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PubMed1990

[Ecological studies on the cercariae of Echinochasmus japonicus].

Results of ecological observation on the cercariae of Echinochasmus japonicus showed that their development in and emergence from Parafossarulus striatulus as well as their activity in infecting the fish host in water were closely related to temperature and light. The number of cercariae emerged varied with the temperature of water. When the infected snails were reared at water temperature of 27-32 degrees C, the germinal cells and germ balls in the daughter radiae multiplied continually into cercariae a majority of which emerged. At water temperature below 24 degrees C, multiplication was hindered and, emergence of cercariae obviously decreased. Cercaria shedding stopped at water temperature of 14 degrees C. The reason why the rate and degree of infection with meta-cercariae of Echinochasmus japonicus in the small-sized species of fresh water fishes were higher than in the large-sized species was that the cercariae were tactic to light and so active in shallow water where they were constantly in contact with smaller fishes. Since the natural infection rate of Parafossa-rulus striatulus was found to be higher (32.2%; 37.3%) during the months of July-October than during the other months, investigations on the first intermediate host should be carried out in early autumn.

Animals↗
PubMed1997

The population dynamics of cercariae of Schistosoma japonicum in Oncomelania hupensis.

The population dynamics and production of cercariae of Schistosoma japonicum in Oncomelania hupensis are reported. The experiments covered the whole life span of positive snails and different intervals of cercariae shedding. The results indicated that two patterns of the dynamics of cercariae shedding had been found in the life span of positive snails. The first was a long-time interval (4-7 days) and progressive decline pattern. The cercariae shedding of positive snails lasted 18-19 weeks in males and for 32-33 weeks (once a week). The second was a short-time interval (1-3 days) and continued release pattern. The cercariae shedding of positive snails lasted for 20-36 days (every day shedding). Shedding cercariae stimulate cercariae development.

Animals↗
PubMed1999

Paramphistomum daubneyi and Fasciola hepatica: influence of temperature changes on the shedding of cercariae from dually infected Lymnaea truncatula.

Dual infections of Lymnaea truncatula with Paramphistomum daubneyi and Fasciola hepatica were performed to determine whether temperature changes in snails (daily water change with spring water at 6 degrees-8 degrees C, which subsequently increased to room temperature at 20 degrees C) would influence snail infection and the production of cercariae by both trematodes. At day 30 post-exposure the surviving snails were individually placed in petri dishes to constitute two groups. Snails from the first group were maintained at a temperature of 20 degrees C, and the water in the petri dishes was changed daily. The protocol was identical for the second group of snails except that the water temperature was 6 degrees-8 degrees C when changed. The interval between exposure and the first shedding of cercariae in snails immersed in cold water for a short period was longer (67-69 days instead of 48-50 days in the 20 degrees C group). In both groups, snails infected only with F. hepatica or P. daubneyi or with both trematodes were detected. In snails infected only with F. hepatica the frequency of cercaria-shedding snails and the total number of metacercariae were significantly greater in the 20 degrees C group. Inversely, in snails infected only with P. daubneyi the frequency of cercaria-shedding snails and the number of metacercariae were significantly greater in the 6 degrees-8 degrees C group. In snails harboring both trematode larval forms, no significant difference in the frequencies of cercaria-shedding snails between the two groups was noted. Metacercariae of both trematodes were obtained from these snails. In the 20 degrees C group, F. hepatica metacercariae were more numerous, whereas in the 6 degrees-8 degrees C group the number of P. daubneyi metacercariae was greater. From these results it appears that greater activity of P. daubneyi cercariae occurs in snails subjected to daily temperature changes (from 6 degrees to 20 degrees C).

Animals↗
PubMed1981

Protein synthesis in vivo by Schistosoma mansoni cercariae.

