Architecture of Paradiplozoon homoion: A diplozoid monogenean exhibiting highly-developed equipment for ectoparasitism

Diplozoidae (Monogenea) are blood-feeding freshwater fish gill ectoparasites with extraordinary body architecture and a unique sexual behaviour in which two larval worms fuse and transform into one functioning individual. In this study, we describe the body organisation of Paradiplozoon homoion adult stage using a combined approach of confocal laser scanning and electron microscopy, with emphasis on the forebody and hindbody. Special attention is given to structures involved in functional adaptation to ectoparasitism, i.e. host searching, attachment and feeding/metabolism. Our observations indicate clear adaptations for blood sucking, with a well-innervated mouth opening surrounded by sensory structures, prominent muscular buccal suckers and a pharynx. The buccal cavity surface is covered with numerous tegumentary digitations that increase the area in contact with host tissue and, subsequently, with its blood. The buccal suckers and the well-innervated haptor (with sclerotised clamps controlled by noticeable musculature) cooperate in attaching to and moving over the host. Putative gland cells accumulate in the region of apical circular structures, pharynx area and in the haptor middle region. Paired club-shaped sacs lying laterally to the pharynx might serve as secretory reservoirs. Furthermore, we were able to visualise the body wall musculature, including peripheral innervation, the distribution of uniciliated sensory structures essential for reception of external environmental information, and flame cells involved in excretion. Our results confirm in detail that P. homoion displays a range of sophisticated adaptations to an ectoparasitic life style, characteristic for diplozoid monogeneans.

[1]  M. Špakulová,et al.  Comparative karyological analysis of four diplozoid species (Monogenea, Diplozoidae), gill parasites of cyprinid fishes , 2011, Parasitology Research.

[2]  T. Hibiya,et al.  On the development of clamps and pairing of diporpae in Diplozoon nipponicum (Monogenea). , 1987 .

[3]  D. Joly Evolutionary Ecology of Parasites: From Individuals to Communities , 2001 .

[4]  R. Wilson,et al.  PROTONEPHRIDIA , 1974, Biological reviews of the Cambridge Philosophical Society.

[5]  A. Maule,et al.  Muscling in on parasitic flatworms. , 1998, Parasitology today.

[6]  N. Lane,et al.  Actin filaments are associated with the septate junctions of invertebrates. , 1988, Tissue & cell.

[7]  M. Gelnar,et al.  Classification and occurrence of abnormally developed Paradiplozoon homoion (Monogenea, Diplozoinae) parasitising gudgeon Gobio gobio. , 2005, Diseases of aquatic organisms.

[8]  D. Halton The surface topography of a monogenean,Diclidophora merlangi revealed by scanning electron microscopy , 2004, Zeitschrift für Parasitenkunde.

[9]  Excretory system of representatives from family Diplozoidae (Monogenea) , 2016, Parasitology Research.

[10]  C. O. Cunningham,et al.  Paradiplozoon homoion Bychowsky & Nagibina, 1959 versus P. gracile Reichenbach-Klinke, 1961 (Monogenea): two species or phenotypic plasticity? , 2002, Systematic Parasitology.

[11]  A. Kohn,et al.  Scanning and transmission electron microscopy of the tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000 (Microcotylidae, Monogenea), parasite of a Brazilian catfish, Hypostomus regani. , 2001, Memorias do Instituto Oswaldo Cruz.

[12]  A. Avenant-Oldewage,et al.  Trace element and metal sequestration in vitellaria and sclerites, and reactive oxygen intermediates in a freshwater monogenean, Paradiplozoon ichthyoxanthon , 2017, PloS one.

[13]  G. Kearn Parasitism and the Platyhelminths , 1997 .

[14]  G. Brennan,et al.  Immunomicroscopical observations on the nervous system of adult Eudiplozoon nipponicum (Monogenea: Diplozoidae). , 2001, International journal for parasitology.

[15]  Sreeurpa Ray,et al.  The Cell: A Molecular Approach , 1996 .

[16]  J. Collins,et al.  An Atlas for Schistosoma mansoni Organs and Life-Cycle Stages Using Cell Type-Specific Markers and Confocal Microscopy , 2011, PLoS neglected tropical diseases.

[17]  M. Koyun,et al.  The molecular and morphometrical description of a new diplozoid species from the gills of the Garra rufa (Heckel, 1843) (Cyprinidae) from Turkey—including a commentary on taxonomic division of Diplozoidae , 2013, Parasitology Research.

[18]  M. Gelnar,et al.  The surface topography of Eudiplozoon nipponicum (Monogenea) developmental stages parasitizing carp (Cyprinus carpio L.) , 2010, Central European Journal of Biology.

[19]  C. O. Cunningham,et al.  Genetic characterization of six species of diplozoids (Monogenea; Diplozoidae) , 2001, Parasitology.

[20]  D. Halton Microscopy and the helminth parasite. , 2004, Micron.

[21]  A. Avenant-Oldewage,et al.  The fluorescent detection of Paradiplozoon sp. (Monogenea: Diplozoidae) attachment clamps' sclerites and integumental proteins. , 2006, The Onderstepoort journal of veterinary research.

