Aneta Yoneva

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.

Ultrastructure of spermiogenesis and mature spermatozoon of Skrjabinoporus merops (Cyclophyllidea, Metadilepididae)

The ultrastructure of the mature spermatozoon and the spermiogenesis of a cestode belonging to the family Metadilepididae is described for the first time. The mature spermatozoon of Skrjabinoporus merops is characterized by twisted peripheral microtubules, the presence of a single crested body, periaxonemal sheath and electron-dense rods, and the absence of intracytoplasmic walls and inclusions (glycogen or proteinaceous granules); no peripheral microtubules where nucleus contacts the external plasma membrane. Four morphologically distinct regions of the mature spermatozoon are differentiated. The proximal part (Region I) contains a single crested body, periaxonemal sheath is absent in some (proximal) sections and is present in others situated closer to the nucleus. The central Region II is nucleated, and is followed by Region III that contains a periaxonemal sheath. The distal pole, Region IV, is characterized by disintegration of the axoneme. Spermiogenesis follows the type III pattern (Bâ and Marchand 1995) although in S. merops a slight flagellar rotation is observed. The differentiation zone is characterized by the absence of striated roots and intercentriolar body; two centrioles are present, one of which gives rise to a free flagellum. The latter rotates and undergoes proximodistal fusion with the cytoplasmic protrusion of the differentiation zone. Spermiological characters of S. merops are similar to those of the families Taeniidae and Catenotaeniidae. The mature spermatozoon differs from those of the Dilepididae (where the metadilepidid species have previously been classified) by the lack of glycogen.

Ultrastructure of spermiogenesis and mature spermatozoon of Angularella beema (Clerc, 1906) (Cestoda, Cyclophyllidea, Dilepididae)

The ultrastructure of the spermiogenesis of a dilepidid cestode species is described for the first time. The spermiogenesis of Angularella beema is characterised by absence of both flagellar rotation and proximodistal fusion. The differentiation zone is surrounded by cortical microtubules and is delimited by a ring of arching membranes. It contains two centrioles, one of which develops the axoneme that grows directly into the elongating cytoplasmic protrusion. This pattern of spermiogenesis was described as the Type IV spermiogenesis of cestodes. Among cestodes, similar pattern of spermiogenesis is known in the family Hymenolepididae and in some representatives of the family Anoplocephalidae. The mature spermatozoon of A. beema consists of five regions differing in their ultrastructural characteristics. It is characterised by the presence of cortical microtubules (spirally arranged at angle of 30–40° to the spermatozoon axis) and a single crested body. There is a periaxonemal sheath in certain parts of the spermatozoon as well as glycogen-like granules between the periaxonemal sheath and the cortical microtubules. The comparisons of the mature spermatozoon of A. beema with those of other two dilepidid species (Dilepis undula and Molluscotaenia crassiscolex) demonstrate some variation within the family: presence of periaxonemal sheath in A. beema and D. undula and its absence in M. crassiscolex; presence of electron-dense rods in D. undula and their absence in A. beema.

Spermiogenesis and sperm ultrastructure of Valipora mutabilis Linton, 1927 (Cestoda, Cyclophyllidea, Gryporhynchidae)

This is the first ultrastructural study of the spermiogenesis and the mature spermatozoon of a cyclophyllidean cestode of the family Gryporhynchidae. The spermiogenesis of Valipora mutabilis begins with the formation of a differentiation zone delimited by arching membranes and containing two centrioles. One of the centrioles develops an axoneme that grows directly into the cytoplasmic protrusion. The other centriole remains situated in a cytoplasmic bud and is subsequently aborted (type IV of cestode spermiogenesis). The mature spermatozoon of V. mutabilis is a filiform cell, tapered at both extremities and lacks mitochondria. The anterior extremity is characterised by the presence of an apical cone and a single helicoidal crested body. The axoneme is of 9 + ‘1’ trepaxonematan pattern, with a periaxonemal sheath. The cortical microtubules are twisted at an angle of about 45° to the spermatozoon axis. The nucleus is electron dense and spirally coiled around the axoneme. The cytoplasm is electron lucent and contains numerous granules of electron-dense material. In contrast to a recent opinion for close phylogenetic relationships, these ultrastructural data demonstrate the distant position between gryporhynchids and the family Taeniidae. The most similar pattern in the ultrastructure of the spermiogenesis and the mature spermatozoon has been described for dilepidids, some hymenolepidids and some anoplocephalids.