Cercariae of Schistosoma mansoni were labeled with [14C]leucine in vivo and their protein studied by one- and two-dimensional electrophoretic fluorography. With molecular weight (Mr) and isoelectric point (pI) as parameters, polypeptides from different stages of cercarial embryogenesis were compared with each other and with those of adult worms. Up to six polypeptides with different isoelectric points were found in a single electrophoretic band of the same Mr. Three polypeptides of high molecular weight were synthesized by cercariae 3 days before escape, and were similar to polypeptides synthesized by adults. The most prominent cercarial polypeptides synthesized three days before emergence (during tail elongation) are two which comigrate electrophoretically with actin and heavy chain of myosin. The rate of actin synthesis decreased at 48 and 24 h, with little or no synthesis in the 4 h preceding emergence. Many polypeptides found in cercariae labeled 4 h before emergence differ in pI and Mr from those synthesized in earlier stages of embryogenesis. From the water in which the cercariae emerge, labeled polypeptides were precipitated which are identical to several found both in shed, washed cercariae and the infected hepatopancreas. Polypeptides of high Mr are not synthesized by cercariae in the period shortly before escape; this stage is represented by polypeptide products with Mr values of 35 000, 53 000 and 58 000.

Animals↗
PubMed1997

Low molecular weight hydrophilic chemicals that attract Echinostoma trivolvis and E. caproni cercariae.

A Petri dish bioassay was used to study chemoattraction of the cercariae of Echinostoma trivolvis and E. caproni in the presence of various bench chemicals prepared as 5 mM solutions in agar. The design compared the attraction of cercariae migrating toward the plugs impregnated with chemicals compared to unimpregnated plugs (controls). Cercariae of both species were significantly attracted to an array of amino acids and to some carbohydrates. Cercariae of E. trivolvis but not E. caproni were attracted to the peptide glutathione. Cercariae of E. trivolvis and E. caproni were repelled from valeric and butyric acids, respectively. Cercariae of E. caproni but not E. trivolvis were repelled from dextrose. Although broad specificity exists for echinostome cercarial penetration in snail intermediate hosts, results of these studies suggest that there are specific differences in cercarial chemoattractants between E. trivolvis and E. caproni.

Amino Acids↗
PubMed2005

Effects of host habitat quality on the viability of Cryptocotyle lingua (Trematoda: Digenea) cercariae.

In an earlier study it was shown that Cryptocotyle lingua cercariae, matured in Littorina littorea from a polluted marine lagoon, displayed slower horizontal swimming rates, and reduced longevity compared to cercariae released by periwinkles from a cleaner environment. This work investigated whether the pollution-induced reduction in swimming rates was due to an inefficient swimming action or the adoption of a less direct swimming path. In addition, cercariae from L. littorea that had been transferred from an 'unpolluted' to a 'polluted' site for 1 month provided information on the speed with which pollutants affect cercariae through their intermediate hosts. Results indicated that, in general, horizontal swimming rates were reduced due to slower swimming rather than disorientation and longer swimming pathways. Effects of host transplantation to a polluted site were clearly evident after 1 month. Evidence suggested that the pollutants accumulated by the cercariae via their first intermediate host affected the neuromusculature associated with swimming performance rather than sensory structures. Bearing in mind the reduced viability of C. lingua cercariae in polluted sites it is assumed that high prevalence of this digenean in gastropods (at such sites) must be due to their continual introduction by infected birds attracted to these habitats from other areas.

Animals↗
PubMed2005

A study of the mechanisms by which the cercariae of Microphallus primas (Jag, 1909) Stunkard, 1957 penetrate the shore crab, Carcinus maenas (L).

This study established the mechanisms by which Microphallus primas cercariae penetrate the crab Carcinus maenas in which they form metacercarial cysts. Light and electron microscopy were used to investigate cercarial features and to follow the fate of cercariae released in close proximity to crabs. It was shown that cercariae were carried in respiratory currents into crabs' branchial chambers where each enveloped itself in a transparent penetration cyst on the gill lamellae. When cercariae were present the number of respiratory current reversals performed by crabs increased. Using an 'artificial branchial chamber' it was possible to observe how cercariae attached to crab gills during breaks in current flow that preceded each current reversal. Inside the penetration cysts the now tail-less larvae used their stylets to pierce holes through which they levered themselves into underlying haemolymph channels in the gills. Histochemical tests demonstrated that the penetration cysts were products of glands in the cercariae and that penetration of the crabs was achieved by mechanical means. The importance of crab respiratory current reversals to the success of cercarial penetration is discussed as it represents the exploitation by a parasite of a host behavioural response to an unrelated stimulus.

