Fasciola hepatica: the ultrastructure of the epithelium of the seminal vesicle, the ejaculatory duct and the cirrus.

The ultrastructure of the seminal vesicle, ejaculatory duct, cirrus sac and cirrus is described. The epithelium of the seminal vesicle consists of a single layer of squamous to cuboidal cells. The apical ends of the cells have thin polymorphic lamellae and long narrow pits, both of which enclose normal spermatozoa. The cells have a moderate amount of GER and Golgi complexes which produce a lucid secretory body. The ejaculatory duct epithelium is composed of cuboidal to columnar cells between or through which project the terminal parts of the ducts of the unicellular prostate glands. The apical surfaces of the epithelia are extended into triangular or filiform projections having thin sinuous lamellae. The cytoplasm contains GER cisternae and Golgi complexes which synthesize a dense ovoid secretion. The cirrus sac and cirrus are covered by a thin modified tegument. The cirrus has many spines and the normal ratio of T1 and T2 type of secretory bodies, whereas the cirrus sac has few spines and the T2 type of secretory body predominates over the T1 type. The significance and possible functions of the structures observed in the three tissues are discussed.     

Fine structure of the cirrus sac and vagina of progenetic Diplocotyle olrikii (Cestoda: Spathebothriidea)

Ultrastructural characteristics of the cirrus sac and vagina of progenetic and monoxenic spathebothriidean tapeworm, Diplocotyle olrikii from the body cavity of Gammarus oceanicus are described. Five loosely arranged muscle layers make up the cirrus sac wall. The nucleated syncytial cytoplasm of the ejaculatory duct's wall has apical lamellae and 3 well-developed muscular layers under epithelim. Numerous prostate ducts pass through syncytial epithelium of it. The numerous prostate glands are localized around cirrus sac. Unique morphology of the prostate granules with an electron-dense core surrounded by a matrix of lower density is revealed for the spathebothriidean tapeworms. The epithelial lining of the cirrus in D. olrikii is nucleated and also is connected to subsurface sunken perikarya. The apical surface of the cirrus is covered with small cone-shaped microtriches. Well-developed 10 closely arranged muscle layers support the cirrus. Three regions of the vagina are distinguished. The distal part of vagina has filamentous microthrix type on the surface similar to tegumental one, as well as middle and proximal regions are covered with small cone-shaped microtriches, reducing in their number to proximal part. Anucleate epithelial lining of vagina is connected to numerous sunken parikarya. The cirrus and vagina epithelial lining has numerous vesicles. A comparison is made of the fine structure of the cirrus sac and vagina of monoxenic and dixenic spathebothriidean species. Ultrastructural data on the cirrus sac and vagina of the Spathebothriidea are compared with those for monozoic and polyzoic cestodes. The ultrastructural spathebothriidean features are discussed.     

Observations on the ultrastructure of the copulatory organ of Archilopsis unipunctata (Fabricius, 1826) (Proseriata,Monocelididae)

The copulatory organ in adult specimens of Archilopsis unipunctata has been studied by transmission electron microscopy.This copulatory organ is of the conjuncta-duplex type with eversible cirrus. The seminal vesicle, lined with a nucleate epithelium, is surrounded by spirally arranged muscles. The fibres are enclosed in a sheath that is continuous with the septum of the bulbus and the basement lamina of the male canal epithelium. Distally to the seminal vesicle the bulbus is filled with the secretory cell-necks of the prostate glands. The male canal shows three different parts: seminal duct, ejaculatory duct and eversible cirrus. At the transition of seminal duct and ejaculatory duct two prostate ducts open into the lumen. The structure of the epithelium lining the different parts of the canal is described. The transition into the cirrus may be recognized by an abrupt change in the thickness, the electron density and the stratification in the basement lamina and by the disappearance of the epithelium absent indeed in the cirrus. The material found inside the cirrus-lumen is different according to the zone considered. The origin of this material and of the cirrus teeth is discussed.Abbreviations ab- apoptotic body - ba- bacteria - bb- basal bodies of cilia - bl- basement lamina - bw- body wall - c- cilia - cb- cell body - cgp- common genital porus - ci- cirrus - cip- cirrus plug - cl- lumen of cirrus - cm- circular muscles - cr- cytoplasmatic remnants - cs- cytoplasmatic sheets - ejd- ejaculatory duct - ep_ej- epithelium of ejaculatory duct - d- desmosomes - f- flagella of spermatozoa - fd- female duct - fp- female porus - gc- golgi complex - gl- glycogen particles - hd- hemidesmosomes - lm- longitudinal muscles - ly- lysosome-like body - m- muscles - m_b- muscles of the bulbus - m_c- muscles of the cirrus - m_c- muscles of the seminal vesicle - mi- mitochondria - ml- microvilli - ms- mesenchyme - n_sd- nuclei of the seminal duct - pd- prostate duct - pg- prostate glands - ri- ribosomes - s- septum - sb- secretory vesicle - sd- seminal duct - sp- spines - sv- seminal vesicle - v- vagina - vd- vas deferens     

