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.).

Spermiogenesis and spermatozoon ultrastructure of Hunterella nodulosa (Cestoda: Caryophyllidea), a monozoic parasite of suckers (Catostomidae) in North America.

Spermiogenesis and ultrastructure of mature spermatozoon of the caryophyllidean cestode Hunterella nodulosa, a parasite of suckers (Catostomidae), have been studied by transmission electron microscopy. This monozoic tapeworm is unique in its mode of attachment and represents the second North American species studied. The process of spermiogenesis of H. nodulosa follows the general pattern already described in other caryophyllideans. The most characteristic feature is the presence of a slight rotation of the flagellar bud, which seems to be a typical character of spermiogenesis in this cestode group. The mature spermatozoon of H. nodulosa is characterized by the presence of one axoneme of 9 + "1" type of the trepaxonematan flatworms surrounded by a semi-arc of cortical microtubules in its anterior extremity, parallel nucleus and cortical microtubules arranged in a parallel pattern, which corresponds to the Type III pattern of cestode spermatozoa according to Levron et al. (2010). Comparison of the present data with those available for other caryophyllideans did not reveal substantial differences, even though they belong to different families, infect different hosts (catostomid, cyprinid and siluriform fishes) and occur in distant zoogeographical regions. This indicates uniformity of the process of sperm formation and spermatozoon ultrastructure in one of the evolutionarily most ancient groups of tapeworms.

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.

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.

Endoparasites of European perch Perca fluviatilis fry: role of spatial segregation.

A total of 246 perch Perca fluviatilis L. fry, 20 to 106 d old from 3 different reservoir subpopulations (epipelagic perch fry, EPF; bathypelagic perch fry, BPF; littoral perch fry, LPF), were examined for parasites. Six species of endoparasites were found: the nematode Camallanus lacustris was the most common, followed by the cestodes Proteocephalus percae, Bothriocephalus claviceps, Glanitaenia osculata and the acanthocephalan Acanthocephalus lucii. All worms were juvenile or immature and were recovered from the intestinal lumen, with the exception of plerocercoids of Triaenophorus nodulosus, which were found in the body cavity or already encysted in the liver (the final site of infection of metacestodes). A marked difference was found in infection rates in the 3 spatially segregated subpopulations of perch fry. Parasites were found almost exclusively in LPF, which were heavily infected (overall prevalence = 30%) compared with the other studied subpopulations. Two species (C. lacustris and T. nodulosus) were found in 1 fish each (prevalence = 3%) in BPF, whereas EPF were uninfected. The species richness and prevalence of parasites in LPF increased from 20-24 d old fry (2 species of parasites; prevalence = 13%) to 106 d old fry (5 species of parasites; prevalence = 80%).

Spermiogenesis and spermatozoon of the tapeworm Ligula intestinalis (Diphyllobothriidea): phylogenetic implications.

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.

Spermatological characters of the aspidogastrean Aspidogaster limacoides Diesing, 1835.

Using transmission electron microscopy, spermiogenesis and the spermatozoon ultrastructure are described in Aspidogaster limacoides (Aspidogastrea, Aspidogastridae), from two cyprinid fishes, Abramis sapa and Blicca bjoerkna, from eastern Slovakia and north-western Russia. Results of the present investigation permit, for the first time in Aspidogastrea, to describe general pattern of spermiogenesis and the organization of the spermatozoon in this presumably the most basal group of parasitic flatworms (Neodermata). Spermiogenesis starts by the formation of a zone of differentiation characterized by long striated rootlets, two centrioles giving rise to flagella, and intercentriolar body composed of different electron-dense plates. Later, a flagellar rotation followed by a proximodistal fusion of the flagella with the cytoplasmic extension takes place. Beside the possession of elements typical of Neodermata, such as two axonemes of the 9 + '1' trepaxonematan pattern, nucleus, mitochondrion and cortical microtubules, the mature spermatozoon of A. limacoides exhibits characteristics unique in Aspidogastrea: (a) a long undulating membrane, (b) an electron-dense zone, and (c) a lamellate body. The terminology and structure of different typical elements of Aspidogastrea are discussed. It is interesting to note that the long undulating membrane and a continuous row of cortical microtubules have already been described in polyopisthocotylean monogeneans, which can be a case of convergence unless future phylogenetic studies confirm relatedness of these groups.

