Host-parasite interface of Paratenuisentis ambiguus (Eoacanthocephala) in naturally infected eel and in laboratory-infected sticklebacks and juvenile carp and rainbow trout

Paratenuisentis ambiguus is described from natural infections in adult eels and from laboratory infections in sticklebacks and juvenile carp and rainbow trout. In captured eels, female worms kept reproducing in the laboratory for at least 1 mo. In the 3 small laboratory hosts female worms did not release eggs and longevity did not exceed 3-30 days. It is concluded that the small fishes were unsuitable final hosts. Worm penetration into the intestinal wall of all hosts was shallow. Thus, the small fishes also proved to be unsuitable paratenic hosts. The worms ruptured the intestinal mucosa and the underlying tunica propria and they often seemed to change their sites of attachment. The proboscides carried an osmiophilic surface coat that seemed to be supported by liquid drops from necrotic host tissue and by osmiophilic material apparently discharged from pores in the worm's proboscis hooks. The coat contained lipids, polysaccharides, and/or proteoglycans and likely other substances. Around the hooks the proboscis tegument harbored conspicuous cisternae of rough endoplasmic reticulum as is typical for cells with secretory function. Mostly, the worms were found with semi-invaginated proboscides. The resulting cavity inside the proboscis seemed to collect lipids and other remnants of host cells from the lesions caused by the worms. Whether the apical hollow might function as a gastric cavity is discussed.     

Description and pathology of Neoechinorhynchus idahoensis n. sp. (Acanthocephala: Neoechinorhynchidae) in Catostomus columbianus from Idaho.

Neoechinorhynchus idahoensis is described from Catostomus columbianus caught in the Salmon River, Stanley Basin, Idaho. The new species is closest to Neoechinorhynchus venustus Lynch, 1936, but is distinguished from it by its smaller and variably structured eggs, anterio-dorsal trunk hump, bent and posteriorly notched proboscis receptacle, and larger proboscis, proboscis receptacle, and hooks. It is distinguished also from 2 other species of Neoechinorhynchus with proboscis hooks in middle and anterior circles about equally large and from 7 other species having lemnisci greatly unequal in length. Histopathology of host tissue showed limited host response exemplified by epithelial damage and hemorrhaging at point of proboscis attachment with subsequent macrophage and other phagocytic cell migration. The proboscis extended through the host epithelium into the submucosa with limited hemorrhaging at the point of attachment. Unorganized collagenous fibers were present. The lumen of the host intestine was obstructed, and compressed villi were present. The trunk of the worm damaged intestinal epithelium near the crypts, causing localized inflammation. The caryophyllaeid cestode Isoglaridacris calentinei Mackiewicz, 1974, was present in concurrent infections of C. columbianus.     

Investing in attachment: evolution of anchoring structures in acanthocephalan parasites

Secure attachment to host tissues is essential for survival and reproduction in parasitic organisms. The production of elaborate attachment structures must be costly, however, and investments in attachment should be approximately proportional to the likelihood that a parasite will be dislodged. In the present study, relative investments in attachment as a function of body size and the type of host used were examined across 138 species of acanthocephalans. These worms live anchored to the intestinal wall of a vertebrate host by inserting their hooked proboscis into host tissues. Taking proboscis volume into account, there is a negative interspecific relationship between the number of hooks borne on the proboscis and their mean length, reflecting a trade‐off between hook number and hook length. This supports the assumption that hooks are costly to produce, because any given species cannot simultaneously maximize both the relative number and relative length of the hooks it produces. There is a positive relationship between total worm size and total hook length, but it is weak, with a slope indicating that, as total body volume increases, total hook length also increases but at a slower rate. Indeed, relative investments in attachment, measured as hook length per unit body volume, decrease as worm size increases. Independently of total body size, investments in hook production are higher in species exploiting endothermic hosts, especially birds, than in those living in ectothermic hosts. Given the greater amounts of food passing through the gut of endotherms, and the richer and denser communities of intestinal parasites that they harbour, they are likely to select for greater investments in attachment. These results support the prediction that investments in attachment are influenced by the probability of being dislodged, and allow comparisons with other groups of intestinal parasites such as cestodes or trematodes. © 2007 The Linnean Society of London, Biological Journal of the Linnean Society, 2007, 90 , 637–645.     

