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.     

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 and cytochemical study of the digestive system in Oxyposthia praedator (Turbellaria,Acoela)

The central parenchyma in Oxyposthia praedator consists of multifunctional cells. These cells digest food material intracellularly, can effect extracellular digestion through release of digestive enzymes by cell lysis and clasmatosis, and synthesize reserve nutritional substances.     

Three new species of Prosorhynchoides (Digenea: Bucephalidae) from Tylosurus gavialoides (Belonidae) in Moreton Bay,Queensland, Australia

We surveyed 30 individuals of Tylosurus gavialoides (Castelnau) (Belonidae) collected from Moreton Bay, Queensland, Australia, and describe three new species of Prosorhynchoides Dollfus, 1929 from them. The new species are morphologically distinct from existing Prosorhynchoides spp. and 28S and ITS-2 ribosomal DNA data further supports our morphological findings. We also conduct the first mitochondrial DNA analysis of species of Prosorhynchoides. The new species from T. gavialoides form a strongly supported clade on the basis of the two ribosomal markers, further supporting the emerging hypothesis that bucephaline clades are strongly associated with host groups. We have not observed any of the new species reported here in over 3500 surveyed individuals of other piscivorous fish in Australia, suggesting that these species are host-specific at least to belonids, if not to only T. gavialoides. Our findings support previous reports that suggest that belonids are exceptional hosts for bucephalids. We predict that further sampling of the numerous other belonid species present in Australian waters, for which nothing is known of the bucephalid fauna, will uncover further bucephalid richness.     

A new species of Prosorhynchoides Dollfus, 1929 (Digenea: Bucephalidae) from Xenentodon cancila Hamilton, 1822 in Mizoram,Northeast India

Prosorhynchoides aspinosus n. sp., a new species of digenean trematode, is described based on morphological, ultrastructural and molecular data from the freshwater fish, Xenentodon cancila Hamilton, 1822, of Tuikum and Tuirial river in Mizoram, Northeast India. It differs from its congeners in the absence of tegumental spines and the length of cirrus sac. These unique morphological characteristics were further supported by the ultrastructural data showing distinct knob-like protrusions in the tegument. Each knob-like protrusion is without cutting plates or spines and appears granulated at higher magnification. The molecular data analyses, along with the sequences of other Prosorhynchoides available in the Genbank, using internal transcribed spacer 2 and 28S rDNA revealed a gap of 4% K2P (Kimura 2 parameter) distance with its closest congener, P. karvei, confirming the novelty of the species, and thus, clearly differentiates the new species from all other valid Prosorhynchoides spp.     

Two new species of Prosorhynchoides (Digenea: Bucephalidae) from Tylosurus crocodilus (Belonidae) from the great barrier reef and French Polynesia

We surveyed 14 individuals of Tylosurus crocodilus Péron & Lesueur 1821 (Belonidae) collected from the waters around Lizard Island and Heron Island, Great Barrier Reef, Queensland, Australia, and the waters around Moorea, French Polynesia. We describe two new species of bucephaline trematodes from them, Prosorhynchoides galaktionovi n. sp. and P. kohnae n. sp. They are morphologically distinct from existing Prosorhynchoides spp., with molecular data from 28S and ITS-2 ribosomal DNA, as well as cox1 mitochondrial DNA, further supporting our morphological findings. Neither species has been observed in other belonid fishes. The new species fall into the clade of species of Prosorhynchoides from belonids previously identified in Australian waters. These findings strengthen the observation that groups of bucephaline species have radiated, at least in part, in tight association with host taxa. There are now five species of Prosorhynchoides known from two belonid species in Australian waters. We, therefore, predict further richness in the nine other belonid species present.     

Alimentary tract of Kowalevskiidae (Appendicularia, Tunicata) and evolutionary implications

The alimentary tract of Kowalevskia tenuis and K. oceanica, the only species of the appendicularian family Kowalevskiidae, was studied both at the light and electron microscope levels and compared with species belonging to the other two families of the class. Kowalevskids show interesting specializations: 1) the pharynx opens on both sides through two opposing spiracles, modified into long ciliated fissures, and possesses an original filtering system of ciliated combs arranged in two pairs of opposing longitudinal rows; 2) the endostyle is absent, its place being taken by a ciliated groove without any glandular cell; 3) posterior to the esophagus, the globular stomach and rectum form a digestive nucleus comprising a few, large cells including two well-developed, specialized valves, cardiac and pyloric; 4) special apical junctions bearing characteristics of both gap and adherens junctions are diffuse along the gut epithelium; 5) the heart is absent. Our data suggest that Kowalevskiidae underwent a high degree of specialization for food filtering and are more closely related to Fritillariidae, with which they share several characters, rather than Oikopleuridae, the latter probably representing the most primitive family of appendicularians.     

