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.     

Tegumental ultrastructure of adult Gynaecotyla squatarolae (Digenea: Microphallidae)

Gynaecotyla squatarolae (Digenea: Microphallidae) adult flukes were recovered from experimental chicks at day 4-6 post-infection and their tegumental ultrastructure was observed with a scanning electron microscopy. They were pyriform in shape, and their anterior halves were concaved ventrally. The whole body surface was covered with tegumental spines, which were wide and 16-17 digitated between oral and ventral suckers. The density of spines and number of digits decreased posteriorly. The oral sucker was subterminal and the excretory pore was at the posterior end of the worm. Two ventral suckers were similar in appearance and protruded near midline of the worm. The genital atrium was dextral to the small ventral sucker. The dorsal surface was covered with tegumental spines, but the spines were sparser than on the ventral surface. On the middle portion of the dorsal surface, a small opening presumed to be the Laurer's canal was seen. From these findings, it has been confirmed that the adult G. squatarolae has unique characteristics in the surface ultrastructure.     

A new species of Opisthioglyphe (Trematoda: Telorchiidae) from gall bladder of turtles in Malaysia

Opisthioglyphe sharmai n. sp. is described from the gall bladder of the Malayan box turtle, Cuora amboinensis, and the black marsh turtle, Siebenrockiella crassicollis, in Malaysia. The new species is morphologically similar to Opisthioglyphe ranae and some other members of the genus parasitic in amphibians and reptiles. Opisthioglyphe sharmai n. sp. is easily differentiated from all other members of the genus by the cirrus sac extending posterior to the ventral sucker, while in all previously known species the cirrus sac is entirely or mostly preacetabular with the base of the structure not reaching beyond mid-line of the ventral sucker. Despite the overall stable morphology, O. sharmai n. sp. is characterized by highly variable arrangement of testes, from tandem to opposite. It is only the second representative of the genus described from turtles and the first species of Opisthioglyphe parasitic in gall bladder, while all previously described members of the genus are parasitic in the intestine of their hosts.     

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.     

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.     

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.     

Surface ultrastructure of Heterophyes nocens (Trematoda: Heterophyidae).

The surface ultrastructure of Heterophyes nocens (Trematoda: Heterophyidae) was studied by scanning electron microscopy(SEM). The adult worms were recovered from experimentally infected cats and from a naturally infected patient. They were leaf-like, ventrally concave, and ovoid or pyriform in shape. Ciliated knob-like sensory papillae (type I) were observed in single or grouped forms on and around the oral sucker, whereas non-ciliated round swellings (type II papillae) were seen on the lip of the ventral sucker. The tegumental spines around the oral sucker were 5-9 pointed, whereas those between the two suckers were 12-17 pointed. Ventrolaterally, three groups of 5-6 type I papillae were located between the oral and ventral suckers, with single ones alternating between them. The genital sucker was protruded or depressed, depending on the contraction state of the flukes, and the gonotyl spine number ranged 50-60. The number of tip points of tegumental spines was decreased posteriorly; finally they became 1-3 pointed. On the dorsal surface, 4 groups of 4-5 type I papillae were symmetrically located on both lateral sides, and the shape and distribution of tegumental spines were similar to those of the ventral surface. Although the tegumental ultrastructure of H. nocens was generally similar to those of other heterophyids, the genital sucker morphology including the number of gonotyl spines and/or the distribution pattern of tegumental spines and sensory papillae were suggested to be the characteristic features of H. nocens.     

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.     

Histopathology, ultrastructure and immunohistochemistry of Coregonus lavaretus hearts naturally infected with Ichthyocotylurus erraticus (Trematoda)

Histopathological, ultrastructural and immunohistochemical investigations were conducted on 26 specimens of powan Coregonus lavaretus (L.) from Loch Lomond (Scotland). The hearts of all 26 powan (15 females and 11 males) investigated harboured metacercariae of the digenean trematode Ichthyocotylurus erraticus (Rudolphi, 1809). The vast majority of metacercariae were located either singly or as an aggregation of white cysts on the surface of the bulbus arteriosus. The intensity of infection ranged from 2 to 200 larvae heart(-1), although the number of metacercariae found on male powan did not exceed 13. Histochemically, the parasite cyst wall gave a strong positive reaction with periodic acid schiff (PAS) and a faint positive signal with Azan-Mallory stain. All the metacercariae cysts were embedded in a granulomatous proliferation of heart epicardium tissue, forming a reactive fibroconnective capsule around the parasite. The capsule enclosing the parasite (produced by the host's reaction to the parasite) measured 13.57 to 90.20 microm (37.43 +/- 3.56) in thickness. Within the capsule wall, eosinophilic granular cells (EGCs), granulocytes, melanocytes and, in some instances, partially degenerated or vacuolated epithelioid cells were observed in close proximity to the cyst wall. Pigment-bearing macrophages were scattered throughout the granulomatous host-tissue reaction and as macrophage aggregates (MAs) within the capsules surrounding parasites. Immunohistochemical tests were applied to infected heart sections using 12 different antisera. Nerve fibres immunoreactive to bombesin, substance P (SP), and atrial natriuretic peptide (ANP) antisera were observed in close proximity to the parasite larvae. The presence of a serotonin-like substance was also observed within host immune-cells surrounding trematode cysts. Large cells of the epicardium were found to be immunoreactive to met-enkephalin and vasoactive intestinal peptide (VIP) antisera but not immunoreactive to anti-protein gene-product 9.5 (PGP9.5) sera.     