Antigenic localities in the tissues of Metagonimus yokogawai observed by immunogoldlabeling method]

In order to determine the antigenic localization in the tissues of the adult Metagonimus yokogawai, immunogoldlabeling method was applied using serum immunoglobulins(IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex(particle size: 12 nm). It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.     

鱊头槽绦虫原尾蚴皮层的超微结构观察

利用透射电镜观察了头槽绦虫 (Bothriocephalusacheilognathi)原尾蚴皮层的超微结构。头槽绦虫原尾蚴的皮层为典型的合胞体结构。皮层表面有浓密的微毛与少量的突起。与成虫相比 ,原尾蚴的微毛长而且粗 ,呈现单态性 ,推测在进入终末宿主的过程中有脱落和再生现象。突起分布在头部与体侧 ,含有较少的电子致密颗粒。在原尾蚴皮层细胞质中观察到三种腺细胞的分泌过程 ,即顶分泌、外分泌和微分泌过程。原尾蚴身体后部观察到尾钩和穿刺腺管道的开口。在核周区和纤维层下可见发育较好的环肌与纵肌。本文还对原尾蚴皮层突起的功能进行了探讨。     

Functions of the tegument of schistosomes: clues from the proteome and lipidome

The tegumental outer-surface of schistosomes is a unique double membrane structure that is of crucial importance for modulation of the host response and parasite survival. Although several tegumental proteins had been identified by classical biochemical approaches, knowledge on the entire molecular composition of the tegument was limited. The Schistosoma mansoni genome project, together with recently developed proteomic and lipidomic techniques, allowed studies on detailed characterisation of the proteins and lipids of the tegumental membranes. These studies identified tegumental proteins and lipids that confirm the function of the tegument in nutrient uptake and immune evasion. However, these studies also demonstrated that compared to the complete worm, the tegument is enriched in lipids that are absent in the host. The tegument is also enriched in proteins that share no sequence similarity to any sequence present in databases of species other than schistosomes. These results suggest that the unique tegumental structures comprise multiple unique components that are likely to fulfil yet unknown functions. The tegumental proteome and lipidome, therefore, imply that many unknown molecular mechanisms are employed by schistosomes to survive within their host.     

Schistosoma mansoni: changes in the outer membrane of the tegument during development from cercaria to adult worm

Hockley D. J. and McLaren D. J. 1973. Schistosoma mansoni: changes in the outer membrane of the tegument during development from cercaria to adult worm. International Journal for Parasitology3: 13–25. The tegumental outer membrane of the cercaria is trilaminate: the adult worm, however, has a seven-layered membrane. Formation of the heptalaminate membrane commences immediately after the cercaria has penetrated the vertebrate host: multilaminate membrane-bounded vacuoles are passed from subtegumental cells into the tegument where they enlarge, join to the outer membrane and open to the exterior. The heptalaminate limiting membrane of the vacuole thus becomes the outer membrane of the tegument. At the same time the original trilaminate tegumental membrane is formed into microvilli which are cast off and thus the cercarial outer membrane is lost. Schistosomula usually have a heptalaminate outer membrane within three hours of penetration. After this time the large vacuoles are replaced by smaller membraneous bodies which presumably contribute to the outer membrane during growth of the schistosomulum. The membraneous bodies are also present in the tegument of the adult worms and there is some evidence that the outer membrane is continually renewed.     

Tegumental changes in adult Schistosoma mansoni harbored in mice treated with artemether

A detailed temporal examination was made of alterations induced by artemether in the tegument of adult Schistosoma mansoni worms using scanning electron microscopy (SEM). Mice infected with S. mansoni cercariae 42 days previously were treated intragastrically with artemether at a single dose of 400 mg/kg. Groups of 3 mice were killed at 24 hr, 72 hr, and 7 days after treatment; the worms were collected by perfusion and examined by SEM. Twenty-four hours after artemether treatment, focal damage to the tubercles on the tegumental surface of male worms was seen. In both male and female worms, there was focal swelling and fusion of tegumental ridges, and sometimes peeling. After 72 hr, the damage to the tegument had increased, especially in female worms, with extensive swelling, fusion, and peeling of the tegumental ridges. In the most severely damaged worms, host leukocytes were seen to be adhered to the damaged tegument. Damage to the oral sucker was also occasionally seen in both male and female worms. Seven days after treatment, the appearance of the tegument had returned to normal in some male and female worms, whereas others still showed apparent damage. The results demonstrate that artemether damages the tegument of adult S. mansoni, and the intensity of damage is more severe in female worms than in males.     