Animals↗
PubMed1999

The influence of temperature on the survival and infectivity of the cercariae of Echinoparyphium recurvatum (Digenea: Echinostomatidae).

Survival and infectivity characteristics are described for cercariae of the echinostome Echinoparyphium recurvatum at 10, 15, 20, 25 and 30 degrees C. Cercarial survival was markedly age- and temperature-dependent, maximum survival time being reduced from 68 h at 10 degrees C to 12 h at 30 degrees C, 50% survival times falling from 47.9 h at 10 degrees C to 8.4 h at 30 degrees C. The mean instantaneous per capita death rate of cercariae increased exponentially with temperature. Cercarial infectivity was also temperature dependent, the main effect of temperature being upon the rate at which infectivity diminished with increasing age of cercariae. Infectivity increased from zero at 10 degrees C to a maximum at 25 degrees C after which it declined to a low level at 30 degrees C. Overall transmission efficiency of cercariae at each experimental temperature was calculated as the ratio of the mean instantaneous per capita death rate (mu) to the mean instantaneous per capita rate of infection (beta). Transmission efficiency of cercariae was found to be maximal at 20 degrees C and at a relatively high level between 10 and 25 degrees C. Transmission efficiency was zero at 10 degrees C and at a very low level at 30 degrees C. These results suggest that the cercariae of E. recurvatum show transmission optima at water temperatures likely to be encountered in natural habitats in Britain and Europe.

Aging↗
PubMed1999

Vaccination of mice with gamma-irradiated Schistosoma japonicum cercariae.

gamma-irradiated cercarial vaccines induce high levels of protection in mice against Schistosoma mansoni infection, however, the same has not been well established for S. japonicum. Here we describe vaccination studies in mice with gamma-irradiated S. japonicum cercariae testing the effectiveness of different irradiation doses, number of vaccinations, and mouse strains. In CBA/Ca mice, a single percutaneous exposure to 500 S. japonicum cercariae previously attenuated by 10, 20, 30, 40 or 50 krad gamma-irradiation induced significant, but comparable levels of protection (34-46%) against challenge infection. In a repeat experiment in C57Bl/6 mice, only groups vaccinated with 10 or 20 krad gamma-irradiated cercariae showed statistically significant, but lower levels of resistance (20-24%). Multiple vaccination of CBA/Ca mice with 500 20 krad gamma-irradiated cercariae did not improve the resistance level (40%). Analysis of IgG responses showed no clear correlation between antibody levels and levels of protection. Western blot analysis suggested that recognition of a 200-kDa antigen might be correlated with protection, that antigens of 42 and 50 kDa may be involved in the protection induced by single vaccination, but that different antigens might be protective in single vs multiple vaccinations. Sera from mice vaccinated with gamma-irradiated cercariae recognized many fewer antigens than more protective sera from mice vaccinated with UV-attenuated cercariae. These results suggest that the mouse may not be a suitable host for studies involving gamma-irradiated S. japonicum vaccines.

Animals↗
PubMed1993

Candidate vaccine antigens identified by antibodies from mice vaccinated with 15- or 50-kilorad-irradiated cercariae of Schistosoma mansoni.

In murine schistosomiasis, the highest levels of resistance to cercarial challenge are obtained by vaccination with radiation-attenuated cercariae. To identify candidate vaccine antigens relevant to the vaccine model, we examined parasite antigens recognized by antibodies from mice vaccinated with irradiated cercariae of Schistosoma mansoni. To optimize recognition of a wide spectrum of antigens, several factors that influence the level of protection in this model were varied; specifically, we examined the effect of (i) single versus multiple vaccinations with irradiated cercariae, (ii) the dose of irradiation (15 or 50 kilorads) administered to the cercariae, and (iii) the genetic background of mouse strains, high-responder (C57BL/6J) versus moderate-responder (CBA/J) mice. We found that the number of vaccinations did not alter antibody specificity but modified the relative antibody titers against particular antigens. The dose of irradiation used to attenuate the immunizing cercariae had a similar effect on antibody titers but in addition influenced antibody specificity. Only mice that had been vaccinated with moderately irradiated cercariae recognized cathepsin B (Sm31) and Sm32. Interestingly, when vaccinated mice of the two strains, C57BL/6J and CBA/J, were compared, differences in antibody responses to particular antigens were observed. Both strains recognized the integral membrane protein Sm23, glutathione S-transferase, and cathepsin B, whereas Sm32 and paramyosin were recognized only by CBA/J mice, and heat shock protein 70 was recognized exclusively by C57BL/6J mice. In this study, we conclusively identified six distinct antigens that are specifically recognized by the humoral immune response of vaccinated mice.

