Céline Levron

Spermatozoa of tapeworms (Platyhelminthes, Eucestoda): advances in ultrastructural and phylogenetic studies.

New data on spermiogenesis and the ultrastructure of spermatozoa of ‘true’ tapeworms (Eucestoda) are summarized. Since 2001, more than 50 species belonging to most orders of the Eucestoda have been studied or reinvestigated, particularly members of the Caryophyllidea, Spathebothriidea, Diphyllobothriidea, Bothriocephalidea, Trypanorhyncha, Tetraphyllidea, Proteocephalidea, and Cyclophyllidea. A new classification of spermatozoa of eucestodes into seven basic types is proposed and a key to their identification is given. For the first time, a phylogenetic tree inferred from spermatological characters is provided. New information obtained in the last decade has made it possible to fill numerous gaps in the character data matrix, enabling us to carry out a more reliable analysis of the evolution of ultrastructural characters of sperm and spermiogenesis in eucestodes. The tree is broadly congruent with those based on morphological and molecular data, indicating that convergent evolution of sperm characters in cestodes may not be as common as in other invertebrate taxa. The main gaps in the current knowledge of spermatological characters are mapped and topics for future research are outlined, with special emphasis on those characters that might provide additional information about the evolution of tapeworms and their spermatozoa. Future studies should be focused on representatives of those major groups (families and orders) in which molecular data indicate paraphyly or polyphyly (e.g. ‘Tetraphyllidea’ and Trypanorhyncha) and on those that have a key phylogenetic position among eucestodes (e.g. Diphyllidea, ‘Tetraphyllidea’, Lecanicephalidea, Nippotaeniidea).

Ultrastructure of spermiogenesis and the spermatozoon of Monorchis parvus Looss, 1902 (Digenea, Monorchiidae), a parasite of Diplodus annularis (Pisces, Teleostei)

Abstract The present paper describes the characteristic ultrastructural features of spermiogenesis and the spermatozoon of Monorchis parvus (Trematoda, Digenea, Monorchiidae). This spermiogenesis is characterized by a flagellar rotation of about 120° and the formation of a novel ultrastructural element: a centriolar extension. It nevertheless follows the general pattern of digeneans. It begins with the formation of a differentiation zone, comprising striated rootlets associated with two centrioles and an intercentriolar body. The mature spermatozoon presents features allowing the distinction between M. parvus and other digenetic trematodes. It possesses an original anterior extremity. The two central elements of the axonemes appear prior to the peripheral doublets. The external ornamentations of the cell membrane are formed parallel to the central elements. Another peculiarity of the male gamete is the presence of two mitochondria. M. parvus, therefore, differs distinctly from other digenetic trematodes.

Spermiogenesis and sperm ultrastructure of the pseudophyllidean cestode Triaenophorus nodulosus (Pallas, 1781)

Spermiogenesis and ultrastructure of the spermatozoon of the pseudophyllidean cestode Triaenophorus nodulosus (Pallas, 1781), a parasite of pike Esox lucius, has been studied by transmission electron microscopy. Spermiogenesis involves firstly the formation of a zone of differentiation with two centrioles associated with striated roots, and an intercentriolar body between them, subsequent growth of the two flagella of unequal length, and a formation of a median cytoplasmic process exhibiting patches of dense material. The nucleus penetrates into spermatid body after flagellar rotation and proximo-distal fusion has started. The mature spermatozoon of T. nodulosus is filiform and contains two axonemes of 9+“1” pattern of the Trepaxonemata, nucleus, cortical microtubules parallel to the spermatozoon axis, and electron-dense granules. The anterior extremity of the gamete contains a single centriole surrounded by numerous electron-dense tubular structures exhibiting spiral arrangement and giving rise to lateral projections, which correspond to the crested body. When the crested body disappears, the spiral pattern of electron-dense tubular structures is changed into a ring, persisting until the centriole of the second axonemes appears. This structure of the crested body of T. nodulosus is unique among the Eucestoda.

Spermiogenesis and sperm ultrastructure of Diphterostomum brusinae (Digenea, Zoogonidae), a parasite of Diplodus annularis (Pisces, Teleostei)

This paper constitutes the first ultrastructural study of spermiogenesis and the spermatozoon of a Digenea belonging to the family Zoogonidae, Diphterostomum brusinae. Spermiogenesis follows the general pattern found in the digeneans. It begins with the formation of a differentiation zone in the spermatid. The two centrioles give rise to flagella. These two flagella undergo a rotation of 90° and fuse with the median cytoplasmic process. The proximo-distal fusion occurs at the level of attachment zones. The mature spermatozoon of D. brusinae presents all the features found in the digeneans gametes: two axonemes, mitochondrion, nucleus and two bundles of parallel cortical microtubules. Nevertheless, several characteristics distinguish the spermatozoon of D. brusinae from other digeneans, in particular the external ornamentations of the plasma membrane and the anterior and posterior extremities. The description of the ultrastructural features of reproduction in this species would contribute to the understanding of the phylogenetic relationships in the Zoogonidae family.

