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Eurytrema coelomaticum: updated morphology of adult worms using advanced microscopy experiments

Published online by Cambridge University Press:  22 January 2020

K.G. Leite Open the ORCID record for K.G. Leite [Opens in a new window] ,
E.J. Lopes-Torres ,
J.G.R. Souza ,
R.H. Neves ,
D.C. Gomes  and
J.R. Machado-Silva
K.G. Leite
Affiliation:
Laboratório de Helmintologia Romero Lascasas Porto, Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Centro Biomédico, Universidade do Estado do Rio de Janeiro, Av. Prof. Manoel de Abreu 444/5° andar, Vila Isabel, 20511-070, Rio de Janeiro, Brazil
E.J. Lopes-Torres
Affiliation:
Laboratório de Helmintologia Romero Lascasas Porto, Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Centro Biomédico, Universidade do Estado do Rio de Janeiro, Av. Prof. Manoel de Abreu 444/5° andar, Vila Isabel, 20511-070, Rio de Janeiro, Brazil
J.G.R. Souza
Affiliation:
Laboratório de Biologia e Parasitologia de Mamíferos Silvestres Reservatórios, FIOCRUZ, Av. Brasil 4365, Manguinhos 21040-900, Rio Janeiro, Brazil
R.H. Neves
Affiliation:
Laboratório de Helmintologia Romero Lascasas Porto, Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Centro Biomédico, Universidade do Estado do Rio de Janeiro, Av. Prof. Manoel de Abreu 444/5° andar, Vila Isabel, 20511-070, Rio de Janeiro, Brazil
D.C. Gomes
Affiliation:
Laboratório de Helmintos Parasitos de Vertebrados, Instituto Oswaldo Cruz, FIOCRUZ Pavilhão Cardoso Fontes 3° andar, Av. Brasil, 4365, Manguinhos, 21040-900, Rio de Janeiro, Brazil
J.R. Machado-Silva*
Affiliation:
Laboratório de Helmintologia Romero Lascasas Porto, Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Centro Biomédico, Universidade do Estado do Rio de Janeiro, Av. Prof. Manoel de Abreu 444/5° andar, Vila Isabel, 20511-070, Rio de Janeiro, Brazil
*
Author for correspondence: J.R. Machado-Silva, E-mail: jrmasilva@gmail.com
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Abstract

Eurytrema coelomaticum is a digenean flatworm of ruminants that is the causative agent of eurytrematosis, a disease of veterinary health concern. Although modern techniques of morphological analysis have provided new insights about the morphology and anatomy of parasitic helminths, most studies on E. coelomaticum adults are based on conventional light microscopy. In the present study, a combined approach using brightfield, fluorescence, confocal and scanning electron microscopies (SEMs), together with the cryofracture technique, have updated morphological data on E. coelomaticum recovered from cattle in Rio de Janeiro State, Brazil. Light microscopy confirmed the presence of several structures present in the current description, such as suckers, pharynx, oesophagus, intestinal bifurcation and the cirrus-sac. Fluorescence stereomicroscopy revealed for the first time the cubic crystal protein inclusions in the forebody, which were further detailed by confocal and SEMs. Confocal microscopy provided detailed information of the muscular architecture associated with the attachment structures (suckers), digestive system (pharynx and oesophagus), egg-forming complex (ovary, Mehlis’ gland and Laurer's canal) and male reproductive system, which are similar to those found in other digenean flukes. SEM images of cryofractured parasites showed mucus and developing eggs within uterine loops. It was demonstrated that the combination of advanced tools generated complementary information, confirming the importance of experimental morphology in parasitology. Therefore, the knowledge of the adult structural organization of E. coelomaticum was improved and this work has contributed to propose new morphological criteria to evaluate the effects of antiparasitic drugs on flukes of medical and veterinary importance.

Keywords

Eurytrema coelomaticummorphologyconfocal microscopyscanning electron microscopycryofracture
Type
Research Paper
Information
Journal of Helminthology , Volume 94 , 2020 , e122
Copyright
Copyright © Cambridge University Press 2020

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Footnotes

*

These authors contributed equally to this work.