Animals↗
PubMedDate not supplied

[A stock of infected snails for the mass breeding of cercariae and schistosomules].

A routine to maintain infected snails Biomphalaria glabrata able to produce 1-2 million cercariae per week and a mean shedding of about 3,500 cercariae per snail was quantitatively described. To a monthly production of about 5.3 and 6.5 million cercariae during the years of 1982 and 1983 it was necessary to expose an average of 1,557 and 1,957 snails, respectively. The efficiency described was maintained by infecting 2,000 snails per month (infectivity index of about 60%). A maximal production of 6,000 cercariae/snail was obtained in the period between 56-70 days after the snail infection by miracidia. By this period, however, 90% of the infected snails were dead. In the summer of 1982-1983 it was observed an impairment on the daily cercarial harvests which was related to ambient temperature increase, and the presence of rotifers in the aquaria water. When the cercariae were transformed into schistosomula in vitro, a yield of 55% schistosomula was obtained, with 7% of tail contamination. Lyophilization of cercariae produced 50.9 +/- 63 g dry weight per 1,000 cercariae.

Animals↗
PubMed2000

Neutral lipids in cercariae, encysted metacercariae, and rediae of Zygocotyle lunata.

High-performance thin-layer chromatography was used to analyze the neutral lipids in the rediae, cercariae, and encysted metacercariae of the paramphistomid trematode Zygocotyle lunata. Visual observations of the chromatograms showed that the most abundant lipid fractions were free sterols and free fatty acids in all larval stages and triacylglycerols in the metacercariae and rediae. The weight of free sterols (x +/- SE) was 120+/-20 ng/cercaria, 56+/-3.8 ng/redia, and 5.9+/-1.5 ng/encysted metacercaria; the weight of triacylglycerols was 13+/-0.88 ng/encysted metacercaria, 6.3+/-0.063 ng/redia, and was not detectable in the cercaria; the weight of free fatty acids was 160+/-17 ng/ cercaria, 76+/-9.1 ng/redia, and 4.2+/-0.46 ng/encysted metacercaria. Oil red O staining of whole larvae showed the presence of neutral lipids in the rediae but not in the cercariae or encysted metacercariae. A dramatic reduction was seen in the quantity of free sterols and free fatty acids in the encysted metacercariae as compared with the cercariae, suggesting that these neutral lipids are used in some way during the transformation from cercaria to metacercaria.

Animals↗
PubMed2003

Relationship of tadpole stage to location of echinostome cercariae encystment and the consequences for tadpole survival.

The effect of echinostome infections on the survival of Rana pipiens tadpoles was examined in relation to developmental stage of tadpoles. Individual tadpoles of Gosner stages 25, 27, 32-33, and 37-39 were exposed to 1 of 4 levels of cercariae (0, 20, 50, or 100). Only tadpoles at stage 25, the earliest stage infected, died within a 5-day experimental period. This stage-specific mortality rate could be explained, in part, by the stage-specific location of encystment of cercariae, which was documented in a separate experiment. In accordance with kidney development, cercariae predominately encysted in the pronephroi during early stages of tadpole development (stages 25 through 31-32) and only in the mesonephroi and associated ducts at later stages (stages 37 through 46). As the mesonephros develops, renal capacity presumably increases. Thus, tadpoles died only when metacercariae concentrated in the functional portion of the kidney with the most limited renal capacity. As tadpoles aged, they also became less susceptible to infections. On average, 69.5% of cercariae that were exposed to stage 25-26 tadpoles successfully encysted. compared with only 8.4% of cercariae exposed to stage 37-38 tadpoles. Exposures of metamorphic frogs (poststage 46) to cercariae revealed that these individuals can become infected with echinostomes. Collectively, our data highlight the host stage-dependent dynamics of tadpole-echinostome interactions.

