Morphological observations of Echinochasmus japonicus cercariae and the in vitro maintenance of its life cycle from cercariae to adults

The cercaria morphology of Echinochasmus japonicus was investigated using light and scanning electron microscopy. Cercariae, liberated from naturally infected snails (Parafossarulus manchouricus), had ovoid bodies and diminutive tails. The cercaria tegument was covered with minute spines. Four type II sensory papillae were observed on the dorsal side of the oral sucker, and type I papillae were distributed on the dorsal tegument surfaces. When cercariae were kept in the same bath as the freshwater fish, Pseudorasbora parva, which were free from trematode infections, parasites encysted only in the gills of fishes at day 4 postinfection (PI). The outermost metacercaria wall was fully formed in host tissues at day 7 PI. Adult worms were recovered from the intestines of rats, chicks, and ducks 28 days after experimental exposure to metacercariae. The head crown of the adult was armed with 24 collar spines, which were interrupted dorsal to the oral sucker, and the species was identified as E. japonicus.     

Surface ultrastructure of Stictodora tridactyla (Trematoda: Heterophyidae) from Kuwait Bay

The surface morphology of Stictodora tridactyla recovered from a kitten that was fed the killifish, Aphanius dispar, naturally infected with the metacercariae was studied using scanning electron microscopy. The body comprised a rounded head, elongate neck, and widely pyriform hind-body. The head bore a circular oral sucker on the ventral side, and concentric rows of peg-like cephalic spines on the dorsal side. The oral sucker was armed with pre-oral spines similar in shape and size to the cephalic spines. The neck and hind-body were densely covered with scale-like multipointed spines, the size and density of which decreased from anterior to posterior parts of the body. Ciliated dome-shaped papillae were found solitarily or as conjugated groups on the head apex and lips of the oral sucker. Non-ciliated dome-shaped papillae were restricted to the lower lip of the oral sucker. The body was devoid of a ventral sucker. The genital opening appeared as a round depression of the tegument at about the anterior third of the body. This is the first record of the occurrence of S. tridactyla in the Arabian Gulf region and A. dispar is a new second intermediate host.     

Brachylaima cribbi (Digenea: Brachylaimidae): scanning electron microscopical observations of the life-cycle stages

Brachylaima cribbi is a recently described species of terrestrial trematode that infects mammals and birds with helicid land snails as its first and second intermediate hosts. The adult worm is 2.5-6.0 mm long by 0.5-0.8 mm wide being a long slender cylindrical worm with oral and ventral suckers in the anterior quarter and genital pore in the posterior quarter. Scanning electron microscopy shows that there is a dense covering of tegumental spines at the anterior end which diminishes towards the posterior extremities of the worm. Development of spines was observed in juvenile and mature adult worms. In young worms 1-3 weeks post infection (wpi) spines appear as buds with a serrated edge each having 1-4 spikes per spine. As the worm ages the spines broaden and by 5 wpi the number of spikes per spine increases to an average of 8.1. The serial development of oral sucker papillae in the cercaria, metacercaria and adult worm was observed with the finding of an elongated papilla with a bifurcated tip on the cercaria becoming a shorter and thicker elongated papilla with a large central stoma on the metacercaria. In the adult worm, this papilla becomes dome-shaped with a small central stoma. For some of these papillae a cilium could be seen extended from the central stoma. Other life-cycle stages illustrated were the hatched egg with an extruded egg membrane minus an operculum and a portion of the branched sporocyst dissected from the digestive gland of the land snail Theba pisana showing a terminal birth pore. Scanning electron microscopy morphological features of the adult worm observed for the first time in a Brachylaima were the unarmed cirrus extended from the genital pore with released sperm present and the Laurer's canal opening visible in tegumental folds on the dorsal surface approximately 300 microm posterior to the genital pore.     

