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A new cercaria and metacercaria of Acanthoparyphium (Echinostomatidae) found in an intertidal snail Zeacumantus subcarinatus (Batillaridae) from New Zealand.

A new 23-collar-spined cercaria and metacercaria are described from intertidal molluscs of the coast of New Zealand. The new cercaria found emerging from the mud snails Zeacumantus subcarinatus (Sowerby) (Prosobranchia: Batillariidae) is characterized mainly by the number and arrangement of the cephalic glands, the size of the suckers, and the size and number of the collar spines. The cercaria encysts in the cockle Austrovenus stutchburyi (Wood) (Bivalvia: Veneridae) which lives in sympatry with the first intermediate host throughout New Zealand. Laboratory infections of cockles by cercariae from naturally infected snails resulted in metacercariae identical to those found in naturally infected cockles. The main features of the cercaria and metacercaria are the presence of a reniform collar with 23 spines, two pairs of small cephalic glands at the oral sucker level, another two pairs of much longer ones posterior to the pharynx, and the excretory vesicle Y-shaped with the main collecting canals extending to the anterior level of the ventral sucker. Each of the main collecting canals had 10-11 pairs of bilateral diverticula between the anterior edge of the ventral sucker and anterior body end. A brief discussion of its possible life cycle and ecology is also provided.

Animals↗

Comparison of the vaccine efficacy of gamma-irradiated Schistosoma japonicum cercariae with the defined antigen Sj62(IrV-5) in pigs.

Development of a vaccine against Schistosoma japonicum which can protect both man and the domestic animal zoonotic reservoirs of infection would be an invaluable tool in attempts to control this infection in those areas in which conventional control methods have failed to break transmission. The pig is a natural host of S. japonicum and because of its anatomical and immunological similarities to humans, it is a potentially valuable host for studies on S. japonicum in particular and schistosomes in general. Radiation-attenuated cercariae are highly effective in inducing immunity in experimental schistosomosis and there are promising reports of partial protection against schistosomes with recombinant-derived individual antigens. In the present study we have set out to establish a protocol for inducing protection with gamma-irradiated cercariae in pigs and to assess the protective capacity of recombinant and naked DNA formulations of Sj62, a 62kDa region of S. japonicum myosin. The corresponding S. mansoni version or Sj62, recombinant IrV-5, has previously been implicated in irradiated vaccine immunity in S. mansoni infections and has been shown to induce high levels of immunity in a variety of hosts. Groups of pigs were immunised three times at 2-week intervals with 2000 cercariae irradiated at 20krad, with Sj62 as a recombinant (rSj62) incorporated in Freund's adjuvant, a micellar preparation, or as a naked DNA construct. Vaccination with irradiated cercariae did not induce significant anti-Sj62 antibody but following intramuscular challenge with 2000 cercariae, the vaccinated pigs showed >95% resistance as assessed by reduced faecal egg output, worm tissue egg burdens and also reduced septal fibrosis. Immunisation with each of the Sj62 formulations induced significant anti-Sj62 antibody responses, the highest titre (>12,800) being with the Freund's preparation but none of the Sj62-immunised groups showed significant resistance to challenge. The data suggest that Sj62 shows little promise as a vaccine candidate for schistosomosis.

Animals↗

Vaccination with irradiated cercariae of Schistosoma mansoni preferentially induced the accumulation of interferon-gamma producing T cells in the skin and skin draining lymph nodes of mice.

