Parasitology Research

, Volume 109, Issue 5, pp 1429–1437 | Cite as

Molecular characterization of Hysterothylacium aduncum (Nematoda: Raphidascaridae) from different fish caught off the Tunisian coast based on nuclear ribosomal DNA sequences

  • Nabil Amor
  • Sarra Farjallah
  • Paolo Merella
  • Khaled Said
  • Badreddine Ben Slimane
Original Paper
First Online:
Received:
Accepted:

Abstract

Larval forms of the genus Hysterothylacium have been previously reported in teleost fish from the North African coasts of central Mediterranean Sea by morphological analysis. In the present study, samples identified morphologically as Hysterothylacium aduncum (n = 62), from Merluccius merluccius, Trachurus mediterraneus and Pagellus erythrinus from different geographical locations of the Tunisian coasts, were genetically characterised by sequences of the first (ITS-1), the 5.8S and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the sequences obtained with those available in public gene databases confirmed that all the samples from the Tunisian coasts belong to a single species, namely H. aduncum. All specimens from the Tunisian coasts showed one indel in position 787 in ITS-2 sequences not reported by any of the previously published sequences from the Atlantic and Pacific Oceans, Adriatic Sea (Mediterranean Sea) and the East Greenland Sea, suggesting the existence of a population-specific pattern exhibiting a low differentiation of this parasite in this area. This is the first molecular characterization of H. aduncum from the Tunisian coasts using ITS rDNA sequences which allows the definition of genetic markers for their unequivocal identification, and provides further biological data on these nematodes in marine fish off the Tunisian coasts, improving the picture of the occurrence of these taxa in the North African coasts of central Mediterranean Sea.

Notes

Acknowledgements

The authors are very grateful to two anonymous referees, whose comments and suggestions were very useful in improving the MS.