Studies on metacercarial excystment in Himasthla leptosoma (Trematoda: Echinostomatidae) and newly emerged metacercariae

Metacercarial cysts of Himasthla leptosoma were subjected to a solution containing bile salts, trypsin and l cysteine at 41°C. The treatment induced intense metacercarial activity and after 20 min metacercariae burst through the cyst walls and emerged. Electron microscopy demonstrated that organisms burst through a small area of cyst wall which was devoid of a layer of lamellae present elsewhere towards the innermost surface. The appearance of ruptured cyst walls indicated that they had been softened by the excystment medium. Newly emerged metacercariae possessed a reniform collar of 29 cephalic spines and these were sometimes withdrawn into pits, presumably by action of a muscle complex in the head region. Sensory papillae were distributed in a bilaterally symmetrical arrangement around the anterior sucker and none were visible on the surface of the ventral sucker. Tegumental spines were found only from a point some distance behind the head collar to the region of the ventral sucker. The most anterior spines were simple and peg-like and they quickly merged posteriorly into more complex palmate forms.     

Redescription and surface ultrastructure of Pygidiopsis macrostomum (Digenea: Heterophyidae)

Pygidiopsis macrostomum Travassos, 1928, a poorly known species originally described from a single specimen from Rattus norvegicus (Erxleben, 1777) in Brazil, is redescribed on the basis of metacercariae from the mesenteries of naturally infected guppies Poecilia vivipara Bloch and Schneider, 1801 (Poeciliidae), and adults obtained from an experimental infection of hamsters. Pygidiopsis macrostomum is characterized by the absence of oral spines, vitellaria extending forward to ventral sucker, uterus reaching pharyngeal level, X-shaped excretory vesicle, and an oral sucker/acetabulum ratio of 1:0.8. The surface ultrastructure shows that the tegument of the metacercaria does not strongly differ from that of adults. The brush-shaped spines of P. macrostomum are similar to those reported for Pygidiopsis summa and Pygidiopsis ardeae, but no differences in spine shape were observed throughout the body.     

Surface ultrastructure of juvenile and adult Acanthoparyphium tyosenense (Digenea: Echinostomatidae)

The tegumental ultrastructure of juvenile and adult Acanthoparyphium tyosenense (Digenea: Echinostomatidae) was observed by scanning electron microscopy. One- to 3-day-old juveniles and 10-day-old adults were harvested from chicks experimentally fed metacercariae from a bivalve, Mactra veneriformis. The juvenile worms were minute, curved ventrally, and had 23 collar spines characteristically arranged in a single row. The lips of the oral sucker had 7 single aciliated sensory papillae and 4 grouped uniciliated sensory papillae. The ventral sucker had 25 aciliated round swellings on its lip. The anterolateral surface between the 2 suckers was densely packed with tongue-shaped tegumental spines, and the ventral surface just posterior to the ventral sucker was covered with peg-like spines. Retractile, peg-like spines were seen on the anterolateral surface, whereas scale-like spines with round tips and broad bases were sparsely distributed posterior to the ventral sucker. The cirrus was characteristically protruding and armed with minute spines. The surface ultrastructure of A. tyosenense was unique, especially in the number and arrangement of collar spines, shape, and distribution of tegumental spines and in distribution of sensory papillae.     

Life cycle and description of a new species of brachylaimid (Trematoda: Digenea) in Spain