The unique ultrastructure of the uterus of the Gyrocotylidea Poche, 1926 (Cestoda) and its phylogenetic implications.

The members of the order Gyrocotylidea are monozoic tapeworms and generally considered to be the most primitive group of the Cestoda in terms of the evolution of this platyhelminth class. As part of a series of ultrastructural studies on Gyrocotyle urna (Wagener, 1852), three regions of the uterus were distinguished. The proximal region of the uterus is characterised by underlying perikarya, the presence of septate junctions within the epithelial wall and two types of specialised outer coverings, lamellae and cilia. The middle, syncytial region of the uterus is covered by short lamellae and marked by a concentration of sunken glandular perikarya (uterine glands). Glandular spheroidal granules (c.0.45 microm in diameter) of moderately dense content and a fine fibrillar texture are liberated by migration through the luminal membrane. The epithelium of the sac-shaped, distal region of the uterine duct is interrupted by cytoplasmic processes of sunken epithelial bodies, covered with lamellae and contains septate junctions. A muscular sphincter surrounds the narrow, terminal region of the distal uterine duct. The ultrastructural pattern of the uterus of the Gyrocotylidea has important discriminating traits (i.e. the presence of sunken perikarya along its entire length, septate junctions within the uterine epithelial cytoplasm of the proximal and distal regions, and cilia on the surface of its proximal region) unique among the Neodermata and which may represent autapomorphic character states for the group.

SEM and TEM study of the armed male terminal genitalia of the tapeworm Paraechinophallus japonicus (Cestoda: Bothriocephalidea).

For the first time, the ultrastructure of the armed cirrus of an echinophallid cestode, Paraechinophallus japonicus (Yamaguti, 1934), has been studied with the use of scanning and transmission electron microscopy. Two sets of eversible copulatory organs (approximately 300 microm in length and approximately 130 microm in width) are present on the dorsal side of each segment near the lateral margin of the strobila. Except for the terminal portion, the cirrus is covered with large spines (up to 40 mircom long, measured from SEM photomicrographs) composed of 2 parts. The basal portion contains a lobed electron-dense outer region that gives way to a reticular meshwork of electron-dense material. The apical region of the spines, composed of a homogeneous, moderately electron-dense matrix, is slightly curved distally. Spines are covered with a cortical zone. Between the spines, the distal cytoplasm is covered with microvilli of about 1.2 microm in length. The wall of the cirrus sac, which is approximately 500 microm long and approximately 250 microm wide, is composed of 2 layers of muscles, i.e., an internal layer of circular muscles and external longitudinal muscles. Microvilli on the cirrus of P. japonicus are reported for the first time in the Cestoda, whereas the spines on the cirrus may represent a synapomorphy of bothriocephalidean cestodes of the Echinophallidae.

Ultrastructure of microtriches on the scolex of Cyathocephalus truncatus (Cestoda: Spathebothriidea).

The scolex surface of the mature spathebothriidean Cyathocephalus truncatus (Pallas, 1781), a parasite of the brown trout Salmo trutta fario L., was studied using scanning and transmission electron microscopy. A particular attention was paid to microtriches, unique structure on the surface of the Cestoda. The scolex of C. truncatus is covered with two types of filiform microtriches (filitriches): aciculate (approximately 3 microm long) and capillate (approximately 10 microm long). Capillate microtriches, which have never been reported in any other spathebothriideans, are described for the first time using transmission electron microscopy. The tegument covered with filiform microtriches only (no spiniform microtriches are present) is typical of cestode groups supposed to be the most basal, e.g., Gyrocotylidea, Spathebothriidea, and Caryophyllidea.

Ultrastructure of the surface structures and secretory glands of the rosette attachment organ of Gyrocotyle urna (Cestoda:Gyrocotylidea).

