Ultrastructure of the rhyncheal apparatus and other structures of the scolex of Grillotia (Christianella) carvajalregorum (Cestoda: Trypanorhyncha).

The scolex ultrastructure was studied in Grillotia (Christianella) carvajalregorum (Cestoda: Trypanorhyncha) using histochemistry and transmission electron microscopy. We show for the first time the presence of scolex glands arranged in two longitudinal acini at the pars vaginalis parenchyma. These glands, along with those scattered in bothrial parenchyma, produce potentially adhesive glycoprotein secretions that are discharged via ducts to the bothrial grooves and apex. A particular type of sensory receptor was found around frontal gland pores, with a possible function in regulating their secretion activity. The internal structure of microtriches varies according to their morphotype and distribution on the scolex, this study providing the first description of the ultrastructure of serrate lanceolate spinitriches. The projections that form serrate margins are an extension of the medulla, differing from similar projections of other spinitriches. The large caps observed in serrate lanceolate spinitriches may reflect their specialization in attachment to and abrasion of intestinal mucosa, while the short caps and large bases of acicular filitriches may reflect their involvement in nutrient absorption. We also describe the rhyncheal apparatus ultrastructure, showing a similar basic structure of tentacular walls than that of other trypanorhynchs. Some differences among species in the number of fibrous layers, composition of the apical cytoplasm and presence of microvilli-like projections were discussed. Finally, our study describes in detail the internal ultrastructure of hollow hooks, evidencing the presence of cytoplasm, mitochondria and fibrils. The location of these fibrils may increase the area of contact surface of hooks on tentacles, possibly allowing for a higher tensile strength than that of solid hooks. We consider that gland location and shape, composition of tentacular wall layers, and hook internal structure may serve as useful characters for the taxonomy and phylogeny of Trypanorhyncha. RESEARCH HIGHLIGHTS: This is the first description of scolex internal ultrastructure in Grillotia carvajalregorum, showing the presence of glands arranged in two longitudinal acini at the pars vaginalis parenchyma, with potentially adhesive functions. The internal ultrastructure of serrate lanceolate spinitriches and acicular filitriches may reflect their specialization in attachment to the host intestinal mucosa and their involvement in nutrient absorption, respectively. Internally, hollow hooks have cytoplasm with mitochondria and fibrils, which are more widely distributed than in solid hooks, possibly increasing their tensile strength.

Pixel by pixel: real-time observation and quantification of passive flotation speeds of three common equine endoparasite egg types.

The efficacy of anthelmintic treatments against populations of endoparasites infecting livestock throughout the world is decreasing. To mitigate this, the use of fecal egg counts is recommended to determine both the necessity, and to ensure the appropriate choice, of anthelmintic treatment. Traditionally, and in order to facilitate easier identification and/or enumeration, samples are analysed after separating eggs from other fecal particulates by exposing them to a solution with a density higher than that of the eggs, but lower than the remaining fecal contents. While many parasite egg flotation protocols exist, little is known about the characteristics of these eggs with respect to their movement through a flotation solution. In this study, we have demonstrated a novel method for the observation and quantification of microscopic (65-100 µm) objects as they experience unassisted flotation. This also represents, to our knowledge for the first time, that the flotation of parasite eggs has been observed and their movement characteristics quantified as they float through solution. Particle tracking and video analysis software were utilised to automatically detect and track the movement of individual eggs as they floated. Three 30 s videos and one 2 min video of each egg type were analysed. If the first 30 s of video were discounted, the differences in mean flotation speed among all videos was statistically significant between egg types (P = 0.0004). Strongyle type eggs (n = 201) moved the fastest with a mean 51.08 µm/s (95% confidence interval: 47.54-54.62). This was followed by Parascaris spp. (n = 131) and Anoplocephala perfoliata eggs (n = 322), with mean speeds of 44.43 µm/s (95% confidence interval: 39.47-49.4) and 31.11 µm/s (95% confidence interval: 29.6-32.61), respectively. This method for evaluating the mean speed of passive flotation may represent a first step towards further optimizing fecal egg flotation and be of interest to parasitologists and veterinary practitioners.

New data on spermiogenesis and trepaxonematan axoneme in basal tapeworms (Cestoda, Caryophyllidea, Lytocestidae) parasitizing cyprinid fishes.

