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Characterization of the carbohydrates of Schistosoma japonicum adult worm, egg and cercaria by analysis of lectin binding and antibody reaction.

Binding of the FITC-labeled lectins, Con A, SBA, WGA, DBA, UEA 1 and RCA 120 were examined on various kinds of cross-sections of S. japonicum adult worms and eggs, and on cercaria specimens. The specificity of the lectin bindings was examined by inhibition test with specific carbohydrates or with unlabeled lectins. Furthermore, the locations of lectin binding in schistosomes were compared with those of the antigen-antibody reactions using the fluorescence technique. It was also studied as to whether lectin binding influenced antigen-antibody reactions. Thus it was found that most lectin binding receptors were located at the same sites, where also FITC-labeled antibodies reacted. But antibodies did not inhibit lectin binding on schistosomes, whereas lectins blocked the antigenic sites to weaken the following antibody reactions on microscope slides. Con A did not show a distinguished pattern on schistosomes, although its carbohydrate specificity was confirmed. On the contrary, the WGA reactions were hardly considered to be carbohydrate specific. PNA and SBA receptors, however, were selectively detected in the egg, testes and adult tegument of schistosomes, corresponding to the sites, where also egg antigenicity was observed. But in the gut of adult schistosomes, in which egg antigenicity was not found, lectin binding with PNA, SBA, UEA 1 and DBA did not occur. Gut reacted only with RCA 120 and Con A. The lectin specificity on tegument of adult schistosomes and on cercaria was similar to that found on egg and testes. The result indicated possibilities of satisfactory purification of schistosomal antigens by using such lectins as PNA, SBA and RCA 120, instead of Con A and WGA, to separate more defined antigenic components.

Animals↗

[Integumentary argentophilic structures in larval stages (miracidium, redia, cercaria) of Fasciola hepatica (Trematoda, Fasciolidae). Comparison with F. gigantica (author's transl)].

Integumentary argentophilic structures of the miracidium and cercaria are described and with those of F. gigantica compared. In the miracidia, the pattern of the organites of the terebratorium and the arrangement of the papillae between the first and second tier of epidermal plates are similar for the two species. However, the disposition of the medio-ventral plate of the second tier allows the miracidia of the two related species to be distinguished statistically. In the cercariae, the chaetotaxy differs between F. hepatica and F. gigantica in two cephalic regions and on the tail (presence of UD papillae in F. hepatica, absence in F. gigantica). The chaetotaxy of a daughter-redia of F. hepatica is also described. There is an important concentration of papillae around the buccal aperture, whereas at the pharyngeal and post pharyngeal levels, papillae are more spaced on the side supposed to be ventral, and scarce on the side considered dorsal.

Animals↗

Murine immunization by cesium-137 irradiation attenuated Schistosoma mansoni cercariae.

Cesium-137, becoming a more readily available ionizing gamma radiation source for laboratory use, was shown to effectively attenuate Schistosoma mansoni cercariae for vaccine production. In parallel comparison studies with the murine model, cesium-137 attenuated cercariae consistently afforded better (P greater than 0.05) protection than did the cobalt-60 prepared vaccine. Dose-response data indicated that the optimal total irradiation with cesium-137 was between 45 and 50 Krad.

Animals↗

Mechanisms of protective immunity against Schistosoma mansoni infection in mice vaccinated with irradiated cercariae. II. Analysis of immunity in hosts deficient in T lymphocytes, B lymphocytes, or complement.

Laboratory mice with genetic or induced immunodeficiencies were vaccinated with irradiated cercariae of Schistosoma mansoni and assayed 4 wk later for their resistance to challenge infection. Athymic nude mice failed to develop immunity to challenge or detectable antibody responses to schistosomula, whereas heterozygote (nu/+) controls and nude mice with thymus grafts displayed highly significant levels of resistance and anti-schistosomulum antibody. Similarly, no resistance or antibody production as observed in vaccinated mice that had been made deficient in B lymphocytes by the injection of anti-mu-chain antisera from birth. In contrast, normal levels of vaccine-induced resistance were observed in mice genetically deficient in the fifth component of complement (C5) and in mice decomplemented before challenge by treatment with cobra venom factor. These results indicate that the resistance to challenge infection induced by irradiated cercariae is both thymus and B lymphocyte dependent and therefore is likely to result from specific immune responses directed against schistosome worms. The data also argue against a role for complement in the effector mechanism of vaccine-induced immunity.

Animals↗

[The chronobiology of the cercaria of Ribeiroia marini (Faust and Hoffman, 1934) parasite of Biomphalaria glabrata: effect of the photoperiod on the rhythm of emergence].

Ribeiroir marini cercariage emerge from the snail-host during the night, according to a circadian rhythm. The photoperiod is the synchronizer of this rhythm. Inversion of the photoperiod is immediately followed by inversion of the periodicity of emergence. No cercaria emerge in continuous light conditions. This inhibitory effect of light exerts its influence on the emergence but not on the production of cercariae. The rhythmic emergence is almost maintained in continous dark, but darkness cannot induce emergence at any time. It is possible that an innate production rhythm interferes with the emergence rhythm.