The surface structures and gland cells of the posterior rosette organ of Gyrocotyle urna Grube et Wagener, 1852, a member of the group presumed to be the most basal of the tapeworms (Cestoda: Gyrocotylidea), was studied by scanning electron and transmission electron microscopy. Surface structures on the outer (oriented away from the intestinal wall) and inner (in contact with the intestinal wall) rosette surfaces differ from each other and represent a transitional form between microvilli and microtriches typical of tapeworms (Eucestoda). The inner surface of the rosette possesses numerous glands. On the basis of the size and electron-density of their secretory granules, three types of unicellular gland cells can be distinguished. The least common type (Type I) is characterized by the production of small, round, electron-dense granules of about 0.3 microm in diameter, whereas another type of secretion (Type II) is formed from homogenous, moderately electron-dense, spheroidal granules of about 0.7 microm in diameter. The most common type of glands (Type III) is recognized by a secretion comprising large, elongate, electron-dense granules of about 1 microm long and 0.5 microm broad. The secretory granules of the three types of the glands are liberated by an eccrine mechanism and the gland ducts open via small pores on the inner rosette surface. The complex of secretory glands of the posterior rosette of G. urna is similar to those in the anterior attachment glands of monogeneans (as opposed to the types of glands present in other helminth groups). However, the tegumental surface structures of Gyrocotyle are supporting evidence for the relationship between the Gyrocotylidea and Eucestoda.

Vitellogenesis and vitelline system in the pseudophyllidean tapeworm Paraechinophallus japonicus: ultrastructural and cytochemical studies.

The vitellogenesis of Paraechinophallus japonicus (Yamaguti, 1934), the first pseudophyllidean tapeworm of the family Echinophallidae studied using transmission electron microscope, is described on the basis of ultrastructural observations of specimens from the benthopelagic fish Psenopsis anomala (Temminck et Schlegel, 1844) (Perciformes: Centrolophidae). The process of vitellogenesis in P. japonicus follows the same general pattern observed in other tapeworms. Five stages of vitellocyte development have been distinguished. The first stage corresponds to immature cells containing ribosomes and mitochondria. The second stage of development is characterized by the appearance of granular endoplasmic reticulum and Golgi complexes, formation of shell globules and lipid droplets at the periphery of the cell cytoplasm. Vitellocyte of the third stage presents accumulation of shell globules and lipid droplets. During the fourth stage, shell globule clusters are formed, and lipid droplets and rosettes of alpha-glycogen are accumulated. Mature vitelline cells are characterized by a great number of lipid droplets with glycogen in the centre of the cytoplasm, whereas shell globule clusters are situated more peripherally. The interstitial tissue of vitelline follicles of P. japonicus is syncytial with long cytoplasmic projections extending between vitelline cells. The presence of a large amount of lipid droplets in the vitelline cytoplasm within the eggs of P. japonicus may be related to egg accumulation in the uterine sac.

Spermatological characters in Diphyllobothrium latum (Cestoda, Pseudophyllidea).

Spermiogenesis and the ultrastructural features of the spermatozoon of Diphyllobothrium latum (Cestoda, Pseudophyllidea) are described using transmission electron microscopy. Spermiogenesis is characterized by the development of two flagella of unequal length that grow asynchronously. When the first growing flagellum starts to rotate, the second one develops. Flagellar rotation is thus asymmetric and asynchronic. It is followed by proximodistal fusion with the median cytoplasmic process. Electron-dense material is present in the apical region of the zone of differentiation in the early stages of spermiogenesis. The intercentriolar body consists of seven plates: three are electron-dense. Four attachment zones occur in the median cytoplasmic process. An atypical arrangement of striated roots was occasionally observed. The mature spermatozoon possesses two axonemes of 9 + "1" trepaxonematan pattern, nucleus, cortical microtubules, electron-dense granules, and lacks mitochondria. The ultrastructure of the anterior extremity of the spermatozoon in D. latum clearly differs from that in the bothriocephalid pseudophyllideans, mainly in the absence of a crested body and a ring of electron-dense tubular structures. The spermatological data support the assumption that the order Pseudophyllidea is formed by two unrelated clades, "Bothriocephalidea" and "Diphyllobothriidea."