Analysis of Variance↗
PubMed2004

Effects of snail size and diet on encystment of Echinostoma caproni cercariae in juvenile Helisoma trivolvis (Colorado strain) and observations on survival of infected snails.

The effects of snail size and diet on encystment of Echinostoma caproni cercariae in juvenile Helisoma trivolvis (Colorado strain) snails were studied. Encystment in neonatal (<1-mm shell diameter) and juvenile (2- to 3-mm shell diameter) snails was compared 24 hr postinfection (PI) after individual exposure of snails of each size to 1, 5, 10, 25, or 50 cercariae. Significantly more cysts were recovered from juvenile snails exposed to 10, 25, or 50 cercariae than from neonatals with comparable exposure. The maximum number of cysts recovered from juveniles exposed to 50 cercariae was 42, compared with a maximum of 15 cysts in neonatals comparably exposed. Size of H. trivolvis was a major factor in determining cyst burden in this planorbid. A diet of either Romaine lettuce leaf or hen's egg yolk did not have a significant effect on the number of cysts recovered at 24 hr PI from juvenile snails exposed to 25 or 75 cercariae. Survival of infected versus uninfected neonatals was also examined for 7 days. Neonatals exposed to 10 cercariae showed a significant decrease in survival at 6 and 7 days PI when compared with uninfected controls.

Analysis of Variance↗
PubMed2004

Effects of copper sulfate toxicity on cercariae and metacercariae of Echinostoma caproni and Echinostoma trivolvis and on the survival of Biomphalaria glabrata snails.

Copper in the form of copper sulfate (CuSO4) decreases the survival of Biomphalaria glabrata snails, but the effects of this molluscicide on Echinostoma caproni and Echinostoma trivolvis, 2 species of digeneans that use B. glabrata as intermediate hosts, are not known. Studies were done on the effects of various concentrations of CuSO4 in artificial spring water (ASW) on the survival and infectivity of E. caproni and E. trivolvis cercariae. Solutions containing 1.0, 0.1, and 0.01% CuSO4 were 100% lethal within 2 hr of exposure for both species. Time to 50% mortality in 0.001% CuSO4 was 8 hr for E. caproni and 16 hr for E. trivolvis; at 24 hr, the controls showed 50 and 65% mortality, respectively. Treatment of cercariae of both species for 0.5 hr in 0.001% CuSO4 had no effect on the ability of cercariae to form normal cysts in juvenile B. glabrata snails. However, treatment with 0.01% CuSO4 for 0.5 hr caused a significant reduction in the ability of cercariae of both species to encyst in snails. Treatment of encysted metacercariae of both species in 0.001% CuSO4 for I hr had no effect on subsequent excystation of these echinostomes in a trypsin-bile salts medium, whereas concentrations of 1.0, 0.1, and 0.01% CuSO4 and 1.0 and 0.1% CuSO4 decreased chemical excystation of E. caproni and E. trivolvis cysts, respectively. Survival studies on the effects of CuSO4 in Locke's solution on chemically excysted metacercariae of both species were also done. Excysted metacercariae of both species were killed by 2 hr in either 0.1 or 0.01% CuSO4 in Locke's solution. However, time to 50% mortality for both species of excysted metacercariae in 0.001% CuSO4 was approximately 5 hr. Time to 50% mortality for the controls was about 12 hr. Survival of juvenile B. glabrata snails was also examined. All B. glabrata snails were dead by 6 hr in 1 and 0.1% CuSO4 in ASW. Biomphalaria glabrata snails showed 50% mortality by about 6 hr in 0.01% CuSO4 and about 80% were still alive at 24 hr in 0.001% CuSO4. All controls were alive at 24 hr, at which time the experiment was terminated. Concentrations greater than 0.001% CuSO4 increased snail mortality, as well as that of the cercariae and excysted metacercariae of E. caproni and E. trivolvis. Our findings suggest that concentrations of copper sufficient to eliminate juvenile B. glabratta snails are also sufficient to kill the cercariae and excysted metacercariae of these digeneans but not the encysted metacercariae, which may be protected by their cyst walls.

Analysis of Variance↗
PubMed2006

Rearing density and susceptibility of Rana pipiens metamorphs to cercariae of a digenetic trematode.

