Parasitology Research

, Volume 118, Issue 5, pp 1393–1402 | Cite as

On the occurrence and molecular identification of Contracaecum larvae (Nematoda: Anisakidae) in Mugil cephalus from Turkish waters

  • Gokmen Zafer PekmezciEmail author
  • Banu Yardimci
Fish Parasitology - Original Paper


Anisakis and Contracaecum species are fish borne zoonotic nematodes. In our previous studies, other larval anisakid and raphidascarid nematodes, Anisakis and Hysterothylacium species, were genetically identified in marine fish from Turkish waters. However, there is no information on molecular identification of larval Contracaecum species in marine fish from Turkey. Therefore, the aim of this study was only to investigate the presence and molecular identification of Contracaecum species in commonly commercialized marine fish from Turkish waters. A total of 475 marine fish, which belong to 21 different species, were sampled from the Aegean (FAO 37.3.1), Mediterranean (FAO 37.3.2), and Black Sea (FAO 37.4.2). The prevalence of Contracaecum L3 larvae in the Aegean Sea was identified as 10% in Mugil cephalus. All Contracaecum L3 larvae were molecularly characterized with RFLP targeting the ITS region and rrnS gene. Moreover, all larvae were analyzed by sequencing of ITS region, rrnS and cox2 gene. All Contracaecum larvae were identified as C. overstreeti based on the cox2 sequence analysis. This is the first report of C. overstreeti larvae in M. cephalus as paratenic and intermediate hosts. Furthermore, the analysis reveals novel information on ITS region. Additionally, the rrnS gene of C. overstreeti was also achieved and deposited in Genbank for the first time. The PCR-RFLP patterns of the ITS region and rrnS gene from C. overstreeti were presented in the present study. Consequently, the presence of C. overstreeti larvae in M. cephalus from the Aegean Sea may also potentially capable of inducing allergic sensitization in humans. 


Contracaecum overstreeti ITS region cox2 rrnS Mugil cephalus  Turkish waters 



We would like to thank Associate Professor Didem Pekmezci (University of Ondokuz Mayis, Turkey) and Oyku Naz Attila (University of Strathclyde, Glasgow, UK) for their help in English proofreading.

Funding information

This study was supported by The Scientific Research Council of the University Ondokuz Mayis, Samsun, Turkey (project number PYO.VET.1902.15.002).

Supplementary material

436_2019_6278_MOESM1_ESM.docx (26 kb)
ESM 1 (DOCX 26.0 kb)