Biomphalaria↗

Skin surface lipids of the domestic chicken, and neutral lipid standards as stimuli for the penetration response of Austrobilharzia variglandis cercariae.

Lipids were extracted from the skin of 2-wk-old domestic chickens using sterile cotton gauze dampened with chloroform:methanol (2:1). Preparative thin-layer chromatography separated the skin lipids into six major fractions: phospholipids, free sterols, free fatty acids, triglycerides, methyl esters, sterol esters. The penetration response of the marine avian schistosome cercaria, Austrobilharzia variglandis, to chicken skin lipid fractions, and to neutral lipid standards, was tested by coating lipids on agar in a Petri dish containing a seawater overlay. All neutral lipids tested produced significantly greater penetration responses than the chloroform control. The phospholipid skin fraction killed cercariae. Lipid from whole chicken skin produced the greatest penetration response, followed by free fatty acids and free sterol skin fractions. Of the standards tested, the whole neutral lipid standard, containing cholesterol, oleic acid, triolein, methyl oleate, and cholesteryl oleate, produced the greatest response, followed by the cholesterol standard and the oleic acid standard.

Animals↗

Cultivation of Schistosoma mansoni in vitro. I. Establishment of cultures from cercariae and development until pairing.

Procedures are described for mass cultivation of Schistosoma mansoni. Cercariae were pooled, concentrated, and axenized through a series of washes in base medium containing antibiotics. Cercariae were sheared through a double-Luer-ended needle connected to two syringes, and tails separated and discarded. Young schistosomules were grown in a medium based upon BME augmented with lactalbumin hydrolysate, glucose, hormones, and other amendments and supplemented with human serum. Human blood cells (Type O) were added to cultures. Intestinal pigment was seen on the 5th day and gut cecal fusion began on day 11. Schistosomules developed steadily to pairing, which was seen first during the 7th wk of culture. Pairing occurred somewhat later in relation to development in vivo.

Animals↗

Chaetotaxy of Psilotrema spiculigerum (Mühling, 1898) Odhner, 1913 (Trematoda: Psilostomatidae) cercariae.

The chaetotaxy of P. spiculigerum mature cercariae was studied. Unlike from the known in the literature up to now, the chaetotaxy of the cercariae was complemented as follows: a - the number and arrangement of the papillae on the dorsal, ventral and lateral surface of the body were first described; b - the arrangement of the papillae on the cephalic region and tail was corrected.

Animals↗

[A comparative study of excretory and sensory apparatus of two cercariae of Diplodiscus parasites of European and African Amphibians (Trematoda, Diploidiscidae) (author's transl)].

The comparative study of the excretory system provides new data on the number of flame cells of two cercariae of Diplodiscus from Poland and Togo but does not allow to distinguish them. On the other hand, the comparison of the superficial sensory apparatus shows significant differences. The cercariae from Togo differ from these of Poland by a much lower number of all the cephalic papillae and papillae of cycle Cv, a higher number of acetabular papillae, and the presence of 9 to 12 pairs of dorsal papillae in the median part of the tail. Therefore the African Diplodicus is not D. subclavatus. We identify it as D. fischthalicus.

Amphibians↗

Morphology and ultrastructure of the redia and pre-emergent cercaria of Opisthorchis viverrini (Trematoda: Digenea) in the intermediate host Bithynia siamensis goniomphalus (Prosobranchia: Bithyniidae).

The intramolluscan stages of the human liver fluke Opisthorchis viverrini were investigated by light, scanning, and transmission electron microscopy. The morphology of the tegument, and the excretory system of the redia and pre-emergent cercaria as well as glandular cells, tail structure, and the eye spots of the cercarial stage are described. Possible environmental adaptations are discussed. The redia is characterized by a tegument which is folded concentrically and covered with densely arranged microvilli. This tegument is composed of a typical syncytium with submerged tegumental cells. The pre-emergent cercaria is characterized by a long tail with prominent fin folds and obliquely striated muscles. The oral sucker is well developed and protrusible, whereas the ventral sucker is faintly developed. Proteinaceous tegumental spines which are arranged transversally and different kinds of presumed sensory structures cover the cercarial body surface. Two well pigmented eye spots with rhabdomeric photoreceptors are located in the anterior part of the body. An extended protonephridial system is connected to a voluminous excretory bladder. The excretory tube opens at the end of the body and does not lead into the tail. Three types of glandular cells could be differentiated according to their vesicles, which are shed into the syncytial layer.

Animals↗

Enhanced efficacy of praziquantel treatment in mice vaccinated with radiation attenuated cercariae of Schistosoma mansoni.

Normal, and vaccinated mice four weeks previously with radiation attenuated cercariae of S. mansoni (500/mouse) were challenged with normal cercariae (150/mouse), then treated one week later with praziquantel (400 mg/kg body weight, orally). Worm burden was determined to calculate the % immunity in all groups under study. Histopathological examination of liver, small and large intestine, spleen and lung was done. Serum IgE level was estimated using the immunoradiometric assay. (IRMA). The % immunity was highest among vaccinated, infected and treated group with minimal pathological changes recorded and highest IgE level. From the data collected, it was found that, the efficacy of praziquantel treatment was enhanced in vaccinated mice and that there was synergistic effect between drug treatment and vaccination when praziquantel was given seven days post-infection (challenge).

