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Echinostoma paraensei and Schistosoma mansoni: adherence of unaltered or modified latex beads to hemocytes of the host snail Biomphalaria glabrata.

Hemocytes derived from a strain (13-16-R1) of Biomphalaria glabrata resistant to Schistosoma mansoni were significantly more likely to bind untreated latex beads than hemocytes from the schistosome-susceptible M line strain. Beads preincubated in 13-16-R1 plasma were more readily bound by both 13-16-R1 and M line hemocytes than beads preincubated in M line plasma. Beads preincubated in plasma derived from snails of either strain infected with the trematode Echinostoma paraensei were more readily bound by hemocytes than beads preincubated in plasma from control snails of the corresponding strain. Plasma from snails exposed to S. mansoni did not have a similar effect. Throughout these experiments, beads receiving a particular treatment were consistently bound at higher rates by 13-16-R1 than M line hemocytes. SDS-PAGE of plasma components eluted from beads revealed differences between treatments, particularly in diffuse bands falling into two groups, of 75-130 and 150-220 kDa. The results indicate that both hemocytes and plasma components from the two host strains differ and identify plasma molecules deserving of additional study as possible modulators of hemocyte effector functions. Also, S. mansoni and E. paraensei provoked different responses in the same host snail.

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Biological methods for the control of freshwater snails.

As a result of the advent of new drugs and diagnostic techniques, the emphasis in the control of schistosomiasis has changed from snail control to chemotherapy for infected individuals. However, chemotherapy does not prevent reinfection and there remains a need to reduce snail densities in human water supplies. In the past, treatment with molluscicides has proved ineffective, expensive and has had environmental drawbacks. Here, Henry Madsen describes research into alternative methods of snail control. As yet, little is known of the predators and parasites of schistosome intermediate hosts, but such agents could be suitable as biocontrol agents. To date, the most promising results have been obtained from experimental introductions of competitive snail species, but this strategy still has its drawbacks under many environmental conditions and the development of a universal method of biological control for the intermediate hosts of schistosomes is still a long way off.

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Adaptation by an urban population of the snail Helix aspersa to a diet contaminated with lead.

Two populations of the garden snail Helix aspersa, from an urban car park and from a semi-rural site, were fed a diet containing 500 microg g(-1) of Pb as PbSO(4). After 2 days, half of each population was removed to a Pb-free diet and half continued on the dosed food, both for 64 days. The snails from the contaminated site had a significantly lower uptake of Pb compared with those from the rural, uncontaminated site. The car park snails also lost Pb more rapidly from their tissues. A second experiment evaluated the effect of a previous exposure to a high Pb diet on Pb uptake. The results suggest that the differences between the two populations are not due to a physiological adaptation, but rather that the car park snails represent an ecotype adapted to a high Pb diet.

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Effects of acidic water on freshwater snails: results from a study of 1000 lakes throughout Norway.

This paper summarizes some results from a monograph which focused on ecological and biogeographical aspects of the biota in Norwegian lakes and rivers with particular reference to the snail fauna. Field studies were made in the course of 20 summers within the period of 1953-1987. Acidic water is the main reason why snails are absent from some 4000 lakes in a low-calcium area in southern Norway ('Acid south'). This area is heavily influenced by acidic precipitation. Snails were not detected in lakes with pH below 5.2. In one lake, three snail species disappeared in parallel with acidification from pH 5.2 to 4.2. The calcium level modified the effect of low pH.

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The effects of starvation and refeeding on egg laying and the synthetic activity of the albumen gland in Bulinus truncatus, a snail vector of urinary schistosomiasis.

