Fisheries Science

, Volume 75, Issue 5, pp 1167–1176 | Cite as

Origin of the diclidophorid monogenean Neoheterobothrium hirame Ogawa, 1999, the causative agent of anemia in olive flounder Paralichthys olivaceus

  • Tomoyoshi Yoshinaga
  • Nobuyuki Tsutsumi
  • Kathryn Ann Hall
  • Kazuo Ogawa
Original Article Biology


In the mid-1990s, Neoheterobothrium hirame suddenly appeared as a new species in olive flounder Paralichthys olivaceus in Japanese coastal waters. Anemia caused by the parasite has prevailed in wild and cultured populations of olive flounder since that time. In this study, to clarify the origin of N. hirame, two Neoheterobothrium species, namely unidentified Neoheterobothrium species (tentatively abbreviated as Neoheterobothrium sp. PL) and N. affine, were collected from Paralichthys lethostigma and Paralichthys dentatus, respectively, off the east coast of North America and compared with N. hirame collected in Japan. No substantial differences were detected in the morphology and DNA sequences of ITS1-5.8S rRNA-ITS2 and mitochondrial cytochrome oxidase subunit I (mt COI) regions between N. hirame and Neoheterobothrium sp. PL. On the other hand, the congeneric N. affine was clearly distinguished from both N. hirame and Neoheterobothrium sp. PL in its longer isthmus and the DNA sequences in ITS1 and mt COI. The absence of differences between N. hirame and Neoheterobothrium sp. PL and the clear difference between both of these and N. affine indicate that N. hirame is conspecific with Neoheterobothrium sp. PL infecting P. lethostigma and that N. hirame was recently introduced from North America to Japan.


Invasion Monogenea Neoheterobothriumaffine Neoheterobothrium hirame Paralichthys dentatus Paralichthys lethostigma Paralichthys olivaceus Parasite 



The authors thank Carter Watterson, Randy Gregory, David Taylor, and the late James Patrick Monaghan, Division of Marine Fisheries, North Carolina Department of Environment and Natural Resources, Morehead City, NC, for their help in collecting southern flounder and summer flounder. The authors also thank Dr. Lichtenfels, National Parasite Collection, U. S. Department of Agriculture, Beltsville, MD, for loaning the museum specimens. This research was partially supported by a Grant-in-Aid for Scientific Research (B) (no. 15405027) from the Japan Society for the Promotion of Science.


