The fine structure of the germinal mass, brood cavity and birth canal of the rediae of the monoxenous hemiuroid digenean Bunocotyle progenetica Chabaud & Buttner, 1959.

Bunocotyle progenetica is a hemiuroid digenean whose sexual adults become fully developed and lay their eggs inside the rediae in the molluscan host. In this study, the fine structure of the germinal mass, brood cavity and birth canal in the B. progenetica rediae was examined using transmission electron and confocal microscopy. The large germinal mass attached to the body wall has a cellular composition typical for this organ. The characteristic traits of this germinal mass are weakly developed supporting tissue and the presence of deep lacunae opening into the brood cavity. These lacunae presumably participate in feeding the deeply lying embryos and facilitate their release into the brood cavity. The germinal mass is also characterized by intensive degeneration of cellular elements, which may represent a mechanism controlling the offspring number, limited in this species by the size of the redial brood cavity. The brood-cavity lining consists of flattened cells bearing lamellar projections and is connected anteriorly with the epithelium of the birth canal. The brood-cavity musculature, which is well developed in other hemiuroid digeneans, is significantly reduced in B. progenetica, most likely because their cystophorous cercariae remain inside the rediae, removing the need for muscle contractions pushing them through the brood cavity. The birth canal comprises three regions distinguished by the structure of the lining and muscle arrangement. The comparison of rediae of B. progenetica with parthenitae of other digeneans has shown that the organization of the redial reproductive apparatus in this species may have been influenced by life-cycle modification.

CHANGES IN SPINATION PATTERNS OVER THE COURSE OF METACERCARIAL DEVELOPMENT OF DIPLOSTOMUM PSEUDOSPATHACEUM NIEWIADOMSKA, 1984 (TREMATODA, DIPLOSTOMIDAE).

Metacercarial development of most Diplostomum species including D. pseudospathaceum occurs in the eye lenses of their fish hosts and is accompanied by radical morphological changes often referred to as metamorphosis. One of the structures undergoing substantial changes in D. pseudospathacewn are tegumental spines. The present study used phalloidin staining and confocal microscopy to examine these changes in D. pseudospathaceum over the course of development from 3-day-old to infective metacercariae. Although the general pattern of spination remained essentially unchanged, most larval spines continued growing in size until late in metacercarial development. From day 10 of development, larval spination was gradually replaced by small incipient definitive spines and the infective metacercariae had only definitive spination. The possible adaptive role of spines in developing metacercariae is discussed.

Morpho-functional specialization of the branching sporocyst of Prosorhynchoides borealis Bartoli, Gibson & Bray, 2006 (Digenea, Bucephalidae).

Sporocysts of Prosorhynchoides borealis were obtained from the marine bivalves Abra prismatica and studied using transmission electron microscopy. The sporocyst body consists of a mass of branching and intertwining hollow tubules that ramify through the host's digestive gland and gonads. This study investigated the ultrastructure of the sporocyst branches which comprise alternate distended areas (brood chambers) with a relatively thin body wall, narrower portions with a thicker body wall (constricted areas) and terminal regions. Pronounced differences between these areas were revealed in the structure of their tegument and body cavity lining, as well as in the cellular composition of the subtegumental layers. Body wall composition in distended areas was consistent with the specialization for cercarial nurture in the brood chambers. The structure of the constrictions suggested a dual role of nutrient absorption and physical separation of adjacent brood chambers. Two types of terminal region were identified, one specialized for the investigation and penetration of host tissues and the other, in which the germinal cells are formed, for cercarial production. The overall structure of the sporocyst branches helps explain why this linear modular system, i.e. brood chambers and constrictions continuously growing into the host tissue, enables the sporocyst's long-term existence and can continuously produce cercariae in numbers comparable with those produced by rediae and/or daughter sporocyst infrapopulations in other digeneans. The origin of the nuclei in the outer tegumental layer of some branching bucephalid sporocysts is also discussed.

An ultrastructural study of the early cercarial development in Prosorhynchoides borealis (Digenea: Bucephalidae) with special reference to formation of the primitive epithelium.

Primitive epithelium and outer tegumental layer formation during early cercarial development was studied in Prosorhynchoides borealis using electron microscopy. It demonstrated that germinal cells freely floating in the sporocyst body cavity divide to give rise to naked cell aggregates. These early embryos are highly irregular in outline and are composed of blastomeres differing in size and structure. In embryos consisting of about 12-14 cells a few (possibly only two) superficial macromeres become concave and produce thin extensions which envelop the embryonic mass before fusing to form a syncytial primitive epithelium. This primitive epithelium forms syncytial connections with underlying embryonic cells. Primordial tegumental cells become apparent in late germinal balls below the primitive epithelium. These cells expand and fuse to give rise to an embryonic nucleated tegument. The embryonic tegument is connected to peripheral embryonic cells by thin cytoplasmic bridges until the basement lamina is formed. Subsequently, the primitive epithelium is shed by the embryos and the nuclei in the embryonic tegument undergo pyknotic degeneration. These results are analysed and compared with data from studies on other trematode species and it is concluded that the primitive epithelium is derived from the embryo in at least the majority of digeneans.

An ultrastructural study of reproduction in the sporocysts of Prosorhynchoides gracilescens (Digenea: Bucephalidae).

