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Gene transcription profile in mice vaccinated with ultraviolet-attenuated cercariae of Schistosoma japonicum reveals molecules contributing to elevated IFN-gamma levels.

Vaccination with ultraviolet-attenuated cercariae of Schistosoma japonicum induced protective immunity against challenge infection in experimental animal models. Our preliminary study on the transcription levels of IFN-gamma and IL-4 in splenic CD4+ T cells revealed that attenuated cercariae elicited predominantly a Th1 response in mice at the early stage, whereas normal cercariae stimulated primarily Th2-dependent responses. Further analysis on the gene profile of the skin-draining lymph nodes demonstrated that the levels of IFN-gamma were significantly higher in vaccinated mice than those in infected mice at day 4, 7 and 14 post-vaccination or post-infection. However, for IL-12 and IL-4, the potent inducers of Th1 and Th2 responses, respectively, as well as IL-10, there were no differences over the course of the experiment between the infected and vaccinated mice. To explore the underlying factors that may potentially contribute to elevated IFN-gamma in vaccinated mice, the mRNA profiles of the skin-draining lymph nodes at day 4 post-exposure were compared using oligonucleotide microarrays. Within the 847 probe sets with increased signal values, we focused on chemokines, cytokines and relevant receptors, which were validated by semi-quantitative RT-PCR. A comprehensive understanding of the immune mechanisms of attenuated cercariae-induced protection may contribute to developing efficient vaccination strategies against S. japonicum, especially during the early stage of infection.

Animals↗

Lacto-N-fucopentaose III (Lewis x), a target of the antibody response in mice vaccinated with irradiated cercariae of Schistosoma mansoni.

Carbohydrates on soluble egg antigens are major epitopes for the antibody responses of patients and mice infected with Schistosoma mansoni. Recently, protective sera of mice vaccinated with irradiated cercariae were shown to recognize carbohydrate epitopes on schistosomal glutathione S-transferase. The present study demonstrates that carbohydrate epitopes are major targets of sera from C57BL/6J and CBA/J mice vaccinated with 15- or 50-kilorad-irradiated cercariae of S. mansoni. Antibody titers to carbohydrate epitopes increased with the number of vaccinations and were considerably higher in C57BL/6J mice than in CBA/J mice. The specificity of this anticarbohydrate response was determined by measuring antibody binding to defined oligosaccharide residues known to be present on the parasite. A predominant target of the humoral anticarbohydrate response of vaccinated mice was lacto-N-fucopentaose III, a molecule relevant for cell trafficking. We observed no binding to its nonfucosylated homolog, lacto-N-neotetraose, or to oligosaccharides present on keyhole limpet hemocyanin. The strongest antibody response to lacto-N-fucopentaose III was observed for C57BL/6J and CBA/J mice repeatedly vaccinated with 15-kilorad-irradiated cercariae, which also achieve the highest levels of protection. Immunoglobulin M was the predominant antibody class binding to lacto-N-fucopentaose III. We conclude that in the irradiated-cercariae vaccine model, C57BL/6J and CBA/J mice produce anticarbohydrate antibodies against various stages of S. mansoni and that the oligosaccharide lacto-N-fucopentaose III is one target of this response. Lacto-N-fucopentaose III and its specific antibodies may profoundly affect host resistance and parasite homing.

Animals↗

[Factors that can affect the breeding and maintenance of infected snails and the production of Schistosoma mansoni cercariae].

Mass production of Schistosoma mansoni cercariae was affected by biological and chemical agents. Rotifers and ostracods, snail predators, were identified in our colony. Rotifers were easily eradicated by washing the aquaria and lettuce with diluted solution of acetic acid. On the other hand, ostracods were difficult to eradicate and led to a high level mortality of infected snails (50-60%). Snails maintained in an incubator at constant temperature and total darkness produced maximum shedding when submitted to brightness and high temperature (about 30 degrees C). The number of cercariae shed was practically the same between pH 5-7. Contaminants such as Cu and Pb added to glass distilled water decreased the cercariae production. In conclusion, laboratory maintenance of large number of infected snails for mass production of cercariae is much simpler and more efficient than the conventional technique with running tap water although a high rate of mortality is observed in the snail colony.

Animals↗

Infectivity of Ornithodiplostomum ptychocheilus and Posthodiplostomum minimum (Trematoda: Diplostomidae) cercariae following exposure to cadmium.

