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Comparative analysis of the Schistosoma mansoni and Schistosoma bovis cercarial production under the influence of praziquantel.

Snails infected with Schistosoma mansoni and S. bovis and fed with a food-Praziquantel mixture stop shedding cercariae for several days. The cercarial production restarts at different levels after stopping treatment: for some Biomphalaria glabrata, the restarting phase of production reaches a high level whereas for other B. glabrata and all of the P. metidjensis, the level of production remains low. Histological studies revealed that at the exact moment of this treatment, there is a total destruction of many nearly mature cercariae whereas the young cercarial embryos and the tegument of the sporocyst remain unharmed. When treatment stops, there is a pronounced proliferation of third-generation sporocysts (Sp III) which invade the available space of the snails' digestive gland.

Animals↗

Discussion of the relationships between Schistosoma and their intermediate hosts, assessment of the degree of host-parasite compatibility and evaluation of schistosome taxonomy.

Studies of the spectrum of the intermediate hosts of schistosomes and the relationships between parasites and intermediate host snails have been carried out with each researcher using different methods. A comparison between compatibility studies has therefore been impossible. The following three basic problems in these kind of studies are discussed in this paper: (1) the standardization of experimental materials such as schistosomes, intermediate and final hosts, (2) the methods involved in the experiments, and (3) the sort of data which must be collected. Standardized methods and materials are described in this paper. The relationships between the genus Schistosoma and the intermediate host snails belonging to the two genera Biomphalaria and Bulinus, and the mechanisms behind the process resulting in various degrees of compatibility between the two involved organisms are reviewed and discussed. Why certain combinations of schistosomes and snails result in the production of cercariae and others do not is still unknown, but there is now some indication of a camouflage behaviour during the intramolluscan stages, e.g., the larvae in the snails cover themselves with snail material and are not recognized as foreign objects. The same mechanism has already been found in the schistosomules in the final host. Much of the confusion in the discussion of the relationships between schistosomes and intermediate hosts is the result of a lack of an objective way to describe and assess the compatibility between the two. Based on a series of experiments with S. haematobium, S. intercalatum, S. bovis, S. mansoni, and several species of possible intermediate hosts, the following index for estimating the degree of compatibility is proposed: the total cercarial production from 100 exposed snails (TCP/100 exposed snails). Seven classes have been suggested: the first one (Class 0) with a TCP/100 exposed snails of zero, is called 'refractory'; the next one (Class I) called 'not very compatible' has a TCP /100 exposed snails between 1 and 10,000; the last group (Class IV) called 'extremely compatible' produced more than 500,001 per 100 exposed snails during the entire lifespan. The ability to use a spectrum of intermediate hosts and the compatibility in the taxonomy of schistosomes is discussed. The results for the different species of schistosomes indicated that each species consists of strains with 'hybrid populations' in between.

Animals↗

Toxicity evaluation of Bayluscide and malathion to three developmental stages of freshwater snails.

Laboratory studies were conducted to assess the conditions under which the use of malathion in ricelands of Cameroon may impact the transmission of schistosomiasis. Helisoma trivolvis and Biomphalaria havanensis were selected as test organisms due to the lack of intermediate snail hosts in the U.S. Using Bayluscide as a reference molluscicidal compound, malathion was tested against snail eggs, juveniles, and adults. Snail eggs were more susceptible to Bayluscide and malathion than juvenile snails which in turn were more susceptible than adult snails. A Bayluscide concentration of 0.200 mg/L caused 100% mortality to adults of both snail species after 24 h exposure. This relatively high toxicity of Bayluscide to freshwater snails is one of the reasons why it has been recommended by the World Health Organization as the molluscicide of choice for control of schistosome-bearing snails. The concentrations of malathion resulting in 100% kill of adult snails after 24 h exposure were 1,200 mg/L for H. trivolvis and 500 mg/L for B. havanensis. After 48 h exposure, these concentrations were reduced to 500 mg/L and 300 mg/L, respectively. Therefore it is expected that the use of malathion for insect control in ricelands of Cameroon may affect the survival of freshwater snails including the intermediate hosts of bilharziasis.

