Folia Parasitologica 71:001 (2024) | DOI: 10.14411/fp.2024.001

Manual for the study of tapeworms (Cestoda) parasitic in ray-finned fish, amphibians and reptiles

Lenta Chervy
Institute of Parasitology, Biology Centre of the Czech Academy of Sciences, České Budějovice, Czech Republic

Based on long-term and often frustrating experiences with the poor quality of tapeworms (Cestoda) collected throughout the world for taxonomic and phylogenetic studies, and considering the increasing obstacles to obtaining new material, a simple, easy-to-use and illustrated methodological guide (manual) is provided. It focusses mainly on key steps in examining hosts, collecting cestodes from poikilothermous vertebrates except elasmobranchs, i.e., from ray-finned fish (Actinopterygii), amphibians and 'reptiles' (a paraphyletic group comprising all sauropsids except birds), and fixing them for subsequent morphological and molecular study. It is proposed that the following methodological points should be followed: (i) ideally only freshly euthanised hosts (not previously frozen) should be used for parasitological examination; (ii) hosts examined should be documented by photographs; host tissue should also be preserved for future genotyping if necessary; (iii) tapeworms should be detached carefully to keep the scolex intact and properly cleaned before fixation; (iv) a small piece of cestode tissue should be always preserved in molecular grade ethanol for DNA sequencing; (v) tapeworms should be fixed as quickly as possible after collecting them and while they are still alive, always using hot (heated) fixatives; this prevents unnatural contraction or deformation and ensures uniform fixation; (vi) each sample (vial) should be properly labelled (a unique code should be given to every cestode sample); (vii) vouchers of sequenced specimens (hologenophores or paragenophores) should always be preserved for identification, and deposited in internationally recognised collections. It is hoped that this guide helps researchers and students to properly process valuable material of cestodes to make it suitable for reliable identification including genotyping and comparative anatomy, which is a prerequisite for any subsequent ecological, biogeographical, phylogenetic life cycle or molecular study.

Keywords: Methodology, field guide, cestodes, host dissection, fixation, morphology, taxonomy, systematics, teleosts, Amphibia, 'Reptilia', Pisces

Received: December 24, 2022; Revised: August 24, 2023; Accepted: October 31, 2023; Published online: January 22, 2024  Show citation

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Chervy, L. (2024). Manual for the study of tapeworms (Cestoda) parasitic in ray-finned fish, amphibians and reptiles. Folia Parasitologica, 71, Article 2024.001. https://doi.org/10.14411/fp.2024.001
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References

  1. Ash A., de Chambrier A., Shimazu T., Ermolenko A., Scholz T. 2015: An annotated list of the species of Gangesia Woodland, 1924 (Cestoda: Proteocephalidea), parasites of catfishes in Asia, with new synonyms and a key to their identification. Syst. Parasitol. 91: 13-33. Go to original source... Go to PubMed...
  2. Barčák D., Alexovič Matiašová A., Čisovská Bazsalovicsová E., Soldánová M., Oros M., Králová-Hromadová I. 2023: An examination of nervous system revealed unexpected immunoreactivity of both secretory apparatus and excretory canals in plerocercoids of two broad tapeworms (Cestoda: Diphyllobothriidea). Parasitology 150: 612-622. Go to original source... Go to PubMed...
  3. Barčák D., Yoneva A., Sehadová H., Oros M., Gustinelli A., Kuchta R. 2019: Complex insight on microanatomy of larval "human broad tapeworm" Dibothriocephalus latus (Cestoda: Diphyllobothriidea). Parasit. Vect. 12: 408. Go to original source... Go to PubMed...
  4. Beveridge I. 2001: The use of life-cycle characters in studies of the evolution of cestodes. In: D.T.J. Littlewood and R.A. Bray (Eds.), Interrelationships of the Platyhelminthes, The Systematics Association Special Volume Series 60. Taylor & Francis, London and New York, pp. 250-256.
  5. Caira J.N., Jensen K. 2014: A digest of elasmobranch tapeworms. J. Parasitol. 100: 373-391. Go to original source... Go to PubMed...
  6. Caira J.N., Jensen K. (Eds.) 2017: Planetary Biodiversity Inventory (2008-2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, Lawrence, KS, 464 pp.