The life cycle of Brachylaima llobregatensis n. sp. (Trematoda: Brachylaimidae) is elucidated. Embryonated, operculated, asymmetric eggs (30.9 x 18.2 microm) are eliminated with feces of natural hosts wood mouse Mus spretus; white-toothed shrew, Crocidura russula; and an experimental host, domestic mouse, Mus musculus var. domesticus. The eggs are ingested by the helicid gastropod Helix (Cornu) aspersa, the only natural and experimental first intermediate host. The miracidium hatches from the egg, infects the snails, and develops into a branched sporocyst in the digestive gland. Microcaudate cercariae emerge from this snail and develop into unencysted metacercariae in the kidney of second intermediate host snails H. (C.) aspersa and Otala punctata (natural hosts) and Theba pisana (experimental host). Ingestion of infected snails leads to the infection of definitive hosts, with the adults inhabiting the middle part of the small intestine. There is a chaetotaxic pattern specific on the acetabular (S(II) 5-6 papillae) and body (papillae absent on P(II)) levels. Three types of cercaria papillae were observed by scanning electron microscopy: argentophilic papillae with fingerlike processes (cephalic, body, and acetabular levels); argentophilic papillae with opening (2 papillae in the M body level); and nonargentophilic dome-shaped papillae (on the cephalic C(II) level, alternating with argentophilic S(II) papillae on the ventral sucker). Suckers are subequal, with the acetabulum located in the posterior part of the anterior third of body. Vitellaria extend from anterior margin of acetabulum to between middle level and anterior margin of anterior testis.     

Tegumental ultrastructures of Echinoparyphium recurvatum according to developmental stages

The present study was performed to observe tegumental ultrastructure of Echinoparyphium recurvatum according to developmental stages. Worms (1, 3, 5 and 15-day old) were recovered from chicks experimentally infected with metacercariae from Radix auricularia coreana. One-day old worms were elongated and ventrally concave, and covered with peg-like tegumental spines except the adjacent areas of the head crown and excretory pore. Type I sensory papillae were distributed on the lip of the oral sucker, and grouped ciliated papillae were around the oral sucker. Peg-like tegumental spines were densely distributed on the anterior surface of the ventral sucker level. The ventral sucker had an aspinous tegument and no sensory papillae. Tegumental spines on the posterior surface of the ventral sucker level were sparsely distributed and disappeared posteriorly. In 3 and 5-day old worms, the tegument around the oral sucker was aspinose and wrinkled concentrically. The ventral sucker had a wrinkled tegument and many bulbous papillae. Type I sensory papillae were distributed between the bulbous papillae. Tegumental spines were spade-shaped with a terminal tip. A total of 45 collar spines including 4 end group ones on both ventral corners was alternately arranged in 2 rows. The 15-day old worms were very stout and their tegumental spines were tongue-shaped without a terminal tip. From the above results, it is confirmed that the surface ultrastructure of E. recurvatum was generally similar to that of other echinostomatid flukes. However, some features, i.e., morphological change of tegumental spines and appearance of sensory papillae on the ventral sucker according to development, and number, shape and arrangement of collar spines, were characteristic, which may be of taxonomic and bioecological significance.     

First record and description of metacercariae of Curtuteria arguinae n. sp. (Digenea: Echinostomatidae), parasite of cockles Cerastoderma edule (Mollusca: Bivalvia) in Arcachon Bay, France

A new Himasthlinae species, Curtuteria arguinae, is described as metacercariae from the cockle Cerastoderma edule (L.), collected at Banc d'Arguin (southwestern France). These metacercariae encysted preferentially in the mantle and also in the foot of cockles. Encysted and chemically excysted metacercariae were studied by light microscopy and scanning electron microscopy, respectively. Excysted metacercariae were elongated and curved ventrally. They bore a 33-spine circumoral collar. Sensory papillae were arranged around the oral sucker and also symmetrically along the ventral surface body, from the collar to the acetabulum. The dorsal and ventral tegument surfaces were densely packed with similar pointed spines. The posterior end of the body was without any spines. Among the Curtuteria species described previously, only Curtuteria haematopodis Smogorjewskaja and Iskova, 1966 had the same number of circumoral collar spines. A 6-yr field survey showed that the cockle population at Banc d'Arguin was subjected to a summer infection of C. arguinae. Curtuteria arguinae phenology of infection is characterized by interannual variability and seasonality (beginning in July-August and maximum in autumn). The first intermediate and final hosts remain unknown.     