Spermatological Characters of Monozoic Tapeworms (Cestoda: Caryophyllidea), Including First Data on a Species from the Indomalayan Catfish

Abstract: The ultrastructure of spermiogenesis and mature spermatozoon in Lytocestus indicus (Cestoda: Lytocestidae) is described; this is the first representative of this group of monozoic, presumably most basal, tapeworms (Eucestoda) from the Indomalayan region to be documented in this manner. Similarly, as in other caryophyllideans, its spermiogenesis involves the formation of a conical differentiation zone with 2 centrioles associated with striated roots and an intercentriolar body. In the course of the process, 1 of the centrioles develops a free flagellum, which fuses with a cytoplasmic protrusion, whereas the other remains oriented in a cytoplasmic bud. Spermiogenesis is also characterized by the presence of electron-dense material in the early stages of spermiogenesis and a slight rotation of the flagellar bud. The mature spermatozoon of L. indicus is a filiform cell tapered at both extremities that lacks mitochondria; its nucleus has parallel disposition to the axoneme and does not reach up to the posterior extremity of the spermatozoon, which is typical for spermatozoa of the type III pattern. The new data confirm that caryophyllideans share the same type of spermiogenesis that is considered to be plesiomorphic in the Eucestoda. The existing information on spermatological ultrastructure of 8 members for 3 of 4 caryophyllidean families from different host groups (cyprinids and catostomids, both Cypriniformes, and mochokids and clariids, both Siluriformes) from 4 zoogeographical regions (Palearctic, Neotropic, Ethiopian, and Indomalayan regions) demonstrates great uniformity in spermiogenesis and sperm ultrastructure, which does not reflect different taxonomic position of the species studied.

First study of vitellogenesis of the broad fish tapeworm Diphyllobothrium latum (Cestoda, Diphyllobothriidea), a human parasite with extreme fecundity

In the present study, the process of vitellogenesis of one of the most prolific organisms, the broad tapeworm, Diphyllobothrium latum, the causative agent of human diphyllobothriosis, was studied for the first time using transmission electron microscopy. Cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for detection of glycogen was applied. Starting from the periphery toward the center of the vitelline follicle four stages of vitellocytes are differentiated: immature vitellocytes, early maturing vitellocytes, advanced maturing and mature vitellocytes. Differentiation into mature vitellocytes involves the formation of shell globule clusters containing shell globules, large amount of saturated lipid droplets and glycogen. A peculiar ultrastructural feature of D. latum vitellogenesis is the presence of lamellar bodies in the cytoplasm of mature vitellocytes. This feature is similar to that present in the closely related caryophyllideans and spathebothriideans. Despite the great similarity observed in the embryonic development of diphylobothriideans, caryophyllideans and spathebothriideans, and the fact that their vitellocytes share a feature not reported from other cestode groups, there are substantial differences in the morphology of vitelline clusters, types, amount and localization of their nutritive reserves.

Spermiogenesis and spermatozoon ultrastructure of the paruterinid cestode Notopentorchis sp. (Cyclophyllidea).

Ultrastructural characters of the spermiogenesis and mature spermatozoon of Notopentorchis sp. (Cestoda, Cyclophyllidea, Paruterinidae), a parasite from Apus affinis (Aves, Apodiformes, Apodidae) from Gabon, are described by means of transmission electron microscopy. Cytochemical analysis for detection of glycogen was applied. Vestigial striated roots associated with the two centrioles are present in the zone of differentiation. The spermiogenesis is characterized by an external growth of free flagellum followed by a proximodistal fusion of the latter with cytoplasmic protrusion, thus, corresponding to the cestode spermiogenesis of the type III pattern described by Bâ and Marchand (Mem. Mus. Natl. Hist. Nat. 166:87–95, 1995). In the final stage of spermiogenesis, a single crested body appears at the base of the forming spermatozoon. The mature spermatozoon of Notopentorchis sp. is filiform and tapering at both extremities. It consists of five regions differing in their ultrastructural characteristics. The anterior extremity of the mature spermatozoon is characterized by the presence of an apical cone and a single crested body. The cytoplasm contains one axoneme of 9 + “1” type of the trepaxonematan Platyhelminthes, a periaxonemal sheath, a layer of twisted cortical microtubules, transverse intracytoplasmic walls, and granules of glycogen. The nucleus is coiled in spiral around the axoneme. The posterior extremity of the spermatozoon is characterized by the presence of electron-dense material. This structural organization corresponds to the morphology of cestode spermatozoon of type VII as defined by Levron et al. (Biol Rev 85: 523-543, 2010). The comparison of the results with those of the two previous studies on paruterinids suggests that several characters of the spermiogenesis and the mature spermatozoon are invariable, i.e. the type III spermiogenesis and the presence of vestigial striated roots, a single crested body, a periaxonemal sheath, and intracytoplasmic walls. The main differences of the sperm cells among members of this family are the lack of dense granules (as in Triaenorhina rectangula) and the presence of electron-dense material in the posterior extremity of the spermatozoon (as in Notopentorchis sp.).