Cytokine response to schistosomula of Schistosoma mansoni was evaluated in the skin of mice during the initial 72 h following infection. These studies showed a significant increase in the levels of IL-4 and IL-10 message in the skin in areas of cercarial penetration. The IL-4 message was detectable in the skin as early as 8 h after infection and the message for IL-10 appeared from 16 h after infection. However, mRNA for IFN-gamma was undetectable in the skin samples for up to 72 h after infection with normal cercariae. In sharp contrast, vaccination with irradiated cercariae induced IFN-gamma and IL-2 responses in the skin within 24 h. Analysis of the cytokine profile of cells isolated from the skin during these early time points showed that T cells are probably not a source of IL-4 or IL-10 in the skin of mice infected with normal cercariae. However, in vaccinated animals, the majority of the IFN-gamma is derived from skin-residing T cells. In vaccinated animals, responses in the skin were mirrored in the skin-draining lymph nodes as well. Analysis of the CD4/CD8 ratio showed a significant decrease in the skin following vaccination suggesting an increase in CD8+ cells. Interestingly however, when vaccinated animals were challenged with normal cercariae, there was a significant reduction in IFN-gamma response in the skin and its draining lymph nodes. These results show that vaccination with irradiated cercariae of S. mansoni, preferentially induce the accumulation of IFN-gamma producing T cells in the skin and skin-draining lymph nodes of mice.

Animals↗

Immunisation of sheep against Schistosoma mattheei using either irradiated cercariae or irradiated schistosomula.

Irradiated cercariae, irradiated schistosomula, or heterologous infections were used to vaccinate sheep against Schistosoma mattheei infection. In the first experiment four doses of 10(4) S. mattheei cercariae irradiated at 6Kr were administered to sheep by percutaneous infection at 4 week intervals. This induced a 74% reduction in a challenge infection compared to control sheep while only 13% protection was achieved in a third group of sheep immunised with normal cercariae of the heterologous parasite S. mansoni. No significant differences were seen in histopathology of the liver of any of the sheep but the pathological changes were more severe in the large and small intestines of sheep vaccinated with the heterologous parasite. In the second experiment with irradiated cercariae only one or two immunising exposures were used. The degree of protection in the adult worm load (9-11%) was not significant and no significant differences were noticed in the pathology of the vaccinated and control animals. In the third experiment four doses of irradiated organisms were used to vaccinate five groups of sheep: 3Kr or 6Kr cercariae were administered by percutaneous infection; 6Kr skin-transformed schistosomula were administered by intra muscular injection; the same 6Kr skin-transformed schistosomula were given by intravenous injection and 6Kr syringe transformed schistosomula were administered by intramuscular injection. The degree of protection (determined as the reduction in worm burden) achieved by these different procedures was respectively 72%, 61%, 77%, 56% and 78%. These results indicate the possibility of making a live vaccine against ovine schistosomiasis and show that effective immunisation is not dependent on the presence of a mature worm infection or on cercarial penetration of the skin by the immunising infection.

Animals↗

Different levels of immunity to Schistosoma mansoni in the mouse: the role of variant cercariae.

Very variable levels of immunity to a second infection with Schistosoma mansoni were recorded in 7 strains of mice, 12--15 weeks following a small primary infection. When 2 or more strains of mice were assayed at the same time, less variation occurred within the experiment than between different experiments. This evidence suggested variation between pools of cercariae as the main cause of variability in immunity. In direct experiments in one strain of mouse, 2 different pools of cercariae stimulated widely different levels of immunity to the same challenge. Conversely, challenge infections drawn from different pools showed different susceptibility to immunity stimulated by the same primary infection. Individual clones of cercariae, from snails infected with single miracidia, showed a high level of susceptibility to immunity stimulated by a small bisexual infection, or were not susceptible at all. Antigenic polymorphism is the most likely explanation for the differences observed between clones of cercariae. However, indirect immunofluorescence showed the presence of at least 1 common antigen on the surface of schistosomula derived from different clones of cercariae and clone-specific antigens have not been detected.

Animals↗

Finding and recognition of the snail intermediate hosts by 3 species of echinostome cercariae.

