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Schistosoma mansoni: egg granuloma reduction in challenged mice after vaccination with irradiated-attenuated cercariae.

Response of Swiss albino mice vaccinated with irradiated-attenuated cercariae or repeatedly infected with normal cercariae of Schistosoma mansoni were known to develop resistance to reinfection. The mean (+/- SD) of the hepatic granuloma numbers and diameters for vaccinated-challenged group (I) were 2 +/- 0.89/1mm2 and 116 +/- 11.5 microns, for repeatedly infected group (II) were 2 +/- 0.63/1mm2 and 2 +/- 0.89/1mm2 and 195 +/- 11.8 microns and for control group (III) were 4 +/- 1.36/1mm2 and 140 +/- 12.3 microns respectively. Mean of the diameter of the granulomas were significantly smaller in the group vaccinated with irradiated cercariae than the other two groups. Most of the granulomas in groups I and II were fibro-cellular while all granulomas in group III were cellular. Predominent cell types of the granulomas were lymphocytes in groups I and II and eosinophils in group III. The incidence of focal hydropic and fatty degenerations, necrosis and prominent kupffer cell hyperplasia were lower in group I. These results supported that granuloma size reduction in all vaccinated animals were apparently effective in sequestering egg toxins and reduced hepatocytes damage. The present study gives encouragement that a vaccine to enhance protection against disease in human schistosomiasis is possible.

Animals↗

Comparison of two strains of Schistosoma mansoni with respect to the sites and kinetics of immune elimination in irradiated cercaria-immunized mice.

A comparison was made of the sites of elimination of NMRI and Mill Hill strains of Schistosoma mansoni in C57BL/6J mice previously immunized with 50 krad gamma-irradiated cercariae of the homologous strain. In the first experiment, the fate of percutaneous challenge infections with 75Se-labeled cercariae was evaluated by autoradiography of tissue squashes and hepatic portal perfusion. For both strains of parasite, migration from skin to lungs was delayed but not reduced in immunized mice relative to controls, with immune elimination taking place at some point after migration to the lungs. In a second experiment, resistance to the Mill Hill strain of S. mansoni was compared in mice challenged by percutaneous infection with cercariae and by intravenous injection with lung schistosomula. Both types of challenge were shown to be vulnerable to immune elimination. We conclude that under the conditions employed in this study, there is no significant difference between the NMRI and Mill Hill strains of S. mansoni in the patterns of migration and elimination, with most or all elimination in both control and immunized mice taking place after migration from the skin.

Animals↗

Identification of Schistosoma mansoni antigens recognized by T cells of C57BL/6 mice immunized with gamma-irradiated cercariae.

Immunization of C57Bl/6 mice with gamma-irradiated Schistosoma mansoni cercariae induces highly significant protection against subsequent challenge with unattenuated living cercariae. The antigens that evoke T cell-mediated immunity in this model are vaccine antigen candidates since T lymphocytes mediate antibody-independent immune responses and control the production of most antibodies. To identify these antigens, spleen cells of mice immunized twice with gamma-irradiated cercariae were tested for proliferative responsiveness and production of interleukins (Il) and interferon-gamma (IFN-gamma) following incubation with separated soluble schistosomular (SSA) and adult worm (SAWA) antigens in T cell western assays. Data obtained at 3, 4, 5, and 6 wk post-secondary immunization in 2 experiments indicated that SSA and SAWA bands of 62-60, 50, and 45 kDa reproducibly elicited T cell proliferation and production of Il-2, Il-4, and IFN-gamma by spleen cells from immunized, but not unimmunized, mice. Bands of 72-68, 29.5, and 28 kDa elicited proliferation and production of Il-2 and Il-4, but not IFN-gamma. Bands of 35-33 and 24 kDa induced either proliferation or Il production at some intervals.

Animals↗

Candidate vaccine antigens that stimulate the cellular immune response of mice vaccinated with irradiated cercariae of Schistosoma mansoni.

Vaccination with radiation-attenuated cercariae confers the highest levels of resistance to challenge infection in experimental schistosomiasis and requires Ag-specific T cells. Therefore, this study aimed to identify specific Ag that stimulate the cellular immune response of mice vaccinated with irradiated cercariae of Schistosoma mansoni. Four experimental groups representing different levels of resistance in the vaccine model (C57BL/6J versus CBA/J mice vaccinated with 15- or 50-krad irradiated cercariae) were compared for in vitro lymphocyte proliferation and lymphokine production. Adult worm extracts fractionated by isoelectric focusing were used as Ag. Lymphocyte proliferation of all groups was limited to three consecutive isoelectric fractions (pH 4.6-6.3). Interestingly, the antibody response of these mice was directed to Ag in the same isoelectric fractions, three of which had previously been identified as paramyosin, heat shock protein 70, and the integral membrane protein Sm23. These Ag as well as two 28 kDa proteins, triosephosphate isomerase and glutathione S-transferase, in purified native or recombinant form or as a synthetic peptide, stimulated lymphocyte proliferation. Lymphocytes of vaccinated C57BL/6J mice generally showed higher levels of proliferation than did CBA/J mice. Interestingly, cells of once-vaccinated mice responded better than did cells of mice vaccinated three times. Lymphokine assays demonstrated that IL-2 and IL-4 was generally reduced after multiple vaccinations and varied qualitatively as well as quantitatively between mouse strains. This study substantiates that the five Ag, paramyosin, heat shock protein 70, triosephosphate isomerase, glutathione S-transferase, and the integral membrane protein Sm23, are important candidates for a defined antischistosomal vaccine.