Suppressing Glucose Transporter Gene Expression in Schistosomes Impairs Parasite Feeding and Decreases Survival in the Mammalian Host

Adult schistosomes live in the host''s bloodstream where they import nutrients such as glucose across their body surface (the tegument). The parasite tegument is an unusual structure since it is enclosed not by the typical one but by two closely apposed lipid bilayers. Within the tegument two glucose importing proteins have been identified; these are schistosome glucose transporter (SGTP) 1 and 4. SGTP4 is present in the host interactive, apical tegumental membranes, while SGTP1 is found in the tegumental basal membrane (as well as in internal tissues). The SGTPs act by facilitated diffusion. To examine the importance of these proteins for the parasites, RNAi was employed to knock down expression of both SGTP genes in the schistosomula and adult worm life stages. Both qRT-PCR and western blotting analysis confirmed successful gene suppression. It was found that SGTP1 or SGTP4-suppressed parasites exhibit an impaired ability to import glucose compared to control worms. In addition, parasites with both SGTP1 and SGTP4 simultaneously suppressed showed a further reduction in capacity to import glucose compared to parasites with a single suppressed SGTP gene. Despite this debility, all suppressed parasites exhibited no phenotypic distinction compared to controls when cultured in rich medium. Following prolonged incubation in glucose-depleted medium however, significantly fewer SGTP-suppressed parasites survived. Finally, SGTP-suppressed parasites showed decreased viability in vivo following infection of experimental animals. These findings provide direct evidence for the importance of SGTP1 and SGTP4 for schistosomes in importing exogenous glucose and show that these proteins are important for normal parasite development in the mammalian host.     

Possible immunological damage to the tegument of Hymenolepis diminuta in mice and rats.

Opaque or darkened areas (DA) of variable size and position occur on Hymenolepis diminuta in mice and rats. In mice DA normally first appear in the neck region of the worm but subsequently they appear elsewhere and increase in number until destrobilation or worm expulsion. The posterior of destrobilated worms is often darkened. In the more immunogenic infections with six cysticercoids there are more DA per worm than in infections with one cysticercoid. DA are areas of the tegument with a homogeneous increase in electron density; abnormal mitochondria; reduced granular endoplasmic reticulum, Golgi complexes and discoidal secretory bodies; and accumulation of lipid droplets. DA disappear from worms maintained for up to 4 h in Hanks' balanced salt solution and can be induced by mechanical damage to the worms. As the numbers of DA increase with the duration and intensity of infection and have similarities with types of cell injury, they are probably sites of worm pathology induced by host immunity.     

Ultrastructural localization of Sm15 and Sm25, two major tegumental adult worm antigens of Schistosoma mansoni.

Sm15 and Sm25 are two of the principal tegumental antigens recognized by antibodies from mice protectively vaccinated with adult worm tegumental membranes and may therefore be potential vaccine candidate antigens. Using antibodies affinity purified from anti-tegumental membrane anti-sera, and antibodies raised against the recombinant antigens, Sm15 and Sm25 were shown to be located specifically in the tegument of adult worms being distributed throughout the syncitium but not associated with the outer membrane.     

The effect of host species on growth and variability of Echinorhynchus salmonis Müller, 1784 (Acanthocephala: Echinorhynchidae), with special reference to the status of the genus

Summary The sex and age (as measured by length) of Echinorhynchus salmonis Müller, 1784 and the host species of this acanthocephalan considerably affected worm body form and size, as well as size of proboscis, proboscis hooks, proboscis receptacle, lemnisci, testes and cement glands. Linear regression analysis indicated that curves describing the growth pattern of these characters by worm length were significantly different as a function of host species. The larger worms recovered from bloater (Coregonus hoyi: Salmonidae) almost invariably showed higher regression coefficient compared to those from smelt (Osmerus mordax: Osmeridae) in all characters. Taxonomic implications of these findings are discussed. Abnormalities in body wall, proboscis hook orientation, lemnisci and cement gland ducts as well as variations in proboscis hook and cement gland numbers are reported, some for the first time. Findings from studies of cement gland pattern invalidate Petrochenko's (1956) splitting up of the genus into three: Echinorhynchus, Metechinorhynchus and Pseudoechinorhynchus. It is proposed that the designation of the last two genera as junior synynoms of the first be accepted. ac]19791205     