  1. 1.
    Michine A (1999) Yoshokujyo oyobi yousei shingyo hirame de hakkensareta kiseichu Neoheterobothrium sp. ni tsuite (On the parasite Neoheterobothrium sp. found in Japanese founder in aquaculture facilities and breeding stocks in a fish hatchery) (in Japanese). Shimaneken Saibaigyogyo Centaa Chosa Houkoku 2:15–23Google Scholar
  2. 2.
    Yoshinaga T, Kamaish T, Segawa I, Kumagai A, Nakayasu C, Yamano K, Takeuchi T, Sorimachi M (2000) Hinketsu hirame no ketuekiseijyo, byorisoshiki oyobi tanseirui Neoheterobothrium hirame no kiseijyokyo (Hematology, histopathology and the monegenean Neoheterobothrium hirame infection in anemic flounder) (in Japanese with English abstract). Fish Pathol 35:131–136Google Scholar
  3. 3.
    Yoshinaga T, Kamaishi T, Segawa I, Yamano K, Ikeda H, Sorimachi M (2001) Anemia caused by challenges with the monogenean Neoheterobothrium hirame in the Japanese flounder. Fish Pathol 36:13–20Google Scholar
  4. 4.
    Yoshinaga T, Kamaishi T, Ikeda H, Sorimachi M (2001) Experimental recovery from anemia in Japanese flounder challenged with the monogenean Neoheterobothrium hirame. Fish Pathol 36:179–182Google Scholar
  5. 5.
    Tanaka T, Shimamoto Y, Takeuchi T (2001) Himame no Neoheterobothrium sho (hinketsusho) ni kansuru kenkyu (Studies on the cause of Anemia caused by Neoheterobothrium hirame against Japanese flounder Paralichthys olivaceus) (in Japanese). Bull Wakayama Res Cent Agric For Fish 3:1–9Google Scholar
  6. 6.
    Mushiake K, Mori K, Arimoto M (2001) Epizootiology of anemia in wild Japanese flounder. Fish Pathol 36:125–132Google Scholar
  7. 7.
    Nakayasu C, Yoshinaga T, Kumagai A (2002) Hematology of anemia experimentally induced by repeated bleeding in Japanese flounder with comments on the cause of flounder anemia recently prevailing in Japan. Fish Pathol 37:125–130Google Scholar
  8. 8.
    Anshary H, Ogawa K, Higuchi M, Fujii T (2001) A study of long-term changes in summer infection levels of Japanese flounder Paralichthys olivaceus with the monogenean Neoheterobothrium hirame in the central Sea of Japan, with an application of a new technique for collecting small parasites from the gill filaments. Fish Pathol 36:27–32Google Scholar
  9. 9.
    Anshary H, Yamamoto E, Miyanaga T, Ogawa K (2001) Infection dynamics of the monogenean Neoheterobothrium hirame among young wild Japanese flounder in the western Sea of Japan. Fish Pathol 37:131–140Google Scholar
  10. 10.
    Sugaya T, Nishioka T, Mori K (2001) Tennen hirame ni kiseisuru Neoheterobothrium no kansenjyokyohaakuchosa (A survey on the infection of Neoheterobothrium hirame in wild Japanese flounder) (in Japanese). Saibaigyogyo Centaa Gijutsuhokoku 1:71–73Google Scholar
  11. 11.
    Tomiyama T, Watanabe M, Ebe K (2004) Infection dynamics of Neoheterobothrium hirame in Japanese flounder in the Joban Sea, eastern Japan. Fish Pathol 39:197–202Google Scholar
  12. 12.
    Yamada T, Motofuji Y (2005) Wakasawan no tennen hirame ni okeru Neoheterobothrium no kansenjyokyo (Infection of Neoheterobothrium hirame in 0+ age wild Japanese flounder in Wakasa Bay) (in Japanese). Saibaigyogyo Centaa Gijutsuhokoku 3:51–56Google Scholar
  13. 13.
    Shirakashi S, Yamada T, Yamada T, Ogawa K (2006) Infection dynamics of Neoheterobothirum hirame (Monogenea) on juvenile olive flounder, Paralichthys olivaceus (Temminck and Schlegel), in coastal waters of Japan. J Fish Dis 29:319–329PubMedCrossRefGoogle Scholar
  14. 14.
    Hayward CJ, Kim JH, Heo GJ (2001) Spread of Neoheterobothrium hirame (Monogenea), a serious pest of olive flounder Paralichthys olivaceus, to Korea. Dis Aquat Org 45:209–213CrossRefGoogle Scholar
  15. 15.
    Yoshinaga T, Kamaishi T, Segawa I (2001) Anemia of Japanese flounder caused by the monogenean Neoheterobothrium hirame. Bull Natl Res Inst Aquac Suppl 5:77–80Google Scholar
  16. 16.
    Shirakashi S, Teruya K, Ogawa K (2008) Altered behaviour and reduced survival of juvenile olive flounder, Paralichthys olivaceus, infected by an invasive monogenean, Neoheterobothrium hirame. Int J Parasitol 38:1513–1522CrossRefGoogle Scholar
  17. 17.
    Piasecki W, Wierzbicka J, Kempter JA (2000) New monogenean species parasitic on gills of bigmouth sole, Hipposlossina stomata Steindachner, 1876 from southern California. Acta Icht Piscat 30:127–134Google Scholar
  18. 18.
    González T, Oliva ME, Acuña EH (2002) Neoheterobothrium chilensis n. sp. (Monogenea: Diclidophoridae), a parasite of the bigeye flounder Hippoglossina macrops (Paralichthyidae: Pleuronectiformes) from northern Chile. J Parasitol 88:337–339PubMedGoogle Scholar
  19. 19.
    Price EW (1943) North American monogenetic trematodes: VI. the family Diclidophoridae (Dichlidophoridea). J Wash Acad Sci 33:44–54Google Scholar