Description and surface topography of the cercaria of Austrobilharzia sp. (Digenea: Schistosomatidae)

The cercaria of Austrobilharzia sp. from the marine prosobranch gastropod Planaxis sulcatus in Kuwait Bay is described. The surface microtopography and pattern of the tegumentary sensory receptors are examined using scanning electron microscopy. The general microtopography of the surface of the cercaria is similar to that previously observed in cercariae of mammalian schistosomes, although differences are recorded in the types, numbers and distribution of the sensory receptors. The study identified more than 13 types of receptors comprising aciliated, uniciliated and for the first time a multiciliated receptor in a strigeid cercaria. The ciliated receptor types differ in the cilium length and structure of the surrounding collar and tegumentary base. The receptor types are site specific: (1) the aciliated and pitlike on the anterior organ-neck region and ventral sucker; (2) the uniciliated with a long flexible cilium with or without collar or a tegumentary base on the body and tail; and (3) the uniciliated with a short rigid cilium and a robust collar and tegumentary base, and the multiciliated with 6 flexible cilia and a high cylindrical collar on the anterior organ tip. The reported SEM information on the sensory receptors may contribute to elucidating their functional role and to establishing morphological characters for the phylogeny of the family Schistosomatidae.     

Tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis]

The tegumental ultrastructure of juvenile and adult Echinostoma cinetorchis (Trematoda: Echinostomatidae) was observed by scanning electron microscopy. Three-day (juvenile) and 16-day (adult) worms were harvested from rats (Sprague-Dawley) experimentally fed the metacercariae from the laboratory-infected fresh water snail, Hippeutis cantori. The worms were fixed with 2.5% glutaraldehyde, processed routinely, and observed by an ISI Korea DS-130 scanning electron microscope. The 3-day old juvenile worms were elongated and ventrally curved, with their ventral sucker near the anterior two-fifths of the body. The head crown was bearing 37-38 collar spines arranged in a zigzag pattern. The lips of the oral and ventral suckers had 8 and 5 type II sensory papillae respectively, and between the spines, a few type III papillae were observed. Tongue or spade-shape spines were distributed anteriorly to the ventral sucker, whereas peg-like spines were distributed posteriorly and became sparse toward the posterior body. The spines of the dorsal surface were similar to those of the ventral surface. The 16-day old adults were leaf-like, and their oral and ventral suckers were located very closely. Aspinous head crown, oral and ventral suckers had type II and type III sensory papillae, and numerous type I papillae were distributed on the tegument anterior to the ventral sucker. Scale-like spines, with broad base and round tip, were distributed densely on the tegument anterior to the ventral sucker but they became sparse posteriorly. At the dorsal surface, spines were observed at times only at the anterior body. The results showed that the tegument of E. cinetorchis is similar to that of other echinostomes, but differs in the number and arrangement of collar spines, shape and distribution of tegumenal spines, and type and distribution of sensory papillae.     

The life-cycle of Petasiger islandicus Kostadinova & Skirnisson, 2007 (Digenea: Echinostomatidae) elucidated with the aid of molecular data

The small planorbid snail Gyraulus cf. laevis (Alder) from Lake Myvatn in Iceland was found to emit large-tailed cercariae with 19 collar spines, and three-spined sticklebacks Gasterosteus aculeatus L. were infected with metacercariae of a species of Petasiger Dietz, 1909. Comparative sequence analysis using ND1 mitochondrial DNA sequences revealed that the rediae and cercariae are conspecific with P. islandicus Kostadinova & Skirnisson, 2007, recently described from an isolated population of the horned grebe Podiceps auritus (L.) at the lake. The redia, cercaria and metacercaria are described and compared with related forms.     