Finding and recognition of snail second intermediate hosts was studied in cercariae of 3 echinostome species. The cercariae of the 3 species accumulated in snail-conditioned water (SCW) with 2 types of orientation mechanisms and responded to different small molecular weight ( < 500 Da) components of SCW. Pseudechinoparyphium echinatum and Echinostoma revolutum cercariae returned by swimming an arc, when swimming in decreasing concentration gradients of SCW (turn-back swimming). The stimulating cues of SCW were identified as hydrophilic organic molecules, probably possessing amino groups. Amino acids contributed to the attractivity of SCW, at least in P. echinatum, but they could not account for the complete attractivity of SCW. Hypoderaeum conoideum were directed chemotactically and swam along increasing concentration gradients of small peptides within SCW, but in decreasing SCW gradients they showed no turn-back swimming. Chemotactic orientation in H. conoideum only started 1 h after emission, which may assist the cercariae to leave the immediate area of their first intermediate host snails and to disperse. Attachments occurred specifically to snail hosts in the 3 species and were stimulated by macromolecular mucus compounds, probably mainly by viscoelastic properties of the mucus. The results of this study show, that host-finding mechanisms and the stimulating host cues of snail invading echinostome cercariae differ considerably from those of schistosome miracidia.

Animals↗

Immunocytochemical study on biologically active neurosubstances in daughter sporocysts and cercariae of Trichobilharzia ocellata and Schistosoma mansoni.

Immunocytochemical techniques applied to sections and whole-mount preparations of cercariae from two species of trematodes, Trichobilharzia ocellata and Schistosoma mansoni, revealed the occurrence of immunoreactivity (IR) to several neurosubstances in the nervous system (NS). Immunostaining was localized in cerebral ganglia, in the main commissure, in anterior and posterior nerve trunks, as well as in a pair of nerve fibres running along the tail. In T. ocellata, immunoreactivity (IR) was observed with antisera raised against: glutamate, FMRFamide, catch-relaxing peptide (CARP), small cardiac peptide B (SCPB), arg-vasotocin (AVT), arg-vasopressin (AVP), and substance P. In S. mansoni antisera raised against glutamate, FMRFamide, CARP, SCPB, alpha-caudodorsal cell peptides (alpha-CDCP), and cholecystokinin (CCK) showed neuronal IR. With the other 51 antisera tested no IR was observed. With anti-APGWamide, IR was observed outside the NS in cells of the wall of the daughter sporocyst and in flame cells of cercariae of T. ocellata. IR to FMRFamide was present in the escape glands of the intrasporocystic cercariae of T. ocellata and S. mansoni. IR to somatostatin was observed in subtegumental parenchymal cells of cercariae of S. mansoni. IR to met-enkephalin was present in cells of the cercarial embryos and in undifferentiated cells in developing cercariae. Trematodes are, together with cestodes, phylogenetically the oldest classes in which glutamate-like material and immunopositivity to a number of neuropeptides isolated from invertebrates has been demonstrated. The results are discussed in relation to immunocytochemical data obtained for other platyhelminths, to endogenous functions of the immunopositive materials, and to their possible role in parasite-host interactions.

Animals↗

Plagiorchis elegans: emergence, longevity and infectivity of cercariae, and host behavioural modifications during cercarial emergence.

We investigated changes in the behaviour of Stagnicola elodes associated with the emergence of Plagiorchis elegans cercariae. Within 15 min of the reduction in light intensity, which triggered the onset of cercarial emergence, infected snails moved to the top of the water column and remained there for 2-3 h. Seventy-nine percent of all cercariae that emerged from the snail did so during this period. Uninfected snails showed no such behavioural changes following the change in light intensity. Cercariae were released in a dense cloud around the snail at the water surface and dispersed passively. Within 3-4 h more than 80% of all cercariae had settled in the bottom 5 cm of the water column. The infectivity of cercariae increased from less than 20% upon emergence from the snail to greater than 75% 4-6 h post-emergence, and then declined steadily to below 5% by 24 h post-emergence. Cercarial longevity was greater than 30 h and exceeded the period of infectivity. This may be related to steadily falling endogenous glycogen levels. A delay in attaining maximum infectivity may represent an adaptive mechanism allowing time for cercarial dissemination, thus reducing superinfection, and subsequent parasite-associated mortality, of second intermediate hosts.