References

Abdelaal, MMO, Brennan, GP, Abdel-Azi, A and Fairweather, I (2016) Ultrastructural changes to the tegumental system and gastrodermal cells of adult Fasciola hepatica following treatment in vivo with a commercial preparation of myrrh (Mirazid). Journal of Helminthology 91, 672–685.CrossRefGoogle Scholar
Adnet, FA, Gonçalves, JP, de Souza, W and Attias, M (2013) A simple and efficient method to observe internal structures of helminths by scanning electron microscopy. Microscopy and Microanalysis 19, 1470–1474.CrossRefGoogle ScholarPubMed
Almeida, F, Oliveira, F, Neves, R, Siqueira, N, Rodrigues-Silva, R, Daipert-Garcia, D and Machado-Silva, JR (2015) Morphometric characteristics of the metacestode Echinococcus vogeli Rausch & Bernstein, 1972 in human infections from the northern region of Brazil. Journal of Helminthology 89, 480–486.CrossRefGoogle ScholarPubMed
Borges, JN, Costa, VS, Mantovani, C, Barros, E, Santos, EG, Mafra, CL and Santos, CP (2017) Molecular characterization and confocal laser scanning microscopic study of Pygidiopsis macrostomum (Trematoda: Heterophyidae) parasites of guppies Poecilia vivipara. Journal of Fish Diseases 40, 191–203.CrossRefGoogle ScholarPubMed
Bray, RA and Justine, JL (2013) Bucephalidae (Digenea) from epinephelines (Serranidae: Perciformes) from the waters off New Caledonia, including Neidhartia lochepintade n. sp. Parasite 20, 56.CrossRefGoogle ScholarPubMed
Bullard, SA, Williams, EH Jr and Bunkley-Williams, L (2012) New genus and species of fish blood fluke (Digenea: Aporocotylidae Odhner, 1912) from stoplight parrotfish, Sparisoma viride (Bonnaterre, 1788), (Labridae: Scarinae) in the Caribbean Sea. Journal of Parasitology 98, 1139–1143.CrossRefGoogle ScholarPubMed
Casanova, JC and Ribas, A (2004) Description of Brachylecithum mackoi n. sp. (Digenea: Dicrocoeliidae) from the European hedgehog, Erinaceus europaeus (Insectivora: Erinaceidae). Journal of Parasitology 90, 793–796.CrossRefGoogle Scholar
Cogswell, AA, Kommer, VP and Williams, DL (2012) Transcriptional analysis of a unique set of genes involved in Schistosoma mansoni female reproductive biology. PLoS Neglected Tropical Diseases 6: e1907.CrossRefGoogle ScholarPubMed
Cohen, SC, Kohn, A and Baptista-Farias, MDF (2001) Scanning and transmission electron microscopy of the Tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000 (microcotylidae, monogenea), parasite of a Brazilian Catfish, Hypostomus regain. Memórias do Instituto Oswaldo Cruz 96, 555–560.CrossRefGoogle Scholar
Colhoun, LM, Fairweather, I and Brennan, GP (1998) Observations on the mechanism of eggshell formation in the liver fluke, Fasciola hepatica. Parasitology 116, 555–567.CrossRefGoogle ScholarPubMed
Conn, DB, Świderski, Z and Miquel, J (2018) Ultrastructure of digenean trematode eggs (Platyhelminthes: Neoophora): a review emphasizing new comparative data on four European Microphalloidea. Acta Parasitologica 63, 1–14.CrossRefGoogle ScholarPubMed
de Souza, W (2011) Técnicas microscopia eletrônica aplicadas às ciências biológicas. 3rd edn.Rio de Janeiro, Sociedade Brasileira de Microscopia, 420 pp.Google Scholar
Faulwetter, S, Vasileiadou, A, Kouratoras, M, Thanos, D and Arvanitidis, C (2013) Micro-computed tomography: introducing new dimensions to taxonomy. Zookeys 263, 1–45.CrossRefGoogle Scholar