Echinochasmus leopoldinae n. sp. (Trematoda: Echinostomatidae) and data on its life-cycle

The echinostomatid trematodeEchinochasmus leopoldinae n. sp. is described on the basis of adults found in the intestine of chicks and mice experimentally infected with metacercariae from the gills of the cichlid fishesCichlasoma urophthalmus andC. synspilum from the Peninsula of Yucatan.E. leopoldinae, which had previously been misidentified asE. zubedakhaname Nasir & Díaz, 1968, is characterised by the presence of 20 collar spines with one angular spine on each side and vitelline follicles confluent in the posterior part of the body. A differential diagnosis ofEchinochasmus species possessing 20 collar spines is provided, and the developmental stages (redia, cercaria, metacercaria and adult) ofE. leopoldinae are described.     

Life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae), a parasite of rats in the Llobregat delta (Spain)

The terrestrial triheteroxenous life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae) is elucidated. Operculated, assymetric, embryonated eggs (25.4 x 12.7 microm) are passed with feces of the natural (Rattus norvegicus and R. rattus) and experimental (albino and wild mice, albino rats, Apodemus sylvaticus, Mus spretus [Muridae] and the golden gerbil) definitive hosts and ingested by the helicid gastropod Pseudotachea splendida, the only natural and experimental first intermediate host. Microcaudate cercariae harbored in branched sporocysts in the digestive gland emerge from this snail and contact P. splendida, Otala punctata, Theba pisana, and Helix (C.) aspersa snails developing into unencysted infective metacercariae in the kidney. Definitive hosts are infected by ingestion of infected snails; the adult parasites inhabit the small intestine. Chaetotaxic cercarial pattern specific at acetabular (S(II) 8-10 papillae) and cephalic (C(III) 13-15 papillae, H 16 papillae) levels. Three types of cercarial papillae are observed by scanning electron microscopy (SEM) and their arrangement is correlated with chaetotaxy for the first time in trematodes: argentophilic papillae with fingerlike process (cephalic, body, and acetabular levels), argentophilic papillae with opening (2 papillae in the M body level), and nonargentophilic dome-shaped papillae (alternated with argentophilic S(II) papillae on the ventral sucker). SEM detected interlacing network of ridges covering the metacercarial body. Adults with multidigitate tegumentary spines were observed by SEM. Subequal suckers; the acetabulum located in the posterior part of anterior fifth of body. Vitellaria extend from between middle level and anterior margin of anterior testis to between middle level and posterior margin of acetabulum. Uterus almost reaches the intestinal bifurcation.     

A histochemical study of the secretory gland cells of Cercaria shikokuensis and their role during development from cercaria to metacercaria.

The roles of secretory glands during the developmental process from an immature cercaria to a metacercaria in Cercaria shikokuensis were studied. Four types of secretory cells were identified in this species. On maturation of the cercaria in redia, the products of ventral gland cells and mucoid gland cells formed a thick surface coat on the mature cercaria, and the products of cephalic gland cells also formed a thin cover on the surface coat. In the process leading to the formation of a metacercaria, the surface coat constituted the outer layer of the cyst, mucoid gland cells secreted mucous substances inside the wall, and then cystogenous gland cells discharged their products to the inner wall. The cyst wall was composed of four layers, and it was thought that the outermost surface layer helped the cyst wall to adhere to the matrix and the intermediate layers helped to put together outer and inner walls.     

Ultrastructure of the ventral sucker of Schistosoma mansoni cercaria

The ventral sucker of Schistosoma mansoni cercaria is a cupshaped structure that is attached to the ventral surface of the organism by a homogeneous connective tissue that surrounds the acetabular glands. The sucker consists of an extensive complex of circular and longitudinal muscles. The longitudinal muscles extend outwoard in a radial pattern to form the cup of the organ. Intermingled with the muscles are nerve bundles and subtegumental cells (cytons). Dendritic nerve fibers connect to sensory papillae which are found on the surface tegument. Two types of sensory papillae are present: a commonly found unsheathed uniciliated papilla, and a previously unidentified tegumental encapsulated structure. Tegument with spines covers the ventral sucker, although the tegumental encapsulated sensory papilla lacks spines. © 1995 Wiley-Liss, Inc.     