Aedes↗

Effects of heavy metal pollution on swimming and longevity in cercariae of Cryptocotyle lingua (Digenea: Heterophyidae).

Effects of heavy metal pollution on the cercariae of the marine trematode Cryptocotyle lingua (Creplin) were studied by measuring horizontal swimming rate (HSR) and longevity. These factors are important for transmission to the next host, a fish. Cercariae released by Littorina littorea (L.) collected from polluted and unpolluted sites were compared. Both HSR and longevity were significantly reduced in cercariae from the polluted environment. Cercarial quality was therefore reduced, directly or indirectly, by development within a metal-accumulating host. Cercariae released by hosts from a clean environment were subjected to nominal concentrations of 2 and 3 mg/l copper, 1 and 2 mg/l zinc, 2 and 5 mg/l iron and 2 and 4 mg/l manganese in artificial seawater. In all cases the HSRs and longevity were reduced. The effect was more pronounced in the higher concentrations. The significant HSR tests indicate that the absorption and effect of metals occurred within 1 min. The cercarial tegument, specialized for absorption in endoparasitic environments, is possibly responsible. Cercariae may therefore be excellent indicator organisms for pollution. The pollution-induced reductions in cercarial quality seem capable of producing transmission failure. Heavy metal pollution could therefore alter parasite populations and communities.

Animals↗

Development of a novel PCR assay capable of detecting a single Schistosoma japonicum cercaria recovered from Oncomelania hupensis.

Location and time-specific variability in Schistosoma japonicum cercarial density has been shown to be high in the mountainous regions of Sichuan Province, China. A polymerase chain reaction (PCR) assay for the detection of schistosome cercariae in these environments would aid in the determination of environmental risk, and the identification of individual-level risk factors. Here the authors present a highly sensitive and specific PCR assay for the detection of S. japonicum cercariae in laboratory samples. As few as 1 and as many as 300 cercariae, from both laboratory and field-collected S. japonicum strains, produced positive amplification results, and repeated assays showed no positive result for S. mansoni nor for non- japonicum cercariae isolated from infected snails collected in Sichuan Province. There was no difference found between the Chinese and Philippine S. japonicum strains. The results presented demonstrate the successful PCR amplification of a target sequence within the SjR2 retrotransposon from samples of S. japonicum cercariae, with the potential for application to natural water samples from endemic areas.

Animals↗

The infection of human skin by schistosome cercariae: studies using Franz cells.

Franz cells (2-chambered, air/fluid phase static diffusion devices, previously used for the study of drugs across viable human skin) are utilized for the first time to investigate the process of infection of human skin by Schistosoma mansoni cercariae. Skin obtained from cosmetic surgery sources was used in the Franz cells to describe the temporal dynamics of the early interaction of cercariae with skin. At 38 degrees C, about 50% of cercariae applied in water to the epidermal surface of the skin were irreversibly attached within 1 min and after 5 min about 85%, were similarly irrecoverable. The technique also provides the means of following the early penetration path of cercariae by histological methods. Franz cell results on the dynamics of attachment/early penetration have been compared with those obtained using artificial skin equivalents and non-human mammalian skin models in the context of the physical and chemical differences between these systems and viable human skin. It is concluded that Franz cells provide a convenient system for directly investigating the early phases of S. mansoni cercariae interaction with human skin.

Animals↗

Penetration of human skin by the cercariae of Schistosoma mansoni: an investigation of the effect of multiple cercarial applications.