Vitellogenesis of diphyllobothriidean cestodes (Platyhelminthes).

The recently erected cestode order Diphyllobothriidea is unique among all tapeworm orders in that its species infect all major groups of tetrapods, including man. In the present paper, the vitellogenesis of representatives of all three currently recognized families of this order was evaluated, based on ultrastructural (transmission electron microscopy) and cytochemical (detection of glycogen) observations. Vitelline follicles of all taxa studied, i.e. Cephalochlamys namaquensis from clawed frogs (Xenopus), Duthiersia expansa from monitors (Varanus) and Schistocephalus solidus that matures in fish-eating birds, contain vitelline cells at various stages of development and interstitial cells. Developing vitellocytes are characterized by the presence of mitochondria, granular endoplasmic reticulum and Golgi complexes involved in the synthesis of shell globules and formation of shell globule clusters. Mature vitellocytes contain lipids and glycogen in different proportions. The most significant differences among the three diphyllobothriidean families were found in the presence or absence of lamellar bodies. Variations of vitelline clusters morphology and types of lipid droplets are described and discussed in relation to the presumed evolutionary history of diphyllobothriideans, which belong to the most basal cestode groups.

Description of embryonic development and ultrastructure in miracidia of Cardiocephaloides longicollis (Digenea, Strigeidae) in relation to active host finding strategy in a marine environment

The functional ultrastructure and embryonic development of miracidia in naturally released eggs of the trematode Cardiocephaloides longicollis were studied using light and transmission electron microscopy. This species has operculated eggs and embryogenesis occurs in the marine environment before an actively infecting ciliated miracidium hatches. Six different developmental stages were identified. The lack of pores in the eggshell indicates its impermeability and the miracidiums dependency on glycogen nutritive reserves, contained in numerous vitellocytes in early embryos. As the development advances, these merge into larger vitelline vacuoles that encircle the miracidium and may aid its hatching. Tissue and primary organ differentiation were observed in advanced stages, i.e., terebratorium, glands, cerebral ganglion, peripheral sensory endings, and eyespots. The anterior part of the body contains a single apical and paired lateral glands, as well as two types of sensory endings, which permit location, adhesion, and penetration of the host. No previous studies describe the embryonic development and ultrastructure of miracidia in strigeids, however, some of the structural features shared with other, well described species with unknown life cycles are emphasised. This study highlights that ultrastructural data have to be interpreted in relation to parasite biology to understand the structural requirements of specific parasite strategies.

The first data on the vitellogenesis of paruterinid tapeworms: an ultrastructural study of Dictyterina cholodkowskii (Cestoda: Cyclophyllidea)

The present study provides the first ultrastructural data of the vitellogenesis in a cestode species of the cyclophyllidean family Paruterinidae, aiming to expand the limited data on the vitellogenesis in cyclophyllidean cestodes and to explore the potential of ultrastructural characters associated with vitellogenesis for phylogenetic and taxonomic studies of this order. The process of vitellocyte formation in Dictyterina cholodkowskii follows the general pattern observed in other tapeworms but exhibits several specific differences in the ultrastructure of vitelline cells. The vitellarium contains vitellocytes at various stages of maturation. The periphery of the vitellarium and the space between maturing vitellocytes are occupied by interstitial cells. Differentiation into mature vitellocytes is characterized by high secretory activity, which involves the development of granular endoplasmic reticulum, Golgi complexes, mitochondria and vitelline globules of various sizes. During vitellogenesis, the progressive fusion of these globules results in the formation of two large membrane-limited vitelline vesicles that eventually fuse into a single large vesicle. Mature vitellocytes are composed of a single vitelline vesicle, a high content of cytoplasmic organelles and have no nucleus. No traces of lipid droplets and glycogen granules are detected in the cytoplasm of mature vitellocytes, which might be related to biological peculiarities of this family, i.e. the release of eggs into environment within the tissues of the paruterine organ, which may serve as a source of nutrients for embryos.