Spermiogenesis and Spermatozoon of the Tapeworm Ligula intestinalis (Diphyllobothriidea): Phylogenetic Implications

Abstract Using transmission electron microscopy, spermiogenesis and the spermatozoon ultrastructural organization are described in Ligula intestinalis (Linnaeus, 1758) (Diphyllobothriidea), a parasite of the great crested grebe Podiceps cristatus (Linnaeus, 1758). Spermiogenesis starts with the differentiation zone of 2 striated rootlets, 2 centrioles giving rise to 2 flagella, and an intercentriolar body. The latter is composed of 5 electron-dense layers separating 4 electron-lucent layers. In the early stages of spermiogenesis, an electron-dense material is present in the apical region of the differentiation zone. Later, the flagella undergo a rotation and fuse with the cytoplasmic extension in a proximo-distal process. The spermatozoon contains 2 axonemes with a 9 + “1” trepaxonematan pattern, the nucleus, the cortical microtubules, and an electron-dense zone. The spermatozoon anterior extremity in L. intestinalis is characterized by the absence of crested bodies and a ring of electron-dense cortical microtubules. Some characters of spermiogenesis and spermatozoon in L. intestinalis confirm the recent splitting of “Pseudophyllidea” into 2 new orders, i.e., Bothriocephalidea and Diphyllobothriidea. The process of spermiogenesis is similar in both orders for the “type I” of spermiogenesis and the presence of electron-dense material. However, the intercentriolar body is clearly more developed in the Diphyllobothriidea than in the Bothriocephalidea. Moreover, these 2 orders seem to differ in the presence or absence of a ring of electron-dense cortical microtubules in the anterior extremity of the spermatozoon.

Spermatozoon ultrastructure of the pseudophyllidean cestode Paraechinophallus japonicus, a parasite of deep-sea fish Psenopsis anomala (Perciformes, Centrolophidae)

This paper provides the first ultrastructural data on the spermatozoon of a pseudophyllidean cestode belonging to the family Echinophallidae, Paraechinophallus japonicus, parasitizing deep-sea fish Psenopsis anomala from Japan. The mature spermatozoon contains two axonemes of the 9+“1” trepaxonematan pattern, nucleus, cortical microtubules, and electron-dense granules. Its anterior extremity exhibits a single electron-dense crested body. One of the most interesting features of the mature spermatozoon is the presence of a ring of tubular structures encircling the axoneme. This character, previously reported in species of the families Bothriocephalidae and Triaenophoridae, may be unique to pseudophyllideans of the group “Bothriocephalidea”, thus supporting the assumption that the order Pseudophyllidea is formed in fact by two unrelated clades; “Bothriocephalidea” and “Diphyllobothriidea”. The posterior extremity contains the posterior part of the nucleus and the disorganized axoneme—a character previously not described in the Eucestoda.

Site influence in parasite distribution from fishes of the Bonifacio Strait Marine Reserve (Corsica Island, Mediterranean Sea)

The spatial distribution of parasite communities from teleost fishes (Diplodus vulgaris, Mullus surmuletus, Pagellus erythrinus, Phycis phycis, Scorpaena scrofa and Symphodus tinca) was studied in the Bonifacio Strait Marine Reserve (Corsica Island) with different protection levels. Canonical correspondence analysis was used to study spatial patterns and to quantify the importance of site influence on parasite communities. To supplement these results, we used the indicator species analysis (Indval) to identify parasite species as bioindicators of a given ecosystem. Moreover, fish spatial distribution in the marine reserve was evaluated in order to link parasite species occurrence and fish abundance. Several parasite species were associated with a specific site, and some of them were considered as site bioindicators, especially in the western part of the protected area (Bruzzi and Moines Islands). However, the relative abundance of fish followed a different spatial pattern. Stronger fish abundance was observed in the eastern part of the reserve (Lavezzi Islands). The parasite species were correlated to sites inside as well outside the reserve. The parasite distribution seemed not to be influenced by the protection status of the site. Furthermore, in our study, most of the parasite species were not linked to a site. The most important factor influencing their distribution was host specificity. This suggested that the parasite life traits history remained a fundamental determinant in the community composition.