Animals↗

Experimental infection of juvenile Biomphalaria glabrata with cercariae of Echinostoma trivolvis.

Infectivity studies were done on laboratory-reared Biomphalaria glabrata juvenile (neonatal) snails exposed to cercariae of Echinostoma trivolvis. Of the 40 snails, 0.7-1.0 mm in shell diameter, which were exposed to a total of 1,000 cercariae for 24 hr, all were infected with 1-29 viable encysted metacercariae per snail. Biomphalaria glabrata, 1-2 mm in shell diameter, were used to determine the effects of an infectious dose of 25 cercariae/juvenile on subsequent snail survival. The median survival time of 24 exposed snails occurred between day 3 and 4 postinfection, at which time more than 90% of 24 control (unexposed) snails were alive. The high mortality in the E. trivolvis-infected B. glabrata juveniles was probably related to cercarial penetration and encystment in the snail tissue, although histological examination of snails after exposure would be required to determine the actual cause of the observed mortality.

Animals↗

[Observation on antibody levels of rabbits infected with single sex cercariae of Schistosoma japonicum].

Nine rabbits infected with male S. japonicum cercariae (worm recovery 15-133, mean 53.1) and 9 rabbits infected with female cercariae (worm recovery 1-102, mean 36.1) were bled periodically until 1 year post-infection. The sera were tested by ELISA, LA and CHR for antibody detection. Results showed that the positive rates were 88.9%, 55.6% and 83.3%, respectively. The duration of positive reaction was 3-51 wk post-infection for ELISA, 4-17 wk for LA and 2-35 wk for CHR and the fluctuated results were obtained in ELISA while positive CHR and LA results usually remained stable. The above-mentioned results suggest that antibody detection methods should not be neglected due to its sensitivity although antigen detection methods are considered more favorable. For the surveillance of schistosomiasis in endemic areas where schistosomiasis is under control, antibody detection methods are still worthy of recommending for case finding of new infections.

Animals↗

Factors affecting the acquisition of Plagiorchis noblei (Trematoda: Plagiorchiidae) cercariae by black fly (Diptera: Simuliidae) larvae and the effect of metacercariae on host survival.

Larvae of 4 species of black flies, Prosimulium mixtum, Simulium vittatum, S. decorum and Stegopterna mutata, were exposed in the laboratory to cercariae of the digenean Plagiorchis noblei in flowing water. Prevalence and intensity of infection of all species varied directly with exposure intensity and decreased with increasing water velocity; prevalence increased with larval size. Infection levels for P. mixtum were higher than for the 3 other species. Mortality among infected larvae of all species was 3 times that of controls. Exposure of black fly larvae to P. noblei cercariae may adversely affect their survival.

Animals↗

In vitro development of Schistosoma mansoni cercariae.

Early cercarial embryos of Schistosoma mansoni developed to swimming cercariae within seven days in vitro, via the same sequence of stages as reported in vivo. The presence of living Biomphalaria glabrata cells (Bge line) was indispensable for proper development. Cultured cercariae were water-sensitive, gave no cercarienhüllen reaction, and appeared to lack the surface glycocalyx. Further development was not detected in mice or hamsters.

Animals↗

Surface ultrastructure of the cercaria of Neodiplostomum seoulense (Trematoda: Neodiplostomidae).

The surface ultrastructure of Neodiplostomum seoulense cercariae was studied by scanning electron microscopy. The cercarial body was cylindrical and its tail bifurcated. Short tegumental spines, with a broad base and single pointed tip, were dense anteriorly but sparse posteriorly on the lateral margins of the body. The acetabulum had stout spikelike spines, centripetally oriented on the inner surface. Sensory papillae, each with a short cilium, were concentrated around the oral sucker; they could be tangoreceptors or chemoreceptors, or both. Many sensory papillae, each with a long cilium, were observed on the tail; these might be rheo- or tangoreceptors. It is suggested that the sensory papillae of N. seoulense cercariae provide for attachment and penetration into the skin of tadpoles, which are the second intermediate host.

Animals↗

In vivo labeling of Schistosoma japonicum cercariae with 35S.

A technique was developed for in vivo labeling of adult Schistosoma japonicum worms using 35S bound to methionine and cysteine. The isotope was applied in an in vivo labeling system in which Oncomelania hupensis snails harboring a patent S. japonicum were individually exposed to the label. Following exposure to cercariae, mice were perfused after 6 wk. For autoradiography and fluorography, the worms were mounted on cardboard, covered with x-ray films, and exposed for 5 wk. With both techniques, images of both male and female worms were obtained in the 1- and 2-microCi groups, but not in the 0.5-microCi group. Fluorography proved more sensitive than autoradiography. Snail survival, labeled cercariae, and worm establishment were not affected by the radiolabeling. 35S was thus found useful for radiolabeling adult S. japonicum worms.

Animals↗