Rostellar apparatus of Fernandezia spinosissima (von Linstow, 1894) (Cestoda, Cyclophyllidea, Davaineidae): microanatomy and fine structure

The rostellar apparatus of Fernandezia spinosissima is examined by light and transmission electron microscopy. It is described as composed of rostellum, pseudoproboscis, and two groups of glandular syncytia, one in the rostellum and the other inwards to the rostellum and the pseudoproboscis. The rostellum is a discoid cushion with muscular walls. There are numerous thin vertical muscular fibres and glandular syncytia inside it. Its tegument has slender microtriches. Peripherally, the rostellum is encircled by thin muscle fibres that protrude anteriorly between rostellar hooks and group posteriorly to form retractors. The rostellar hook guards are associated with a complicated network of muscles and nerves. The pseudoproboscis is a thick-walled ring around the apical part of the scolex adjacent to the rostellum. Its tegument has microtriches with modified spines that are interpreted as accessory spines. The walls of the pseudoproboscis possess a loose structure of parallel muscular fibres and glandular processes. The glandular syncytia, interpreted as modified tegumental perikarya, have basophilic cytoplasm and stain positively for protein and carbohydrate. They form cellular processes, which protrude to reach to the tegument. These results provide structural details characterising one of the rostellar types recognized in the order Cyclophyllidea, i.e. the davaineid rostellar apparatus.     

Tegumental ultrastructure of the juvenile and adult Himasthla alincia (Digenea: Echinostomatidae)

The tegumental ultrastructure of juvenile and adult Himasthla alincia (Digenea: Echinostomatidae) was observed by scanning electron microscopy. One-, 5- (juveniles) and 20-day-old worms (adults) were harvested from chicks experimentally fed metacercariae from a bivalve, Mactra veneriformis. The juvenile worms were elongated and curved ventrally. The head crown bore 31 collar spines, arranged in a single row. The lip of the oral sucker had 12 paired, and 3 single type I sensory papillae, and the ventral sucker had about 25 type II sensory papillae. The anterolateral surface between the two suckers was densely packed with tegumental spines with 4-7 pointed tips. The adult worms were more elongated and filamentous, and had severe transverse folds over the whole body surface. On the head crown and two suckers, type I and II sensory papillae were more densely distributed than in the juvenile worms. Retractile brush-like spines, with 8-10 digits, were seen on the anterolateral surface, whereas claw-shaped spines, with 2-5 digits, were sparsely distributed posteriorly to the ventral sucker. The cirrus characteristically protruded out, and was armed with small spines distally. The surface ultrastructure of H. alincia was shown to be unique among echinostomes, especially in the digitation of its tegumental spines, the distribution of sensory papillae and by severe folds of the tegument.     

Structure of the copulative apparatus of Sobolevicanthus gracilis (Cestoda: Cyclophyllidea)

Fine structure of genital atrium, Furman body, distal region of vagina and sac cirrus were investigated. The broad polymorphism of surface microstructures (microtriches) in various parts of copulative apparatus has been revealed. The constitution of prostate glands, being the modified cytons of syncitial epithelium of the intrabursal section of spermaduct wall, is described. Based on obtained and reference data, a comparative morphological analysis of copulative apparatus organization in lowest and highest cestodes was carried out. The hypothesis suggests that evolution of its organization was connected with the reduction of the muscular elements and simultaneous improvement of their managing, their isolation from surrounding parenchyma and development of supporting connective tissue elements. It is shown, that Fuhrmann's body has its own developed muscular system armed with power microtriches and being capable to evaginate into the distal region of vagina. It is supposed that S. gracilis obtains of two different ways of copulation in: by means of cirrus that executes the cross fertilization; and by means of the additional copulative formation--Furman body, which guarantees the self fertilization of worms.     