The effect of cadmium (Cd) exposure on infectivity of cercariae of 2 trematode species (Ornithodiplostomum ptychocheilus and Posthodiplostomum minimum) to their second intermediate fish host was evaluated. Individual fathead minnows (Pimephales promelas) were exposed to cercariae that had been treated with Cd solutions at concentrations of 0, 2, 20, or 200 microg/L for 2 hr. Two weeks later, the numbers of encysted metacercariae in the brain (O. ptychocheilus) and body cavity (P. minimum) of the fish were evaluated. ANOVA analyses indicated a strong negative effect of Cd concentration on cercaria infectivity. The species x Cd concentration interaction was not significant, indicating that the magnitude of Cd-induced reduction in infectivity was similar between O. ptychocheilus and P. minimum. The results show that short-term exposure to Cd, even at low concentrations, interferes with transmission processes that affect the recognition and penetration of cercariae, the migration and survival of metacercariae within the second intermediate host, or both.

Animals↗

Complement-mediated tail degradation of Neodiplostomum seoulense cercariae.

The furcocercus cercariae of Neodiplostomum seoulense (Digenea: Neodiplostomidae) penetrate the skins of tadpoles and shed their tails. The speculated mechanism of this tail loss was physical efforts required to produce a vigorous zigzag motion during skin penetration; no other mechanism has been proposed. We examined the relationship between the host serum and cercarial tail loss. Cercariae of N. seoulense were collected from experimentally infected Segmentina hemisphaerula, and lots of 300 cercariae were cultured in medium 199 contained several types of sera. Cercarial tail degradation was induced in all media, but all the cercariae cultured except those cultured in media containing fetal bovine serum (FBS) died within 48 hr. After 72 hr cultivation in media containing FBS, cercarial tail degradation was induced in 67.0%; in continuous cultivation 13.3% of larvae survived for 7 days. Tail degradation did not occur in the absence of serum and when serum was heat inactivated at 56 degrees Celsius for 30 min. The addition of 20 mM ethylenediaminetetraacetic acid (EDTA) blocked cercarial tail degradation completely. Moreover, the addition of 20 mM MgCl2 restored tail degradation blocked by EDTA. These results suggest that the alternative complement pathway is related with the N. seoulense cercarial tail degradation induced by serum.

Animals↗

Preparation of mouse monoclonal antibodies and evidence for a host immune response to the preacetabular gland proteinase of Schistosoma mansoni cercariae.

Forty-six monoclonal antibodies were produced against the preacetabular gland secretions of Schistosoma mansoni cercariae by two different immunization protocols. These antibodies were of both the IgG and the IgM classes. One IgM monoclonal antibody (Ia4D6) was further characterized. It was specific to the cercarial stage by ELISA and showed specific binding to the 30,000 Mr proteinase in crude cercarial secretions by Western blot analysis. Preincubation of this antibody with purified cercarial proteinase resulted in inhibition of proteolytic activity, and it mediated complement-dependent cytotoxicity to cercariae in vitro. Immunoperoxidase staining of cercariae localized this antibody to vesicles visible within the preacetabular glands and their secretory ducts, and to secreted material. ELISA and Western blot analysis also showed that sera from infected mice and patients with schistosomiasis reacted with the cercarial proteinase. These studies demonstrate that a proteinase secreted into the host by invading cercariae is immunogenic and provide a monoclonal antibody probe for further characterization of its structure and function.

Animals↗

Evaluation of N,N-diethyl-m-toluamide (DEET) as a topical agent for preventing skin penetration by cercariae of Schistosoma mansoni.

The effect of skin application of N,N-diethyl-m-toluamide (DEET) on the penetration and migration behavior of cercariae of Schistosoma mansoni was evaluated in vitro and in vivo in a mouse model. These studies showed that DEET at concentrations of 7.5% or higher was 100% effective in immobilizing and killing cercariae of S. mansoni in vitro. Ultrastructural studies on such DEET-exposed cercariae showed transformative and degenerative changes involving both tegument and deeper parenchymal structures. Fatal tissue lesions were evident as early as 5 min postexposure to DEET, and became more extensive with increasing exposure time. Cutaneous application of DEET (as a pure chemical in isopropanol or as a commercial insect repellent preparation) was more than 99% effective in preventing entry of S. mansoni cercariae into the mouse tail skin. Radiolabeling and tracer studies confirmed that 7.5% DEET applied to the skin prior to infection was highly effective in preventing schistosomular migration to the lungs.

Administration, Topical↗

A longifurcate distome cercaria from Bulinus truncatus snails in the Jordan Valley, Jordan.

Bulinus truncatus snails collected from water bodies of the South Shuna region, north of the Dead Sea, were found infected with a pharyngeal longifurcate distome cercaria. This new form of cercaria is named Cercaria bulini I. It has three pairs of penetration glands, 12 pairs of flame cells, and lacks the intestinal cecae. This cercaria is an active swimmer and develops within an elongated sporocyst. Other details on the morphology, behavior, and development of C. bulini I are presented.