References

  1. Abollo E, Gestal C, Pascual S (2001) Anisakis infestation in marine fish and cephalopods from Galician waters: an updated perspective. Parasitol Res 87:492–499PubMedCrossRefGoogle Scholar
  2. Anderson TJC, Blouin MS, Beech RN (1998) Population biology of parasitic nematodes: application of genetic markers. Adv Parasitol 41:219–283PubMedCrossRefGoogle Scholar
  3. Avise JC (2000) Phylogeography: the history and formation of species. Harvard University Press, CambridgeGoogle Scholar
  4. Balbuena JA, Karlsbakk E, Kvenseth AM, Saksvik M, Nylund A (2000) Growth and emigration of third-stage larvae of Hysterothylacium aduncum (Nematoda: Anisakidae) in larval herring Clupea harengus. J Parasitol 86(6):1271–1275PubMedGoogle Scholar
  5. Blouin MS, Yowell CA, Courtney CH, Dame JB (1995) Host movement and the genetic structure of populations of parasitic nematodes. Genetics 141:1007–1014PubMedGoogle Scholar
  6. Blouin MS, Yowell CA, Courtney CH, Dame JB (1998) Substitution bias, rapid saturation, and the use of mtDNA for nematode systematics. Mol Biol Evol 15:1719–1727PubMedGoogle Scholar
  7. Bullini L, Arduino P, Cianchi R, Nascetti G, D'Amelio S, Mattiucci S, Paggi L, Orecchia P, Plotz J, Berland B, Smith JW, Brattey J (1997) Genetic and ecological research on Anisakid endoparasites of fish and marine mammals in the Antarctic and Arctic-Boreal Regions. In: Battaglia B, Valencia J, Walton DW (Eds.), Antarctic Communities: Species, Structure and Survival. Cambridge University Press 464Google Scholar
  8. Criscione CD, Poulin R, Blouin MS (2005) Molecular ecology of parasites: elucidating ecological and microevolutionary processes. Mol Ecol 14:2247–2257PubMedCrossRefGoogle Scholar
  9. D'Amelio S, Mathiopoulos KD, Santos CP, Pugachev ON, Webb SC, Picanço M, Paggi L (2000) Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: Ascaridoidae) defined by polymerase chain reaction-based restriction fragment length polymorphism. Int J Parasitol 30:223–226PubMedCrossRefGoogle Scholar
  10. D'Amelio S, Barros NB, Ingrosso S, Fauquier DA, Russo R, Paggi L (2007) Genetic characterization of members of the genus Contracaecum (Nematoda: Anisakidae) from fish-eating birds from west-central Florida, USA, with evidence of new species. Parasitology 134:1041–1051PubMedCrossRefGoogle Scholar
  11. Daschner A, Pascual CY (2005) Anisakis simplex: sensitization and clinical allergy. Curr Opin Allergy Clin Immunol 5:281–285PubMedCrossRefGoogle Scholar
  12. Farjallah S, Ben Slimane B, Blel H, Amor N, Said K (2006) Anisakid parasites of two forkbeards (Phycis blennoides and Phycis phycis) from the eastern Mediterranean coasts in Tunisia. Parasitol Res 100:11–17PubMedCrossRefGoogle Scholar
  13. Farjallah S, Ben Slimane B, Busi M, Paggi L, Amor N, Blel H, Said K, D'Amelio S (2008a) Occurrence and molecular identification of Anisakis spp. from the North African coasts of Mediterranean Sea. Parasitol Res 102(3):371–379PubMedCrossRefGoogle Scholar
  14. Farjallah S, Merella P, Ingrosso S, Rotta A, Ben Slimane B, Garippa G, Said K, Busi M (2008b) Molecular evidence for the occurrence of Contracaecum rudolphii A (Nematoda: Anisakidae) in shag Phalacrocorax aristotelis (Linnaeus) (Aves: Phalacrocoracidae) from Sardinia (western Mediterranean Sea). Parasitol Int 57:437–440PubMedCrossRefGoogle Scholar
  15. George-nascimento M, Llanos A (1995) Microevolutionary implications of allozymic and morphometric variations in sealworms Pseudoterranova sp. (Ascaridoidea: Anisakidae) among sympatric hosts from the Southeastern Pacific Ocean. Int J Parasitol 25:1163–1171PubMedCrossRefGoogle Scholar
  16. Hillis DM, Dixon MT (1991) Ribosomal DNA: molecular evolution and phylogenetic inference. Q Rev Biol 66:411–453PubMedCrossRefGoogle Scholar
