Magdaléna Bruňanská

Spermiogenesis in the pseudophyllid cestode Eubothrium crassum (Bloch, 1779).

Abstract. Spermiogenesis of the pseudophyllidean tapeworm Eubothrium crassum has been described by the aid of transmission electron microscopy for the first time. Initially, early spermatids form a distal cytoplasmic protrusion, a differentiation zone containing a small electron-dense, apically oriented region. Out of this region, two centrioles with rootlets develop. The centrioles become orientated in the same plane with the appearance of an intercentriolar body. Now, the long axes of the rootlets are parallel with each other and with the long axis of the nucleus. Two flagella of subsequently unequal length are formed very rapidly. Simultaneously, a median cytoplasmic process (MCP) develops distal to the flagella. Two arching membranes appear at the base of the differentiation zone. Each flagellum, still being in contact with an intercentriolar body, rotates to a position parallel with the MCP. The nucleus migrates very rapidly into the MCP at this stage. Subsequently, the two flagella fuse with the MCP. Finally, the basal bodies with the rootlets detach from the flagella, the intercentriolar body changes its structure and spermatids are pinched off from a condensing residual cytoplasm at the level of the arching membranes.

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 CYATHOCEPHALUS TRUNCATUS (PALLAS, 1781) KESSLER, 1868 (CESTODA: SPATHEBOTHRIIDEA)

Spermiogenesis and sperm ultrastructure of adult Cyathocephalus truncatus, a member of presumably basal group of “true” cestodes (Eucestoda), have been examined for the first time by using transmission electron microscopy. The process of sperm formation corresponds in basic pattern to that of the Pseudophyllidea. In addition, the 2 pairs of electron-dense attachment zones are present in median cytoplasmic process of C. truncatus. However, mature spermatozoa of C. truncatus differ significantly from those of the pseudophyllideans, especially in the morphology of the proximal and distal spermatozoon extremities. The proximal extremity of the mature spermatozoon lacks a crested body, which is present in more derived cestodes and some pseudophyllideans. The distal end of the mature spermatozoa exhibits different morphology in the gametes from testes and those from receptaculum seminis. New for the Eucestoda is a finding that a lateral cytoplasmic extension creates the distal end of the spermatozoa from testes, resembling sperm of some Monogenea and Digenea. In contrast, the distal extremity of the spermatozoa from receptaculum seminis contains only a nucleus. Despite the above-mentioned peculiarities, the ultrastructural data on sperm/spermiogenesis suggest close relationships of the Spathebothriidea and Pseudophyllidea.

Vitellogenesis in two spathebothriidean cestodes

Vitellogenesis in two spathebothriidean tapeworms, dixenous adult Cyathocephalus truncatus and monoxenous progenetic Diplocotyle olrikii, has been examined using transmission electron microscopy and cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for glycogen. Each vitelline follicle consists of vitellocytes at various stages of development and one irregularly shaped interstitial cell. Projections of the interstitial cell enclose the vitellocytes and extend as a cytoplasmic sheath on the follicular periphery. An outer thin fibrous layer (= extracellular lamina) covers the cytoplasmic sheath in C. truncatus, but lacks in D. olrikii. Maturing and mature vitellocytes contain vitelline material in the form of single small shell globules that gradually fuse and give rise to the large shell globule clusters. Morphology of shell globule clusters differs slightly in both species. In addition, single “lamellar” granules are present in the cytoplasm of vitellocytes of C. truncatus, but not in D. olrikii. Both electron lucent and electron dense lipid droplets are present in the vitellocytes of C. truncatus, whereas only electron dense lipids occur in D. olrikii. A single lipid droplet turns up occasionally in the nuclei of some of the vitellocytes of C. truncatus. The ultrastructural features of vitellogenesis in spathebothriideans resemble those reported previously in “lower” cestodes, especially in pseudophyllideans.

Spermiogenesis in the proteocephalidean cestode Proteocephalus torulosus (Batsch, 1786)

Spermiogenesis of the proteocephalidean cestode Proteocephalus torulosus (Batsch, 1786) was examined for the first time using transmission electron microscopy. Spermiogenesis begins with the formation of a distal cytoplasmic protrusion, a differentiation zone, at the periphery of the early spermatid. This differentiation zone is lined with cortical microtubules and contains two centrioles aligned along the same axis. Subsequently, each centriole is associated with the striated root and the intercentriolar body appears between them. A flagellar bud arises from each centriole, growing later as a free flagellum. Simultaneously, a median cytoplasmic process (MCP) develops distally to the flagella. The two flagella, which are of unequal length, become longer and rotate towards the MCP. At this stage, two arching membranes appear at the base of the differentiation zone. The nucleus elongates and when both flagella are fused with the MCP, the nucleus subsequently migrates into the MCP. Finally, the advanced spermatids detach from a condensing residual cytoplasm at the level of the arching membranes.