Spermiogenesis and spermatozoon of the tapeworm Parabothriocephalus gracilis (Bothriocephalidea): Ultrastructural and cytochemical studies

Spermiogenesis and spermatozoon ultrastructure of the tapeworm Parabothriocephalus gracilis were described using transmission electron microscopy (TEM). Spermiogenesis is characterized by the formation of a zone of differentiation with two centrioles associated with striated rootlets, and an intercentriolar body between them. The two flagella undergo a rotation of 90° until they become parallel to the median cytoplasmic extension with which they fuse. Electron-dense material is present in the apical region of the zone of differentiation in the early stages of spermiogenesis. This electron-dense material is characteristic for the orders Bothriocephalidea and Diphyllobothriidea. The mature spermatozoon contains two axonemes of the 9 + ‘1’ trepaxonematan pattern, nucleus, parallel cortical microtubules and electron-dense granules of glycogen. The anterior extremity of the spermatozoon exhibits a single helical electron-dense crested body 130 nm thick. One of the most interesting features is the presence of a ring of cortical microtubules surrounding the axoneme. This character has been reported only for species of the order Bothriocephalidea and may be unique in this cestode group.

Ultrastructure of spermiogenesis and mature spermatozoon of Breviscolex orientalis (Cestoda: Caryophyllidea).

Spermiogenesis and spermatozoon ultrastructure of the caryophyllidean cestode Breviscolex orientalis Kulakovskaya, 1962, first member of the family Capingentidae studied, a parasite of cyprinid fish Abbottina rivularis, are described using transmission electron microscopy. Spermiogenesis in B. orientalis follows the Type II pattern described by Bâ and Marchand (Mém Mus Natl Hist Nat 166:87–95, 1995) for cestodes. It begins with the formation of a zone of differentiation containing a large nucleus and a pair of centrioles. The centrioles are separated from one another by an intercentriolar body composed of three electron-dense layers. Each centriole is associated with typical striated roots. At the beginning of the spermiogenesis, an electron-dense material is observed in the apical region of the differentiation zone. During the initial stage of spermiogenesis, one of the centrioles gives rise to a free flagellum, which then rotates and undergoes proximodistal fusion with the cytoplasmic protrusion of the differentiation zone. The mature spermatozoon of B. orientalis corresponds to the Type III pattern described by Levron et al. (Biol Rev 85:523–543, 2010). It is characterized by the absence of mitochondrion and crested body. Five regions of the mature spermatozoon are differentiated. The main ultrastructural characteristics are: one axoneme of 9 + “1” trepaxonematan pattern, cortical microtubules and nucleus. The comparison of the spermiogenesis of B. orientalis with those of the other caryophyllidean species demonstrates some variation within the order relative to the presence and morphology of the intercentriolar body, the presence of slight rotation of the flagellar bud and a complete proximodistal fusion of the free flagellum with a cytoplasmic protrusion.

Spermatological characters of bothriocephalideans (Cestoda) inferred from an ultrastructural study on Oncodiscus sauridae and Senga sp.

Spermiogenesis and ultrastructure of the spermatozoon of two bothriocephalidean cestodes, Oncodiscus sauridae from the lizardfish Saurida nebulosa Valenciennes, 1850 and Senga sp. from the eel Mastacembelus armatus (Lacepède, 1800), have been studied using transmission electron microscopy. Spermiogenesis included the formation of a zone of differentiation, where two centrioles associated with the striated rootlets occur. An intercentriolar body composed of one thick central electron-dense plate and two thinner plates on each side appears between two centrioles. Two flagella of unequal length grow and undergo a vertical rotation and proximodistal fusion with the median cytoplasmic process. Subsequently, the nucleus penetrates into the median cytoplasmic extension. The electron-dense material in the early stages of spermiogenesis is characteristic for the apical region of the differentiation zone. This electron-dense material is typical for basal tapeworms, e.g., Bothriocephalidea, Caryophyllidea, Diphyllobothriidea, and Spathebothriidea. The mature spermatozoon of O. sauridae and Senga sp. is filiform and possesses two axonemes of the 9 + “1” trepaxonematan pattern, a nucleus, cortical microtubules, and electron-dense granules. The anterior part of the gamete contains a single electron-dense crested body. The most interesting character found is the presence of a ring of cortical microtubules encircling the axoneme in the anterior part of the spermatozoon. This feature has been detected only for species of the order Bothriocephalidea and may represent a synapomorphy of these tapeworms. A classical pattern for spermatological characters (spermiogenesis of type I with dense material in early stages and sperm of type II with a characteristic ring of cortical microtubules in the anterior part) in Bothriocephalidea is discussed.