1. The effects of starvation (for 1, 2, 3, 6, 9 and 12 days, respectively) and refeeding (12 days starvation and 1, 2 and 3 days refeeding, respectively) on egg laying and albumen gland activity in the freshwater snail, Bulinus truncatus were studied. 2. The egg laying of starving snails rapidly decreased and ceased by day 6 of starvation. Egg laying was restored 24 hr after refeeding. The recorded decrease in albumen gland wet weight was proportional to the starvation periods. The DNA contents of the glands of the different experimental groups was not statistically different from the controls. 3. Albumen gland synthetic activity expressed as 14C-glucose incorporation into galactogen/microgram DNA and 3H-amino acids into total protein was determined. The glands showed an abrupt decrease in synthetic activity after 1 day of starvation and gradually decreased further until days 9-12. The decrease in activity of the glands was more rapid than that of egg laying. Upon refeeding, the activity of the glands recovered rapidly, simultaneous with the increase in wet weight and egg laying. 4. In conclusion, there is a correlation between egg laying and the in vitro activity of albumen glands. The results show a short-term effect of starvation on the fecundity of the snails. Such studies could be useful in field studies as well as snail control by applying molluscicides under optimal conditions.

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The sugar chemoreception niches of Bulinus (physopsis) globosus (Morelet) and Bulinus rohlfsi (Clessin), the two most important snail hosts of Schistosoma haematobium (Bilharz) in west Africa.

1. The behavioural responses of two species of freshwater pulmonate snails [Bulinus (P.) globosus and Bulinus rohlfsi] to sugar gradients were investigated by means of diffusion olfactometers. 2. Both snail species proved to be very discriminating in their responses. Of the 17 sugars tested, 35.3%, namely D(-)glucuronic, maltotriose, maltose, cellobiose, D(-)arabinose, D(+)mannose proved to be statistically significant attractants or arrestants to B. rohlfsi. Only 23.5% of these sugars (maltotriose, maltose), D(+) mannose and D(+) xylose were significant attractants or arrestants to B. (P.) globosus. 3. Glucuronic acid was a significant repellent to B. rohlfsi but none of the sugars was a repellent to B. (P.) globosus. 4. The results are compared with those obtained for other snail species and their relevance to the ecology and control of the snails are discussed.

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Survival of snails and characteristic lesions of Fasciola hepatica infection in four European species of Lymnaea.

The survival of snails and characteristic organ lesions were investigated in four different age categories (new-born, 1 and 2 mm shell height, adults) and in four species of Lymnaea (L. glabra, L. palustris, L. peregra ovata, L. truncatula) infected by Fasciola hepatica. Mortalities were higher in infected groups on Days 30 and 45 post-infection (+42% and +45%, respectively) than in controls. Lesions depended on the species and age of the Lymnaea. Epithelial necrosis of the kidney was recorded in L. glabra, L. palustris and L. p. ovata, whereas intralamellar deposits were found in the same organ of L. truncatula. All categories of L. truncatula had evolutive infections (snails with living parthenitae); the young snails of other species became infected, but the adults did not, as estimated at Day 45 post-infection. The lesions were related to the intensity of evolutive infections: necrosis of the digestive gland was found in moderate to low infections and, conversely, gonadal atrophy with epithelial necrosis was found in highly infected snails.

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A neotropical snail host of Schistosoma mansoni introduced into Africa and consequences for the schistosomiasis transmission Biomphalaria tenagophila in Kinshasa (Democratic Republic of Congo).

Malacological surveys carried out in the early 1970s in water bodies of the Kinshasa area, Lower Zaire (Democratic Republic of Congo), showed the appearance of a Biomphalaria species which was identified as Biomphalaria camerunensis. In 1976, other surveys confirmed the presence of the species in several sites and showed numerous infected snails with Schistosoma mansoni, demonstrating for the first time an active transmission of the parasite responsible of the intestinal schistosomiasis in this area. The most recent malacological sampling was carried out by one of us in 1994 in Mangungu River and revealed the presence of apparently the same snail species. However, conchological, anatomical and molecular studies showed that this snail may be considered as an introduced neotropical species, B. tenagophila. To our knowledge, this is the second example of the introduction of a neotropical snail host of schistosomes into Africa.