Cercaria caribbea LVIII Cable, 1963 (Digenea: Cyathocotylidae) in the Republic of Korea and its surface ultrastructure

Cercaria caribbea LVIII Cable, 1963 (Digenea: Cyathocotylidae) was detected from a brackish water gastropod species (Cerithideopsilla cingulata) in a coatal area of Shinan-gun, Jeollanam-do (Province), the Republic of Korea, and its surface ultrastructure was studied using a scanning electron microscope. The cercariae were found freely swimming or enveloped within daughter sporocysts when the snail host was mechanically broken. They were morphologically characterized by a linguiform and ventrally concave body, a long and bifurcated tail, and the presence of a holdfast (=tribocytic) organ posterior to the ventral sucker. On the whole ventral and dorsal surfaces, peg-like tegumental spines were densely distributed. Around the oral sucker, several sensory papillae, each with a short cilium, were distributed, and on the tail, sensory papillae, each with an extensively long cilium, were observed. This is the first record describing a cyathocotylid cercaria from a brackish water gastropod in the Republic of Korea.     

An ultrastructural study of the cuticle in the marine annelid Heterodrilus (Tubificidae, Clitellata)

The ultrastructure of the cuticle in four species of the marine Heterodrilus (H. paucifascis, H. pentcheffi, H. flexuosus, H. minisetosus) is investigated with transmission electron microscopy. The noncellular cuticle consists of several parts; closest to the epidermis is a thick zone of collagen fibers embedded in a matrix. The matrix continues outside the fiber zone, forming a layered epicuticle. The external surface of the epicuticle is covered by evenly distributed, membrane-bound bodies, termed epicuticular projections. The epicuticular projections have their longitudinal axis perpendicular to the surface of the cuticle and are attached to the surface by either the surrounding membrane itself or by short pedestals. Microvilli, extensions from the epidermal cells, penetrate and sometimes pass completely through the cuticle. There is interspecific variation in the morphology of the cuticle. The four studied species differ in the arrangement of the collagen fibers, from irregularly distributed fibril bundles to orthogonally arranged fiber layers, as well as in the number and density of layers in the epicuticle. One of the studied species, H. paucifascis, shows intraspecific variation, which is associated with sample locality. The Bahamian specimens of H. paucifascis have four layers in the epicuticle, club-shaped epicuticular projections, and collagen fibers forming a less defined orthogonal grid, while the Belizean specimens have three layers in the epicuticle, epicuticular projections with a bulging part at midlevel, and a distinct orthogonal grid. Based on these findings the variation in the morphology of the cuticle appears to be dependent on both phylogenetic constraints, and functional and environmental factors.     

Microscopic anatomy and ultrastructure of the digestive system of Natatolana obtusata (Vanhöffen, 1914) (Crustacea, Isopoda)

The scavenging cirolanid isopod Natatolana obtusata was caught using primarily meat‐baited traps at a station in the Weddell Sea at Kap Norvegica. The hindguts of most specimens were densely stuffed with meat from the traps. The anatomy and the ultrastructure of different sections of the digestive system are described. The large and extremely expandable mouth opens into a short oesophagus which extends posteriorly into a small stomach. Lateralia and primary filter can be observed directly when the mouth opening is widened to its extreme. The anterior hindgut is extremely variable in its lumen and very distensible, depending on the nutritional condition. The cuticle of the hindgut is colonized by fungi. The length of the midgut glands varies with the nutritional conditions. Morphology and anatomy of the digestive system of Natatolana obtusata are discussed with the background of the lifestyle of the species and compared with other taxa.     