Ribeiroia marini: Surface ultrastructure of redia,cercaria, and adult

Rediae, cercariae, and adults of Ribeiroia marini were examined using a scanning electron microscope to determine the types of tegumental sensory structures and their locations. Sensory structures were observed among numerous tegumental folds in the area immediately surrounding the mouth of the rediae. These sensory structures are similar in appearance, location and fine structure to sensory structures described from the anterior tips of rediae known to be predacious on the sporocysts of Schistosoma mansoni. These uniciliated structures may function as chemoreceptors to aid the redia in migration through snail tissue. Five types of sensory structures bearing one, two, or multiple cilia were distinguishable on the cercariae. These structures were located on and around the oral sucker, dorsal and ventral body surfaces and on the tail. They may be used by the cercariae to locate the intermediate host fish and to find suitable sites within the lateral line scales for encystment. The ventral surface of the adult fluke is covered with spines and shows an absence of sensory structures on the general body surface. Sensory structures were seen in the area surrounding the oral and ventral suckers. The extended cirrus organ has a folded tegument, but lacks spines or sensory structures.     

Description of Bucephalus anguillae n. sp. (Trematoda: Bucephalidae), a parasite of the eel Anguilla anguilla (Anguillidae) from a brackish water lagoon of the Adriatic Sea

The digenetic trematode Bucephalus anguillae n. sp. is described from the intestine of eel, Anguilla anguilla L., originating from a brackish water fish farm on the Italian coast of the Adriatic Sea. The new taxon is 1 of 12 Bucephalus species characterized by an anterior rhynchus surrounded by 7 tentacular appendages, each when fully protruded with 2 prongs. Scanning electron microscopy reveals, for the first time in a Bucephalus species, the crescent-shaped, unspined field located between the rhynchus and the dorsal tentacles. A comparison of B. anguillae n. sp. with 11 congeneric species revealed its remarkable similarity with B. polymorphus Baer, 1827; however, the new species has a larger cirrus sac, larger pharynx, vitelline gland fields not extending the level of pharynx, ovary located in the pharyngeal area rather than fairly posterior to pharynx, smaller testes, relatively wider rhynchus, and tegumental armature comprising slightly larger spines. Multivariate discriminant analyses confirmed a differentiation of B. anguillae from populations of B. polymorphus; the combination of 4 variables, namely cirrus sac length, pharynx width, cirrus sac width, and rhynchus width yielded a total separation of compared species.     

Lesions induced by complement in vitro on the protoscoleces of Echinococcus multilocularis: a study by electron microscopy.

Changes in the ultrastructure of the tegument of Echinococcus multilocularis protoscoleces during complement-mediated lysis in vitro was studied using transmission and scanning electron microscopy. It was found that the total disintegration of protoscoleces by complement proceeds through formation of ‘tegumental bubbles’ and disruption of the external plasma membrane. This sequence of events was evident in the appearance of numerous loose membrane fragments and vesicles, the lifting of the external unit membrane of the microtriches and the release of organelles from the distal cytoplasm. Subsequent events, such as the appearance of a ‘fuzzy’ coat and disruption of the basement membrane, were probably due to autolysis.     

Surface ultrastructure of the adult stage of Acanthotrema felis (Trematoda: Heterophyidae)

The surface ultrastructure of Acanthotrema felis (Trematoda: Heterophyidae) adults, recovered from a kitten experimentally infected with the metacercariae, was observed using a scanning electron microscope. The worm was leaf-like, ventrally concave and covered with scale-like multi-pointed tegumental spines. The spines on the anterior surface were short but broad, and had 10-12 pointed tips. The cytoplasmic processes protruded around the spines, like pockets for the spines. The ventrogenital opening was crescent, or kidney-shaped, and had protuberances with minute spines on its surrounding tegument. The spines on the posterior surface were long, but narrow, with 6-8 pointed tips. The cytoplasmic processes on this tegument were ridge-like, and elevated along the row of the spines. The surface ultrastructure of A. felis is generally similar to that of other heterophyid flukes, but some features are characteristic, and may be of taxonomic and bio-ecological significance.     