Animals↗

[Morphology and biology of cystophorous cercariae from the Bering sea].

Three new species of cystophorous cercariae, Cercaria appendiculata Pelseneer, 1906, C. octocauda Tschubrik, 1952 and C. saccocaudata Tschubrik, 1966, are described from the gastropod Tectonatica clausa (Broderip et Sowerby). C. saccocaudata is a larva of trematodes of the genus Lecithaster. The development of parthenites of this species carries out within one summer season. Parthenites of C. appendiculata and C. octocauda are capable to survive unfavourable winter conditions and produce cercariae in the next summer. The above three species were found to eject cercariae through the ejaculatory tube which is an important adaptation of chemiurids to the infection of the second intermediate hosts.

Larva↗

Scanning electron microscopical studies on the tegument of adult worms of Schistosoma mansoni originating from ultraviolet-irradiated and non-irradiated cercariae.

The surface topography and ultrastructural changes of Schistosoma mansoni worms developed from ultraviolet-irradiated cercariae were compared with those from non-irradiated cercariae. The tegument of worms developed from irradiated cercariae showed a variety of changes including the occurrence of oedemata and most of the tubercles being torn at the tip and having lost their spines on the dorsal surface of some male worms. In females, parts of the tegument were devoid of spines and surface lesions were present. The range and extent of these changes differed not only among individual worms but also between different regions of individual worms resulting in generalized deformities in those worms originating from irradiated cercariae.

Animals↗

Identification of sex in Schistosoma mansoni cercariae.

A simple method for determining the sex of larval stages of Schistosoma mansoni involved cytological demonstration of sex chromatin (heterochromatin) in the interphase and prophase nuclei of cercariae. Cerariae from unimiracidial infections were fixed, and permanent slides were stained with thionin, Giemsa, or fuchsin. On examination, a given cercaria showed either 1 or 2 sex chromatin bodies in the nuclei, the female with 1 and the male with 2 bodies. The results of observation on the sex of cercariae from the slides were matched with those of worms obtained from mice injected with the corresponding batch of cercariae. The accuracy of cytologic sex identification was greater than 80% in these experiments. The sex chromatin in both adult and larval stages was in the form of large, dark purplish bodies usually situated towards the periphery of the nuclei and could be easily distinguished from the other chromatin granules present in the nuclei.

Animals↗

A feasible and effective technique for the detection of Schistosoma japonicum cercaria-infested water.

OBJECTIVE: To develop a new technique for detecting Schistosoma japonicum (S. japonicum) cercariae so as to predict the distribution of area with high transmission potential and to control acute infection of schistosoma. METHODS: A C-6 film was established and used to adhere the cercariae on water surface and compared with a few other conventional detecting techniques. RESULTS: The average rate of the cercaria-recovery was 40% in laboratory and field using C-6 film, which was 20 times higher than that of traditional nylon-tissue adsorbing and precipitating method. Either in some areas where oncomelania were present or in some places where the snail had been under control, the technique could detect the cercariae. CONCLUSION: The technique can be characterized by a feasible, economical, promptly accessible method, especially for field application.

Animals↗

Conditions for labeling of Schistosoma mansoni cercaria with technetium-99m.

Study of the labeling of Schistosoma mansoni cercaria with technetium-99m (99mTc) at room temperature (25 degrees C) and 37 degrees C shows that the incorporation of radioactivity in this cercaria increases with the increase in stannous chloride concentration, reaching a constant value threshold at 130.00 microM. Strong binding of the 99mTc was obtained since the radioactivity was not washed out. The characteristic motion of the cercaria, labeled at 25 degrees C and 37 degrees C, was only modified at the concentration of 1300 microM for both temperatures, showing that the methodology here described can be applied to living structures. With this technique it is possible to obtain a cercariae suspension labeled with a radionuclide that is very inexpensive and readily available. Furthermore, it is a gamma emitter with a photon energy of 140 keV that would permit one to make scannings of the infected animals and causes less of an environmental impact and present fewer radioactive disposal problems than the long lived radionuclides.

Animals↗

[Morphology and chaetotaxy of Sanguinicola armata cercaria (Trematoda: Sanguinicolidae)].

A morphological description of Sanguinicola armata cercaria is given. This larva belongs to the group of brevifurcate and apharingeal cercariae. Mouth opening is on ventral surface of the anterior organ. The body and branches of the furca bear a membrane. The body/tail ratio is much more than in other cercariae of the family Sanguinicolidae. The gland apparatus of the larva is represented by several types of secretory cells. The penetration glands form a group of 5 pairs of cells on the ventral side. Their ducts form a single bunch directed anterior, go through the anterior organ and open terminally on the proboscis, which is able to turn outside. Other gland cells apparently represent specialized cytones of the tegument. Excretory formula: 2[(1 + 1) + (1 + 1)] = 8. The external surface of the larva bears a small number of sensilla. Some similarities of the families Sanguinicolidae and Schistosomatidae in the cheatotaxy of cercaria have been noticed.