Monozoic caryophyllidean cestodes, intestinal parasites of cyprinid fishes, represent a group of tapeworms with an unclear evolutionary history. As spermatology may provide phylogenetically important data, the spermiogenesis and ultrastructure of the mature spermatozoon have been investigated using an integrative approach combining transmission electron microscopy, cytochemistry and electron tomography in Khawia rossittensis (Szidat, 1937). The process of spermatid formation is accompanied by the presence of ultrastructural characters not described in traditional models of spermiogenesis, e.g., apical electron-dense material, the two striated roots situated unusually opposite each other, branching of typical striated roots, an intercentriolar body comprising five electron-dense and four electron-lucent layers, rotation of both free flagella and flagellar buds to the median cytoplasmic process at 90°, and a complete proximodistal fusion. The synchronous rotation of both flagellar buds and growing free flagella is an evolutionarily linked pattern favouring the hypothesis that the Caryophyllidea are not ancestral but are secondarily derived from polyzoic forms. Electron tomography analysis has revealed a unique feature of two helicoidal tubular structures in the central electron-dense core of the axoneme of mature spermatozoon. These data provide new insights into the architecture of the 9 + '1' axoneme, which is shared by male gametes of all trepaxonematan Platyhelminthes.

Raillietiniaisis in a Toddler From Hawaii: A Case of Mistaken Tapeworm Identity.

We present the first reported case from the United States of human infection with the intestinal cestode Raillietina, a parasite primarily of rodents and poultry. As the differential diagnosis between the morphologically similar Dipylidium caninum was challenging, we discuss the biology, epidemiology, and clinical management of this rare parasite.

Echinococcus multilocularis Leuckart, 1863 (Taeniidae): new data on sperm ultrastructure.

The present study establishes the ultrastructural organisation of the mature spermatozoon of Echinococcus multilocularis, which is essential for future research on the location of specific proteins involved in the sperm development in this species and also in Echinococcus granulosus. Thus, the ultrastructural characteristics of the sperm cell are described by means of transmission electron microscopy. The spermatozoon of E. multilocularis is a filiform cell, which is tapered at both extremities and lacks mitochondria. It exhibits all the characteristics of type VII spermatozoon of tapeworms, namely a single axoneme, crested bodies, spiralled cortical microtubules and nucleus, a periaxonemal sheath and intracytoplasmic walls. Other characteristics observed in the male gamete are the presence of a >900-nm long apical cone in its anterior extremity and only the axoneme in its posterior extremity. The ultrastructural characters of the spermatozoon of E. multilocularis are compared with those of other cestodes studied to date, with particular emphasis on representatives of the genus Taenia. The most interesting finding concerns the presence of two helical crested bodies in E. multilocularis while in the studied species of Taenia, there is only one crested body. Future ultrastructural studies of other species of the genus Echinococcus would be of particular interest in order to confirm whether or not the presence of two crested bodies is a characteristic of this genus.

Vitellogenesis of diphyllobothriidean cestodes (Platyhelminthes).

The recently erected cestode order Diphyllobothriidea is unique among all tapeworm orders in that its species infect all major groups of tetrapods, including man. In the present paper, the vitellogenesis of representatives of all three currently recognized families of this order was evaluated, based on ultrastructural (transmission electron microscopy) and cytochemical (detection of glycogen) observations. Vitelline follicles of all taxa studied, i.e. Cephalochlamys namaquensis from clawed frogs (Xenopus), Duthiersia expansa from monitors (Varanus) and Schistocephalus solidus that matures in fish-eating birds, contain vitelline cells at various stages of development and interstitial cells. Developing vitellocytes are characterized by the presence of mitochondria, granular endoplasmic reticulum and Golgi complexes involved in the synthesis of shell globules and formation of shell globule clusters. Mature vitellocytes contain lipids and glycogen in different proportions. The most significant differences among the three diphyllobothriidean families were found in the presence or absence of lamellar bodies. Variations of vitelline clusters morphology and types of lipid droplets are described and discussed in relation to the presumed evolutionary history of diphyllobothriideans, which belong to the most basal cestode groups.

Larval helminths in the invasive American brine shrimp Artemia franciscana throughout its annual cycle.