Effect of gluten on the digestive tract and fat body of Telodeinopus aoutii (Diplopoda)

Adult specimens or larvae of invertebrates used as food for vertebrates are often maintained close to gluten so they might become vectors for cereal proteins. However, the tissues and internal organs can respond differently in animals with different feeding habits. The midgut epithelium might be a first and sufficient barrier preventing uptake and effects of gluten on the whole body, while the fat body is the main organ that accumulates different xenobiotics. Good models for such research are animals that do not feed on gluten-rich products in their natural environment. The project's goal was to investigate alterations in the midgut epithelium and fat body of the herbivorous millipede Telodeinopus aoutii (Diplopoda) and analyze cell death processes activated by gluten. It enabled us to determine whether changes were intensified or reversed by adaptive mechanisms. Adult specimens were divided into control and experimental animals fed with mushrooms supplemented with gluten and analyzed using transmission electron microscopy, flow cytometry, and confocal microscopy. Two organs were isolated for the qualitative and quantitative analysis: the midgut and the fat body. Our study of the herbivorous T. aoutii which does not naturally feed on gluten containing diet showed that continuous and prolonged gluten feeding activates repair processes that inhibit the processes of cell death (apoptosis and necrosis) and induce an increase in cell viability.     

The digestive gland of Viviparus ater (Mollusca, Gastropoda, Prosobranchia): an ultrastructural and histochemical study

The digestive gland of Viviparus ater was studied using histochemical and ultrastructural methods. Only one cell type was observed in the tubule epithelium of the gland. The cells are involved in an endocytotic process mediated by clathrin-coated vesicles and in the intracellular digestion of food materials (thus they can be regarded as digestive cells). The different stages of digestion and exocytotic extrusion of residual bodies into the tubule lumen were shown by electron microscopy. Very few, small mucocytes are scattered among the digestive cells. Calcium concretions, glycogen-containing cells and endocrine cells are scattered in the area of connective tissue present among the digestive tubules.     

Morphological and ultrastructural characterization of the alimentary canal in larvae of Streltzoviella insularis (Staudinger) (Lepidoptera: Cossidae)

Streltzoviella insularis (Staudinger) is an important tree‐boring pest, that primarily damages Sophora japonica (Linnaeus) and Ginkgo biloba (Linnaeus), as well as other common species, at great economic cost to the urban landscape construction industry in China. In the present study, the alimentary canal morphology of S. insularis was observed using light microscopy, and its ultrastructure was investigated by scanning and transmission electron microscopy. The foregut of S. insularis can be divided into the pharynx, esophagus, crop, proventriculus, and cardiac valve. The well‐developed crop forms the longest section of the foregut. It is able to store large amounts of food and is lined with a monolayer of epithelial cells. Many sclerotized microspines occur on the surface of the anterior intima and there are dense spines on the posterior intima of the proventriculus. Epithelial cells of the midgut include columnar cells, goblet cells, and regenerative cells, but endocrine cells are absent. The hindgut consists of the pyloric valve, ileum, and rectum. There is no clear distinction between the ileum and colon. The intima surface of the pyloric valve carries many microspines, whereas the intestinal wall of the rectum is thin with well‐developed rectal pads. The rectal epithelial cells form a squamous monolayer. A cryptonephric excretory system is located in the hindgut. There are six spiral Malpighian tubules, in which a cellular layer on a basement membrane encloses a lumen. These results will provide the basis for further studies of the structure and function in S. insularis larvae.     

Prosorhynchoides gracilescens (Rudolphi, 1819) (Digenea: Bucephalidae) from Lophius piscatorius L. is a species complex: a redescription of this species (sensu stricto) from the western Mediterranean and the description of P. borealis n. sp. from the northern North-East Atlantic

The bucephalid digenean Prosorhynchoides gracilescens (Rudolphi, 1819) is a common intestinal parasite of the angler fish Lophius piscatorius in European marine waters. Detailed studies of new material collected off the coasts of both Marseilles and Corsica in the western Mediterranean, and comparison with museum material from the northern North-East Atlantic, indicated that the accepted concept of this species comprises two distinct taxa. The Mediterranean form occurs in relatively small numbers and has small eggs, a large rhynchus and a vitelline distribution that finishes well short of the rhynchus, whereas the NE Atlantic form, which comprises the majority of records, occurs often in large numbers, has larger eggs, a relatively smaller rhynchus and a vitelline distribution that tends to reach the rhynchus. Since the type-material is from the Mediterranean and resembles the Mediterranean form, there was little option but to consider the latter as P. gracilescens (sensu stricto) and to describe the NE Atlantic form as a new species, for which the name P. borealis n. sp. is coined to reflect its northerly distribution. Both species are described and figured in detail, and a table and illustration of the diagnostic features are presented. In addition, some discussion is included on the distribution of these two species and whether P. gracilescens (s. str.) might be a relict species, and on non-European records of P. gracilescens (sensu lato).     

