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.

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Ceske Budejovice Czech Republic

Zobrazit více v PubMed

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. DOI

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. DOI

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. DOI

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.

Caira J.N., Jensen K. 2014: A digest of elasmobranch tapeworms. J. Parasitol. 100: 373-391. DOI

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.

Caira J.N., Jensen K., Barbeau E. (Eds.) 2023: Global Cestode Database. World Wide Web electronic publication. http://www.tapewormdb.uconn.edu/

Caira J.N., Scholz T., Georgiev B.B. 2006: Cestode systematics and phylogeny move toward. Syst. Parasitol. 65: 171-174. DOI

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. DOI

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. DOI

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. DOI

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.

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. DOI

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. DOI

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.

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. DOI

Chervy L. 2002: The terminology of larval cestodes or metacestodes. Syst. Parasitol. 52: 1-33. DOI

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. DOI

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. DOI

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. DOI

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. DOI

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. DOI

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. DOI

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. DOI

Gibson D.I. 1984: Technology as applied to museum collections: the collections, fixation and conservation of helminths. Syst. Parasitol. 6: 241-255. DOI

Hewitson T.D., Darby I.A. 2010: Histology Protocols. Humana Press, New York, 229 pp. DOI

Humason G.L. 1979: Animal Tissue Techniques. Fourth Edition. W.H. Freeman and Company, San Francisco, 661 pp.

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. DOI

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. DOI

Jones A. 1990: Techniques for hand-sectioning thick-bodied platyhelminths. Syst. Parasitol. 15: 211-218. DOI

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. DOI

Khalil L.F., Bray R.A., Jones A. (Eds.) 1994: Keys to the Cestode Parasites of Vertebrates. CAB International, Wallingford, 751 pp. DOI

Kiernan J.A. 2008. Histological and Histochemical Methods: Theory and Practice. Fourth Edition. Scion Publishing, Banbury, UK, 606 pp.

Kuchta R., Choudhury A., Scholz T. 2018: Asian fish tapeworm: the most successful invasive parasite in freshwaters. Trends Parasitol. 34: 511-523. DOI

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. DOI

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.

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. DOI

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.

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. DOI

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. DOI

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. DOI

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.

Malmberg G. 1957: About the occurrence of Gyrodactylus on Swedish fish. Sodra Sveriges Fiskeriforening Yearbook 1956: 19-76. (In Swedish.)

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. DOI

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. DOI

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. DOI

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. DOI

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. DOI

Palm H.W. 2004: The Trypanorhyncha Diesing, 1863. PKSPL Press, Bogor, 710 pp.

Palm H.W., Bray R.A. 2014: Marine Fish Parasitology in Hawaii. Westarp & Partner Digitaldruck, Hohenwarsleben, 303 pp.

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. DOI

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. DOI

Pritchard M.H., Kruse G.O.W. 1982: The Collection and Preservation of Animal Parasites. University of Nebraska Press, Lincoln, Nebraska, 141 pp.

Prudhoe S., Bray R.A. 1982: Platyhelminth Parasites of the Amphibia. British Museum (Natural History), London and Oxford University Press, Oxford, 217 pp.

Ross L.G., Ross B. (Eds.) 2008: Anaesthetic and Sedative Techniques for Aquatic Animals. Third Edition, Blackwell Publishing, Oxford, 222 pp. DOI

Rozario T., Newmark P.A. 2015: A confocal microscopy-based atlas of tissue architecture in the tapeworm Hymenolepis diminuta. Exp. Parasitol. 158: 31-41. DOI

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. DOI

Schmidt G.D. 1986: CRC Handbook of Tapeworm Identification. CRC Press, Inc., Boca Raton, Florida, 675 pp.

Scholz T., Bray R.A., Kuchta R., Řepová R. 2004: Larvae of gryporhynchid cestodes (Cyclophyllidea) from fish: a review. Folia Parasitol. 51: 131-152. DOI

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. DOI

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. DOI

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. DOI

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.

Scholz T., Kuchta R. 2022: Fish tapeworms (Cestoda) in the molecular era: achievements, gaps and prospects. Parasitology 149: 1876-1893. DOI

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. DOI

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.

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.

Yamaguti S. 1959: Systema Helminthum, vol. II. The Cestodes of Vertebrates. Keigaku Publishing House, Tokyo, 860 pp.

Yoneva A., Scholz T., Bruňanská M., Kuchta R. 2015: Vitellogenesis of diphyllobothriidean cestodes (Platyhelminthes). Compt. Rend. Biol. 338: 169-179. DOI

Yoneva A., Scholz T., Kuchta R. 2018: Comparative morphology of surface ultrastructure of diphyllobothriidean tapeworms (Cestoda: Diphyllobothriidea). Invertebr. Biol. 137: 38-48. DOI

Revision of Isoglaridacris (Cestoda: Caryophyllidea), parasites of suckers (Catostomidae) in North America: molecular phylogeny and description of four new species

Machine learning models accurately predict clades of proteocephalidean tapeworms (Onchoproteocephalidea) based on host and biogeographical data

Three new species of Ophiotaenia La Rue, 1911 (Cestoda: Proteocephalidae) from dipsadine snakes (Squamata: Colubridae) in Ecuador

Diversity and biology of Spirometra tapeworms (Cestoda: Diphyllobothriidea), zoonotic parasites of wildlife: A review