Animals↗

A trap for the detection of schistosome cercariae.

A simple trap for schistosome cercariae has been developed using clear nail varnish as a matrix and linoleic acid as stimulant affixed to a 75 x 25-mm glass slide. Schistosoma mansoni cercariae will attach to the surface and easily can be visualized either stained or unstained. The trap works well under laboratory and field conditions where the retrieval rate varies from 30% to 100%. Under laboratory conditions there appears to be some form of affinity for cercariae to the surface.

Animals↗

Filtration, centrifugation and mouse exposure for the detection of schistosome cercariae in water.

Three methods for detection of schistosome cercariae in water were studied under laboratory and simulated field conditions in Egypt. Water samples tested by filtration and centrifugation quantitatively reflected the density of cercariae in moderately turbid canal water (30 Nephelometry Turbidity Unites, NTU). When tested in waters of increasing turbidity, the centrifuge gave more consistent results than the filtration system. The centrifugation technique was more efficient in processing the water samples and took almost half the time of filtration. Mouse exposure was useful in detecting cercariae, but was not quantitative. However, there was a significant improvement in worm recovery when mice were more close together in the floatation device. Mouse exposure was also expensive and cumbersome to use. It appears from the present results that the technique of choice for further cercariometric work under Egyptian conditions may be centrifugation.

Animals↗

The effects of lead, cadmium, and mercury on the mortality and infectivity of Schistosoma mansoni cercariae.

The effects of lead, cadmium, and mercury chloride salts on survival and infectivity of Schistosoma mansoni cercariae were examined. The LC 50 for 5 hr exposure was approximately 500 microM for lead, 2.5 microM for cadmium, and 6 microM for mercury. These toxicity figures are significantly higher than the levels of these heavy metals reported to occur in the natural waters of Egypt where schistosome transmission occurs. Infectivity of cercariae surviving exposure to metals at the LC 50 was reduced to approximately one-half that of control mice infected with unexposed cercariae.

Analysis of Variance↗

Patterns of argentophilic papillae of Schistosoma rodhaini and S. mansoni cercariae.

Argentophilic papillar patterns on cercariae of Schistosoma rodhaini from Kenya and S. Africa, and S. mansoni from Kenya and Brazil were compared with each other and with earlier results on cercariae of a Puerto Rican stock. The basic total number of papillae of both species is the same (62), and the basic numbers of papillae in the various groups are the same: dorsal, 20; ventral, 18; lateral, 10 on each side; and acetabular, 4. The cercariae of these 2 species differ, however, in patterns of papillae, chiefly the dorsal patterns, and can be distinguished from each other.

Animals↗

[Attempt to determine by chaetotaxy the genus of a microphallid cercaria parasiting Cerithium meditteraneum Deshayes (author's transl)].

Chetotaxy of a cercaria parasite of Cerithium meditteraneum is studied. Sensory receptors C III, St V, St DL, AID, PI and U, are similar to those of Maritrema linguilla and Maritrema subdolum. However in the three cercariae, the ventral, dorsal and lateral papillae, except AID and PI (V, L, D) are different. The chetotaxy of this cercariae shows that they belong to the genus Maritrema. Investigations of the biological cycle are under way to verifie this hypothesis.

Animals↗

Transcriptomic analysis of eggs, rediae and cercariae reveal stage-specific adaptations in the rumen fluke Calicophoron daubneyi.