Spermatological characters of the pseudophyllidean cestode Bothriocephalus scorpii (Müller, 1776).

Spermiogenesis of Bothriocephalus scorpii (Cestoda, Pseudophyllidea) includes an orthogonal development of two flagella, followed by a flagellar rotation and a proximo-distal fusion with the median cytoplasmic process. The fusion occurs at the level of four attachment zones. The presence of dense material in the apical region of the differentiation zone in the early stage of spermiogenesis appears to be a characteristic feature for the Pseudophyllidea. The mature spermatozoon possesses two axonemes of 9+"1" pattern of the Trepaxonemata, nucleus, cortical microtubules, electron-dense granules and crested body. The anterior part of the gamete exhibits a centriole surrounded by electron-dense tubular structures arranged as incomplete spiral. When the crested body disappears, the electron-dense tubular structures are arranged into a ring encircling the axoneme. The electron-dense tubular structures and their arrangement appear to be a specific feature for the clade "Bothriocephalidea". The organization of the posterior extremity of the gamete with the nucleus is described for the first time in the Pseudophyllidea.

Spermatological characters of monozoic tapeworms (Cestoda: Caryophyllidea), including first data on a species from the Indomalayan catfish.

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.

Morphology and taxonomic status of two little-known nematode species parasitizing North American fishes.

Examination of some freshwater and brackishwater (estuarine) fishes in South Carolina in October 2009 yielded, in addition to other parasites, 2 little-known nematode species identified as Dichelyne fastigatus Chandler, 1935 (Cucullanidae), from the red drum, Sciaenops ocellatus (Linnaeus), from an estuary, and Rhabdochona ovifilamenta Weller, 1938 (Rhabdochonidae), from the shorthead redhorse, Moxostoma macrolepidotum (Lesueur), from Lake Moultrie. Light and scanning electron microscopy (the latter used for the first time for these species) made it possible to describe several important, but previously unreported, taxonomic features in D. fastigatus, such as the location of the excretory pore and deirids, the shape of deirids and a gubernaculum, the shape and size of eggs, the presence of precloacal ventral oblique muscle bands, and 11 pairs of caudal papillae and a pair of phasmids. It distinctly differs from the most similar Dichelyne cotylophora (Ward and Magath, 1917), a parasite of North American freshwater percids, in the number and arrangement of postanal papillae and by a markedly elevated cloacal region. Records of Dichelyne lintoni Barreto, 1922, from S. ocellatus probably concern D. fastigatus. Examination of R. ovifilamenta revealed a high degree of morphologic and biometric variability in this species. Based on our analysis, Rhabdochona laurentiana Lyster, 1940 , Rhabdochona milleri Choquette, 1951, and Rhabdochona catostomi Kayton, Kritsky, and Tobias, 1979, are synonymized with R. ovifilamenta Weller, 1938, typically a parasite of North American catostomids.

Cestodes (Caryophyllidea) of the stinging catfish Heteropneustes fossilis (Siluriformes: Heteropneustidae) from Asia.

The stinging catfish Heteropneustes fossilis (Bloch) (Siluriformes: Heteropneustidae) has been reported to harbor as many as 19 species of caryophyllidean tapeworms (Cestoda) of 11 genera in tropical Asia (Indomalayan zoogeographical region). However, a critical review of the species composition has shown that only 1 species, Lucknowia fossilisi Gupta, 1961 (Lytocestidae), is a specific parasite of H. fossilis. Three other species, Djombangia penetrans Bovien, 1926 (syn., Djombangia caballeroi Sahay and Sahay, 1977 ), Pseudocaryophyllaeus ritai Gupta and Singh, 1983 (syn. Pseudocaryophyllaeus lucknowensis Gupta and Sinha, 1984 ), and Pseudocaryophyllaeus tenuicollis (Bovien, 1926) Ash, Scholz, Oros and Kar, 2011 (syn. P. mackiewiczi Gupta and Parmar, 1982 ), were found only once. Lucknowia fossilisi is redescribed on the basis of new material collected in West Bengal and voucher specimens from Maharashtra, India. A total of 9 species of Capingentoides, Lucknowia, Lytocestus, Pseudoadenoscolex, Pseudocaryophyllaeus, Pseudoheteroinverta, and Sukhapatae are newly synonymized with L. fossilisi and previous synonymies of 9 other species, proposed by Hafeezulah (1993), are confirmed. Generic diagnosis of Lucknowia Gupta, 1961 is amended. In addition, 1 species of Pseudobatrachus and 2 species of the monotypic genera Pseudoneckinverta and Sudhaena are invalidated as nomina nuda.