Animals↗

Effect of topical agents on cercariae of Schistosoma mansoni.

DEET (N, N-diethyl-m-toluamide) is one of the reliable and most widely used insect repellents. The present work was planed to evaluate the effect of free DEET, controlled release DEET and white precipitate ointment on the viability of cercariae of S. mansoni in-vitro. They were also topically applied to mice to study their efficiency in preventing cercarial skin penetration. Free DEET and controlled release DEET formula caused immobilization and death of cercariae within twenty and five minutes respectively. The number of adults detected after application of free DEET and white precipitate ointment to mice skin prior to infection were significantly lower than the control group. When controlled release DEET was applied no adults could be detected indicating failure of cercariae to enter through the skin. This was confirmed by histopathological study of the liver which was free of granuloma. Scanning electron microscopy revealed tegumental changes in cercariae exposed to both free DEET and controlled released DEET. So, topical application of any of the three chemicals was effective in controlling S. mansoni infection. The best was with controlled release DEET.

Administration, Topical↗

Effects of Solanum nigrum leaves water extract on the penetration and infectivity of Schistosoma mansoni cercariae.

A crude water extract of Solanum nigrum leaves was used as a chemical attenuate to Schistosoma mansoni cercariae prior to infection of Swiss female mice. Cercariae were exposed to 2.5, 5, 7.5 and 10 mg/l concentrations of the extract for 30 min. The effect on the ability of cercariae to penetrate mice skin, as well as, effect on schistosome worm burden after 8 weeks of infection were measured. The observed reduction of cercarial penetration was significant at 7.5 and 10 mg/l concentrations (p < 0.001). The mean number of worm burden declined from 28.5 worms/ mouse in untreated group to 4.4 worms/mouse with 7.5 mg/l treatment (p < 0.01). At a concentration of 10 mg/l, mice had no adult worm. The cercarial infectivity, as measured by the proportion of worms recovered in relation to the number of cercariae administrated, decreased with the increase in the extract concentration and was significant at a concentration of 7.5 mg/l (p < 0.01). The number of schistosome eggs in hepatic tissue decreased in treated mice. The reduction in egg count (per gram liver) was significant at 5 mg/l (p < 0.05) and 7.5 mg/I (p < 0.001). The treatment with Solanum water extract had no effect on female fecundity. These data point to Solanum as a promising agent for the control of schistosomiasis.

Animals↗

[Are nasal Trichobilharzia cercariae potential threath to human health?].

During late summer in 1995 to 1997, repeated outbreaks of maculopapular skin eruptions were observed on legs of children after wading in the pond of the Family Park in Reykjavík, Iceland. Investigation, starting in autumn 1997, revealed that the causative agent was a previously undescribed schistosome cercaria of the genus Trichobilharzia, shed by Radix peregra, the only snail occurring in the pond. This was the first report of swimmer's itch in Iceland. Infection experiments with cercariae from the pond have revealed adult worms and eggs of a Trichobilharzia in the nasal area of ducklings (Anas platyrhynchos f.dom.) 18-23 days p.i., and schistosomula in the spinal cord of BALB/c mice 3, 6 and 10 days p.i. Moreover, a mouse killed 3 days p.i. also had schistosomula in the lungs. During the prepatent period the infected ducklings had neuromotoric symptoms and gross pathology revealed petechiae in the nasal cavity. The results indicate that the cercaria responsible for swimmer's itch in Iceland is a nasal schistosome. Furthermore, adults of two visceral Trichobilharzia species have been found in Icelandic whooper swans Cygnus cygnus. As schistosomula of both nasal and visceral Trichobilharzia species are able to develop and migrate for several days in a non-specific mammalian host, humans are warned to expose themselves to Trichobilharzia cercariae in Iceland.

English Abstract↗

[Opinions on the ways of reaching the eyeball by cercaria of the Diplostomum genus].

The views on the ways of migration of Diplostomum cercaria from body surface to fish eyeball, among them about the ways of parasites dislocation through the tissues, beginning from the penetration to host body--was presented. The hypothesis on passive in-coming transport of cercaria by the circulatory system of fish was questioned; on the other hand it has been argued active penetration of parasites through tissues, according to the hypothesis of cercaria migration stimulated by the respective concentration of carbonate anhydrasis in host. It has been stated that migrations of cercaria to fish eyeball may be different depending on the penetrations place of parasite on body host.

Animals↗