Rumen flukes, particularly the trematode Calicophoron daubneyi, are emerging parasites of livestock in Europe, yet transcriptomic insights into their environmental and intermediate host stages remain limited. Here, we present a comprehensive transcriptomic analysis of eggs at three distinct developmental stages (freshly excreted, early developmental and eye-spot stages), as well as rediae and cercariae, of C. daubneyi. High-quality RNA-sequencing (RNA-seq) datasets revealed both shared and stage-specific transcriptional profiles with each developmental stage exhibiting its own distinct expression pattern. Subsequent GO-Term enrichment analyses revealed that fully embryonated eggs in eye-spot-stage especially upregulated genes related to cilia assembly, movement and motility, reflecting preparation for miracidial hatching and host-seeking behavior. Rediae showed enhanced transcription of genes involved in diverse metabolic and biosynthetic processes, supporting rapid asexual proliferation within the snail intermediate host. Cercariae exhibited predominant upregulation of genes associated with signal transduction and energy metabolism, indicating the adaptation to its changing environmental conditions. These findings provide the first transcriptomic insights into the biology of C. daubneyi outside the definitive host, reveal molecular mechanisms underlying development, transmission and adaptation to a changing environment and identify stage-specific genes as potential targets for interventions aimed at disrupting the parasites life cycle and controlling rumen flukes in the future.

Animals↗

Cyton II: a subtegumental cell type in the cercaria of Schistosoma mansoni.

In Schistosoma mansoni cercaria, an aggregate of subtegumental cells is found in a small, dorsoanterior area of the body (middivision). These cells are nestled between two laterally positioned flame cells and the muscle that delimits the anterior end of the body, and the anterior end of the central ganglion. This highly amorphous cell type, designated as cyton II, has a heterochromatic nucleus and a cytoplasm that is elaborated into coarse, tortuous processes. Its cytoplasm contains ribosomes, mitochondria, sparse amounts of endoplasmic reticulum, and two types of circular-to-oval concentric membranous bodies. One type has an electron-dense core and measures 200-250 nm on the short axis, and the other is completely membranous and measures 100-125 nm on the short axis. The cell body of cyton II communicates with the tegument that covers a small, dorsoposterior area of the anterior organ (oral sucker); however, we could not confirm a tegumental connection with the body division. When cercariae transform into schistosomules, the concentric membranous bodies of cyton II migrate into the anterior organ's tegument via cytoplasmic processes of the cell. The major function of previously described cells that have similar membranous bodies is to supply additional membranes to the outer tegument during development into an adult worm. A multilaminated outer membrane is an adaptation to the survival of the schistosomule and adult worm in the bloodstream of the vertebrate host (Hockley amd McLaren ['73]). The presence of membranous bodies from cyton II in the tegument does not confirm that this cell type participates in the formation of multilaminated membranes. Its precise function remains to be determined.

Animals↗

Schistosoma mansoni: fractionation and characterization of the glycocalyx and glycogen-like material from cercariae.

The glycocalyx (GCX) that covers schistosomal cercariae is a complex molecule that has immunomodulating properties. Here, we purified milligram amounts of GCX using Anguilla lectin which binds to the GCX covering the cercarial body and tail. Typically, 10 million cercariae were extracted with phenol, dialyzed, and chromatographed on a Sepharose 2B-CL column. An average of 39 mg of total carbohydrate eluted near the void volume from which 31 mg of glycogen-like material was further separated by lectin affinity chromatography. Its identity was established by compositional analysis, sensitivity to amylase digestion, and its nuclear magnetic resonance spectrum. The lectin-bound GCX was eluted with 0.1 M fucose with a final yield of 5.3 mg carbohydrate. Fucose composed 40% of the total GCX carbohydrate with lesser but approximately equal amounts of galactose, glucosamine, and galactosamine present. NMR data indicated that the amino sugars were N-acetylated. Glucose was also present but in varying amounts in different preparations of GCX. Oligosaccharides were released from GCX by hydrazinolysis and separated by electrophoresis after reductive amination to 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS). Bands comigrating with standards containing 11, 12, 16, and 17 sugar residues were detected. Thus, the GCX is a complex structure composed of oligosaccharides, probably linked to a peptide.

Amino Sugars↗

Schistosoma mansoni: interleukin-1 increases phagocytosis and superoxide production by hemocytes and decreases output of cercariae in schistosome-susceptible Biomphalaria glabrata.

Decreases in the number of Schistosoma mansoni cercariae released from susceptible M-line Biomphalaria glabrata were detected following injection with the recombinant human cytokine, interleukin-1. No differences in either the time post-exposure at which shedding began or the percentage of snails shedding cercariae were detected between interleukin-1 injected, heat-inactivated interleukin-1 injected, or sham injected controls. However, sham injected and heat-inactivated interleukin-1 injected snails maintained significantly higher (approximately three-fold) levels of cercarial production compared to interleukin-1 injected snails over 8 weeks of cercarial shedding. Injection of interleukin-1 into schistosome-susceptible (M-line) and resistant (13-16-R1) strains of B. glabrata increased hemocyte phagocytosis of target particles and phagocytosis stimulated O2- production in both snail strains at 24 hr postexposure to the parasite. Resistant 13-16-R1 snails maintained, on average, 2.4 times the number of O2- producing phagocytic cells than did M-line susceptible snails, indicating that the incomplete abrogation of cercarial shedding in M-line snails may be due to an inadequate number of activated circulating effector cells in these snails. These data strongly support the contention that the evolutionarily conserved cytokine, interleukin-1, or a molecule in snail plasma with interleukin-1-like immunospecificity, biological activity, and function plays a significant role in the maintenance of susceptibility or resistance to S. mansoni infection in B. glabrata. Finally, these data also supply evidence for the evolutionary conservation of the function and role of interleukin-1, O2-, and antioxidant defense mechanisms in this host-parasite relationship.