Egg formation and the early embryonic development of Aspidogaster limacoides Diesing, 1835 (Aspidogastrea: Aspidogastridae), with comments on their phylogenetic significance.

Ultrastructural aspects of the early embryonic development of the aspidogastrean Aspidogaster limacoides are described and their phylogenetic implications discussed. Whereas the proximal regions of the uterine lumen usually contain unembryonated eggs or eggs with early embryos, the posterior or distal regions of the uterus are filled with eggs containing a fully-developed cotylocidium. The eggs of A. limacoides can be classified as polylecithal due to the presence of numerous vitellocytes which accompany each fertilized oocyte or ovum during egg formation. The results of the study are described in details under six headings: (1) general characteristics of the intrauterine eggs; (2) eggshell and operculum formation; (3) unembryonated eggs; (4) zygote formation and early cleavage divisions; (5) embryonic envelope formation; and (6) early degeneration or apoptosis of some blastomeres. The late differentiation of the operculum, possible functions of GER-bodies, and the early degeneration of vitellocytes and some blastomeres in this species are compared, drawn and discussed with corresponding observations reported for other parasitic Platyhelminthes. The most important differences are apparent in the number of egg envelopes and their mode of formation in A. limacoides compared with previous reports for both digeneans and cestodes. The results of the present TEM study indicate that the three macromeres, resulting from two cleavage divisions, take part in the formation of a single embryonic outer envelope in A. limacoides, and that this takes place at a very early stage of embryogenesis. Their fusion results in the formation of a single continuous cytoplasmic layer surrounding the early embryo, which is composed of only a small number of undifferentiated blastomeres. The early separation of the macromeres may indicate an equal cleavage pattern. These results suggest that the systematic position of the Aspidogastrea among the Platyhelminthes still remains somewhat equivocal, and indicate the need for more studies on the embryonic development, larval morphogenesis and molecular phylogeny for the elucidation of the relationships between this enigmatic group and related taxa.

Vitellogenesis of basal trematode Aspidogaster limacoides(Aspidogastrea: Aspidogastridae).

The vitellogenesis of the trematode Aspidogaster limacoides (Aspidogastrea: Aspidogastridae), a parasite of cyprinid fishes, is described here using transmission electron microscopy. Four different stages of vitellocytes are differentiated: immature vitellocytes, early maturing vitellocytes, advanced maturing vitellocytes and mature vitellocytes. The process follows the same general pattern already described in other free-living neoophorans and parasitic flatworms (i.e. Trematoda, Monogenea and Cestoda): differentiation into mature vitelline cells involves the development of mitochondria, granular endoplasmic reticulum, Golgi complexes, lipid droplets and shell-globules. Mature vitellocytes of A. limacoides are composed of numerous shell-globule clusters, few lipid droplets and glycogen granules. They differ from those of another aspidogastrean Rugogaster hydrolagi in that they possess numerous globules tightly packed and by the presence of only one type of vitelline material. The interstitial tissue of vitelline follicles of A. limacoides contains a peripheral nucleus and long cytoplasmic projections extending between vitelline cells. Since aspidogastreans are considered as an archaic group of parasitic flatworms and thus have a strategic phylogenetic position, future works needs to pay special attention to the ultrastructural and chemical composition of mature vitellocytes within this basal group of trematodes.

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).