[22]  A. Avenant‐Oldewage,et al.  Soft Tissue Digestion of Paradiplozoon vaalense for SEM of Sclerites and Simultaneous Molecular Analysis , 2015, The Journal of parasitology.

[23]  C. O. Cunningham,et al.  IDENTIFICATION OF EUROPEAN DIPLOZOIDS (MONOGENEA, DIPLOZOINAE) BY RESTRICTION DIGESTION OF THE RIBOSOMAL RNA INTERNAL TRANSCRIBED SPACER , 2004, The Journal of parasitology.

[24]  Giovanni Strona,et al.  Three-Dimensional Imaging of Monogenoidean Sclerites by Laser Scanning Confocal Fluorescence Microscopy , 2006, The Journal of parasitology.

[25]  An ultrastructural study of the surface and attachment structures of Paradiplozoon homoion (Bychowsky & Nagibina, 1959) (Monogenea: Diplozoidae) , 2017, Parasites & Vectors.

[26]  L. A. Vøllestad,et al.  The relationship between developmental instability of gudgeon Gobio gobio and abundance or morphology of its ectoparasite Paradiplozoon homoion (Monogenea) , 2007 .

[27]  P. Ramasamy,et al.  Scanning and transmission electron microscopic studies of the surface of Vallisia indica (Monogenea, Polyopisthocotylea) , 1987 .

[28]  M. M. El-Naggar,et al.  Scanning electron microscope observations on the monogenean skin parasite Macrogyrodactylus congolensis [Prudhoe, 1957] Yamaguti, 1963 , 2003 .

[29]  B. Koubková,et al.  Ethanol-fixed material used for both classical and molecular identification purposes: Eudiplozoon nipponicum (Monogenea: Diplozoidae) as a case parasite species , 2010, Parasitology Research.

[30]  K. Buchmann,et al.  Interactions between monogenean parasites and their fish hosts. , 2002, International journal for parasitology.

[31]  A. Maule,et al.  Monogenean neuromusculature: some structural and functional correlates. , 1998, International journal for parasitology.

[32]  L. A. Vøllestad,et al.  Asymmetries in the attachment apparatus of a gill parasite , 2007 .

[33]  P. Horák,et al.  Surface carbohydrates of Eudiplozoon nipponicum pre- and post-fusion , 2004, Journal of Helminthology.

[34]  Z. Zdráhal,et al.  Major acid endopeptidases of the blood-feeding monogenean Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae) , 2016, Parasitology.

[35]  J. Bovet Contribution à la morphologie et à la biologie de "Diplozoon paradoxum" V. Nordmann, 1832 , 1965 .

[36]  J. D. Smyth The physiology of trematodes , 1968 .

[37]  K. Lyons The epidermis and sense organs of the monogenea and some related groups. , 1973, Advances in parasitology.

[38]  D. C. Gomes,et al.  A new description of the reproductive system of Schistosoma mansoni (Trematoda: Schistosomatidae) analyzed by confocal laser scanning microscopy , 2004, Parasitology Research.

[39]  G. Kearn,et al.  Ultrastructural observations on the tegument and associated structures of the monogenean Cichlidogyrus halli typicus (Price & Kirk, 1967) Paperna, 1979. , 1991, International journal for parasitology.

[40]  G. Brennan,et al.  Ultrastructure of the surface structures of Allodiscocotyla diacanthi (Polyopisthocotylea: Monogenea) from the gills of the marine teleost fish, Scomberoides tol. , 1995, International journal for parasitology.

[41]  E. Dzika,et al.  Ultrastructure of the digestive tract of Paradiplozoon homoion (Monogenea) , 2015, Parasitology Research.

[42]  M. Benovics,et al.  A novel type I cystatin of parasite origin with atypical legumain-binding domain , 2017, Scientific Reports.

[43]  T. Fortoul,et al.  Visualization and 3D Reconstruction of Flame Cells of Taenia solium (Cestoda) , 2011, PloS one.

[44]  M. Gelnar,et al.  Investigation of Paradiplozoon homoion (Monogenea, Diplozoidae) life cycle under experimental conditions. , 2007, Parasitology international.

[45]  K. Lyons Sense organs of monogenean skin parasites ending in a typical cilium , 1969, Parasitology.

[46]  G. Brennan,et al.  Ultrastructure of the surface structures and haptor of Empleurosoma pyriforme (Ancyrocephalinae; Monopisthocotylea: Monogenea) from the gills of the teleost fish Therapon jarbua , 2000, Parasitology Research.

[47]  Aspects of the morphology of the juvenile life stages of Paradiplozoon ichthyoxanthon Avenant-Oldewage, 2013 (Monogenea: Diplozoidae) , 2014, Acta Parasitologica.

[48]  M. Gelnar,et al.  Eudiplozoon nipponicum in focus: monogenean exhibiting a highly specialized adaptation for ectoparasitic lifestyle , 2011, Parasitology Research.

[49]  A. V. Nordmann Mikrographische Beiträge zur Naturgeschichte der wirbellosen Thiere , 1832 .