Figueira, GF, Oliveira, VH, Taroda, A, Alfieri &, AA and Headley, SA (2014) Molecular characterization of Eurytrema coelomaticum in cattle from Paraná, Brazil. Revista Brasileira de Parasitologia Veterinária 23, 383–386.CrossRefGoogle ScholarPubMed
Franco-Acuña, DO, Pinheiro, J, Oliveira-Menezes, A, Brandolini, SV, DaMatta, RA and de Souza, W (2011) Light and scanning electron microscopy of sporocysts of Eurytrema coelomaticum (Giard et Billet, 1892) Looss, 1907. Veterinary Parasitology 177, 72–78.CrossRefGoogle ScholarPubMed
Gonçalves, JP, Oliveira-Menezes, A, Maldonado, A Jr, Carvalho, TMU and de Souza, W (2016) Ultrastructural and cytochemical characterization of T1 and T2 secretory bodies from the tegument of Echinostoma paraensei. Micron 80, 59–65.CrossRefGoogle ScholarPubMed
Hanna, REB (2015) Fasciola hepatica: histology of the reproductive organs and differential effects of triclabendazole on drug-sensitive and drug-resistant fluke isolates and on flukes from selected field cases. Pathogens 4, 431–456.CrossRefGoogle ScholarPubMed
Hanna, REB, Moffetta, D, Forstera, FI, Trudgettb, AG, Brennanb, GP and Fairweatherb, I (2016) Fasciola hepatica: a light and electron microscope study of the ovary and of the development of oocytes within eggs in the uterus provides an insight into reproductive strategy. Veterinary Parasitology 221, 93–103.CrossRefGoogle ScholarPubMed
Hildebrand, J, Okulewicz, J and Popiolek, M (2007) A new dicrocoeliid from the bank vole Clethrionomys glareolus (Rodentia: Microtidae) from Poland. Journal of Parasitology 93, 151–154.CrossRefGoogle ScholarPubMed
Ilha, MR, Loretti, AP and Reis, AC (2005) Wasting and mortality in beef cattle parasitized by Eurytrema coelomaticum in the State of Paraná, Southern Brazil. Veterinary Parasitology 133, 49–60.CrossRefGoogle Scholar
Jiraungkoorskul, W, Sahaphong, S, Tansatit, T, Kangwanrangsan, N and Pipatshukiat, S (2005) Eurytrema pancreaticum: the in vitro effect of praziquantel and triclabendazole on the adult fluke. Experimental Parasitology 111, 172–177.CrossRefGoogle ScholarPubMed
Krupenko, DY (2014) Muscle system of Diplodiscus subclavatus (Trematoda: Paramphistomida) cercariae, pre-ovigerous, and ovigerous adults. Parasitology Research 113, 941–952.CrossRefGoogle ScholarPubMed
Krupenko, DY (2019) Oral sucker in Digenea: structure and muscular arrangement. Zoomorphology 138, 29–37.CrossRefGoogle Scholar
Krupenko, DY and Dobrovolskij, AA (2015) Somatic musculature in trematode hermaphroditic generation. BMC Evolutionary Biology 15, 189.CrossRefGoogle ScholarPubMed
Kumar, D, McGeown, JG, Reynoso-Ducoing, O, Ambrosio, JR and Fairweather, I (2003) Observations on the musculature and isolated muscle fibres of the liver fluke, Fasciola hepatica. Parasitology 127, 457–473.CrossRefGoogle ScholarPubMed
Lopes-Torres, EJ, de Souza, W and Miranda, K (2013) Comparative analysis of Trichuris muris surface using conventional, low vacuum, environmental and field emission scanning electron microscopy. Veterinary Parasitology 196, 409–416.CrossRefGoogle ScholarPubMed
Lopes-Torres, EJ, Maldonado, A Jr, Anjos, DH, de Souza, W and Miranda, K (2015) Redescription of Spirura guianensis (Nematoda: Spiruridae) from a rare South American Gracile Opossum. Acta Tropica 150, 87–93.CrossRefGoogle Scholar
