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The intrauterine polylecithal eggs of the spathebothriidean cestode Didymobothrium rudolphii (Monticelli, 1890) were examined by means of transmission electron microscopy (TEM). Each unembryonated egg is composed of a fertilised oocyte or ovum and several vitelline cells, all surrounded by a newly formed shell. The lumen of the proximal uterus is packed with unutilised vitelline material and eggs at different stages of shell formation. In the proximal region of the uterus, the fertilised oocytes, initially surrounded by dense, discontinuous islands of eggshell material and containing long axonemes of spermatozoa in their cytoplasm, were frequently observed. Sperm axonemes also remain in the oocyte cytoplasm of eggs surrounded by a thick electron-dense shell until the sperm nucleus is transformed into a male pronucleus. Despite the fact that the two-pronuclei stage and cell divisions within the eggs of D. rudolphii were never observed, individual eggs containing several blastomeres of different sizes were seen in the middle and distal regions of the uterus. This provides indirect evidence that at least a few initial cleavage divisions must take place in the intrauterine eggs and direct evidence that the early embryonic development of D. rudolphii starts in utero. The several vitellocytes present in each egg contain nutritive reserves for the developing embryos; these are composed mainly of numerous lipid droplets and a moderate amount of glycogen. In the eggs containing early embryos composed of several blastomeres, the cytoplasm of the degenerating vitellocytes exhibits the presence of so-called ‘foci of cytoplasmic degradation’, which appear to be involved in the autolytic process of the vitellocyte cell components and inclusions, such as a high accumulation of lipids and glycogen. This progressive degeneration of the vitellocytes, considered as an example of programmed cell death or apoptosis, likely contributes towards the resorption of nutritive reserves by the developing embryo. Some of the results of this study are commented upon in relation to the affiliation of the spathebothriideans with other lower cestode groups.
Ultrastructural and cytochemical characteristics of GER-bodies observed in the vitellocyte cytoplasm of the intrauterine eggs of the caryophyllidean cestode Wenyonia virilis are described. In this species GER-bodies were observed only in the cytoplasm of vitellocytes, surrounded by a newly formed egg-shell. They are composed of spherical areas of condensed, electron-dense cytoplasm which contains concentrically arranged parallel lamellae of granular endoplasmic reticulum (GER), forming characteristic balls of different sizes. Each GER-body is surrounded by numerous free ribosomes, polyribosomes, α-glycogen rosettes and large mitochondria. Results of cytochemical analysis by means of PATSC-SP test for polysaccharides indicated that glycogen is absent within the GER-bodies, however, a strongly positive reaction was observed only in large aggregations of α-glycogen rosettes and β-glycogen particles, localised usually near GER-bodies.
Ultrastructural evidence for early intraurerine embryonic development of Wenyonia virilis is presented. At the initial stage of egg formation, the fertilized oocyte or ovum is surrounded by numerous vitellocytes and newly formed eggshell. Individual vitellocytes undergo progressive fusion into a vitelline syncytium. During cleavage divisions, three types of blastomeres are formed: macromeres, mesomeres and micromeres. Two large macromeres contain a large nucleus with spherical nucleolus and numerous small heterochromatin islands dispersed in moderately electron-dense nucleoplasm. The granular cytoplasm shows a few large mitochondria. Medium-sized mesomeres contain a spherical nucleus with numerous heterochromatin islands, adjacent to the nuclear envelope, and a prominent electron-dense nucleolus. Their nuclei are embedded in granular cytoplasm with a few large and numerous small mitochondria and Golgi complexes. The small micromeres are characterized by presence of spherical nucleoli with large areas of highly condensed heterochromatin and a few islands of granular electron-lucent nucleoplasm. Their granular cytoplasm shows a few small lipid droplets and several spherical mitochondria. Majority of micromeres give rise to the hexacanth but many of them also undergo degeneration or apoptosis. Both mesomeres and macromeres are engaged in the formation of the oncospheral envelopes. The outer envelope is formed by a fusion of two macromeres whereas the inner envelope originates from a fusion of mesomeres. The intrauterine eggs of W. virilis usually contain an embryo at the early preoncopheral phase of development and possesses three primary envelopes: (1) thick eggshell; (2) thin cytoplasmic layer of the outer envelope and (3) inner envelope. Based on embryonic development, egg type and life-cycle characteristics, caryophyllideans tend to show closer affinities to spathebothriideans than to the former pseudophyllideans.
Ultrastructural aspects of the advanced embryonic development and cotylocidial morphogenesis of the aspidogastrean Aspidogaster limacoides are described. The posterior or distal regions of the uterus are filled with eggs containing larvae at advanced stages of morphogenesis and fully-formed cotylocidia. Various stages and organs of this larva are described in detail, including the aspects of the developing and fully-differentiated cotylocidium, the body wall (tegument and musculature), glandular regions and the protonephridial excretory system. Blastomere multiplication by means of mitotic divisions takes place simultaneously with the degeneration or apoptosis of some micromeres; this frequently observed characteristic is compared and discussed in relation to corresponding reports for other neodermatans. During the advanced stages of the embryonic development of A. limacoides, the vitelline syncytium disappears and the size of the embryo increases rapidly. Evident polarization of the differentiating larva was observed; towards one pole of the egg, cytodifferentiation of the mouth, surrounded by the oral sucker and cephalic glands, takes place, whereas, towards the opposite pole, differentiation of the posterior sucker (incipient ventral disc) occurs. The oral and posterior suckers are formed from numerous embryonic cells which have differentiated into myocytes. The central part of the oral sucker undergoes invagination and forms the future pharynx and intestine. Fully-developed cotylocidia of A. limacoides have a neodermatan type of tegument, flame cells and two types of glandular structures. These results suggest a sister relationship between the Aspidogastrea and the Digenea, although the systematic position of aspidogastreans in relation to other platyhelminth taxa remains somewhat equivocal.
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