  17. Hillis DM, Bull JJ (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Syst Biol 42:182–192Google Scholar
  18. Hu M, D'Amelio S, Zhu X, Paggi L, Gasser R (2001) Mutation scanning for sequence variation in three mitochondrial DNA regions for members of the Contracaecum osculatum (Nematoda: Ascaridoidea) complex. Electrophoresis 22:1069–1075PubMedCrossRefGoogle Scholar
  19. Ishikura H (1989) General survey of Anisakis and Anisakiasis in Japan. In: Ishikura H, Namiki M (eds) Gastric Anisakiasis in Japan. Springer–Verlag, Tokyo, pp 3–11Google Scholar
  20. Kellermanns E, Klimpel S, Palm HW (2007) Molecular identification of ascaridoid nematodes from the deep-sea onion-eye grenadier (Macrourus berglax) from the East Greenland Sea. Deep Sea Res Oceanogr Res Paper 54:2194–2202CrossRefGoogle Scholar
  21. Klimpel S (2005) Distribution of nematodes of the family Anisakidae in commercially important fish species from the central and northern North Sea. Zeitschrift fur Fischkunde 7:161–168Google Scholar
  22. Klimpel S, Ruckert S (2005) Life cycle strategy of Hysterothylacium aduncum to become the most abundant anisakid fish nematode in the North Sea. Parasitol Res 97:340–343CrossRefGoogle Scholar
  23. Klimpel S, Seehagen A, Palm HW, Rosenthal H (2001) Deep-water metazoan fish parasites of the world. Logos Verlag, Berlin, 316Google Scholar
  24. Klimpel S, Palm HW, Seehagen A (2003) Metazoan parasites and food composition of juvenile Etmopterus spinax (L., 1758) (Dalatiidae, Squaliformes) from the Norwegian Deep. Parasitol Res 89:245–251PubMedCrossRefGoogle Scholar
  25. Klimpel S, Palm HW, Busch MW, Kellermanns E, Ruckert S (2006) Fish parasites in the Arctic deep-sea: poor diversity in meso-/bathypelagial vs. heavy parasite load in a demersal fish. Deep Sea Res Oceanogr Res Paper 53:1167–1181CrossRefGoogle Scholar
  26. Klimpel S, Kleinertz S, Hanel R, Rückert S (2007) Genetic variability in Hysterothylacium aduncum, a raphidascarid nematode isolated from sprat (Sprattus sprattus) of different geographical areas of the northeastern Atlantic. Parasitol Res 101:1425–1430PubMedCrossRefGoogle Scholar
  27. Koie M (1993) Aspects of the life cycle and morphology of Hysterothylacium aduncum (Rudolphi, 1802) (Nematoda, Ascaridoidea, Anisakidae). Can J Zool 71:1289–1296CrossRefGoogle Scholar
  28. Li AX, D'Amelio S, Paggi L, He F, Gasser RB, Lun ZR, Abollo E, Turchetto M, Zhu XQ (2005) Genetic evidence for the existence of sibling species within Contracaecum rudolphii (Hartwich, 1964) and the validity of Contracaecum septentrionale (Kreis, 1955) (Nematoda: Anisakidae). Parasitol Res 96:361–366PubMedCrossRefGoogle Scholar
  29. Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452PubMedCrossRefGoogle Scholar
  30. Martin-Sánchez J, Paniagua I, Valero A (1998) Contribution to the knowledge of Hysterothylacium aduncum through electrophoresis of the enzymes glucose phosphate isomerase and phosphoglucomutase. Parasitol Res 84:160–163Google Scholar
  31. Mattiucci S, Nascetti G (2008) Advances and trends in the molecular systematics of Anisakid nematodes, with implications for their evolutionary ecology and host–parasite co-evolutionary processes. Adv Parasitol 66:47–148PubMedCrossRefGoogle Scholar
  32. Mattiucci S, Cianchi R, Paggi L, Arduino P, Margolis L, Brattey J, Webb SC, D'Amelio S, Orecchia P, Bullini L (1997) Genetic and ecological data on the Anisakis simplex complex, with evidence for a new species (Nematoda, Ascaridoidea, Anisakidae). J Parasitol 86:401–416CrossRefGoogle Scholar
  33. Mattiucci S, Paggi L, Nascetti G, Abollo E, Webb SC, Pascual S, Cianchi R, Bullini L (2001) Genetic divergence and reproductive isolation between Anisakis brevispiculata and Anisakis physeteris (Nematoda: Anisakidae). Int J Parasitol 31:9–14PubMedCrossRefGoogle Scholar