On the morphology of Acanthostomum spiniceps (Looss, 1896) and A. absconditum (Looss, 1901) (Digenea: Cryptogonimidae: Acanthostominae) with particular reference to the juvenile stage

The morphology of juvenile and adult stages of Acanthostomum spiniceps and A. absconditum, from bagrid fish of the river Nile in Egypt, was studied with both light and scanning electron microscopy. In early juveniles, circumoral spines are absent and the entire body surface is covered with tegumental spines. Late juveniles show gradual differentiation of the circumoral tegument into a collar of spines associated with a reduction in density of tegumental spines at the posterior extremity of the body. Genital primordia appear when juveniles are about 1.75 mm long. The distributions of tegumental spines on adult A. spiniceps and A. absconditum are similar. Spines are denser on the dorsal and ventral surfaces of the anterior and middle parts of the body and less dense towards the posterior end. The tegumental fold surrounding the ventral sucker of A. absconditum has spines while the fold of A. spiniceps lacks them. The most important morphological features differentiating both species are the number of circumoral spines, body shape, ratio of body length to width, sucker sizes, and the presence or absence of spines on the ventral sucker.     

<Emphasis Type="Italic">Petasiger islandicus</Emphasis> n. sp. (Digenea: Echinostomatidae) in the horned grebe <Emphasis Type="Italic">Podiceps auritus</Emphasis> (L.) (Aves: Podicipedidae) from Iceland

Petasiger islandicus n. sp. is described and figured from a demographically isolated population of the horned grebe Podiceps auritus auritus (L.) in Lake Myvatn (Iceland). This new species belongs to the group of species with 19 collar spines which possess a large elongate-oval cirrus-sac, well-developed pars prostatica and massive bulb-like cirrus. Within this group, P. islandicus appears most similar to P. oschmarini Kostadinova & Gibson, 1998, a form with similar body dimensions described from the same host, but differs in having a larger head collar, collar spines, oral sucker, pharynx, testes and sucker-width ratio, and a smaller cirrus-sac, cirrus and eggs. Two Nearctic species resemble P. islandicus in general morphology but differ as follows: P. pseudoneocomense Bravo-Hollis, 1969 has a larger body and collar width, notably shorter collar spines, smaller testes and sucker-width ratio, and a shorter but much wider cirrus-sac which is also smaller relative to the ventral sucker and almost entirely anterior to it; and P. caribbensis Nassi, 1980 has a smaller body, shorter collar spines and a seminal vesicle which is small in relation to the cirrus-sac, vitelline fields reaching anteriorly to the level of the genital pore and the intestinal bifurcation is located more anteriorly.     

The life cycle of Australapatemon magnacetabulum (Digenea: Strigeidae) from Northwestern Argentina

The life cycle of Australapatemon magnacetabulum Dubois, 1988 was resolved experimentally. Planorbid snails Biomphalaria tenagophila (d'Orbigny, 1835) collected in a small pond at the confluence of the San Lorenzo and Arias Rivers, near Salta City, Province of Salta, Argentina, were found to be shedding furcocercous cercariae possessing 4 pairs of penetration glands, 1 pair of unpigmented eyespots, 6 pairs of flame cells in the body, and 1 pair in the tail stem. Metacercariae were found encysted in naturally, and experimentally, exposed leeches Helobdella adiastola Ringuelet, 1972, Helobdella triserialis (Blanchard, 1849), Haementeria eichhorniae Ringuelet 1978, and Haementeria sp., and within their sporocysts in naturally infected planorbid intermediate hosts. Sexually mature adults were recovered from domestic chicks and a duck 8-28 days postexposure by metacercariae from leeches. The identification of the species was based upon the characteristic large ventral sucker and a genital cone, crossed by a hermaphroditic duct with internal folds, occupying approximately a 1/4 to 1/5 of the hindbody.