  7. Caira J.N., Jensen K., Barbeau E. (Eds.) 2023: Global Cestode Database. World Wide Web electronic publication. http://www.tapewormdb.uconn.edu/
  8. Caira J.N., Scholz T., Georgiev B.B. 2006: Cestode systematics and phylogeny move toward. Syst. Parasitol. 65: 171-174. Go to original source... Go to PubMed...
  9. de Chambrier A. 2001: A new tapeworm from the Amazon, Amazotaenia yvettae n. gen., n. sp. (Eucestoda: Proteocephalidea) from the siluriform fishes Brachyplatystoma filamentosum and B. vaillanti (Pimelodidae). Rev. Suisse Zool. 108: 303-316. Go to original source...
  10. de Chambrier A., Alves P.V., Schuster R.K., Scholz T. 2021: Ophiotaenia echidis n. sp. (Cestoda: Proteocephalidae), from the saw-scaled viper, Echis carinatus sochureki (Stemmler) (Ophidia: Viperidae), one of the world's deadliest snakes, from the United Arab Emirates. Int. J. Parasitol. Parasit. Wildlife 14: 341-354. Go to original source... Go to PubMed...
  11. de Chambrier A., Coquille S.C., Brooks D.R. 2006: Ophiotaenia bonneti sp. n. (Eucestoda: Proteocephalidea), a parasite of Rana vaillanti (Anura: Ranidae) in Costa Rica. Folia Parasitol. 53: 125-133. Go to original source... Go to PubMed...
  12. de Chambrier A., Paulino R.C. 1997: Proteocephalus joanae sp. n. (Eucestoda: Proteocephalidea), a parasite of Xenodon neuwiedi (Serpentes: Colubridae) from South America. Folia Parasitol. 44: 289-296.
  13. de Chambrier A., Pinacho-Pinacho M., Hernández-Orts J., Scholz T. 2017b: A new genus and two new species of proteocephalidean tapeworms (Cestoda) from cichlid fish (Perciformes: Cichlidae) in the Neotropics. J. Parasitol. 103: 83-94. Go to original source... Go to PubMed...
  14. de Chambrier A., Rego A.A., Mariaux J. 2004: Redescription of Brooksiella praeputialis and Goezeella siluri (Eucestoda: Proteocephalidea), parasites of Cetopsis coecutiens (Siluriformes) from the Amazon and proposition of Goezeella danbrooksi sp. n. Rev. Suisse Zool. 111: 111-120. Go to original source...
  15. de Chambrier A., Scholz T., Mariaux J., Kuchta R. 2017a: Onchoproteocephalidea I Caira, Jensen, Waeschenbach, Olson & Littlewood, 2014. In: J.N. Caira and K. Jensen (Eds.), Planetary Biodiversity Inventory (2008-2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, Lawrence, Kansas, USA, pp. 251-277.
  16. de Chambrier A., Vaucher C., Renaud F. 1992: Etude des caractères morpho-anatomiques et des flux géniques chez quatre Proteocephalus (Cestoda: Proteocephalidae) parasites de Bothrops jararaca au Brésil et description de trois espèces nouvelles. Syst. Parasitol. 23: 141-156. Go to original source...
  17. Chervy L. 2002: The terminology of larval cestodes or metacestodes. Syst. Parasitol. 52: 1-33. Go to original source... Go to PubMed...
  18. Chervy L. 2009: Unified terminology for cestode microtriches: a proposal from the International Workshops on Cestode Systematics in 2002-2008. Folia Parasitol. 56: 199-230. Go to original source... Go to PubMed...
  19. Conn D.B., Swiderski Z. 2008: A standardised terminology of the embryonic envelopes and associated developmental stages of tapeworms (Platyhelminthes: Cestoda). Folia Parasitol. 55: 42-52. Go to original source... Go to PubMed...
  20. Cooper D.W. 1988: The preparation of serial sections of platyhelminth parasites, with details of the materials and facilities required. Syst. Parasitol. 12: 211-229. Go to original source...
