Ultrastructure of the intrauterine eggs of Didymobothrium rudolphii (Monticelli, 1890) (Cestoda, Spathebothriidea, Acrobothriidae) and its phylogenetic implications

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.

Cestoda and monogenea of some teleost fishes off the Mudanya Coast (Sea of Marmara, Turkey)

The following helminths are reported from the Sea of Marmara off the Mudanya coast. Cestoda: Didymobothrium rudolphii in Solea solea, Clestobothrium crassiceps in Merluccius merluccius, tetraphyllidean larvae in Gobius niger, G. cobitis, M. merluccius, Eutrigla gurnardus, S. solea, Scorpaena scrofa and Progrillotia dasyatidis in G. niger. Monogenea: Trochopus pini in E. gurnardus, Ligophorus confusus in Liza ramada, Tetraonchoides paradoxus in Uranoscopus scaber, Microcotyle mugilis in L. ramada and Anthocotyle merluccii in M. merluccius.

Cryptic species of Didymobothrium rudolphii (Cestoda: Spathebothriidea) from the sand sole, Solea lascaris, off the Portuguese coast, with an analysis of their molecules, morphology, ultrastructure and phylogeny

SUMMARYDidymobothrium rudolphii (Cestoda: Spathebothriidea) was collected seasonally from the sand sole, Solea lascaris, off the northern, central and southern areas of the Portuguese coast. Morphological and molecular analyses were conducted in order to examine the possible existence of cryptic species and to facilitate the circumscription of their morphological boundaries. Data were compared between D. rudolphii specimens from each of the 3 geographical areas and 4 seasons, and principal components analysis of 18 morphological characters was used to detect differences. Two distinct genotypes were present with sequence divergences of 1·9% and 2·1% in the large subunit (lsrDNA) and second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA), respectively. The less common ‘central’ genotype was present only off the central area from summer to winter, whereas the ‘common’ genotype was present throughout the year off the northern and southern areas, but only during spring in the central area. No sequence variation was found within each genotype. The presence of 2 distinct genetic entities was supported by morphological analyses, which showed the ‘central’ genotype specimens to be more slender and elongate, although morphometric ranges overlapped considerably for most characters. Molecular phylogenetic analysis of 4 of the 5 known genera of the Spathebothriidea showed Spathebothrium to be the earliest branching lineage and the 2 genotypes of Didymobothrium formed a sister group to Cyathocephalus. The concordance of genetic differences with variation in host diet according to season and locality could account for sympatric speciation occurring in the central region of the Portuguese coast.

Ultrastructure of the ovary, ovicapt and oviduct of the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890)

The ultrastructural details are presented of the ovary, ovicapt and oviduct of the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890) from the intestine of the sand sole Solea lascaris. Oogonia, maturing oocytes and mature oocytes are surrounded by a syncytial interstitial cytoplasm, one of the distinctive traits of which is the presence of numerous myelin-like bodies. Oocyte inclusions comprise cortical granules and a small number of lipid droplets. The thickened, nucleated epithelium of the ovicapt lacks any apical structure and is a prolongation of the narrow ovarian epithelium. The muscular sphincter of the ovicapt is formed by a band of longitudinal muscles and bands of radial muscles at right angles to the longitudinal layer, and numerous myocytes surround the ovicapt wall. The oviduct of D. rudolphii is subdivided into three regions: (1) the proximal oviduct; (2) the fertilization chamber — the region distal to the point of entry of the duct from the seminal receptacle; and (3) the ovovitelline duct — the region distal to the point of entry of the duct from the vitelline reservoir. A comparative analysis is made between the structures of the ovary, ovicapt and oviduct of D. rudolphii and those of two other spathebothriideans, Cyathocephalus truncatus and Diplocotyle olrikii, with a discussion of ultrastructural traits that might be used as taxonomic criteria within the order Spathebothriidea.

Vitellocyte ultrastructure in the cestode Didymobothrium rudolphii (Monticelli, 1890): possible evidence for the recognition of divergent taxa within the Spathebothriidea

In the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890) the fine structure of the vitellocytes at different stages of their development within the vitelline follicles, vitelline ducts and uterus was studied for the first time using transmission electron microscopy. The vitellocyte inclusions of D. rudolphii are shell globule clusters containing tightly packed shell globules associated with a matrix of moderate electron density, glycogen granules, large electron-lucent lipid droplets (up to 3 μm in diameter), and, occasionally, a lipid droplet may occur in the nucleus of the vitellocytes. The diameter of the clusters ranges from 0.4 to 2.5 μm, the number of shell globules in the clusters varies from 8 to 45, and the size of the globules ranges from 0.12 to 0.25 μm and they are of approximately homogeneous sizes within a cluster. Most vitellocyte lipid droplets have a heterogeneous configuration with a ‘cavity’ inside them when they are within vitelline ducts and intrauterine eggs. Vitellocytes of the eggs contain dark concentric bodies and lipid droplets. The interstitial tissue has a syncytial structure. The morphological parameters of the diameter and shape of shell globule clusters, arrangement of shell globules in clusters, number and diameter of globules within clusters, types of lipid droplets and presence of dark concentric bodies are compared with those of two other spathebothriidean genera, Cyathocephalus and Diplocotyle. The comparative data demonstrate that vitelline material morphology has unique features in three spathenothriidean genera and may be used as evidence for the recognition of separate taxa.