from the remaining one specimen has a new haplotype H5 that is close to H2 (Table 2). Hemiculter leucisculus is natively distributed in Korea, China, Vietnam, Far-East Russia, and Taiwan (Wu 1964) but has established as alien species in Uz-bekistan, Afghanistan, Iran, Kazakhstan, Turkmeni-stan, Iraq, and Azerbaijan (Mustafayev et al. 2015; Wang et al. 2016). The present collection in Japan represents a new country record for H. leucisculus as an alien fish. As many cyprinids are imported as live fishing bait from China to Japan (Saito et al. 2011), the species is likely to be included in those imported fishes and actually found as an ornamental fish at pet shops in Japan. Under these situations, the Japanese population of the species may have originated from release of some imported individuals by hobbyists or their accidental escape from aquaria.Hemiculter leucisculus is a typical r-selected spe-cies and has the high invasive potential (Wang et al. 2016). During our sampling, some individuals of the species were observed to jump out of the water: a similar behavior was previously reported in China to be associated with the spawning of the species (Wu et al. 1979). No information is available on the establishment of the species in Japan, but it is neces-sary to monitor its abundance and reproduction, es-Table 2.  Five haplotypes of CO1 from Hemiculter leucisculus. H1 to H4 were reported by Mustafayev et al. (2015), and H5 is a newly found haplotype.Variable siteHaplotype 9 237 300 399 432 462 468 513 531H1 G G A G G G A C TH2 A T AH3 A T G A A G TH4 A T G AH5 A T A A G− 19−New record of Hemiculter leucisculus in Japan Masato Nitta, Koichiro Kawai and Kazuya Nagasawapecially in the stream sampled and adjacent waters.AcknowledgementsWe thank the students of the laboratories of Aqua-culture and Benthos Ecology, Hiroshima University, for their assistance during the study. This study was partially supported by JSPS KAKENHI grands (no. 15J05777 to M. N. and no. 15K07527 to K. N.)ReferencesBasilewsky, S. 1855. Ichthyographia Chinae borea-lis. N. mém. Soc. Nat. Moscou, 10: 215–263, 9 pls.Berg, L. S. 1909. Ichthyologia Amurensis. Zapiski Imperatorskoy Akad. Nauk, 24(9): 1–270. (In Russian).Hubbs, C. L. & K. F. Lagler. 2004. Fishes of the Great Lakes Region. Revised Edition. xxxii, 276 pp. University of Michigan Press, Bloomfield Hills.Mustafayev, N. J., Ibrahinmov, Sh. R. & Levin, B. A. 2015. Korean sharpbelly Hemiculter leucisculus (Basilewsky, 1855) (Cypriniformes, Cyprinidae) is a new species of Azerbaijan fauna. Rus. J. Biol. Invasions, 6: 252–259.Oshima, M. 1923. [On the distribution of Taiwanese freshwater fish, and geographical relationship with neighboring areas]. Dobutsugaku Zasshi, 35: 1–49. (In Japanese).Saito, H., Niwa, N., Kawai, K. & Imabayashi, H. 2011. Current state of aquatic animals sold as sport fishing bait in western Japan. Bull. Hiro-shima Univ. Mus., 3: 45–57. (In Japanese with English abstract).Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. 2013. MEGA6: Molecular Evolu-tionary Genetics Analysis version 6.0, Mol. Biol. Evol., 30: 2725–2729.Vasil’eva, E. D. & Kozlova, M. S. 1988. On the tax-onomy of the sawbellies of the genus Hemiculter (Cyprinidae) of the Soviet Union. Vopr. Ikhtiol., 28: 883–895. (In Russian with English title).Wang, T., Jakovlić, I., Huang, D., Wang, J.-g. & Shen, J.-z. 2016. Reproductive strategy of the invasive sharpbelly, Hemiculter leucisculus (Basilewsky 1855), in Erhai Lake, China. J. Appl. Ichthyol., 32: 324–331.Ward, R. D., Zemlak, T. S., Innes, B. H., Last, P. R. & Hebert, P. D. N. 2005. DNA barcoding Aus-tralia’s fish species. Phil. Trans. R. Soc. B, 360: 1847–1857.Wu, X.-w. 1964. [The Cyprinid Fishes of China, vol. 1]. Shanghai Scientific & Technical Publishers, Shanghai. (Translated from Chinese to Japanese by Nakajima T. & Kobayakawa, M. 1980. 346 pp. Tatara Shobō, Yonago).Wu, X.-w., Yang, G.-r., Yue, P.-q. & Huang, H.-j. 1979. [Economic Fauna Sinica of China, Fresh-water Fishes, Second Edition]. iv, 153 pp., 38 pls. Science Press, Beijing. (In Chinese).(Received January 19, 2017; Accepted August 18, 2017)− 20−New record of Hemiculter leucisculus in Japan

First Japanese record of the sharpbelly Hemiculter leucisculus (Basilewsky, 1855) (Cypriniformes: Cyprinidae) from Okayama Prefecture, western Honshu

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First Japanese record of the sharpbelly Hemiculter leucisculus (Basilewsky, 1855) (Cypriniformes: Cyprinidae) from Okayama Prefecture, western Honshu

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