  21. Cribb T.H., Bray R.A. 2010: Gut wash, body soak, blender, and heat-fixation: approaches to the effective collection, fixation and preservation of trematodes of fishes. Syst. Parasitol. 76: 1-7. Go to original source... Go to PubMed...
  22. Efford I.E., Tsumura K. 1969: Observations on the biology of the trematode Megalodiscus microphagus in amphibians from Marion Lake, British Columbia. Am. Midl. Nat. 82: 197-203. Go to original source...
  23. Fernandes I.M., Bastos Y.F., Barreto D.S., Lourenço L.S., Penha J.M. 2017: The efficacy of clove oil as an anaesthetic and in euthanasia procedure for small-sized tropical fishes. Braz. J. Biol. 77: 444-450. Go to original source... Go to PubMed...
  24. Fyler C.A. 2007: Comparison of microthrix ultrastructure and morphology on the plerocercoid and adult scolex of Calliobothrium cf. verticillatum (Tetraphyllidea: Onchobothriidae). J. Parasitol. 93: 4-11. Go to original source... Go to PubMed...
  25. Gibson D.I. 1984: Technology as applied to museum collections: the collections, fixation and conservation of helminths. Syst. Parasitol. 6: 241-255. Go to original source...
  26. Hewitson T.D., Darby I.A. 2010: Histology Protocols. Humana Press, New York, 229 pp. Go to original source...
  27. Humason G.L. 1979: Animal Tissue Techniques. Fourth Edition. W.H. Freeman and Company, San Francisco, 661 pp.
  28. Jackson J.A., Tinsley R.C. 2001: Host-specificity and distribution of cephalochlamydid cestodes: Correlation with allopolyploid evolution of pipid anuran hosts. J. Zool. 254: 405-419. Go to original source...
  29. Jensen K., Bullard S.A. 2010: Characterization of a diversity of tetraphyllidean and rhinebothriidean cestode larval types, with comments on host associations and life-cycles. Int. J. Parasitol. 40: 889-910. Go to original source... Go to PubMed...
  30. Jones A. 1990: Techniques for hand-sectioning thick-bodied platyhelminths. Syst. Parasitol. 15: 211-218. Go to original source...
  31. Justine J.-L., Briand M.J., Bray R.A. 2012: A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies. Parasitol. Res. 111: 341-351. Go to original source... Go to PubMed...
  32. Khalil L.F., Bray R.A., Jones A. (Eds.) 1994: Keys to the Cestode Parasites of Vertebrates. CAB International, Wallingford, 751 pp. Go to original source...
  33. Kiernan J.A. 2008. Histological and Histochemical Methods: Theory and Practice. Fourth Edition. Scion Publishing, Banbury, UK, 606 pp.
  34. Kuchta R., Choudhury A., Scholz T. 2018: Asian fish tapeworm: the most successful invasive parasite in freshwaters. Trends Parasitol. 34: 511-523. Go to original source... Go to PubMed...
  35. Kuchta R., Kołodziej-Sobocińska M., Brabec J., Młocicki D., Sałamatin R., Scholz T. 2021: Sparganosis (Spirometra) in Europe in the molecular era. Clin. Infect. Dis. 72: 882­-890. Go to original source... Go to PubMed...
  36. Kuchta R., Scholz T. 2017: Bothriocephalidea Kuchta, Scholz, Brabec & Bray, 2008. In: J.N. Caira and K. Jensen (Eds.), Planetary Biodiversity Inventory (2008-2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, Lawrence, Kansas, USA, pp. 29-45.
  37. Kuchta R., Scholz T., Vlčková R. Říha M., Walter T., Yuniar A.T., Palm H.W. 2009: Revision of tapeworms (Cestoda: Bothriocephalidea) from lizardfish (Saurida: Synodontidae) from the Indo-Pacific region. Zootaxa 1977: 55-67. Go to original source...
  38. Lamothe-Argumedo Damborenea C., García-Prieto L., Lunaschi L.I., Osorio-Sarabia D. 2010: Guide to Helminthological Collections of Latin America. Publicaciones Especiales del Instituo de Biología 22, Universidad Nacional Autónoma de México, México, D.F., Mexico, 61 pp.