Antigenic localities in the tissues of the young adult worm of Paragonimus westermani using immunogold labeling method

In order to observe the antigenic localization in the tissues of the young adult Paragonimus westermani, immunogold labeling method was applied using serum immunoglobulins(IgG) of the dog which infected with isolated metacercariae from Cambaroides similis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size; 12 nm). It was observed by electron microscopy at each tissues of the worm. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were predominantly labeling on the epithelial lamela and lumen of caecum. The above finding showed that antigenic materials in young adult worm tissue were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells.     

Megalodiscus temperatus: Scanning electron microscopy of the tegumental surfaces

The tegumental surface of Megalodiscus temperatus forms cobblestonelike areas with rows of indentations encircling the worm. This pattern merges in several areas into folds and ridges, some of which represent the musculature of the posterior sucker and genital pore. Papillae surrounding the base of the oral sucker appear as two types: one with a bulb-like base and a short apical knob; the second typified by a hair-like structure (cilium?) of variable length projecting from a pit. From their location on the oral sucker and the resemblance to previously described structures, they are presumed to be sensory receptors. A circle of papillae on the closed posterior sucker was also observed. Rod-shaped bacteria were seen adhering to some of the worms observed but they were not found to be in any special association with the tegument or in any specific areas of the worm's surface.     

Induction cues for tegument formation during the transformation of Schistosoma mansoni cercariae

Adult schistosomes are parasitic blood flukes that have a continuous double lipid bilayered membrane surrounding the entire worm. This tegumental membrane is synthesised during invasion of the vertebrate host by free-swimming infectious forms called cercariae. As cercariae invade their final hosts they lose their tails and encounter a changing environment that includes altered temperature, sugar concentration and osmolarity. We have identified a glucose transporter protein designated SGTP4 that is found exclusively in the outer adult tegument and on membranous vesicles within the tegumental cytoplasm. By using immunofluorescence analysis to monitor the appearance and distribution of SGTP4 we can track the process of new tegumental membrane formation and examine the cues that trigger this developmental pathway. Cercariae in water do not transform their tegument while those incubated in rich medium do so rapidly. We have examined which of the many constituents of rich medium are responsible for triggering this transformation. Incubation in a solution of moderate osmolarity (120 mOsM PBS) is sufficient by itself to trigger tegument transformation, albeit at a slower rate relative to incubation in rich medium. Adding either glucose (to 100 mM) to the solution or increasing the temperature of incubation (from 22 degrees C to 37 degrees C) further increased the rate of tegument biogenesis. The introduction of glucose together with an increase in the incubation temperature further accelerated the process, suggesting that these factors act synergistically to promote transformation rates. The critical nature of osmolarity in inducing the process is highlighted by the fact that transformation proceeds as efficiently in 360 mOsM alone as it does in rich medium. While the fatty acids linolenic acid (cis-9, cis-12, cis-15-octadecatrienoic acid at 1 mM) and capric acid (Decanoic acid, at 0.1 mM) have both been proposed to stimulate tegumental transformation, we show that neither promotes the morphogenesis of a normal schistosomulum tegument. The schistosomicide praziquantel (to 1 mM) has no detectable effect on new tegument formation.     

Schistosoma mansoni: Electron probe X-ray microanalysis of the elemental composition of the tegument and subtegumental tissues of adult worms

The elementary composition [Na, Mg, P, S, Cl, K, Ca and Fe] of the tegument, tegumental spines, and subtegumental tissues of adult male and female Schistosoma mansoni have been determined by electron probe X-ray microanalysis of unfixed, freeze-dried cryosections. Statistical analysis of the results suggests that there are distinct differences in the elemental composition of the tissues both between and within individual male and female worms, and between male and female worms in general. In particular, there were significant variations in the elemental contents of the tissues between individual male and female worms, which may reflect differences in the physiology and/or metabolic state of the worms. Significant differences in the elemental composition of the various tissues examined within individual worms were also found. In general, in both male and female worms, there were significantly higher elemental levels in the tegument, as opposed to the subtegumental tissues. The elemental composition of the tegumental spines in both male and female worms differed from that of the tegumental cytoplasm, although the differences in the elemental composition between spines from male and female worms reflected the differences in the elemental content between the teguments themselves. Differences in the elemental composition of the tissues between male and female worms were also found, with the female tegument containing significantly higher elemental levels (with the exception of Cl) than the male tegument. In particular, the tegument of female worms contained higher levels of calcium and, in relatively small areas, isolated calcium-containing granules. This higher tegumental calcium level in female worms may reflect a higher calcium demand by sexually mature female worms due to the presence, within the mature vitelline cells, of calcium-containing corpuscles and the production of large numbers of eggs.     