One of the best examples of rapid displacement of native species by an invader is the eradication of native Artemia salina and A. parthenogenetica in the Mediterranean by the introduced American A. franciscana. Previous studies based on sampling from limited time periods suggest that the success of the American species as a competitor may be due partly to different parasite burden, since native Artemia spp. have high cestode infection rates regulating their density. The aim of this study is to test the hypothesis that the helminth infection in A. franciscana in its invasive range is low throughout its annual life cycle. Samples of A. franciscana were collected every second month from La Tapa saltern (Andalusia) during one year. Five helminth species were recorded: cestodes Flamingolepis liguloides, F. flamingo, Gynandrotaenia stammeri (all flamingo parasites), Eurycestus avoceti (a shorebird parasite) and larval spirurids of the Acuariinae (the first record of nematodes in Artemia). The overall infection rate was low, with total prevalence 5.9% and prevalence of individual parasite species between 0.2 and 3.2%. The mean abundance of helminths was 0.005-0.155 (av. 0.068), 5-13 times lower than in native congeners. Waterbird counts indicate that the low infection rates cannot be explained by lack of definitive hosts. The results are consistent with the hypothesis that helminths have no regulating effect on the invasive brine shrimp in the Mediterranean. The replacement of the native populations by the invader can be partially explained by a competition mediated by parasites/predators through a differential impact on host fitness.

Coprologically diagnosing Anoplocephala perfoliata in the presence of A. magna.

Current copro-diagnostic tests for Anoplocephala perfoliata show high variation in their sensitivity and given the morphological similarity of Anoplocephala spp. eggs, this could be related to the presence of Anoplocephala magna alone or co-existing with A. perfoliata. In the present study, coprology was significantly more sensitive (p<0.01) at detecting A. magna than A. perfoliata. This difference was independent of the parasite burden and was greater when testing was limited to horses with mature or gravid tapeworms. A. magna infection was strongly linked to young horses (≤ 2 years). The eggs of A. magna are smaller. Using 15 and 70 µm cut-offs for oncosphere diameter and the major shell bisector length, respectively, the eggs of A. perfoliata were identified with 100% sensitivity, 97% specificity and 98% sensitivity, 84% specificity. The use of these two morphometric variables would therefore be useful for the copro-identification of A. perfoliata in countries where both species coexist.

Cytocomposition of the vitellarium in Khawia sinensis Hsü, 1935 (Cestoda, Caryophyllidea, Lytocestidae): another caryophyllidean species with lamellar bodies and lipids.

The vitellarium of the invasive caryophyllidean tapeworm Khawia sinensis Hsü, 1935 from carp Cyprinus carpio L. was examined by means of transmission electron microscopy and cytochemical staining for glycogen with periodic acid-thiosemicarbazide-silver proteinate (PA-TSC-SP). A vitellarium consists of numerous follicles of irregular size that are interconnected by a net of vitelline ducts. Vitelline follicles are composed of vitelline cells at various stages of development that are interconnected by interstitial tissue. Vitelline follicles are surrounded by a cytoplasmic sheath associated with an intercellular matrix. Extensive development of the granular endoplasmic reticulum and Golgi complexes are both involved in the production of shell globules/shell globule clusters and characterise cytodifferentiation of vitellocytes. Nuclear and nucleolar transformation lead to the formation and storage of intranuclear glycogen, a feature specific for the Caryophyllidea. Newly observed within the mature vitellocytes of Khawia sp. is the presence of lamellar bodies and a few lipid droplets. These cytoplasmic inclusions first occur in the mature cells within the follicles and persist in the vitelline cells within vitelloducts and intrauterine eggs. Two types of lamellar bodies are detected: regular lamellar-structured body and irregular lamellar-structured body. None of the lamellar bodies are membrane bound. Results of the present study indicate that the formation of lamellar bodies may be closely related to the endoplasmic reticulum or shell globule clusters. Some of the shell globule clusters are transformed into lamellar body clusters. Ultrastructural features of vitellocytes in K. sinensis are compared with those of other monopleuroid, polypleuroid, and strobilated cestodes.

Evidence of apoptosis in Raillietina echinobothrida induced by methanolic extracts of three traditional medicinal plants of Northeast India.

The therapeutic benefits of medicinal plants in terms of anthelmintic properties are known since time immemorial in India, particularly among natives of the Northeast India. However, only sporadic and scarce reports on scientific validation of these plants are available. The present study was conducted on the cestode Raillietina echinobothrida, to establish whether the anthelmintic activity of Potentilla fulgens, Alpinia nigra and Millettia pachycarpa was mediated by apoptosis or not. Light microscopic observation following MTT assay revealed the highest percentage of inhibition of viability among the worms by methanol extract of M. pachycarpa (89.33%), followed by A. nigra (65%) and P. fulgens (37%). Ultrastructural observations revealed swelling of mitochondria, disruption of mitochondrial membrane, vacuolization of mitochondria, appearance of apoptotic bodies in the cytoplasm, disintegration of nuclear membrane and nucleolus were very common throughout the tegument. DAPI stained specimens showed typical morphology of apoptosis, like nuclear condensation and fragmentation in the extracts treated parasites. A decrease in mitochondrial membrane potential was also recorded in the treated groups. Confirmatory TUNEL assay and DNA fragmentation assay of the extracts treated parasites also confirmed the apoptotic nature of cell death and is concluded to be responsible for paralysis and death of the parasite.