  34. Mattiucci S, Paggi L, Nascetti G, Portes Santos C, Costa G, Di Beneditto AP, Ramos R, Argyrou M, Cianchi R, Bullini L (2002) Genetic markers in the study of Anisakis typica (Diesing, 1860): larval identification and genetic relationships with other species of Anisakis Dujardin, 1845 (Nematoda: Anisakidae). Syst Parasitol 51:159–170PubMedCrossRefGoogle Scholar
  35. Mattiucci S, Nascetti G, Dailey M, Webb SC, Barros NB, Cianchi R, Bullini L (2005) Evidence for a new species of Anisakis Dujardin, 1845: morphological description and genetic relationships between congeners (Nematoda: Anisakidae). Syst Parasitol 61:157–171PubMedCrossRefGoogle Scholar
  36. Mattiucci S, Contadini R, Paoletti M, Nascetti G (2006) Relazioni genetiche tra specie del genere Contracaecum (Railliet et Henry, 1912), parassiti di pinnipedi, ed aspetti coevolutivi ospite-parassita. Abstracts XVI Congresso Societa' Italiana di Ecologia 98Google Scholar
  37. Mattiucci S, Abaunza P, Damiano S, Garcia A, Santos MN, Nascetti G (2007) Distribution of Anisakis larvae identified by genetic markers and their use for stock characterization of demersal and pelagic fish from European waters: an update. J Helminthol 81:117–127PubMedCrossRefGoogle Scholar
  38. Mattiucci S, Paoletti M, Webb SC (2009) Anisakis nascettii n. sp. (Nematoda: Anisakidae) from beaked whales of the southern hemisphere: morphological description, genetic relationships between congeners and ecological data. Syst Parasitol 74(3):199–217PubMedCrossRefGoogle Scholar
  39. Mattiucci S, Paoletti M, Marcer F, Gazzonis A, Manfredi MT, Fernandez AJ, Gonzales F, Nascetti G (2010) Genetic heterogeneity within Anisakis physeteris (sensu lato) (Nematoda: Anisakidae) from sperm whales, Physeter macrocephalus, from Mediterranean Sea (Apulian coast) and Atlantic Ocean (Canaries coast). Parassitologia 52:357Google Scholar
  40. McCarthy J, Moore TA (2000) Emerging helminth zoonoses. Int J Parasitol 30:1351–1360PubMedCrossRefGoogle Scholar
  41. Navone GT, Sardella NH, Timi JT (1998) Larvae and adults of Hysterothylacium aduncum (Rudolphi, 1802) (Nematoda: Anisakidae) in fishes and crustaceans in the South West Atlantic. Parasite 5:127–136PubMedGoogle Scholar
  42. Paggi L, Nascetti G, Cianchi R, Orecchia P, Mattiucci S, D'Amelio S, Berland B, Brattey J, Smith JW, Bullini L (1991) Genetic evidence for three species within Pseudoterranova decipiens (Nematoda, Ascaridida, Ascaridoidea) in the North Atlantic and Norwegian and Barents Seas. Int J Parasitol 21:195–212PubMedCrossRefGoogle Scholar
  43. Paggi L, Nascetti G, Webb SC, Mattiucci S, Cianchi R, Bullini L (1998) A new species of Anisakis Dujardin, 1845 (Nematoda: Anisakidae) from beaked whale (Ziphiidae): allozyme and morphological evidence. Syst Parasitol 40:161–174CrossRefGoogle Scholar
  44. Paggi L, Mattiucci S, Gibson DI, Berland B, Nascetti G, Cianchi R, Bullini L (2000) Pseudoterranova decipiens species A and B (Nematoda: Ascaridoidea): nomenclatural designation, morphological diagnostic characters and genetic markers. Syst Parasitol 45:185–197PubMedCrossRefGoogle Scholar
  45. Palm HW, Klimpel S, Bucher C (1999) Checklist of metazoan fish parasites of German coastal waters. Ber Inst Meereskd Kiel 307:1–148Google Scholar
  46. Petter AJ, Maillard C (1988) Larves d'Ascarides parasites de poissons en Méditerranée occidentale. Bull Mus Hist Nat Paris 4(10):347–369Google Scholar
  47. Pontes T, D'Amelio S, Costa G, Paggi L (2005) Molecular characterization of larval anisakid nematodes from Marine fishes of Madeira by a PCR-based approach, with evidence for a new species. J Parasitol 91:1430–1434PubMedCrossRefGoogle Scholar