Schistosoma mansoni: topochemical features of cercariae, schistosomula, and adults

The teguments of developing and mature cercariae, recently transformed, and 1-wk-old schistosomula and adult worms were examined for the ultrastructural location of macromolecular carbohydrates and polyelectrolytes. The surface of mature cercariae within sporocysts and cercariae released from the snail is covered by a filamentous coat which reacts with cytochemical reagents for the demonstration of vicinal glycols, but neither the coat nor the surface of the tegument plasmalemma binds cationic colloidal iron at low pH.Upon penetrating mammalian skin, the cercaria sheds its surface coat; the tegument surface of newly transformed schistosomula, older schistosomula and adult worms stains en bloc with acidic colloidal iron, as does the tegument plasmalemma of mature cercariae if the overlying filamentous coat is first removed by physicochemical means. The cercarial coat thus serves to mask anionic groups at the surface of the tegument plasmalemma which become functionally exposed after penetration of the mammalian host. The distribution of colloidal iron binding sites coincides with those for the carbohydrate-complexing phytohemmagglutnin, concanavalin A, which suggests that these membrane-fixed anions are acid mucopolysaccharides, glycoproteins or glycolipids. Carbohydrate-containing material was also localized within membrane-bound vesicles of the tegument matrix and perikarya of developing cercariae and postcercarial schistosomes, suggesting that surface mucosubstances contributing to the tegument glycocalyx of these worms are elaborated, at least in part, by the tegument itself.     

Immunological damage to Hymenolepis diminuta following a challenge infection in C57 mice

Immunological damage to Hymenolepis diminuta following a challenge infection in C57 mice. International journal for Parasitology17: 795–803. Ultra-structural observations were made on the scpleces of Hymenolepis diminuta from 10-worm 2° infections in C57 mice to determine whether the immune stress causing reduced establishment, stunting of growth and early rejection was accompanied by significant damage to the worm fine structure. In almost all of the worms recovered there was extensive damage to tissues throughout the scolex, even in those examined 24 h post-secondary-infection (p-2°-i). Many of the muscle, tegumental and parenchyma cells were completely disrupted and dispersed, leaving only isolated strands of cytoplasm and cell debris. A number of excretory ducts were breached, and large and apparently empty spaces were evident throughout the scolex tissues. There was an increase in lipid accumulation in the tegumental syncytium, underlying cytons and in the suckers. Extensive vacuolation also occurred in the sucker tegument. Although some scoleces showed certain regions to be apparently undamaged, close examination revealed a fine structure consistent with enhanced metabolic activity. The structural abnormalities observed are discussed in relation to host immunity.     

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.     

Hymenolepis diminuta: ultrastructural abnormalities in worms from C57 mice

Ultrastructural studies, including stereological analyses of micrographs, have been made of five-worm primary infections of Hymenolepis diminuta from C57 mice to determine whether the immune destrobilation/rejection process was accompanied by significant changes in the fine structure of the scolex tegument. Destrobilation/rejection of worms occurred from Days 9-12 after infection. For the first 5 days after infection, the scolex tegument showed no detectable differences in ultrastructure compared with that of "control" worms from either Wistar rats or immunosuppressed C57 mice. By Day 6, large lipid deposits were observed in the tegument and associated musculature of worms from untreated C57 mice. Further, worms recovered from Days 6-8 after infection also showed increased activity of the Golgi apparatus, GER, and mitochondria of the tegument, resulting in increased numbers of discoidal secretory bodies. Concomitant with destrobilation/rejection from Day 9 was a drop in the number of secretory bodies, an increase in autophagic activity throughout the tegument, and a blistering of the tegument surface plasma membrane. The possible functional significance of the results is discussed in relation to host immunity.