Spermatological characters of the caryophyllidean cestode Khawia sinensis Hsü, 1935, a carp parasite

Spermatological characters of the caryophyllidean tapeworm Khawia sinensis (Lytocestidae), a carp parasite, have been investigated for the first time. The process of the spermatozoon formation corresponds in basic pattern to that of Khawia armeniaca (Bruňanská and Poddubnaya, Parasitol Res 99: 449–454, 2006). The presence of dense material at the early stages of spermiogenesis, the intercentriolar body, flagellar rotation (free flagellum plus flagellar bud), and a complete proximodistal fusion have been reported. In addition, it was shown that the developing spermatids may exhibit a free flagellum distally. The unipartite young spermatid and/or mature spermatozoon of K. sinensis consist of one axoneme of the 9 + “1” trepaxonematan pattern, nucleus, and cortical microtubules. These components are situated in cytoplasm, which is either electrolucent (young spermatid) or moderately electrodense (mature spermatozoon). The crested body is absent. The proximal extremity of the male gamete of K. sinensis contains a centriole and a few (up to six) cortical microtubules arranged in a semicircle. The number of cortical microtubules increases distally and reaches maximum (up to 15) in the middle part of the filiform cell, which contains the nucleus. The posterior extremity of the mature spermatozoon exhibits elements of the disorganized axoneme. The ultrastructural characteristics of spermiogenesis and the mature spermatozoon of K. sinensis support basal position of the Caryophyllidea (and Spathebothriidea) relative to other eucestode orders.

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.

Spermiogenesis in the caryophyllidean cestode Khawia armeniaca (Cholodkovski, 1915).

Spermiogenesis of the monozoic cestode Khawia armeniaca (Cholodkovski, 1915), a parasite of Capoeta capoeta sevangi (De Filippi, 1865) (Pisces: Cyprinidae), has been investigated by means of transmission electron microscopy for the first time. The present results show that the type of spermiogenesis in K. armeniaca shares the main features described previously in the pseudophyllideans and/or spathebothriideans: apical dense material in the zone of differentiation in the early stages of spermiogenesis, rotation of flagellum and flagellar bud, proximodistal fusion, and penetration of the nucleus into the spermatid body after the fusion of the free flagellum with the MCP has started. However, only one axoneme and a simple intercentriolar body represent presumably more evolved characters, typical for “higher” tapeworms. Ultrastructural data of spermiogenesis in K. armeniaca may support the hypothesis that the Caryophyllidea are secondarily derived from an ancestor closely allied with the Pseudophyllidea.

Ultrastructure and cytochemistry of vitellogenesis in Wenyonia virilis Woodland, 1923 (Cestoda, Caryophyllidea).

Vitellogenesis in Wenyonia virilis was examined by transmission electron microscopy (TEM), including the cytochemical detection of glycogen at the ultrastructural level with the periodic acid-thiosemicarbazide-silver proteinate (PA-TSC-SP) technique. Mature vitelline follicles have cells in various stages of development, progressing from immature cells of gonial type near the periphery of the follicle to maturing and mature vitellocytes towards the centre. Maturation is characterized by: (1) increase in cell volume; (2) increase in nuclear surface area restoring the N/C (nucleo-cytoplasmic) ratio; (3) nucleolar transformation; (4) extensive development of parallel cisternae of GER, the shell-protein producing units; (5) development of Golgi complexes, engaged in shell-granule/shell-globule formation and packaging; (6) synthesis and storage of glycogen in the cytoplasm; (7) simultaneous, independent formation and storage of intranuclear glycogen; (8) continuous fusion of small shell-granules into larger shell-globules and fusion of these into large shell-globule clusters with a progressive increase in the number and size of the latter; and (9) disintegration of GER in the medial layer of vitellocyte cytoplasm, degenerative changes and accumulation of glycogen and shell-globule clusters within the cytoplasm. The functional significance of numerous shell-globule clusters and the relatively small amount of nuclear and cytoplasmic glycogen is analysed. Unlike vitellogenesis of other caryophyllids, the nuclear glycogen of mature vitellocytes in W. virilis is randomly dispersed in the nucleoplasm and never forms a high central accumulation, the so-called “nuclear vacuole”. The nutritive function of vitellocytes appears greatly reduced in W. virilis, a fact perhaps related to the intrauterine development of the early embryos. The ultrastructure of vitellogenesis in W. virilis is compared with that in other lower cestodes, both monozoic and polyzoic. Conclusions concerning interrelationships of the vitellogenesis pattern of the ultrastructural cytochemistry of mature vitellocytes of W. virilis to intrauterine embryonation, absence of uterine glands and an extensive uterus characteristic for this species, are drawn and discussed.

Spermatological characters of the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890)

Spermiogenesis and the spermatozoon ultrastructure of the adult Didymobothrium rudolphii (Spathebothriidea) have been examined using transmission electron microscopy for the first time. The dense material is present in the apical region of the differentiation zone in the early stage of spermiogenesis, similarly as in other basal tapeworms examined to date. The orthogonal development of the two flagella is followed by a flagellar rotation and their proximodistal fusion with the median cytoplasmic process (MCP). The two pairs of electron-dense attachment zones in the MCP mark the lines where the proximodistal fusion of MCP with two axonemes takes place. D. rudolphii exhibits polymorphism of the intercentriolar body during spermiogenesis. The mature spermatozoon possesses the two axonemes of 9 + “1” trepaxonematan pattern, nucleus, cortical microtubules (CMs), and electron-dense granules. The anterior extremity of the gamete lacks a crested body and exhibits a centriole surrounded by a semiarc of electron-dense tubular structures. The two parallel rows of the CMs have been found in the proximal part of the two-axoneme region of the spermatozoon for the first time in the Eucestoda. The posterior extremity of the gamete exhibits pattern of the disorganized axoneme. The ultrastructural features of the sperm/spermiogenesis support a view about the close relationships of the Spathebothriidea and Diphyllobothriidea and the basal position of the Spathebothriidea within the Eucestoda.