The ultrastructure of the spermatozoon of Neochasmus sp. (Cryptogonimidae, Digenea, Trematoda) and its phylogenetic significance

Jamieson B. G. M. and Daddow L. M. 1979. The ultrastructure of the spermatozoon of Neochasmus sp. (Cryptogonimidae, Digenea, Trematoda) and its phylogenetic significance. International Journal for Parasitology12: 547–559. The typical spermatozoon is filiform, monopartite and internally strongly asymmetrical, with a mean length of 254 μm. Its two axonemes are wholly incorporated though not deeply. The six major regions, in anteroposterior (proximal-distal) sequence, are: acrosomoid, prenuclear, uniaxonemal, nuclear, intermediate (amitochondrial), posterior mitochondrial and posterior amitochondrial regions. Centrioles and flagellar rootlets, present in the spermatid, are absent from the spermatozoon. The right axoneme (axial unit) originates more anteriorly than the left and anteriorly bears an acrosome-like projection (acrosomoid). Spinose regions carrying epiplasmalemmal projections are reported for the first time in the Digenea. Dimorphism exists as replacement of the typical posterior mitochondrial and amitochondrial regions by an amitochondrial region characterized by asymmetrical distribution of spines around one axoneme. Excepting known schistosomatid sperm, the digenean sperm is diagnosed by the combination of three characters: cortical microtubules, two fully incorporated 9 + 1 axonemes and presence of mitochondria although no one of these characters is limited to the group. Subtle intertaxon variations occur.     

Serological and Bacteriological Investigation of Mycobacterium ranae (fortuitum)

Twenty strains of a species of fast growing Mycobacterium, some named M. fortuitum and others said to be similar to the strain of M. peregrinum, in the possession of the National Collection of Type Cultures, were examined serologically and bacteriologically. They were all found to be of the same species, but they were divisible into four serotypes (I to IV), on a basis of three variable antigens, which do not exactly correspond to the subspecies described in previous reports. The strains varied in the number of years that they had been kept in culture and were of widely divergent origin. With the exception of serotype II strains, which all came from sputa from Welsh miners, the serotypes were independent of years in culture or site of origin. Based on bacteriological and serological identity and chronological priority, we consider that M. ranae (Kuster) Bergey et al. should be reinstated in place of M. fortuitum Cruz. A modified scheme for identification of the species is proposed, which would not exclude any of the four serotypes.     

Comparative cytogenetics of opisthorchid species (Trematoda, Opisthorchiidae)

In the present study karyotypes and chromosomes of five species of the family Opisthorchiidae (Opisthorchis felineus (Rivolta, 1884), O. viverrini (Poirier, 1886), Metorchis xanthosomus (Creplin, 1846), M. bilis (Braun, 1893), and Clonorchis sinensis (Cobbold, 1875)) were compared. Karyotypes of O. felineus, M. xanthosomus, M. bilis and C. sinensis consist of two pairs of large meta- and submetacentrics and five pairs of small chromosomes (2n = 14). The karyotype of O. viverrini is 2n = 12, which indicates a fusion of two chromosomes of opisthorchid ancestral karyotype. Analysis of mitotic and meiotic chromosomes was performed by heterologous in situ hybridization of microdissected DNA probes obtained from chromosomes 1 and 2 of O. felineus and chromosomes 1 and 2 of M. xanthosomus. Results of chromosome staining (C- and AgNOR-banding) and FISH of telomeric probes and ribosomal DNA probe on opisthorchid chromosomes were used for chromosome comparison. Data on chromosome number in opisthorchid species were also discussed.     

Surface ultrastructure of the developmental stages of Heterophyopsis continua (Trematoda: Heterophyidae).

Ultrastructural observations were made on the tegument of juvenile and adult stages of Heterophyopsis continua using scanning electron microscopy. On the surface posterior to the ventral sucker, the tegumental processes were bandlike in the metacercariae, cobblestonelike in the flukes 2 days postinfection (PI), and velvety at 3 days PI. The anterior surface between the oral and ventral suckers of the metacercariae was packed densely with tegumental spines having a 10- to 14-pointed tip. In flukes 6 days PI, the number of points increased to 15-17. The tegumental spines immediately behind the ventral sucker on the metacercariae surface possessed 5-7 points; posteriorly the points were reduced in size and in number. Ciliate sensory papillae (type I), as single or clumped forms of 2 or 3, were abundant around the oral and ventral suckers of metacercaria and adult worms. The clumped papillae appeared bilaterally symmetrical on the dorsal and ventral surfaces. The ciliate papillae may function in tango-, rheo-, and/or chemoreception. On the lip of the ventral sucker, 6-7 aciliate domed papillae (type II) were arranged in an equidistant manner. At 2 days PI each type II papilla became a clumped form having 2 or 3 papillae. Type II papillae may function as tango- and/or pressure-receptors. The structure and distribution of papillae suggest that the ventral sucker likely functions as a holdfast organ and the oral sucker as a probing organ involved in feeding.