  20. 20.
    Yoshinaga T, Tsutsumi N, Shima T, Kamaishi T, Ogawa K (2001) Experimental infection of southern flounder Paralichthys lethostigma with Neoheterobothrium hirame (Monogenea: Diclidophoridae). Fish Pathol 36:237–239Google Scholar
  21. 21.
    Ogawa K (1999) Neoheterobothrium hirame sp. nov. (Monogenea: Diclidophoridae) from the buccal cavity wall of Japanese flounder Paralichthys olivaceus. Fish Pathol 34:195–201Google Scholar
  22. 22.
    Iwashita M (2000) Gyoruikisei Pseudodactylogyrus aka tanseirui no shubunka ni kansuru kenkyu (Phylogenetic study of the species of the subfamily Pseudodactylogyrinae) (in Japanese). PhD Thesis, University of Tokyo, TokyoGoogle Scholar
  23. 23.
    Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and clustal X version 2.0. Bioinformatics 23:2947–2948PubMedCrossRefGoogle Scholar
  24. 24.
    Desdevises Y, Jovelin R, Jousson O, Morand S (2000) Comparison of ribosomal DNA sequences of Lamellodiscus spp. (Monogenea, Diplectanidae) parasitising Pagellus (Sparidae, Teleostei) in the North Mediterranean Sea: species divergence and coevolutionary interactions. Int J Parasitol 30:741–746PubMedCrossRefGoogle Scholar
  25. 25.
    Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  26. 26.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  27. 27.
    Hebert PD, Cywinska A, Ball SL, deWaard JR (2003) Biological identification through DNA barcodes. Proc Biol Sci 270:313–321PubMedCrossRefGoogle Scholar
  28. 28.
    Hebert PD, Ratnasingham S, deWaard JR (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc Biol Sci 270:96–99CrossRefGoogle Scholar
  29. 29.
    Littlewood DTJ, Rohdes K, Clough KA (1997) Parasite speciation within or between host species? Phylogenetic evidence from site-specific polystome monogeneans. Int J Parasit 27:1289–1297CrossRefGoogle Scholar
  30. 30.
    Hansen H, Bakke TA, Bachmann L (2007) DNA taxonomy and barcoding of monogenean parasites: lessons from Gyrodactylus. Trends Parasitol 23:363–367PubMedCrossRefGoogle Scholar
  31. 31.
    Hayward CJ (2005) Monogenea polyopisthocotylea. In: Rohde K (ed) Marine parasitology. CSIRO, Collingwood, pp 55–63Google Scholar
  32. 32.
    Nagibina LF (1953) Heterobothrium affinis (Linton) and its position in the system of monogenetic trematodes of the family Diclidophoridae Fuhrmann (in Russian). Trudy Zool Inst Akad Nauk SSSR 13:137–144Google Scholar
  33. 33.
    Yamaguti S (1963) Systema Helminthum. Monogenea and Aspidocotylea, vol 4. Interscience, New YorkGoogle Scholar
  34. 34.
    Mamaev UL (1987) O sistematićeskom polożenii roda Neoheterobotrium Price, 1943 (Monogenea, Diclidophoridae) w svjazi s opisaniem novogo vida N. syacii sp. n. [On the systematic position of the genus Neoheterobotrium Price, 1943 (Monogenea, Diclidophoridae) in connection with the description of a new species N. syacii sp. n.] (in Russian with English abstract). Parazitologiya 21:69–73Google Scholar
  35. 35.
    Taraschewski H (2006) Hosts and parasites as aliens. J Helminthol 80:99–128PubMedCrossRefGoogle Scholar
  36. 36.
    Meinila M, Kuusela J, Zietara MS, Lumme J (2004) Initial steps of speciation by geographic isolation and host switch in the salmonid pathogen Gyrodactylus salaris (Monogenea: Gyrodactylidae). Int J Parasitol 34:515–526PubMedCrossRefGoogle Scholar
  37. 37.
    Kirk RS (2003) The impact of Anguillicola crassus on European eels. Fish Manag Ecol 10:385–394CrossRefGoogle Scholar
  38. 38.
    Burreson EM, Ford SE (2004) A review of recent information on the haplosporidia, with special reference to Haplosporidium nelsoni (MSX disease). Aquat Living Resour 17:499–517CrossRefGoogle Scholar
  39. 39.
    Kamaishi T, Yoshinaga T (2003) Detection of Haplosporidium nelsoni in Pacific oyster Crassostrea gigas in Japan. Fish Pathol 37:193–195Google Scholar
  40. 40.
    Elston RA, Farley CA, Kent ML (1986) Occurrence and significance of bonamiasis in European flat oysters Ostrea edulis in North America. Dis Aquat Org 2:49–54CrossRefGoogle Scholar
  41. 41.
    Shirakashi S, Nishioka T, Ogawa K (2009) Neoheterobothrium hirame (Monogenea) alters the feeding behaviour of juvenile olive flounder, Paralichthys olivaceus. Fish Sci 75:121–128CrossRefGoogle Scholar

Copyright information

© The Japanese Society of Fisheries Science 2009

Authors and Affiliations

  • Tomoyoshi Yoshinaga
    • 1
  • Nobuyuki Tsutsumi
    • 1
    • 2
  • Kathryn Ann Hall
    • 1
    • 3
  • Kazuo Ogawa
    • 1
  1. 1.The Graduate School of Agricultural and Life SciencesThe University of TokyoTokyoJapan
  2. 2.Nippon Institute for Biological ScienceTokyoJapan
  3. 3.Sessile Marine InvertebratesQueensland MuseumSouth BrisbaneAustralia

Personalised recommendations