  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefGoogle Scholar
  2. Berland B (1961) Nematodes from some Norwegian marine fishes. Sarsia 2:1–50CrossRefGoogle Scholar
  3. Cannon LRG (1977) Some larval ascaridoids from south-eastern Queensland marine fishes. Int J Food Microbiol 7:233–243Google Scholar
  4. 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 134:1041–1051CrossRefGoogle Scholar
  5. Deardorff TL, Overstreet RM (1980) Contracaecum multipapillatum (= C. robustum) from fishes and birds in the northern Gulf of Mexico. J Parasitol 66:853–856CrossRefGoogle Scholar
  6. Dezfuli BS, Manera M, Bosi G, DePasquale JA, D’Amelio S, Castaldelli G, Giari L (2016) Anguilla anguilla intestinal immune response to natural infection with Contracaecum rudolphii A larvae. J Fish Dis 39:1187–1200CrossRefGoogle Scholar
  7. EFSA (2010) Scientific opinion of the panel on biological hazards on risk assessment of parasites in fishery products. EFSA J 8(4):1543CrossRefGoogle Scholar
  8. Farjallah S, Merella P, Ingrosso S, Rotta A, Slimane BB, Garippa G, Said K, Busi M (2008) 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 57:437–440CrossRefGoogle Scholar
  9. Farris JS, Källersjö M, Kluge AG, Bult C (1994) Testing significance of incongruence. Cladistics 10:315–319CrossRefGoogle Scholar
  10. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefGoogle Scholar
  11. Fontenelle G, Knoff M, Verícimo MA, São Clemente SC (2018) Epicutaneous sensitization with nematode antigens of fish parasites results in the production of specific IgG and IgE. J Helminthol 92:403–409CrossRefGoogle Scholar
  12. Froese R, Pauly D (2017) FishBase, Accessed 19 Dec 2017
  13. Garbin LE, Mattiucci S, Paoletti M, González-Acuña D, Nascetti G (2011) Genetic and morphological evidences for the existence of a new species of Contracaecum (Nematoda: Anisakidae) parasite of Phalacrocorax brasilianus (Gmelin) from Chile and its genetic relationships with congeners from fish-eating birds. J Parasitol 97:476–492CrossRefGoogle Scholar
  14. Garbin LE, Mattiucci S, Paoletti M, Diaz JI, Nascetti G, Navone GT (2013) Molecular identification and larval morphological description of Contracaecum pelagicum (Nematoda: Anisakidae) from the anchovy Engraulis anchoita (Engraulidae) and fish-eating birds from the Argentine North Patagonian Sea. Parasitol Int 62:309–319CrossRefGoogle Scholar
  15. Giriskin AO, Alasonyalilar-Demirer A, Giriskin O (2012) A case of Contracaecum sp. (Ascaridida: Anisakidae) infection in Dalmatian Pelican (Pelecanus crispus). Kafkas Univ Vet Fak Derg 18:227–229Google Scholar
  16. Gutiérrez-Galindo JF, Osanz-Mur AC, Mora-Ventura MT (2010) Occurrence and infection dynamics of anisakid larvae in Scomber scombrus, Trachurus trachurus, Sardina pilchardus, and Engraulis encrasicolus from Tarragona (NE Spain). Food Contr 21:1550–1555CrossRefGoogle Scholar
  17. Hillis DM, Bull JJ (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Syst Biol 42:182–192CrossRefGoogle Scholar
  18. Iglesias L, Gómez del Prado MC, Adroher FJ, Valero A (1998) Floridosentis elongatus y Contracaecum sp., parásitos de Mugil cephalus en Baja California Sur, México, First Virtual Congress about Pharmacy (IVCP), Universidad de Granada, Spain, C-068Google Scholar
  19. Iglesias L, Benítez R, Adroher FJ, Valero A (2011) Helminth infection in Mugil incilis from Cartagena de Indias, Colombian Caribbean coast. Helminthologia 48:36–40CrossRefGoogle Scholar
  20. Im KI, Shin HJ, Kim BH, Moon SI (1995) Gastric anisakiasis cases in Cheju-do, Korea. Korean J Parasitol 33:179–186CrossRefGoogle Scholar
  21. Ishikura H, Takahashi S, Sato N, Matsuura A, Nitto H, Tsunokawa M, Kikuchi K (1996) Epidemiology of anisakidosis and related human diseases and studies on parasites infecting marine mammals, fishes and squids. Bull Mar Biomed Inst Sapporo Med Univ Sch Med 3:23–37Google Scholar
  22. Jabbar A, Fong RW, Kok KX, Lopata AL, Gasser RB, Beveridge I (2013) Molecular characterization of anisakid nematode larvae from 13 species of fish from Western Australia. Int J Food Microbiol 161:247–253CrossRefGoogle Scholar
  23. Keskin E, Koyuncu CE, Genc E (2015) Molecular identification of Hysterothylacium aduncum specimens isolated from commercially important fish species of Eastern Mediterranean Sea using mtDNA cox1 and ITS rDNA gene sequences. Parasitol Int 64:222–228CrossRefGoogle Scholar