It has previously been postulated that L-arginine emitted by penetrating Schistosoma mansoni cercariae serves as an intraspecific signal guiding other cercariae to the penetration site. It was suggested that penetrating in groups offers a selective advantage. If this hypothesis is correct and group penetration at one site on the host offers an advantage, it would follow that at such a site, successive groups of cercariae would be able to penetrate skin in either greater numbers or at a faster rate. This prediction was tested by the use of an in vitro model of cercarial penetration based on the Franz cell and using human skin. It was demonstrated that there was no increase in the percentage of cercariae able to penetrate the skin with subsequent exposures. Consequently, it seems unlikely that the release of L-arginine by cercariae during penetration could have evolved as a specific orientation system based on a selective advantage offered by group penetration.

Animals↗

Comparison of Schistosoma mansoni irradiated cercariae and Sm23 DNA vaccines.

Immunization with defined antigens is generally less effective at inducing host protection against experimental infection with Schistosoma mansoni than vaccination with attenuated infective cercariae. We predicted that quantitative and/or qualitative differences existed between the immune responses generated to attenuated cercariae and those induced by defined antigens. Thus, we compared immune responses typically associated with protection in the murine model between animals vaccinated with attenuated cercariae and mice immunized with DNA encoding Sm23, a schistosome integral membrane protein that has previously been shown to confer protection. Mice vaccinated three times with attenuated cercariae demonstrated higher levels of protection than Sm23-vaccinated animals but spleen cells from Sm23 DNA vaccinated mice produced significantly higher levels of schistosome antigen-specific IFN-gamma. Both vaccines induced similar levels of Sm23-specific antibody and post-challenge dermal inflammation. However, the pulmonary inflammatory responses following challenge were much less pronounced in DNA immunized animals compared to those receiving irradiated cercariae. Thus, although Sm23 DNA vaccination effectively induced parasite-specific IFN-gamma and antibody responses, it failed to evoke other critical responses needed for optimal vaccine efficacy.

Animals↗

Behavioural and physiological aspects of swimming in cercariae of the digenetic trematode, Proterometra macrostoma.

1. Cyclical swimming behaviour of the cercariae of the digenetic trematode, Proterometra macrostoma, involves a highly regular alternating swim-sink sequence. During periods of swimming the cercariae are propelled upward through the water by alternating lateral contractions of the tail. Following each burst of swimming the cercariae passively drift downward. 2. Suction electrode recordings from the tail during swimming reveal that a single biphasic potential precedes each contraction in a burst. 3. The site of initiation of the rhythmic activity is the transverse band at the junction of the furcae and body of the tail. Sensory feedback does not play a major role in maintenance of the rhythm. 4. A burst of swimming can be initiated by tactile stimulation of the tail. At no time is a cercaria refractory to tactile stimulation. 5. The tail of P. macrostoma cercariae is an autonomous locomotor organ specialized for the brief free-living period between emergence from the snail host and infestation of the primary host.

Action Potentials↗

A new monorchiid cercaria (Digenea) parasitising the purple clam Amiantis purpurata (Bivalvia: Veneridae) in the Southwest Atlantic Ocean, with notes on its gonadal effect.

An unnamed microcercous cercaria (Digenea: Monorchiidae), a parasite of Amiantis purpurata (Lamarck, 1818) (Bivalvia: Veneridae) and its corresponding metacercaria from the province of Buenos Aires and the Patagonian coast of the Southwest Atlantic Ocean, are described. The cercaria described in this paper differs from the three other monorchiid microcercous cercariae, i.e., Lasiotocus minutus (Manter, 1931), Lasiotocus elongatus (Manter, 1931), and Cercaria caribbea XXXVI Cable, 1956, mainly because of the extension of the excretory vesicle and the location of the ventral sucker. Cercariae artificially extracted from sporocysts encyst in a dish and form metacercariae enveloped by a gelatinous sac with two prolongations, which are used to adhere to the substratum. The monorchiid described in this paper has a life cycle similar to those of L. minutus and L. elongatus, although the adult stage of the present species is still unknown. Their larvae are similar in morphology and have venerid clams as their first hosts. The presence of a monorchiid larva is reported for the first time in the Southern Hemisphere. Its monthly prevalence rates, ranging from 0 to 25% (mean: 8.3%), are given from the Patagonian coast. The infection seems to cause castration as it was observed that during March through to May, when most gametes were produced in uninfected individuals, 81% of the infected individuals did not produce gametes.