Animals↗

Toxicity evaluation of ammonium sulphate and urea to three developmental stages of freshwater snails.

Studies were performed to evaluate the toxic effects of ammonium sulphate and urea (chemical fertilizers currently applied in ricelands of Cameroon) against eggs, juveniles, and adults of two species of freshwater snails (Helisoma trivolvis and Biomphalaria havanensis). Results obtained from ammonium sulphate tests indicated 24-h LC50 values of 558 mg/L and 669 mg/L for eggs; 393 mg/L and 526 mg/L for juveniles, and 701 mg/L and 657 mg/L for adults of H. trivolvis and B. havanensis, respectively. Similar analysis with urea revealed LC50 values of 14,241 mg/L and 13,532 mg/L for eggs; 18,255 mg/L and 24,504 mg/L for juveniles and 30,060 mg/L and 26,024 mg/L for adults of H. trivolvis and B. havanensis, respectively. Following 48 h exposure, the concentrations of ammonium sulphate killing 100% of snails were 1,250 mg/L and 1,000 mg/L for the adults of H. trivolvis and of B. havanensis, respectively. Those of urea were computed to be 25,000 mg/L for H. trivolvis and 35,000 mg/L for B. havanensis. In rice culture in Cameroon, these fertilizers are applied at doses of 100 kg/ha (ammonium sulphate) and of 150 kg/ha (urea); hence, the above found concentrations lethal to snails appeared to be 10 to 13 times (ammonium sulphate) and to be 165 to 235 times (urea) higher assuming an average water depth of 10 cm in these ricefields. Therefore, the use of ammonium sulphate and urea as chemical fertilizers in ricelands of the Republic of Cameroon might adversely affect the survival of freshwater snails only in the case of spills or of stressful environmental conditions. Under normal laboratory conditions, both chemicals show a low molluscicidal activity with urea being about 25 to 35 times less potent than ammonium sulphate.

Ammonium Sulfate↗

Fermentation and toxin studies of the molluscicidal strains of Bacillus brevis.

Strain SS86-4 was one of 40 Bacillus brevis strains shown to be molluscicidal to the schistosomiasis snail vector Biomphalaria glabrata. When grown in mB4 medium in 2-L fermentors, SS86-4 was molluscicidal only if fructose or phenylalanine was present in the medium. This is reminiscent of secondary fermentation factor effects, in this case an antioxidant effect. In vivo proteases also were capable of reducing molluscicidal activity. The molluscicidal toxin has an LC50 of 1 microgram toxin protein ml-1 (approx. 1 p.p.m.) and may be described as a small proteinaceous, heat-stable, oxygen-sensitive entity associated with the particulate portion of the cell wall fraction of B. brevis that is formed prior to sporulation. Initial information indicates that its HPLC signature shows major peaks at 148.37 and 163.96 s and consists of two bands of approximately 5.3 kDa and 8.7 kDa on PAGE gel.

Animals↗

Physiologic studies of snail-schistosome interactions and potential for improvement of in vitro culture of schistosomes.