Animals↗

ICAM-1 and iNOS expression increased in the skin of mice after vaccination with gamma-irradiated cercariae of Schistosoma mansoni.

Host responses to migrating schistosomula of Schistosoma mansoni were compared in the skin of naive, multiply infected, or vaccinated (with gamma-irradiated cercariae) mice during the first 72 hr after cercarial penetration. Cellular response to the migrating parasite was minimal in the skin of naive mice for up to 72 hr after infection. In sharp contrast, the multiply infected or vaccinated animals exhibited a marked inflammatory response in the skin as early as 8 hr after cutaneous penetration of the challenge cercariae. This early inflammatory response in the skin of sensitized animals was characterized by a significant increase in the number of infiltrating cells, predominantly mononuclear cells and neutrophils. Increased exudation of serum proteins was also present in the skin of sensitized animals in areas of cercarial challenge. A time course of analyses revealed that mononuclear cell numbers increased significantly in the skin of vaccinated animals as early as 60 min after a challenge infection and continued to be present at a significantly higher level up to 72 hr after challenge. Peak neutrophil responses occurred in the skin at 24 hr (in multiply infected animals) and at 48 hr (in vaccinated animals) after a challenge infection. Along with the massive cellular infiltration there was an increased tissue expression of ICAM-1 and mRNA for iNOS in the skin of sensitized animals. Further analysis showed that in sensitized animals increased ICAM-1 expression was predominantly found on endothelial cells lining dermal capillaries, especially in areas around schistosomular migration and on cells that surrounded schistosomula in the dermis. In naive animals, however, a similar infection did not induce any ICAM-1 expression or iNOS production in the skin. Thus, an ICAM-1 mediated early accumulation of mononuclear cells in the skin and local production of nitric oxide may be important for the initial cutaneous inflammatory immune responses to migrating schistosomula of S. mansoni in vaccinated animals. On the contrary, in naive animals a potential parasite-induced suppression of ICAM-1 may play an important role in reducing cellular reaction in the skin and consequently help the parasite evade immune responses in the skin.

Animals↗

On Schistosoma margrebowiei Le Roux, 1933: the morphology of the egg, miracidium and cercaria, the compatibility with species of Bulinus, and development in Mesocricetus auratus.

The morphology of the egg, miracidium, and cercaria of Schistosoma margrebowiei are described. The compatibility of S. margrebowiei with species of Bulinus has been examined. The parasite develops well in the diploid, tetraploid and octoploid snails of the truncatus/tropicus complex with overall infection rates of 38.1%, 30.1% and 29.1% respectively, and in reticulatus group snails (Bulinus wrighti) with an infection rate of 45.8% of those snails surviving the prepatent period. Only two species of the forskali complex (Bulinus bavayi, Aldabra; Bulinus beccarii, South Arabia) are slightly compatible, and snails of the africanus group are incompatible. The overall worm return from 24 hamsters exposed individually to 100 cercariae was 39.8%; 82.1% of the worms were paired, the remainder unpaired. The growth of the paired worms was recorded from 28-60 days. The prepatent period in hamsters in 33 days, in sheep 38 days. The mean egg production was 837 eggs/day in infections ranging from 33 to 60 days. Most eggs (80.9%) were deposited in the intestine, and only 18.2% were deposited in the liver. The parasite is pathogenic in hamsters, and peak death rate occurred in the 50-60 day group infections which coincided with the period of peak egg production.

Animals↗

[The attachment response of the cercaria of Schistosoma mansoni. Influence of natural substrates and temperature (author's transl)].

The behaviour of the cercaria, following contact with different animal and human substrates during swimming, has been studied. Four patterns of behavior related to the intensity of stimulation can be distinguished: 1. backward reaction, 2. stopping, 3. forward reaction, 4. attachment (Figs. 2 and 3). Mechanical stimulation of the cercaria by touching or water turbulence, prior to contact with the substrate has an insignificant effect on attachment. Both attachment and the first step in penetration (crawling over the substrate) are stimulated by a rise in temperature (Figs. 4 and 5) and the chemical properties of the substrate. However, the steps in penetration may possibly be stimulated by chemical components and perceived by receptors different from those involved in attachment.

Animals↗