Lopes-Torres, EJ, Girard-Dias, W, Mello, WN, Simoes, RO, Pinto, IS, Maldonado, A Jr, de Souza, W and Miranda, K (2019) Taxonomy of Physaloptera mirandai (Nematoda: Physalopteroidea) based in three-dimensional microscopy and phylogenetic positioning. Acta Tropica 195, 115–126.CrossRefGoogle ScholarPubMed
Machado-Silva, JR, Lanfredi, RM and Gomes, DC (1997) Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by scanning electron microscopy. Memórias do Instituto Oswaldo Cruz 92, 647–653.CrossRefGoogle ScholarPubMed
Machado-Silva, JR, Pelajo-Machado, M, Lenzi, HL and Gomes, DC (1998) Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by confocal laser scanning microscopy. Memórias do Instituto Oswaldo Cruz 93, 303–307.CrossRefGoogle ScholarPubMed
Mair, GR, Maule, AG, Shaw, C and Halton, DW (1998) Muscling in on parasitic flatworms. Parasitology Today 14, 73–76.CrossRefGoogle ScholarPubMed
Mair, GR, Maule, AG, Day, TA and Halton, DW (2000) A confocal microscopical study of the musculature of adult Schistosoma mansoni. Parasitology 121, 163–170.CrossRefGoogle ScholarPubMed
Maldonado, A Jr, Vieira, GO and Lanfredi, RM (2003) Echinostoma luisreyi n. sp. (Platyhelminthes: Digenea) by light and scanning electron microscopy. Journal of Parasitology 89, 800–808.CrossRefGoogle ScholarPubMed
Marques, JS, Rocha, BM, Manso, PPA and D’Ávila, S (2017) New insights on the morphology of a digenean parasite (Digenea: Brachylaimidae, Brachylaima mazzantii (Travassos, 1927) using confocal laser scanning microscopy. Zoosystema 39, 449–462.CrossRefGoogle Scholar
Neves, RH, Pereira, MJ, de Oliveira, RM, Gomes, DC and Machado-Silva, JR (1998) Schistosoma mansoni Sambon, 1907: morphometric differences between adult worms from sympatric rodent and human isolates. Memórias do Instituto Oswaldo Cruz 93, 309–312.CrossRefGoogle ScholarPubMed
Neves, RH, de Lamare, BC, Machado-Silva, JR, Carvalho, JJ, Branquinho, TB, Lenzi, HL, Hulstijn, M and Gomes, DC (2005) A new description of the reproductive system of Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by confocal laser canning microscopy. Parasitology Research 95, 43–49.CrossRefGoogle Scholar
Pereira, CAJ, Oliveira, LLS, Coaglio, AL, Santos, FSO, Cezar, RSM, Mendes, T, Oliveira, FLP, Conzensa, G and Lima, WS (2016) Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro. Veterinary Parasitology 15, 160–166.CrossRefGoogle Scholar
Pinheiro, J, Franco-Acuña, DO, Oliveira-Menezes, A, Brandolini, SV, de Souza, W and DaMatta, RA (2011) Ultrastructure of the sporocysts of Eurytrema coelomaticum (Giard Et Billet, 1892) Looss, 1907. Veterinary Parasitology 182, 201–212.CrossRefGoogle ScholarPubMed
Pinheiro, J, Franco-Acuña, DO, Oliveira-Menezes, A, Brandolini, SV, DaMatta, RA and de Souza, W (2012) New insight into the morphology of Eurytrema coelomaticum (Trematoda, Dicrocoeliidae) cercariae by light, scanning, and transmission electron microscopies. Parasitology Research 111, 1437–1445.CrossRefGoogle ScholarPubMed
Pinheiro, J, Fanco-Acuña, DO, Oliveira-Menezes, A, Brandolini, SVPB, Adnet, FAO, Lopes Torres, EJ, Miranda, FJB, de Souza, W and DaMatta, RA (2015) Additional study of the morphology of eggs and miracidia of Eurytrema coelomaticum (Trematoda). Helminthologia 52, 244–251.CrossRefGoogle Scholar