Ultrastructure of vitellogenesis and vitellocytes in the trypanorhynch cestode Aporhynchus menezesi, a parasite of the velvet belly lanternshark Etmopterus spinax.

This is the first TEM examination of vitellogenesis in the cestode Aporhynchus menezesi, a parasite of the velvet belly lanternshark Etmopterus spinax and a member of a little-studied trypanorhynch family, the Aporhynchidae. The synthetic activity of vitellocytes plays two important functions in the developmental biology of cestodes: (1) their shell-globules serve in eggshell formation; and (2) their accumulated reserves of glycogen and lipids represent a food source for the developing embryo. In A. menezesi, vitelline follicles consist of cells at various stages of development, from peripheral, immature cells of the gonial type to mature cells towards the centre of the follicle. These stages are: (I) immature; (II) early differentiation; (III) advanced maturation; and (IV) mature. Gradual changes involved in this process occur within each stage. Vitellogenesis involves: (1) an increase in cell volume; (2) the development of a smooth endoplasmic reticulum and an accelerated formation and accumulation of both unsaturated and saturated lipid droplets, along with their continuous enlargement and fusion; (3) the formation of individual ß-glycogen particles and their accumulation in the form of glycogen islands scattered among lipid droplets in the cytoplasm of maturing and mature vitellocytes; (4) the rapid accumulation of large, moderately saturated lipid droplets accompanied by dense accumulations of ß-glycogen along with proteinaceous shell-globules or shell-globule clusters in the peripheral layer during the advanced stage of maturation; (5) the development of cisternae of granular endoplasmic reticulum that produce dense, proteinaceous shell-globules; (6) the development of Golgi complexes engaged in the packaging of this material; and (7) the progressive and continuous enlargement of shell-globules into very large clusters in the peripheral layer during the advanced stage of maturation. Vitellogenesis in A. menezesi, only to some extent, resembles that previously described for four other trypanorhynchs. It differs in: (i) the reversed order of secretory activities in the differentiating vitellocytes, namely the accumulation of large lipid droplets accompanied by glycogenesis or ß-glycogen formation during early differentiation (stage II), i.e. before the secretory activity, which is predominantly protein synthesis for shell-globule formation (stage III); (ii) the very heavy accumulation of large lipid droplets during the final stage of cytodifferentiation (stage IV); and (iii) the small number of ß-glycogen particles present in mature vitellocytes. Ultracytochemical staining with PA-TCH-SP for glycogen proved positive for a small number of ß-glycogen particles in differentiating and mature vitellocytes. Hypotheses, concerning the interrelationships of patterns of vitellogenesis, possible modes of egg formation, embryonic development and life-cycles, are commented upon.

Revisiting caryophyllidean type of spermiogenesis in the Eucestoda based on spermatozoon differentiation and ultrastructure of Caryophyllaeus laticeps (Pallas, 1781).

Spermiogenesis of the monozoic cestode Caryophyllaeus laticeps (Pallas, 1781) (Caryophyllidea: Caryophyllaeidae), a parasite of Abramis brama (Pisces: Cyprinidae), has been investigated by means of transmission electron microscopy and cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for glycogen. The process of spermatozoon formation corresponds in the basic pattern to that of Khawia armeniaca as described by Brunanská and Poddubnaya (Parasitol Res 99:449-454, 2006). The dense material at the early stages of spermiogenesis, an intercentriolar body, the formation of a free flagellum and flagellar bud, the penetration of the nucleus into the spermatid body when the fusion of the free flagellum with the median cytoplasmic process has started, and a complete proximodistal fusion have been determined. In contrast to previous data on the caryophyllidean type spermiogenesis, the latter more recent observations show the presence and the rotation of a free flagellum and a flagellar bud. This pattern indicates clearly a derived stage of spermiogenesis in the Caryophyllidea, when the second shorter flagellum is greatly reduced comparing with that in the Spathebothriidea, Diphyllobothriidea, or Bothriocephalidea, thus forming a flagellar bud. The flagellar bud occurs in all stages of spermiogenesis and represents an evolved character in the Caryophyllidea. The mature spermatozoon of C. laticeps consists of one axoneme of the 9 + "1" trepaxonematan structure, parallel cortical microtubules, and a nucleus. Cell components are situated in a moderately electrondense cytoplasm, containing glycogen in the principal regions (II, III, and IV) of the spermatozoon. A crested body is absent. Similarities and differences between spermatozoa of caryophyllideans as well as other Eucestoda are discussed.