  24. Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874CrossRefGoogle Scholar
  25. 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 66:47–148CrossRefGoogle Scholar
  26. Mattiucci S, Cianchi R, Nascetti G, Paggi L, Sardella N, Timi JT, Webb SC, Bastida R, Rodríguez D, Bullini L (2003) Genetic evidence for two sibling species within Contracaecum ogmorhini Johnston & Mawson, 1941 (Nematoda: Anisakidae) from otariid seals of boreal and austral regions. Syst Parasitol 54:13–23CrossRefGoogle Scholar
  27. Mattiucci S, Paoletti M, Olivero-Verbel J, Baldiris R, Arroyo-Salgado B, Garbin L, Navone G, Nascetti G (2008a) Contracaecum bioccai n. sp. from the brown pelican Pelecanus occidentalis (L.) in Colombia (Nematoda: Anisakidae): morphology, molecular evidence and its genetic relationship with congeners from fish-eating birds. Syst Parasitol 69:101–121CrossRefGoogle Scholar
  28. Mattiucci S, Paoletti M, Webb SC, Sardella N, Timi JT, Berland B, Nascetti G (2008b) Genetic relationships among species of Contracaecum Railliet & Henry, 1912 and Phocascaris Höst, 1932 (Nematoda: Anisakidae) from pinnipeds inferred from mitochondrial cox2 sequences, and congruence with allozyme data. Parasite 15:408–419CrossRefGoogle Scholar
  29. Mattiucci S, Paoletti M, Solorzano AC, Nascetti G (2010) Contracaecum gibsoni n. sp. and C. overstreeti n. sp. (Nematoda: Anisakidae) from the Dalmatian pelican Pelecanus crispus (L.) in Greek waters: genetic and morphological evidence. Syst Parasitol 75:207–224CrossRefGoogle Scholar
  30. Mattiucci S, Cipriani P, Paoletti M, Nardi V, Santoro M, Bellisario B, Nascetti G (2015) Temporal stability of parasite distribution and genetic variability values of Contracaecum osculatum sp. D and C. osculatum sp. E (Nematoda: Anisakidae) from fish of the Ross Sea (Antarctica). Int J Parasitol Parasites Wildl 4:356–367CrossRefGoogle Scholar
  31. Merella P, Garippa G (2001) Metazoan parasites of grey mullets (Teleostea: Mugilidae) from the Mistras Lagoon (Sardinia-western Mediterranean). Sci Mar 65:201–206CrossRefGoogle Scholar
  32. Nadler SA, Hudspeth DS (2000) Phylogeny of the Ascaridoidea (Nematoda: Ascaridida) based on three genes and morphology: hypotheses of structural and sequence evolution. J Parasitol 86:380–393CrossRefGoogle Scholar
  33. Nagasawa K (2012) The biology of Contracaecum osculatum sensu lato and C. osculatum A (Nematoda: Anisakidae) in Japanese waters: a review. Biosph Sci 51:61–69Google Scholar
  34. Olivero-Verbel J, Baldiris-Ávila R, Arroyo-Salgado B (2005) Nematode infection in Mugil incilis (Lisa) from Cartagena Bay and Totumo marsh, north of Colombia. J Parasitol 91:1109–1112CrossRefGoogle Scholar
  35. Ozturk T, Yesil A (2018) Metazoan parasite fauna of the red mullet, Mullus barbatus ponticus Essipov, 1927 in the Sinop coasts of the Black Sea. Turk J Fish Aquat Sci 18:153–160CrossRefGoogle Scholar
  36. Pekmezci GZ (2014) Occurrence of Anisakis simplex sensu stricto in imported Atlantic mackerel (Scomber scombrus) represents a risk for Turkish consumers. Int J Food Microbiol 185:64–68CrossRefGoogle Scholar
  37. Pekmezci GZ, Bolukbas CS, Gurler AT, Onuk EE (2013) Occurrence and molecular characterization of Hysterothylacium aduncum (Nematoda: Anisakidae) from Merlangius merlangus euxinus and Trachurus trachurus off the Turkish coast of Black Sea. Parasitol Res 112:1031–1037CrossRefGoogle Scholar
  38. Pekmezci GZ, Onuk EE, Bolukbas CS, Yardimci B, Gurler AT, Acici M, Umur S (2014a) Molecular identification of Anisakis species (Nematoda: Anisakidae) from marine fishes collected in Turkish waters. Vet Parasitol 201:82–94CrossRefGoogle Scholar
  39. Pekmezci GZ, Yardimci B, Onuk EE, Umur S (2014b) Molecular characterization of Hysterothylacium fabri (Nematoda: Anisakidae) from Zeus faber (Pisces: Zeidae) caught off the Mediterranean coasts of Turkey based on nuclear ribosomal and mitochondrial DNA sequences. Parasitol Int 63:127–131CrossRefGoogle Scholar
  40. Peter AJ, Maillard C (1988) Larves d’ascarides parasites de poisons en Mediterranee occidentale. Bull Mus Natl Hist Nat 10(sect. A):347–369Google Scholar