  48. Shih HH (2004) Parasitic helminth fauna of the cutlass fish, Trichiurus lepturus L., and the differentiation of four anisakid nematode third-stage larvae by nuclear ribosomal DNA sequences. Parasitol Res 93:188–195PubMedCrossRefGoogle Scholar
  49. Shih HH, Jeng MS (2002) Hysterothylacium aduncum (Nematoda: Anisakidae) infecting a herbivorous fish, Siganus fuscescens, off the Taiwanese coast of the northwest Pacific. Zool Stud 41:185–189Google Scholar
  50. Szostakowska B, Myjak P, Kur J (2002) Identification of anisakid nematodes from the Southern Baltic Sea using PCR-based methods. Mol Cell Probes 16:111–118PubMedCrossRefGoogle Scholar
  51. Takahashi S (1998) Anisakidosis: global point of view. In: Ishikura H, Aikawa M, Itakura H, Kikuchi K (eds) Host response to international parasitic zoonoses. Springer, Tokyo, pp 109–120Google Scholar
  52. 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
  53. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTALW: 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
  54. Valentini A, Mattiucci S, Bondanelli P, Webb SC, Mignucci-Giannone AA, Colom-Llavina MM (2006) Genetic relationships among Anisakis species (Nematoda: Anisakidae) inferred from mitochondrial cox2 sequences, and comparison with allozyme data. J Parasitol 92:156–166PubMedCrossRefGoogle Scholar
  55. Zhang L, Hu M, Shamsi S, Beveridge I, Li H, Xu Z, Li L, Cantacessi C, Gasser RB (2007) The specific identification of anisakid larvae from fishes from the Yellow Sea, China, using mutation scanning-coupled sequence analysis of nuclear ribosomal DNA. Mol Cell Probes 21:386–390PubMedCrossRefGoogle Scholar
  56. Zhu XQ, D'Amelio S, Gasser RB, Yang TB, Paggi L, He F, Lin RQ, Song HQ, Ai L, Li AX (2007) Practical PCR tools for the delineation of Contracaecum rudolphii A and Contracaecum rudolphii B (Ascaridoidea: Anisakidae) using genetic markers in nuclear ribosomal DNA. Mol Cell Probes 21:97–102PubMedCrossRefGoogle Scholar
  57. Zhu XQ, D'Amelio S, Hu M, Paggi L, Gasser RB (2001) Electrophoretic detection of population variation within Contracaecum ogmorhini (Nematoda: Ascaridoidea: Anisakidae). Electrophoresis 22:1930–1934PubMedCrossRefGoogle Scholar
  58. Zhu XQ, D'Amelio S, Paggi L, Gasser RB (2000) Assessing sequence variation in the internal transcribed spacers of ribosomal DNA within and among members of the Contracaecum osculatum complex (Nematoda: Ascaridoidea: Anisakidae). Parasitol Res 86:677–683PubMedCrossRefGoogle Scholar
  59. Zhu XQ, D'Amelio S, Palm HW, Paggi L, George-Nascimento M, Gasser RB (2002) SSCP-based identification of members within the Pseudoterranova decipiens complex (Nematoda: Ascaridoidea: Anisakidae) using genetic markers in the internal transcribed spacers of ribosomal DNA. Parasitology 24:615–623Google Scholar
  60. Zhu XQ, Gasser RB, Podolska M, Chilton N (1998) Characterization of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. Int J Parasitol 28:1911–1921PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Nabil Amor
    • 1
  • Sarra Farjallah
    • 1
    • 2
  • Paolo Merella
    • 3
  • Khaled Said
    • 1
  • Badreddine Ben Slimane
    • 4
  1. 1.Unité de Recherche: Génétique, Biodiversité et Valorisation des Bioressources UR/09-30, Institut Supérieur de Biotechnologie de MonastirMonastirTunisia
  2. 2.Unité de Recherche: Génétique, Biodiversité et Valorisation des Bioressources UR/09-30, Institut Supérieur de Biotechnologie de MonastirMonastirTunisia
  3. 3.Sezione di Parassitologia e Malattie Parassitarie Dipartimento di Biologia AnimaleUniversità degli Studi di SassariSardaigneItaly
  4. 4.Institut Supérieur des Sciences et Technologies de l’Environnement de Borj EssedriaBorj EssedriaTunisia

Personalised recommendations