Prosorhynchoides gracilescens (Rudolphi, 1819) (Digenea: Bucephalidae) from Lophius piscatorius L. is a species complex: a redescription of this species (sensu stricto) from the western Mediterranean and the description of P. borealis n. sp. from the northern North-East Atlantic

The bucephalid digenean Prosorhynchoides gracilescens (Rudolphi, 1819) is a common intestinal parasite of the angler fish Lophius piscatorius in European marine waters. Detailed studies of new material collected off the coasts of both Marseilles and Corsica in the western Mediterranean, and comparison with museum material from the northern North-East Atlantic, indicated that the accepted concept of this species comprises two distinct taxa. The Mediterranean form occurs in relatively small numbers and has small eggs, a large rhynchus and a vitelline distribution that finishes well short of the rhynchus, whereas the NE Atlantic form, which comprises the majority of records, occurs often in large numbers, has larger eggs, a relatively smaller rhynchus and a vitelline distribution that tends to reach the rhynchus. Since the type-material is from the Mediterranean and resembles the Mediterranean form, there was little option but to consider the latter as P. gracilescens (sensu stricto) and to describe the NE Atlantic form as a new species, for which the name P. borealis n. sp. is coined to reflect its northerly distribution. Both species are described and figured in detail, and a table and illustration of the diagnostic features are presented. In addition, some discussion is included on the distribution of these two species and whether P. gracilescens (s. str.) might be a relict species, and on non-European records of P. gracilescens (sensu lato).     

The neuroactive substances and associated muscle system in Rhipidocotyle campanula (Digenea,Bucephalidae) from the intestine of the pike Esox lucius

We report about the muscular system and the serotonergic and FMRFamidergic components of the nervous system of the Bucephalidae trematode, Rhipidocotyle campanula, an intestinal parasite of the pike. We use immunocytochemical methods and confocal scanning laser microscopy (CLSM). The musculature is identified by histochemical staining with fluorescently labeled phalloidin. The body wall musculature of R. campanula contains three layers of muscle fibres – the outer thin circular, intermediate longitudinal and inner diagonal muscle fibres running in two opposite directions. The digestive system of R. campanula possess of a well-developed musculature: radial, longitudinal and circular muscle elements are detected in the pharynx, circular and longitudinal muscle filaments seen in the oesophagus, and longitudinal and the circular muscle fibres were found in the intestinal wall. Specific staining indicating the presence of actin muscle filaments occurs in the cirrus sac localized in the posterior body region. The frontal region of anterior attachment organ, the rhynchus, in R. campanula is represented by radial muscle fibres. The posterior part of the rhynchus comprise of radial muscles forming the organ's wall, and several strong longitudinal muscle bundles. Serotonergic and FMRFamidergic structures are detected in the central and peripheral compartments of the nervous system of R. campanula, that is, in the paired brain ganglia, the brain commissure, the longitudinal nerve cords, and connective nerve commissures. The innervations of the rhynchus, pharynx, oesophagus and distal regions of the reproductive system by the serotonergic and FMRFamidergic nervous elements are revealed. We compare our findings obtained on R. campanula with related data for other trematodes.     

Morphogenesis of the hydrogenosome: An ultrastructural study

Summary— The morphogenesis of hydrogenosomes in several trichomonad species (Tritrichomonas foetus, Trichomonas vaginalis, Tritrichomonas suis, Trichomonas gallinae, Tritrichomonas augusta and Monocercomonas sp) was investigated by transmission electron microscopy of thin sections and freeze-fracture replicas of whole cells or the isolated organelle. Close proximity, and even continuity, between endoplasmic reticulum and hydrogenosomes was observed. Structures were seen connecting hydrogenosomes to each other and to cytoplasmic structures. Morphological evidence is presented showing that in all the trichomonads here studied, hydrogenosomes, like mitochondria, may divide by two distinct processes: segmentation and partition. In the segmentation process, the hydrogenosome grows, becoming enlongated with the appearance of a constriction in the central portion. Microfibrillar structures appear to help the furrowing process, ending with a total fission of the organelle. In the partition process, the division begins by an invagination of the inner hydrogenosome membrane, forming a transversal septum, separating the organelle matrix into two compartments. We suggest that myelin-like structures seen either in close contact with or in the vicinity of the hydrogenosomes may be a source of membrane lipids for hydrogenosome growth.     