  39. Levron C., Bruňanská M., Kuchta R., Freeman M., Scholz T. 2006: Spermatozoon ultrastructure of the pseudophyllidean cestode Paraechinophallus japonicus, a parasite of deep-sea fish Psenopsis anomala (Perciformes, Centrolophidae). Parasitol. Res. 100: 115-121. Go to original source... Go to PubMed...
  40. Levron C., Miquel J., Oros M., Scholz T. 2010: Spermatozoa of tapeworms (Platyhelminthes, Eucestoda): advances in ultrastructural and phylogenetic studies. Biol. Rev. 85: 523-543. Go to original source... Go to PubMed...
  41. Levron C., Scholz T., Vancová M., Kuchta R., Conn D.B. 2016: Ultrastructure of embryonated eggs of the cestode Gyrocotyle urna (Gyrocotylidea) using cryo-methods. Zoomorphology 135: 279-289. Go to original source...
  42. Lichtenfels J.R., Pritchard M.H. 1982: A guide to the parasite collections of the world. Special Publication of the American Society of Parasitologists 1982: 1-79.
  43. Malmberg G. 1957: About the occurrence of Gyrodactylus on Swedish fish. Sodra Sveriges Fiskeriforening Yearbook 1956: 19-76. (In Swedish.)
  44. Naylor G.J.P., Caira J.N., Jensen K., Rosana K.A.M., Straube N., Lakner C. 2012a: Elasmobranch phylogeny: a mitochondrial estimate based on 595 species. In: J.C. Carrier, J.A. Musick and M.R. Heithaus (Eds.), The Biology of Sharks and Their Relatives. CRC Press, Boca Raton, Florida, pp. 31-56. Go to original source...
  45. Naylor G.J.P., Caira J.N., Jensen K., Rosana K.A.M., White W.T., Last P.R. 2012b: A DNA sequence-based approach to the identification of shark and ray species and its implications for global elasmobranch diversity and parasitology. Bull. Am. Mus. Nat. Hist. 367: 1-262. Go to original source...
  46. Neitemeier-Duventester X., Bick A., Theisen S., Palm H.W. 2022: Confocal laser scanning microscopy (CLSM) as a new tool for morphological characterisation of both newly collected and museum voucher specimens of the Trypanorhyncha Diesing, 1863 (Platyhelminthes: Cestoda). Folia Parasitol. 69: 026. Go to original source... Go to PubMed...
  47. Neves R.H., de Lamare Biolchini C., Machado-Silva J.R., Carvalho J.J., Branquinho T.B., Lenzi H.L., Hulstijn M., Gomes D.C. 2005: A new description of the reproductive system of Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by confocal laser scanning microscopy. Parasitol. Res. 95: 43-49. Go to original source... Go to PubMed...
  48. Paddock S.W. 1998: An introduction to confocal imaging. In: S.W. Paddock (Ed.), Confocal Microscopy: Methods and Protocols 122. Methods in Molecular Biology. Humana, Totowa, New Jersey, pp. 1-34. Go to original source...
  49. Palm H.W. 2004: The Trypanorhyncha Diesing, 1863. PKSPL Press, Bogor, 710 pp.
  50. Palm H.W., Bray R.A. 2014: Marine Fish Parasitology in Hawaii. Westarp & Partner Digitaldruck, Hohenwarsleben, 303 pp.
  51. Pleijel R., Jondelius U., Norlinder E., Nygeren A., Oxelman B., Schander C., Sundberg P., Thollesson M. 2008: Phylog­enies without roots? A plea for the use of vouchers in molecular phylogenetic studies. Mol. Phylogenet. Evol. 48: 369-371. Go to original source... Go to PubMed...
  52. Poddubnaya L., Kuchta R., Scholz T. 2020: Ultrastructural patterns of the excretory ducts of basal neodermatan groups (Platyhelminthes) and new protonephridial characters of basal cestodes. Parasit. Vect. 13: 442. Go to original source... Go to PubMed...
  53. Pritchard M.H., Kruse G.O.W. 1982: The Collection and Preservation of Animal Parasites. University of Nebraska Press, Lincoln, Nebraska, 141 pp.
  54. Prudhoe S., Bray R.A. 1982: Platyhelminth Parasites of the Amphibia. British Museum (Natural History), London and Oxford University Press, Oxford, 217 pp.