Despite extensive efforts to develop suitable media for rearing the intramolluscan stages of schistosomes, successful in vitro culture of these parasites remains elusive. Recent 31P NMR studies demonstrated that the levels of free phospholipids, particularly phosphatidylcholine, in the digestive gland of the snail, Biomphalaria glabrata, were dramatically reduced when the host was infected with Schistosoma mansoni. It was speculated that absorption of host phosphatides may be an important source of membrane phospholipid precursors and fatty acids for developing sporocysts and cercariae. During the present investigations, B. glabrata was maintained on a high fat diet of egg yolk, and the lipid composition of control uninfected and infected snails examined by 31P and 13C NMR. In addition, the levels of host hemolymph metabolites, including glucose and urea, considered as indicators of parasite nutrient uptake, were monitored. The lipid level of snails fed egg yolk was greatly increased, and hosts developed patient infections in approximately half the time of infected snails maintained on lettuce. The composition of the free phospholipids accumulated in the tissues of B. glabrata fed egg yolk were the same as those previously reported in the cercarial stage of S. mansoni. Moreover, the fatty acids of S. mansoni and those reported here in the neutral lipids and free phosphatides in the host tissues were similar. Uninfected snails maintained on lettuce had higher hemolymph levels of glucose than those reared on egg yolk, and infected hosts on egg yolk had significantly lower levels of hemolymph urea. Beta-hydroxybutyrate was the principal hemolymph metabolite in snails fed egg yolk, but was not detected in snails maintained on lettuce.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗

Molluscicidal activity and clinico-pathological effect of Agave lophantha.

Dry powder and different extracts of Agave lophantha were tested against Biomphalaria alexandrina. The results showed that the butanol extract has high molluscicidal activity. The activity of the dry powder has been found to be stable under the effect of some simulated field conditions. Also the toxicological effect of the plant on mice was tested through determination of certain parameters such as total protein, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase enzymes as well as histopathological study on liver and kidney.

Animals↗

IMP synthesis using immobilized adenosine (phosphate) deaminase.

Inosinic acid (IMP) has been prepared by the deamination of adenosine monophosphate (AMP) with an immobilized adenosine (phosphate) deaminase extracted from the snail Biomphalaria glabrata. The enzyme has been immobilized in polyacrylamide beads. The preparation and characterization of this system are described.

Adenosine Deaminase↗

The life cycle of Clinostomum complanatum Rudolphi, 1814 (Digenea, Clinostomidae) on the floodplain of the high Paraná river, Brazil.

The life cycle of the digenetic trematode Clinostomum complanatum was studied on the floodplain of the high Paraná river, Brazil. Sampling was done between June 1999 and June 2000. The mollusc Biomphalaria peregrina was the first intermediate host with a prevalence of infection of 0.75%. The fish Loricariichthys platymetopon, Hoplosternum littorale, Parauchenipterus galeatus, Hoplias malabaricus and Loricaria sp. were second intermediate hosts, with the prevalence varying from 2.45% ( H. malabaricus) to 60.8% ( L. platymetopon). The birds Ardea cocoi, Phalacrocorax brasilianus, Egretta alba and Egretta thula were definitive hosts, with the prevalence ranging between 5.5% ( E. thula) and 95% ( A. cocoi). The high prevalences in birds, especially in A. cocoi, suggest some kind of mechanism facilitating the predation of infected fish. The importance of the various hosts is discussed, and it is concluded that the floodplain of the high Paraná river has suitable conditions for the maintenance of the life cycle of C. complanatum.

Animals↗

Schistosoma mansoni miracidial host-finding: species specificity of an Egyptian strain.

The effect of snail-conditioned water (SCW) from Biomphalaria alexandrina, a pigmented and an albino strain of B. glabrata, and Lymnaea stagnalis on the host-finding behavior of miracidia of two Brazilian and one Egyptian strain of Schistosoma mansoni was studied. Miracidia of the Egyptian strain significantly preferred their suitable host B. alexandrina versus the other snail species with their behavior patterns of host location and their responses after contact with the host. However, miracidia of both Brazilian strains did not differentiate between SCW from three of the snail species; only the pigmented B. glabrata elicited weaker responses. The releasing cues of SCW for miracidial host-finding phases are macromolecular glycoconjugates. An analysis of SCW by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotting, and subsequent carbohydrate detection showed that the band patterns of glycoconjugates differed significantly among the four snail strains. Therefore, differing chemical characteristics of the signaling glycoconjugates could be the basis for the observed host specificity in miracidial host-finding.