An ultrastructural investigation of the testes and spermiogenesis in Myobia murismusculi (Schrank) (Acari,Actinedida: Myobiidae)

Summary

The stages of spermiogenesis in Myobia murismusculi were investigated on the basis of ultrastructural analysis of both the testes and the female organs: receptaculum seminis and seminal duct. The walls of the testes consist of a thin epithelial layer. Germ and secretory cells lie free in the lumen of the testes. In the early stages of differentiation, both cell types represent clusters of sister cells joined by intercellular bridges. Each secretory cell contains prominent RER and Golgi complex, which produce single dense granule. Growing gradually the granule fills the whole volume of the cell's cytoplasm. Mature secretory cells disintegrate and the secretory product discharges into the testicular lumen. The germ cells are represented by the early, the intermediate and the late spermatids as well as the immature sperm (prospermia). Neither spermatogonia nor meiotic figures were observed in adult males. As spermiogenesis starts, numerous narrow invaginations of the outer membrane (peripheral channels) develop on the cell surface. They form a wide circumferential network connected to pinocytotic vesicles. Owing to the secretory activity of the Golgi complex, a large acrosomal granule is formed in the early spermatids. A long acrosomal filament runs along the intranuclear canal. Nuclear material condenses and forms two spherical bodies of different electron density. The lighter one can be observed until the stage of the late spermatids, when the nuclear envelope almost completely disappears. The electron-dense nuclear body transforms into a definite chromatin body, which is observed in the mature sperm as a cup-shaped structure. The late spermatids are characterized by the presence of a large electronlucent vacuole, which seems to be unique for the process of spermiogenesis in Actinedida. After the spermia enter the female genital tract, the peripheral channels disappear as well as the vacuole. The cells form long amoeboid arms with a special microtubular layer underneath the plasma membrane. The chromatin body is encircled by a large acrosomal granule of complex shape provided by long extensions running deep into the cytoplasm. The cytoplasm contains no organelles except for a group of unmodified mitochondria in the post-nuclear region. The main characteristics of the Myobia spermiogenesis are discussed with regard to other actinedid mites.     

The female genital system of Ooperipatellus decoratus (Onychophora, Peripatopsidae): an ultrastructural study

The female genital system of the oviparous peripatopsid Ooperipatellus decoratus consists of an ovary, oviducts equipped with receptacula seminis and additional pouches, uteri, and a vagina. It is examined using transmission and scanning electron microscopy. The ovary is made up of paired ovarian tubes united anteriorly and posteriorly and differentiated into a sterile dorsal part and a fertile ventral part with exogenous oocytes. Fertilization presumably occurs in the oviducts once the oocytes pass the receptaculum seminis. Although the receptacula seminis have been reported to occur in juvenile O. decoratus females only, the present study reveals that they are present in adult females as well. Their wall consists of a cuboidal epithelium covered with a thin collagen-muscle layer. The additional pouches are projections of the oviducts facing the receptacula seminis. They are distally closed to the haemocoel by a flattened epithelium and lack external muscle cells. A thin collagen layer is only found proximally. The uteri are characterized by a columnar epithelium with folded cell membranes allowing extension of the uteri, thus facilitating the passage of the large uterine eggs towards the vagina. Another dominating feature of the uteri is a distally increasing secretory production, which probably contributes to chorion development. Cilia occurring along the entire length of the uteri are considered to assist in the transport of eggs towards the vagina.     

Ultrastructure of the anterior alimentary tract of a monogenean, Diclidophora merlangi

The anterior alimentary tract of Diclidophora merlangi is composed of a complex series of morphologically distinct epithelia interconnected by septate desmosomes and penetrated by the openings of numerous unicellular glands. The mouth and buccal cavity are lined by an infolding of modified body tegument, distinguished by uniciliate sense receptors, buccal gland openings, and in the buccal region by a dense, spiny appearance. The prepharynx is covered by an irregularly folded epithelium and, for part of its length, by the luminal cytoplasm of the prepharyngeal gland cells. The epithelium is syncytial and pleiomorphic, and regional variation in structure is common. A separate epithelium invests the lips of the pharynx and its free surface is greatly amplified by numerous, dense lamellae of varying dimensions. The lip epithelium is continuous with cytoplasmic processes of cells located external to the pharynx. A further, distinct epithelium borders the pharynx lumen and is composed of discrete cytoplasmic units connected by short septate desmosomes. The oesophagus is lined by a modified caecal epithelium, lacking haematin cells, and, in places, is perforated by the openings of oesophageal gland cells; it is continuous with the syncytial connecting tissue of the gut caeca.