  55. Ross L.G., Ross B. (Eds.) 2008: Anaesthetic and Sedative Techniques for Aquatic Animals. Third Edition, Blackwell Publishing, Oxford, 222 pp. Go to original source...
  56. Rozario T., Newmark P.A. 2015: A confocal microscopy-based atlas of tissue architecture in the tapeworm Hymenolepis diminuta. Exp. Parasitol. 158: 31-41. Go to original source... Go to PubMed...
  57. Ruedi V., de Chambrier A. 2012. Pseudocrepidobothrium ludovici sp. n. (Eucestoda: Proteocephalidea), a parasite of Phractocephalus hemioliopterus (Pisces: Pimelodidae) from Brazilian Amazon. Rev. Suisse Zool. 119: 137-147. Go to original source...
  58. Schmidt G.D. 1986: CRC Handbook of Tapeworm Identification. CRC Press, Inc., Boca Raton, Florida, 675 pp.
  59. Scholz T., Bray R.A., Kuchta R., Řepová R. 2004: Larvae of gryporhynchid cestodes (Cyclophyllidea) from fish: a review. Folia Parasitol. 51: 131-152. Go to original source... Go to PubMed...
  60. Scholz T., de Chambrier A., Kudlai O., Tkach V.V, McAllister C.T. 2023: A global survey of tapeworms (Cestoda: Proteocephalidae) of 'true' frogs (Amphibia: Ranidae), including a tabulated list of all proteocephalids parasitising amphibians. Folia Parasitol. 70: 009. Go to original source... Go to PubMed...
  61. Scholz T., Choudhury A., McAllister C.T. 2022: A young parasite in an old fish host: a new genus for proteocephalid tapeworms (Cestoda) of bowfin (Amia calva) (Holostei: Amiiformes), and a revised list of its cestodes. Int. J. Parasitol. Parasit. Wildl. 18: 101-111. Go to original source... Go to PubMed...
  62. Scholz T., Choudhury A., Uhrová L., Brabec J. 2019: The Proteocephalus species-aggregate in freshwater centrarchid and percid fishes of the Nearctic region (North America). J. Parasitol. 105: 798-812. Go to original source... Go to PubMed...
  63. Scholz T., Kuchta R. 2017: Amphilinidea Poche, 1922. In: J.N. Caira and K. Jensen (Eds.), Planetary Biodiversity Inventory (2008-2017): Tapeworms from Vertebrate Bowels of the Earth. University of Kansas, Natural History Museum, Special Publication No. 25, Lawrence, Kansas, USA, pp. 21-28.
  64. Scholz T., Kuchta R. 2022: Fish tapeworms (Cestoda) in the molecular era: achievements, gaps and prospects. Parasitology 149: 1876-1893. Go to original source... Go to PubMed...
  65. Scholz T., Tavakol S., Luus-Powell W. 2020: First adult cyclophyllidean tapeworm (Cestoda) from teleost fishes: host-switching beyond tetrapods in Africa. Int. J. Parasitol. 50: 561-568. Go to original source... Go to PubMed...
  66. Scholz T., Vanhove M.P.M., Smit N., Jayasundera Z., Gelnar M. (Eds.) 2018: A Guide to the Parasites of African Freshwater Fish. ABC Taxa 18. CEBioS, Royal Belgian Institute of Natural Sciences, Brussels, 424 pp.
  67. Ulmer M.J. 1950: A critique of methods for the measurement of parasitic worms. Papers of the Michigan Acad. Sci., Arts Lett. 36: 149-151.
  68. Yamaguti S. 1959: Systema Helminthum, vol. II. The Cestodes of Vertebrates. Keigaku Publishing House, Tokyo, 860 pp.
  69. Yoneva A., Scholz T., Bruňanská M., Kuchta R. 2015: Vitellogenesis of diphyllobothriidean cestodes (Platyhelminthes). Compt. Rend. Biol. 338: 169-179. Go to original source... Go to PubMed...
  70. Yoneva A., Scholz T., Kuchta R. 2018: Comparative morphology of surface ultrastructure of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea). Invertebr. Biol. 137: 38-48. Go to original source...