Cercariae of many digenean trematodes target particular developmental stages of their hosts. For some digeneans that are parasites of amphibians, infection appears timed to host metamorphosis. The success and timing of metamorphosis is itself affected by a number of factors, including host density. We investigated the degree to which rearing density of Rana pipiens larvae influenced time to metamorphosis and snout-to-vent length and mass at metamorphosis, as well as establishment of cercariae of the trematode Manodistomum syntomentera Stafford, 1905. As expected, individuals metamorphosed later, were smaller, and weighed less at metamorphosis if they were reared under intermediate to high densities compared with low densities, in experimental outdoor mesocosms. Cercariae establishment was higher in smaller metamorphs that took longer to metamorphose within the low-density treatment. Additionally, cercariae establishment was lower in larvae from the low-density tanks compared with larvae from the intermediate- to high-density tanks. However, more tadpoles had failed to metamorphose in the intermediate to high rearing densities by the time cercariae were no longer available from natural collections of first intermediate hosts, Physa spp. Larval amphibians under crowded conditions should experience increased susceptibility to trematode establishment in nature, but only if they metamorphose within the time period when cercariae are still available.

Analysis of Variance↗
PubMed2002

[Preliminary study of cercaria antigen of Schistosoma japonicum before and after ultraviolet irradiation].

OBJECTIVE: To study the changes in the constituents of the cercaria antigen of Schistosoma japonicum before and after ultraviolet irradiation. METHODS: The cercaria of Schistosoma japonicum were exposed to ultraviolet light (UV) irradiation at a dose of 400 mgrW/cm2 for 1 min, and the UV-irradiated cercaria antigen (UVCA) and normal cercaria antigen (NCA) were simultaneously analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. RESULTS: At least 2 antigens with relative molecular mass (Mr) of 212 000 and 82 000 were identified in UVCA but not in NCA by SDS-PAGE analysis, and the concentrations of the antigens with Mr of 116 000, 26 000 and 16 000 in UVCA were significant higher than those in NCA. On the other hand, the antigenic molecule with Mr of 67 000 in NCA was recognized by serum from pigs vaccinated with UV-attenuated cercariae, but not by serum from pigs with Schistosoma japonicum infection. Antigens with Mr of 79 000 and 94 000 were apparently more strongly reactive with the former porcine serum than with the latter. CONCLUSION: The results suggest that all the novel antigens arising from or increased by UV exposure, or antigens specifically recognized by serum from pigs vaccinated by UV-attenuated carcariae may be the principal factors in the highly protective immunity provoked by irradiated cercariae.

Animals↗
PubMed1975

Oxidative enzymes in the development of Fasciola hepatica L. V. Activity of oxidases and dehydrogenases in the Cercaria and Metacercaria.

The object of the study was the investigation of the occurrence and localization of oxidative enzymes in the 4th and 5th larval stages of the liver fluke, i. e. in the cercaria and metacercaria. The following enzymes were detected: cytochrome oxidase, peroxidase, NADH and NADPH tetrazolium reductases/diaphorases) as well as succinate, isocitrate, malate, lactate, alpha-glycerophosphate, glyceraldehyde phosphate, glucose-6-phosphate, 6-phosphogluconate, beta-hydroxybutyrate, L-glutamate and alcohol dehydrogenases. The occurrence and localization of the enzymes were investigated histochemically in the cercaria still unreleased from the snail tissues, in the free natatorial cercaria, and in the encysted specimen, i.e. metacercaria. Among the enzymes studied only peroxidase was found to be absent from the cercaria and metacercaria, the latter larva being deprived of alcohol and L-glutamate dehydrogenases as well. The aerobic path, i.e. the Krebs cycle, was ascertained as the principal mode of obtaining energy in the free natatorial cercaria and metacercaria, and glycolysis as the main energy path for the undetached larva. The analysis of the above-mentioned oxidative enzymes was the basis for the description, within the available range, of the function and metabolism of individual organs of cercaria in different periods of its development. In the recapitulation the author discusses the effect of the parasitic way of life of the larval forms of Fasciola hepatica on their energy metabolism.

Alcohol Oxidoreductases↗