Animals↗

Production of Schistosoma mansoni cercariae by Biomphalaria glabrata from a focus in Belo Horizonte, Minas Gerais.

The snail density, levels of infection and the monthly production of Schistosoma mansoni cercariae by Biomphalaria glabrata were determined in a focus of Barreiro de Baixo (Belo Horizonte, MG, Brazil). During a period of 38 months (1984 to 1987) 5,366 snails were collected of which 324 (6.03%) were infected with S. mansoni. The total number of cercariae shed was 5,667,312. Each snail shed an average of 17,422 cercariae during the time that it was under study in the laboratory. The greatest longevity of infected snails was 218 days. Natural cure was observed in 42 (12.9%) of the infected specimens about 130 days after collection. The average snail density in the focus during the period of study was 16.3 snails per scoop. The shedding of cercariae by snails collected from the field was compared with laboratory bred specimens infected in mass with the LE strain of S. mansoni from Belo Horizonte. The laboratory infected snails shed an average of 6,061 cercariae each, a value 2.8 times less than the field specimens due to a shorter life span. The prevalence of schistosomiasis in the focus was 14.3%.

Animals↗

Influence of temperature on development of Schistosoma mansoni female cercariae in Biomphalaria glabrata.

In these experiments the ratio of male to female S. mansoni larvae in B. glabrata from Belo Horizonte and Ribeirão das Neves Minas Gerais, Brazil, either reared in laboratory or collected in the field, varied from 1:1 to 1:1.3 or 1.4:1. Cercariae of LE strain of Schistosoma mansoni, shed by 39 snails maintained at 25 +/- 0.5 degrees C were used to infect mice on a weekly basis. Subsequent perfusion resulted in 76.6% male and 23.4% female worms. The cercariae produced by 32 infected snails maintained at 27 +/- 0.5 degrees C were inoculated into mice and produced 43.4% male and 56.6% female worms (p < 0.05). Cercariae eliminated by snails collected in Barreiro and Ressaca, Belo Horizonte, during hot months, produced 45.7 to 47.7% male and 52.3 to 54.3% female worms. A lower number of cercariae shed by snails collected in Gorduras, Belo Horizonte, at 20 +/- 3.0 degrees C, produced 51.6% male and 48.4% female worms. Thus, in this region the infection of vertebrate hosts with S. mansoni cercariae would be more severe in the summer due to the higher level of parasites and the number of eggs.

Animals↗

Reappraisal of experimental infections with cercariae and schistosomula of a Brazilian strain of Schistosoma mansoni in mice.

The present investigation involves a reevaluation of previous results obtained after experimental infection of Swiss Webster mice with cercariae and schistosomula of the Schistosoma mansoni LE strain maintained under laboratory conditions. Three experimental groups of mice were considered: the animals of the first group were percutaneously (ring method) infected with cercariae, those of the second were subcutaneously inoculated with cercariae and the mice of the third were inoculated by the same route with schistosomula transformed in vitro. The data obtained so far indicated that the most effective method of infection is the subcutaneous injection with schistosomula, with a mean adult worm burden recovery of 54.1% when compared to the abdominal percutaneous and subcutaneous routes of infection with cercariae, in which the values were 36.7% and 32.4%, respectively. This suggests that, in experimental infections of SW mice with a LE S. mansoni strain, the skin is to be considered an effective attrition site in the percutaneous route, whereas in the case of inoculation with cercariae, a small amount of larvae fails to be transformed into viable schistosomula, possibly due to skin phase avoidance. A brief discussion about attrition sites and elimination of larval S. mansoni worms in mice is presented.

Animals↗