Agar↗

Development, hatching, and infectivity of Echinostoma caproni (Trematoda) eggs, and histologic and histochemical observations on the miracidia.

Light microscopy studies were done on the eggs and miracidia of the intestinal trematode Echinostoma caproni. Eggs were obtained by homogenizing adult worms derived from ICR mice in artificial spring water (ASW). To study egg development and to obtain miracidia, eggs were incubated in ASW in either a light or a dark environment for 10 days at 28 degrees C. Egg development was characterized and staged at 2-day intervals from day 0 to day 10 postembryonation. No significant difference in development was seen in eggs maintained under conditions of light or darkness. Eggs maintained in darkness for 10 days and exposed to incandescent light produced a large synchronous hatch of miracidia within 3 h of exposure to light. As expected, miracidia used to expose Biomphalaria glabrata snails produced patent infections at 5-7 weeks postinfection (p.i.). Embryonated eggs aged from 0 to 10 days that had been fed to B. glabrata snails also produced infections within 4 weeks of infection. Miracidial longevity studies showed that 50 newly hatched miracidia maintained at either 4 degrees, 12 degrees, 22 degrees, or 38 degrees C lived for 6, 28, 14, and 5 h, respectively. Histology studies were done on whole miracidia stained in 0.01% neutral red and in Schneider's acetocarmine. Histochemistry studies of miracidia stained with alcian blue (pH 2.5) showed the presence of acid mucopolysaccharides in the epidermal plates; miracidia treated with periodic acid-Schiff plus 0.5% malt diastase showed the presence of glycogen in the body and in the epidermal/subepidermal region. Miracidia stained with Lillie's Oil Red O showed the presence of sparse neutral fat droplets in the body.

Animals↗

Application of synchrotron-radiation-based computer microtomography (SRICT) to selected biominerals: embryonic snails, statoliths of medusae, and human teeth.

Synchrotron-radiation-based computer microtomography (SRmicroCT) was applied to three biomineralised objects First, embryonic snails of the freshwater snail Biomphalaria glabrata, second, rhopalia (complex sense organs) of the medusa Aurelia aurita, and third, human teeth. The high absorption contrast between the soft tissue and mineralised tissues, i.e. the shell in the first case (consisting of calcium carbonate) and the statoliths in the second case (consisting of calcium sulphate hemihydrate), makes this method ideal for the study of biomineralised tissues. The objects can be non-destructively studied on a micrometre scale, and quantitative parameters like the thickness of a forming a snail shell or statolith crystal sizes can be obtained on a length scale of 1-2 mum. Using SRmicroCT, the dentin-enamel border can be clearly identified in X-ray dense teeth.

Animals↗

Schistosoma rodhaini: intramolluscan larval development, migration and replication processes of daughter sporocysts.

The larval development of Schistosoma rodhaini in Biomphalaria glabrata presents two features of interest. The first concerns the migration of daughter sporocysts: only a fraction of the daughter sporocysts produced by the mother sporocyst migrate directly towards the digestive gland of the snail and become cercariogenous; the others remain in the anterior region of the snail where the majority transforms into small sporocystogenous sporocysts. The second point concerns the replication processes: only direct replication of daughter sporocysts was observed in sporocysts in a prehepatic localization. The demographic processes involved in the development of S. rodhaini are compared with other species of Schistosoma and interpreted as an adaptive character for better exploitation of the snail host.

Animals↗

Field trials to control schistosome intermediate hosts by the plant molluscicide Ambrosia maritima L. in the Senegal River Basin.

Several field experiments were carried out in Senegal to evaluate the molluscicidal activity of Ambrosia maritima L. Applications of 400 mg L-1 of dry leaves of the plant in creeks of the Lampsar river did result in a reduction of Bulinus spp of 54-56% 2 weeks after treatment. Treatment of a closed irrigation canal caused a reduction of the density of Biomphalaria pfeifferi up to 77% 2 weeks after the application of 150 mg L-1 and 300 mg L-1. In both trials, however, a quick increase in the snail density was observed after treatment. It is concluded that the large amounts of crude plant material required to achieve snail reductions of less than 80% compromise the use of A. maritima in schistosomiasis control programmes.

