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Original Article

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea

Sohyun Won1, Bae Keun Park2, Baek Jun Kim1,†, Hye Won Kim1, Jun Gu Kang1, Tae Seo Park3, Hong Yul Seo3, Ye Eun3, Ki Gyoung Kim3, Joon Seok Chae1
The Korean Journal of Parasitology 2014;52(2):169-175.
Published online: April 18, 2014

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Corresponding author (jschae@snu.ac.kr)

†Current address: Department of Climate and Ecology, National Institute of Ecology, Seocheon 325-813, Korea.

• Received: August 31, 2013   • Revised: January 13, 2014   • Accepted: January 24, 2014

© 2014, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea
Korean J Parasitol. 2014;52(2):169-175.   Published online April 18, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.2.169
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Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea
Korean J Parasitol. 2014;52(2):169-175.   Published online April 18, 2014
DOI: https://doi.org/10.3347/kjp.2014.52.2.169
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Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea
Image Image Image Image Image
Fig. 1 Dock-Sil Mountain (▴, altitude 639 m) in Gageo-do (Island) (125°7"E, 34°4"N), Korea. Gageo-do is at Shinan-gun, Jeollanam-do (Province) and Korea's south-westernmost Island. Dock-Sil Mountain is the highest peak in Shinan-gun. (A) The geographical map of Asia. (B) The location of Mokpo (•), Heuksan-do (⋆), and Gageo-do, Korea. Gageo-do is 136 km distant from Mokpo and 70 km from Heuksan-do. (C) A map showing the topography of the Gageo-do and Dock-sil Mountain (▴).
Fig. 2 Terrestrial leeches collected from Gageo-do, Korea. (A) 10 min after feeding blood on a human leg. (B) 30 min after feeding on a human leg. (C, D) 70% alcohol-fixed terrestrial leeches after feeding.
Fig. 3 Results of PCR amplification of COI and 18S rRNA genes from the Gageo-do land leech. (A) COI gene PCR amplicons (710 bp). (B) 18S rRNA gene amplicons (1.2 kb). Out of the total 1.8 kb 18S rRNA, PCR amplifications were conducted in 2 independent parts which are overlapped each other with approximately 1.2 kb in length; M, Elpis 100 bp marker; 1, 2, 3, and 4; Gageo land leeches sample.
Fig. 4 A phylogenetic tree illustrating the genetic relationship of Haemadipsoid leeches using the neighbor-joining method implemented in Clustal X algorithm with Mega 4.0. 18S rRNA gene sequences of Haemadipsa rjukjuana obtained from Mt. Dock-Sil in Gageo-do, Shinan-gun, Jeollanam-do, Korea compared to previous reference sequences. Bold letters are sequences identified in this study.
Fig. 5 Phylogenetic tree for COI gene showing the taxonomic status of haemadipsoid leeches using the neighbor-joining method implemented. COI gene sequences of Haemadipsa rjukjuana obtained from Mt. Dock-Sil in Gageo-do, Shinan-gun, Jeollanam-do, Korea. Bold letters are sequences identified in this study. H. rjukjuana Gageo 1 indicates COI type A and H. rjukjuana Gageo 27 indicates COI type B.

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Fig. 5

Molecular Identification of Haemadipsa rjukjuana (Hirudiniformes: Haemadipsidae) in Gageo Island, Korea
Target genes Name of PCR primers primer sequences (5´-3´) PCR product size References Nuclear 18S rRNAa A AACCTGGTTGATCCTGCCAGT 1.2 kb Apakupakul et al. [15] Y CAGACAAATCGCTCC C CGGTAATTCCAGCTC 1.2 kb B TGATCCTTCCGCAGGTTCACCT Mitochondrial CO1 LCO 1490 GGTCAACAAATCATAAAGATATTGG 710 bp Folmer et al. [16] HCO 2198 TAAACTTCAGGGTGACCAAAAAATCA Taxon Locality GenBank accession no.
18S rNRA COI Haemadipsa rjukjuana Gageo (In this study) Gageo Islands, Korea KC524508a - Haemadipsa rjukuana Gageo 1 (In this study) Gageo Islands, Korea - KC524509a Haemadipsa rjukjuana Gageo 27 (In this study) Gageo Islands, Korea - KC524510a Haemadipsa rjukjuana L00112A Taiwan - HQ322438 Haemadipsa rjukjuana L00115A Taiwan - HQ322443 Haemadipsa rjukjuana L00098A Ryukyu Islands, Japan - HQ322462 Haemadipsa hainana voucher L00153A Hainan Island, China - HQ322473 Haemadipsa picta voucher L00151A Taiwan - HQ322470 Haemadipsa rjukjuana isolate HARY Taiwan HQ203097 HQ203174 Haemadipsa limuna isolate HACH China HQ207191 HQ203169 Haemadipsa montana isolate HZKI Nepal HQ203105 HQ203182 Haemadipsa ornata isolate HPISUM Sumatra HQ203101 HQ203178 Haemadipsa trimaculosa isolate HAKY Thailand HQ203095 HQ203172 Haemadipsa interrupta Thailand EU100069 EU100091 Haemadipsa sumatrana Borneo AY425464 AY425446 Haemadipsa sylvestris Vietnam AF116005 AF003266 Chtonobdella bilineata Australia AF116006 AF003267 Chtonobdella whitmani Australia EU100065 EU100087 Idiobdella seychellensis Seychelle Islands EU100070 EU100094 Malagabdella fallax Madagascar EU100071 EU100096 Diestecostoma mexicana Mexico EU100068 EU100089 Xerobdella lecometi Slovenia AF099947 EU100099 Aliolimnatis michaelseni Congo AF116010 AF116029 Hirudo medicinalis France AY786464 EU100093
Table 1. Nucleotide sequences of PCR primers and conditions for amplification of 18S rRNA and COI genes
Table 2. Collection localities and GenBank accession numbers used for phylogenetic analyses of haemadipsoid leeches in this study

GenBank accession numbers of H. rjukjuana Gageo which were acquired in this study are contemporary numbers, and the record is processing to release to the public database.

Table 1.

Table 2.

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