The germinal development in Prosorhynchoides gracilescens sporocysts was studied using electron microscopy. The germinal cells proliferated and developed within multiple floating germinal masses located in the terminal portions of sporocyst branches. The germinal masses were composed of supporting and germinal cells. Supporting cells possessed numerous flattened extensions that spread around and between developing germinal cells to form three-dimensional mesh network, which maintained the integrity of the germinal mass. Morphological evidences of close interactions between supporting and germinal cells were numerous gap junctions between the two cell types and phagocytosis of small fragments of germinal cells cytoplasm by supporting cells. The germinal cells displayed structural differences that seemed to reflect their sequential developmental changes. These changes included (1) cell growth and increase of organelles number, (2) dispersion of nuclear chromatin and increase of nucleolus size, (3) polarization of the cell, (4) appearance of specific structures such as nuage and laminated inclusions. The germinal cells left the germinal masses to finish their differentiation in the body cavity and then cleaved to give rise to cercarial embryos. Ultrastructural features of the germinal elements of P. gracilescens sporocysts are discussed in the light of existing controversy concerning the nature of the germinal sacs reproduction.

An ultrastructural study of alimentary tract development in the cercariae of Diplostomum pseudospathaceum (Digenea: Diplostomidae).

Morphogenesis of the alimentary canal in developing Diplostomum pseudospathaceum cercariae has been studied using electron microscopy. The foregut primordium appears in early cercariae as a cellular cord. Later, the lumen develops within the foregut primordium, and its cells give rise to the cellular epithelium, limiting this lumen. During subsequent development, the lateral plasma membranes, separating the cells, disappear from the primary foregut epithelium as do nuclei and most of cellular organelles. These events seem to progress in several steps, and eventually, the foregut lining becomes the thin anucleate syncytial layer. In late cercariae, this layer becomes connected with nucleated cytons, producing secretory inclusions. Each of two caecal branches appears to arise from a row of large cuboid cells. The primordial gastrodermal cells are involved in synthetic and secretory activity and give rise to numerous secretory inclusions. These inclusions release their contents into the cavities which develop between adjacent primordial caecal cells. The intercellular cavities gradually increase in size and fuse to eventually form a single caecal lumen. In mature cercariae, the large caecal lumen packed with electron-dense secretory material is limited by a thin cellular gastrodermis, displaying no secretory features.

An ultrastructural study of excretory system development in the cercariae of Prosorhynchoides gracilescens (Rudolphi, 1819) and Prosorhynchus squamatus Odhner, 1905 (Digenea, Bucephalidae).

The ultrastructure of the developing excretory system of Prosorhynchoides gracilescens and Prosorhynchus squamatus cercariae is described. The development pattern was similar in both species. In early embryos the two main collecting tubes were composed of a layer of cells which were wrapped around the lumen. Later, the tubes fused and the excretory epithelium of the fusion zone and that of the lateral caudal ducts became a syncytium. The collecting tubes in the cercarial body retained their cellular organization. As the tails grew, additional excretory pores were formed in the tail stem where thickened portions of the caudal duct epithelium contacted the surface tegument. Following this, the distal portions of the lateral caudal ducts lost contact with the primary excretory pores and progressively degenerated. Excretory atrium development started with differentiation of secretory active cytons peripheral to the fusion zone. These cells gave rise to cytoplasmic extensions that penetrated the fusion zone wall to eventually form a continuous cytoplasmic layer. This layer eventually replaced some of the fusion zone excretory epithelium and became the lining of the excretory atrium. The anterior end of the fusion zone differentiated into an excretory bladder and a short posterior portion gave rise to the caudal vesicle.

An ultrastructural study of the cercarial excretory system in Bucephaloides gracilescens and Prosorhynchus squamatus.

The ultrastructure of the flame cells, capillaries, collecting tubes, excretory bladder, excretory atrium, caudal vesicle, lateral caudal ducts and excretory pores of cercariae of Bucephaloides gracilescens (Rudolphi, 1819) Hopkins, 1954 and Prosorhynchus squamatus Odhner, 1905 (Digenea: Bucephalidae) is described. Both species are essentially similar except for some details. The terminal parts of the protonephridia have all the structural features that are typical of trematodes. The collecting tubes in the cercarial body are composed of cells that are wrapped around the lumen. The main collecting tubes are joined to the excretory bladder syncytium by septate junctions. Features of P. squamatus excretory bladder epithelium indicate that it is involved in secretory activity, but this is not the case in B. gracilescens. In both species the luminal surface of the excretory bladder epithelium is increased by lamellae, and the basal plasma membrane forms invaginations. In the bladder syncytium of P. squamatus both apical lamellae and basal invaginations are more developed and mitochondria are also more numerous. The excretory atrium is lined by a syncytium with nucleated cytons located in the surrounding parenchyma. The atrium lining is not continuous with the body tegument and possesses specific secretory inclusions and a thick glycocalyx. Septate junctions connect the atrium syncytium to the excretory bladder epithelium at its anterior end and to the syncytial excretory epithelium lining the caudal vesicle and the lateral caudal ducts at its posterior. In the excretory pores the caudal duct syncytium is joined to the tegument by septate desmosomes.