Animals↗

Schistosoma mansoni: long-term maintenance of clones by microsurgical transplantation of sporocysts.

Schistosoma mansoni sporocysts originally derived from monomiracidially infected Biomphalaria glabrata snails were serially transplanted into the cephalopedal sinus of anesthetized snails by the microsurgical implantation of fragments of parasitized hepatopancreas and ovotestis. Three to six passages each of five male and five female clones were maintained for as long as 2.0 years. Of the recipient snails which survived surgery, 87% released cercariae, usually beginning 5-7 weeks after surgery. The percentage of snails which released cercariae increased with successive passages. The mean survival time of surgically infected snails after cercarial emergence began was 9.2 +/- 0.5 weeks, nearly the same as that of miracidially infected snails. Longevities of snails infected with male or female clones were similar. Recipient snail size and age did not influence cloning success. Beginning 5 weeks from the onset of cercarial emergence large numbers of cercariae (a mean of 3900/snail from male clones and 1300/snail from females) were obtained during each shedding period. These results clearly demonstrate that the microsurgical transplantation of sporocysts is a practical means of maintaining and expanding populations of genetically homogeneous schistosomes (clones).

Animals↗

Schistosoma mansoni: relationship between cercarial production levels and snail host susceptibility.

Two populations of Biomphalaria glabrata snails differing slightly in their susceptibility to Schistosoma mansoni infection showed dramatic differences in cercarial output per snail. Exposed to five or more miracidia, snails from a group with a 90-100% susceptibility rate (Group A) produced nearly twice the number of cercariae as those from a group with a 70-80% susceptibility rate (Group B). Exposure of individual snails to known numbers of miracidia resulted in higher numbers of primary (mother) sporocysts in Group A snails than in Group B snails. However, monomiracidial exposure of snails from both groups resulted in equivalent numbers of cercariae produced per positive snail, indicating that, once established, all primary sporocysts possess a similar reproductive potential. Morphometric analysis of serially sectioned 9-day-old primary sporocysts supported this conclusion; the size of the primary sporocysts and the size and numbers of secondary (daughter) sporocysts within each primary sporocyst were comparable in snails from both groups. The data indicate cercarial production in this system is regulated prior to, and/or during, early development of the primary sporocyst.

Animals↗

Identification of putative egg-laying hormone containing neuronal systems in gastropod molluscs.

Of gastropod molluscs, only in the Aplysiidae and the Lymnaeidae have the genes encoding the respective egg-laying hormones been cloned and the neurons controlling egg laying and egg-laying behavior been identified. Immunocytochemistry, using antibodies raised against alpha-CDCP (one of the neuropeptides encoded on the egg-laying hormone gene of Lymnaea stagnalis), identified neurons in various species of gastropods. In the basommatophoran snail, Biomphalaria glabrata, large and small neurons were observed in areas of the central nervous system similar to where immunoreactive cells exist in L. stagnalis, i.e., in the cerebral and pleural ganglia. In the stylommatophoran snail (Helix aspersa) and the slug (Limax maximus), large immunopositive neurons occur in the visceral and right parietal ganglia. In L. maximus, small immunoreactive neurons were found in the cerebral ganglia while in H. aspersa similar cells were observed intermingled with the large cells in the visceral and right parietal ganglia. Similar to the situation in L. stagnalis, in the female part of the reproductive tract of B. glabrata, L. maximus, and A. californica, but not in H. aspersa, neurons and/or fiber tracts are present. The results indicate that egg-laying hormone precursor molecules of gastropod molluscs are phylogenetically closely related. The alpha-CDCP antiserum may allow the identification of hitherto unknown egg-laying regulating systems of gastropod molluscs.

Amino Acid Sequence↗