Classical and molecular cytogenetics of Khawia sinensis (Cestoda: Caryophyllidea), invasive parasite of carp, Cyprinus carpio.

Chromosomes of the invasive tapeworm Khawia sinensis (Caryophyllidea), the specific parasite of common carp, were analyzed by means of conventional Giemsa staining and using fluorescent DAPI and YOYO-1 dyes, silver staining, and fluorescent in situ hybridization (FISH) with 18S rDNA probe. The karyotype is composed of eight pairs of metacentric and telocentric chromosomes (2n = 16, n = 3m + 5t, TCL = 42.54 µm). Constitutive heterochromatin was located at pericentromeric regions of all pairs, except for the largest metacentric pair (no. 1), which possessed no DAPI-positive band. FISH with rDNA probe revealed that both homologues of chromosome pair no. 6 carry a cluster of ribosomal arrays, which were located interstitially close to the centromere. Present results are compared with previous cytogenetic data on Khawia spp., and comments are made on the karyotypes with respect to their phylogenetic links.

Vitellogenesis in the cestode Atractolytocestus huronensis Anthony, 1958 (Caryophyllidea: Lytocestidae).

Vitellogenesis in the parthenogenetic caryophyllidean tapeworm Atractolytocestus huronensis Anthony, 1958, parasitizing the carp Cyprinus carpio L., has been examined using light (LM) and transmission electron microscopy and cytochemical staining with periodic acid-thiosemicarbazide-silver proteinate for glycogen. It was found that formation of vitellocytes shares the same basic patterns in lower tapeworms with parthenogenetic and normal reproduction. Each vitelline follicle of A. huronensis consists of vitellocytes at various stages of development and an interstitial tissue. Projections of the interstitial tissue surround each vitellocyte and extend as a cytoplasmic sheath on the follicular periphery. In contrary to other Eucestoda, interstitial tissue of A. huronensis includes numerous electron-dense vesicles of various sizes. Maturing and mature vitellocytes contain vitelline material in the form of single small shell globules, which may gradually fuse and give rise to the large shell globule clusters. Shell globule clusters are of two types in A. huronensis. Additional vitelline material is represented by single "lamellar" granules and glycogen in the cytoplasm of mature vitellocytes. The ultrastructural features of vitellogenesis and/or vitellocyte in A. huronensis are compared with that in other caryophyllideans and/or Neodermata.

Do glial cells exist in the nervous system of parasitic and free-living flatworms? An ultrastructural and immunocytochemical investigation.

It is still unclear whether flatworms have specialized glial cells. At present there are no special methods available for the identification of glial cells in flatworms. The aim of this research was to carry out detailed investigations of the CNS in two species ofcestodes, and to get an idea whether these cells may fit into the concept of glia. Three types of glial cells have been found in Grillotia erinaceus: (1) fibroblast-like cells in the cerebral ganglion (CG); (2) glial cells in bulbar nerves with filaments and laminar cytoplasm; (3) a 3rd type of cells forms multilayer envelopes in the main cords (MC); also they make contacts with the excretory epithelium. To demonstrate the existence of glial cells, an immunocytochemical and ultrastructural investigation of Ligula intestinalis was undertaken. Intensive S100b-like immunoreaction (IR) was found in the GG and in the MC. IR-varicosities were mostly located asymmetrically on the MC, and no IR was found in neuropiles. Small glial cells were found on the surface of the MC; they have oval nuclei and dense cytoplasm with slim processes going around the neuropile and enclosing neurons. Long junctions are seen between cell processes but with neurons they usually possess juxtaposition contacts. Glial cells lack vesicles or synapse-like structures. Intensive S100b-like-IR has been shown in the CNS of cestodes for the first time. Results from ultrastructural research support the immunocytochemical date.

Cellular organization of the oncosphere of Mosgovoyia ctenoides (Cestoda: Anoplocephalidae).