Fasciola gigantica: Immunolocalization of 28.5 kDa antigen in the tegument of metacercaria and juvenile fluke

Specific monoclonal antibody (MoAb) to 28.5 kDa tegumental antigen (TA) was used to localize this antigen in the tissues of metacercariae, newly excysted juvenile (NEJ), 1, 3, 5, and 7-week-old juveniles of Fasciola gigantica by using indirect immunofluorescence, immunoperoxidase and immunogold techniques. Both indirect immunofluorescence and immunoperoxidase detections showed that this antigen was concentrated in the tegument particularly in its outer rim, tegumental cells and their processes as well as epithelial linings of the oral sucker. Unlike adult F. gigantica, it was not detected in spermatogenic cells in the testes, cells of Mehlis’gland, oocytes within the ovary, and ovum within the egg of parasites. At the ultrastructural level, the immunogold labeling showed deposit of gold particles specifically in G₂ tegumental granules and on the surface membrane. Thus, this 28.5 kDa antigen is expressed in the tegument and associated structures of juvenile parasites, and it could be a major component of the G₂ granules which are shown to fuse with the surface membrane and contribute material to replace the casted-off membrane. This process is the replenishment and turnover of the surface membrane to prevent the attachment of the host immune effector cells.     

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.     

Ultrastructure of the Tegumental Basement Membrane of Fasciola hepatica(Trematoda)

The fine–structural characteristics of the basement membrane of the tegument of F. hepatica were examined following extraction fixations and tannic acid infiltration. The basement membrane was shown to consist of three layers: lamina lucida, lamina densa, and lamina reticularis. The lamina densa appeared amorphous and homogeneous with tannic acid impregnation. The lamina reticularis appeared as a dense network of 10–12 nm fibrils. Anchoring fibrils cross this layer and form loops. Along their length they contact hemidesmosomes of muscles, thus connecting muscle to muscle and to tegument. The tegument/basement membrane contact is enhanced by extensions of the lamina densa into infoldings of the tegumental basal membrane. Where tegumental spines reach the basement membrane, the contact is reinforced by hemidesmosomes that connect to anchoring fibrils reaching toward the underlying muscles. The basement membrane thus seems to be a complex structure integrating the distal tegumental layer with underlying tissues and transducing muscle contractions to the tegument and its spines.     

Tegument and apical end organ fine structure in the metacestode and adult Proteocephalus ambloplitis

The tegument of plerocercoid and adult P. ambloplitis was examined. Differences in tegument structure existed between these two stages. Plerocercoids of P. ambloplitis lacked extensive vacuolization and unicellular gland cells characteristic of adult tegument. Plerocercoid microtriches were short and conoid; adult microtriches were lenticular with an extended, whip-like shaft. An inclusion, not previously reported from proteocephalid cestodes, is described. Adult tegument had ducts, originating from underlying unicellular glands, extending through the distal cytoplasm and opening to the exterior between microtriches.

The apical end organ cavity of P. ambloplitis contained numerous labyrinth-like spherical bodies. These structures appeared to be synthesized and secreted into the end organ by a thin cellular lining of the end organ. This lining was composed of discrete, filamentous cells believed to be modified subtegumental cell bodies. Spherical structures identical to those observed within the end organ cavity occurred within this cellular lining. The spherical bodies may be associated with enzymes necessary for tissue migration by the metacestode.     

Electron microscope studies of Fasciola hepatica III. Fine structure of the prostate gland.

An ultrastructural study of the prostate gland of Fasciola hepatica shows it to be composed of numerous unicellular glands. These gland cells contain an extensive granular endoplasmic reticulum (GER) system parts of which are intimately associated with septum-like invaginations of the plasma membrane extending almost to the nucleus. Also associated with the GER are many Golgi complexes which secrete large electron-lucid carbohydrate-rich secretory vesicles. The secretion passes up the gland ducts along with a very dense granular and fibrillar material. The ducts have a peripheral microtubular skeleton and are tightly bound to the epithelium of the ejaculatory duct by septate desmosomes. Secretory vesicles are stored in the expanded ends of the ducts where they pass through the ejaculatory epithelium and their content is discharged by the bursting of their limiting membrane.     