Pinto, HA and de Melo, AL (2016) Comments on human eurytremiasis in Brazil. World Journal of Experimental Medicine 6, 55–57.CrossRefGoogle Scholar
Radio, S, Fontenla, S, Solana, V, et al. (2018) Pleiotropic alterations in gene expression in Latin American Fasciola hepatica isolates with different susceptibility to drugs. Parasite Vectors 11, 56.CrossRefGoogle ScholarPubMed
Roberts, JR, Orelis-Ribeiro, R, Dang, BT, Halanych, KM and Bullard, SA (2016) Blood flukes of Asiatic softshell turtles: revision of Coeuritrema Mehra, 1933 (Digenea: Schistosomatoidea) and a new species infecting Chinese softshell turtles, Pelodiscus sinensis (Trionychidae), from Vietnam. Folia Parasitologica 6, 63.Google Scholar
Sakamoto, T and Oikawa, T (2007) Cubic crystal protein inclusions in the neodermis of the pancreatic fluke, Eurytrema pancreaticum and Eurytrema coelomaticum. Parasitology Research 101, 1393–1399.CrossRefGoogle ScholarPubMed
Šebelová, S, Stewart, MT, Mousley, A, Fried, B, Marks, NJ and Halton, DW (2004) The musculature and associated innervation of adult and intra molluscan stages of Echinostoma caproni (Trematoda) visualised by confocal microscopy. Parasitolology Research 93, 196–206.CrossRefGoogle Scholar
Skírnisson, K, Kolářová, L, Horák, P, Ferté, H and Jouet, D (2012) Morphological features of the nasal blood fluke Trichobilharzia regenti (Schistosomatidae, Digenea) from naturally infected hosts. Parasitology Research 110, 1881–1892.CrossRefGoogle ScholarPubMed
Souza, JGR, Garcia, JS, Manso, PPA, Neves, RH, Maldonado, A Jr and Machado-Silva, JR (2011) Development of the reproductive system of the Echinostoma paraensei in Mesocricetus auratus analyzed by light and confocal scanning laser microscopy. Experimental Parasitology 128, 341–346.CrossRefGoogle ScholarPubMed
Souza, JGR, Garcia, JS, Gomes, APN, Machado-Silva, JR and Maldonado, A Jr (2017) Comparative pattern of growth and development of Echinostoma paraensei (Digenea: Echinostomatidae) in hamster and Wistar rat using light and confocal laser canning microscopy. Experimental Parasitology 183, 41–49.CrossRefGoogle Scholar
Srivastava, M and Misra-Bhattacharya, S (2015) Overcoming drug resistance for macro parasites. Future Microbiology 10, 1783–1789.CrossRefGoogle ScholarPubMed
Taylor, J (2008) Cryofracture or freeze-fracture, a method to expose internal tissue surfaces and cell surfaces for viewing in the scanning electron microscope. Microscopy Today 16, 56–58.CrossRefGoogle Scholar
Terasaki, K, Itagaki, T, Shibahara, T, Noda, Y and Moriyama-Gonda, N (2001) Comparative study of the reproductive organs of Fasciola Groups by Optical Microscope. Journal of Veterinary Medical Science 63, 735–742.CrossRefGoogle ScholarPubMed
Travassos, L (1944) Revisão da família Dicrocoeliidae, Odhner, 1910. Monografias do Instituto Oswaldo Cruz. 2, 1–357.Google Scholar
Travassos, L, Teixeira, JF and Kohn, A (1969) Trematódeos do Brasil. Memórias do Instituto Oswaldo Cruz 67, 1–886.Google Scholar
Womble, MR, Cox-Gardiner, SJ, Cribb, TH and Bullard, SA (2015) First record of Transversotrema Witenberg, 1944 (Digenea) from the Americas, with comments on the taxonomy of Transversotrema patialense (Soparkar, 1924) Crusz and Sathananthan, 1960, and an updated list of its hosts and geographic distribution. Journal of Parasitology 101, 717–725.CrossRefGoogle ScholarPubMed