The ultrastructure of the infective oncosphere of the cestode Mosgovoyia ctenoides (Anoplocephalidae) is described. The surface of the infective oncosphere is covered by a thin cytoplasmic layer of tegument connected by a narrow cytoplasmic process with the binucleate subtegumental cell, situated deeper in the body. Below the basal matrix of the cytoplasmic layer of the tegument are situated wide bands of the peripheral, somatic musculature responsible for body movements. The 3 pairs of hooks and their muscles form a complex hook muscle system, responsible for coordinated hook action. Five major types of cells have been distinguished: (1) a binucleate subtegumental cell, (2) a binucleate penetration gland, (3) 2 nerve cells, (4) numerous somatic cells, and (5) about 6 germinative cells. The approximate number of cells is 24 (26 nuclei, including 2 syncytial structures). The results of this study, when compared with other published reports from other cestode taxa, support previous hypotheses that the progressive reduction of oncosphere cells is an adaptive feature in cestode evolution.

Anthelmintic efficacy of Flemingia vestita (Fabaceae): alteration in the activities of some glycolytic enzymes in the cestode, Raillietina echinobothrida.

The crude root-peel extract of Flemingia vestita, genistein and praziquantel were tested against some selected glycolytic enzymes--hexokinase (HK), phosphofructokinase (PFK), phosphoenolpyruvate carboxykinase (PEPCK), pyruvate kinase (PK), lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and malic enzyme (ME)--of the fowl tape worm, Raillietina echinobothrida. Following exposure to the various treatments, the activities of HK, PFK, PEPCK and LDH increased by 33-39%, 41-125%, 44-49% and 55-67%, respectively, and that of PK decreased by 14-26% in the parasite at the time of paralysis. The MDH and ME activities of the tissue homogenate were also found to be higher by 22-43% and 28-59%, respectively, in the treatments. However, whereas the activity of both cytosolic and mitochondrial MDH increased by 33-58% and 43-73%, respectively, the cytosolic ME activity showed an increase of 33-39%, and there was no significant enhancement in the mitochondrial ME activity. Histochemically, the enhancement in the activities of HK, LDH and MDH was clearly discernible. The enhanced glycolytic activity seems to be a function of anthelmintic stress caused by the phytochemicals.

Ultrastructure of the tegument of the anoplocephalid cestode Mosgovoyia ctenoides (Railliet, 1890) Beveridge, 1978.

The tegument of the mature proglottids of M. ctenoides was examined by means of TEM. The tegument of this species consists of two layers: (1) the external cytoplasm, and (2) the tegumental perikarya situated in the cortical parenchyma. The tegument surface is covered by typical microtriches. The anucleated external layer of cytoplasm is rich in vesicles of different shape and electron-density, but it lacks mitochondria. Large pore canals penetrate the external cytoplasmic layer. This layer is separated from the perikarya by a basal lamina, being connected with the tegument cell bodies by cytoplasmic bridges. The granular cytoplasm of perikarya contains typical cell organelles such as mitochondria, GER, Golgi complexes, free rybosomes, numerous vesicles and lipid droplets inclusions. The large nuclei of the perikarya with prominent nucleoli frequently contain large intranuclear, highly osmiophilic lipid droplets.

Ultrastructure of the spermatozoon of the pseudophyllidean cestode Eubothrium crassum (Bloch, 1779).

The fine structure of the mature spermatozoon of the pseudophyllidean tapeworm Eubothrium crassum, a parasite of salmonid fishes, has been studied by transmission electron microscopy for the first time. The mature spermatozoon of E. crassum is filiform and tapered at both extremities. It contains two axonemes of unequal length showing the 9 + "1" pattern of Trepaxonemata. The anterior extremity exhibits a crested body 50-100 nm thick. It spirals around the outside of the anterior region of the spermatozoon. The nucleus is electron-dense, exhibiting a fibrous appearance in its middle (the broadest) region. The cortical microtubules are of two types and are situated parallel to the spermatozoon axis. The cytoplasm is slightly electron-dense and contains numerous electron-dense granules in region II of the spermatozoon. A ring of electron-dense, centred microtubules surrounds the axoneme, together with the underlying ring of moderately electron-dense, subjacent submicrotubular material in region V. The anterior and posterior extremities of the spermatozoon lack cortical microtubules and contain a single axoneme. Our results reveal several peculiarities, in which the spermatozoon of E. crassum differs from those of other pseudophyllidean cestodes.