Fasciola gigantica: Histology of the digestive tract and the expression of cathepsin L

The digestive tract of Fasciola gigantica is composed of the oral sucker, buccal tube, pharynx, esophagus, and caecum. The tegumental-type epithelium lines the first four parts of the digestive tract while the caecal-type epithelium lines the remaining parts from the caecal bifurcation. The caecal-epithelial cells are classified into 3 types according to their staining properties and ultrastructural characteristics, as related to the amount of food contents in the caecal lumen. All caecal-type epithelial cells synthesize and secrete cathepsin L, a major group of enzymes in the digestive tract, as detected by in situ hybridization and immunolocalization. Moreover, the secreted cathepsin L is also adsorbed on the outer surface of the tegument and the glycocalyx coating of the surface of the tegument, whereas the tegumental cells and tegumental syncytium covering the parasite’s body and lining the proximal part of the digestive tract exhibit no in situ hybridization signal and immunostaining for cathepsin L.     

Ultrastructure of Gonionchus australis (Xyalidae,Nematoda)

Observations are reported on the ultrastructure of the buccal cavity, body cuticle, spermatids, spermatozoa, male genitalia, and caudal glands of Gonionchus australis. The buccal cuticle is a continuation of the pharyngeal cuticle. Anteriorly it is secreted by arcade tissue and overlaps the mouth rim; laterally it forms longitudinal tooth ridges. The non-annulated cephalic cuticle differs sharply from the remainder of the body wall cuticle. The cortical and basal zones become much thinner, while a largely structureless, lucent median zone expands to fill the bulk of the lips and lip flaps. Spermatids possess fibrous bodies, multimembrane organelles, mitochondria, and compact chromatin. The spermatozoa of G. australis resemble those of most other nematodes by the absence of the nuclear envelope and presence of fibrous bodies, mitochondria, and compact chromafin. The ejaculatory duct possesses microvilli. Two ejaculatory glands lie beside the duct. Two neurons are located within each spicule and each part of the paired gubernaculum. Caudal gland nuclei are large, with dispersed chromatin. The ducts of all three caudal glands are filled with secretory vesicles.     

An electron microscope study of the tegument of Hunterella nodulosa Mackiewicz and McCrae, 1962 (cestoidea: Caryophyllidea)

Ultrastructural observations using transmission and scanning electron microscopy reveal the tegument to be basically similar to that of other cestodes. The syncytial distal cytoplasm is devoid of organelles except for rod-shaped bodies, believed to be secretory vesicles, and lamellated bodies which probably contribute the raw material for new microtriches. There is evidence that these vesicles originate from the Golgi found in the sub-cuticular cells.Three types of microtriches are described: typical ones with well-developed spines, ones with short filaments instead of spines, and ones with no spines. Microtriches with spines are found only on the anterior part of the worm and may serve to anchor the worm. Microtriches on the posterior have no spines and are believed to be primarily involved in the absorption of nutrients. Between these two regions there is a transitional zone where all three types of microtriches can be found. In general the microtriches are quite uniformly distributed throughout the surface of the worm. The presence of cestodarian-like microtriches raises interesting evolutionary questions.Histochemical tests localized acid and alkaline phosphatase activity on various parts of the tegument, as well as on host intestine, while acian blue tests showed that acid mucopolysaccharide levels correspond with the concentration of the tegument vesicles.     

Phaneropsolus spinicirrus n. sp. (Digenea: Lecithodendriidae), a human parasite in Thailand

Phaneropsolus spinicirrus n. sp., collected in a postpraziquantel treatment stool of a 44-yr-old woman from Kalasin Province in northeastern Thailand, is described. It is the second species of the genus Phaneropsolus Looss, 1899, found to parasitize a human host. It differs from the first species, Phaneropsolus bonnei Lie, 1951, in the presence of a short spinose cirrus and the structure and distribution of tegumental spines. The new species is most similar to Phaneropsolus perodictici Goodman and Panesar, 1986, isolated from the potto, Perodicticus potto, in Uganda because digeneans of both species have numerous spines on the cirrus. However, the new species differs from P. perodictici in having more conspicuous spines, the genital pore at the posterior border of the oral sucker or lateral to the pharynx, a body and internal organs of larger size, conical spines on the tegument, an ovoid to bilobed ovary and a large V-shaped excretory bladder. Phaneropsolus spinicirrus is distinguishable from other previously reported Phaneropsolus by possessing a spinose cirrus.