  41. Pulleiro-Potel L, Barcala E, Mayo-Hernández E, Muñoz P (2015) Survey of anisakids in commercial teleosts from the western Mediterranean Sea: infection rates and possible effects of environmental and ecological factors. Food Control 55:12–17CrossRefGoogle Scholar
  42. Reiczigel J, Rózsa L (2005) Quantitative Parasitology 3.0. BudapestGoogle Scholar
  43. Rózsa L, Reiczigel J, Majoros G (2000) Quantifying parasites in samples of hosts. J Parasitol 86:228–232CrossRefGoogle Scholar
  44. Salati F, Meloni M, Cau M, Angelucci G (2013) Presence of Contracaecum spp. in teleosts cultured and fished in Sardinia. Vet Parasitol 196:382–387CrossRefGoogle Scholar
  45. Schaum E, Müller D (1967) Die Heterocheilidiasis. Eine Infektion des Mensches mit Larven von Fisch Ascariden. Dtsch Med Wochenschr 92:2230–2233CrossRefGoogle Scholar
  46. Shamsi S, Butcher AR (2011) First report of human anisakidosis in Australia. Med J Aust 194:199–200Google Scholar
  47. Shamsi S, Suthar J (2016) A revised method of examining fish for infection with zoonotic nematode larvae. Int J Food Microbiol 227:13–16CrossRefGoogle Scholar
  48. Shamsi S, Gasser R, Beveridge I, Shabani AA (2008) Contracaecum pyripapillatum n. sp. (Nematoda: Anisakidae) and a description of C. multipapillatum (von Drasche, 1882) from the Australian pelican, Pelecanus conspicillatus. Parasitol Res 103:1031–1039CrossRefGoogle Scholar
  49. Shamsi S, Eisenbarth A, Saptarshi S, Beveridge I, Gasser RB, Lopata AL (2011a) Occurrence and abundance of anisakid nematode larvae in five species of fish from southern Australian waters. Parasitol Res 108:927–934CrossRefGoogle Scholar
  50. Shamsi S, Gasser RB, Beveridge I (2011b) Mutation scanning-coupled sequencing of nuclear ribosomal DNA spacers as a tool for the specific identification of different Contracaecum (Nematoda: Anisakidae) larval types. Mol Cell Probes 25:13–18CrossRefGoogle Scholar
  51. Shamsi S, Steller E, Chen Y (2018a) New and known zoonotic nematode larvae within selected fish species from Queensland waters in Australia. Int J Food Microbiol 272:73–82CrossRefGoogle Scholar
  52. Shamsi S, Turner A, Wassens S (2018b) Description and genetic characterization of a new Contracaecum larval type (Nematoda: Anisakidae) from Australia. J Helminthol 92:216–222CrossRefGoogle Scholar
  53. Simsek E, Ciloglu A, Yildirim A, Pekmezci GZ (2018) Identification and molecular characterization of Hysterothylacium (Nematoda: Raphidascarididae) larvae in bogue (Boops boops L.) from the Aegean Sea, Turkey. Kafkas Univ Vet Fak Derg 24:525–530Google Scholar
  54. Swofford DL (2002) PAUP*: Phylogenetic Analysis Using Parsimony (*and Other Methods), version 4.0b10. Sinauer Associates, SunderlandGoogle Scholar
  55. Szostakowska B, Fagerholm HP (2007) Molecular identification of two strains of third-stage larvae of Contracaecum rudolphii sensu lato (Nematoda: Anisakidae) from fish in Poland. J Parasitol 93:961–964CrossRefGoogle Scholar
  56. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680CrossRefGoogle Scholar
  57. Valles-Ríos ME, Ruiz-Campos G, Galavíz-Silva L (2000) Prevalencia e intensidad parasitaria en Mugil cephalus (Pisces: Mugilidae), del Río Colorado, Baja California, México. Rev Biol Trop 48:495–501Google Scholar
  58. Valles-Vega GI (2011) Supervivencia de larvas III de Contracaecum multipapillatum sensu lato (Nematoda: Anisakidae) de lisas (Pisces: Mugilidae) en diferentes medios culinarios y aspectos sobre su ciclo de vida en La Paz, B.C.S. BSc thesis, Universidad Autónoma de Baja California Sur, La PazGoogle Scholar
  59. Valles-Vega GI, Molina-Fernández D, Benítez R, Hernández-Trujillo S, Adroher FJ (2017) Early development and life cycle of Contracaecum multipapillatum sl from a brown pelican Pelecanus occidentalis in the Gulf of California, Mexico. Dis Aquat Org 125:167–178CrossRefGoogle Scholar
  60. Vidal-Martínez VM, Osorio-Sarabia D, Overstreet RM (1994) Experimental infection of Contracaecum multipapillatum (Nematoda: Anisakinae) from Mexico in the domestic cat. J Parasitol 80:576–579CrossRefGoogle Scholar
  61. Zhu X, Gasser RB, Podolska M, Chilton NB (1998) Characterization of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. Int J Parasitol 28:1911–1921CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Aquatic Animal Diseases, Faculty of Veterinary MedicineOndokuz Mayis UniversitySamsunTurkey

Personalised recommendations