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Thin layer chromatographic analysis of glucose and maltose in estivated Biomphalaria glabrata snails and those infected with Schistosoma mansoni.

Thin layer chromatography was used to analyze the glucose and maltose concentrations of the digestive gland-gonad complex (DGG) of uninfected-estivated Biomphalaria glabrata snails and estivated B. glabrata patently infected with Schistosoma mansoni. All snails were estivated in a most chamber at a relative humidity of 98+/-1% and a temperature of 23+/-1 degrees C for 14 days. Carbohydrates were extracted from the DGG with 70% aqueous ethanol, and extracts were analyzed on silica gel preadsorbent plates using ethyl acetate-glacial acetic acid-methanol-water (60:15:15:10) mobile phase, alpha-naphthol-sulfuric acid detection reagent, and quantification by densitometry. The concentrations of glucose and maltose were significantly reduced in both uninfected-estivated snails and infected-estivated snails.

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Specific inhibitors of mitogen-activated protein kinase and PI3-K pathways impair immune responses by hemocytes of trematode intermediate host snails.

To characterize molecular mechanisms regulating snail cellular immune responses, the contributions of mitogen-activated protein kinases (MAPKs) and phosphatidylinositol 3-kinase (PI3-K) were examined in hemocytes of the trematode intermediate host snails Biomphalaria glabrata and Lymnaea stagnalis. Simultaneous measurement of phagocytosis/encapsulation and H2O2 production by hemocytes in the presence or absence of specific signal transduction inhibitors was used to assess the role of extracellular-signal regulated kinases 1 and 2 (ERK1/2), p38, JNK and PI3-K. Hemocyte spreading was significantly reduced in a dose-dependent manner by the ERK inhibitor, PD098059, and by wortmannin, a potent PI3-K inhibitor. The JNK inhibitor, SP600125, and the p38 kinase inhibitor, SB203580, had no effect on hemocyte spreading. Sheep red blood cell phagocytosis was significantly impaired by PD098059, SP600125, and SB203580. Hydrogen peroxide production during phagocytosis was severely inhibited by PD098059. Additionally, PD098059, but not the other inhibitors, significantly impaired the cellular encapsulation of trematode larvae and H2O2 production during encapsulation. These results suggest that MAPK and PI3-K signal transduction pathways play a pivotal role in the immune responses of snail hemocytes. PI3-K and ERK appear to strongly regulate cell motility. ERK, JNK and p38 contribute to phagocytosis-mediated signal transduction. ERK also play a major role in oxidative burst activation and the encapsulation of trematode larvae by snail hemocytes.

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Alterations in carbohydrates and the protein metabolism of the harmful freshwater vector snail Lymnaea acuminata induced by the Euphorbia tirucalli latex extract.

To know the short- as well as long-term effect of aqueous latex extracts of Euphorbia tirucalli on carbohydrate and protein metabolism, the snail Lymnaea acuminata was exposed to sublethal doses of 0.37 and 0.55 mg/L for a 24-h and 0.20 and 0.31 mg/L for a 96-h exposure period. Significant (P<0.05) alterations in the glycogen, pyruvate, lactate, total protein, and free amino acid level, as well as in the activity of enzyme lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase, and alanine aminotransaminase were observed in the nervous, hepatopancreatic, and ovotestis tissues of the freshwater vector snail L. acuminata exposed to sublethal doses of E. tirucalli latex extract. The alterations in all biochemical parameters were significantly (P<0.05) time and dose dependent. After the 7th day of the withdrawal of treatment, there was significant (P<0.05) recovery in glycogen, pyruvate, lactate, total protein, and the free amino acid level and in the activity of the lactic dehydrogenase, succinic dehydrogenase, cytochrome oxidase, protease, aspartate aminotransaminase and alanine aminotransaminase enzymes in all three of the studied tissues of the snail, which supports the view that the plant product is safe for use as a molluscicide for the control of harmful freshwater vector snails in the aquatic environment.

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The snail (Biomphalaria glabrata) genome project.

In 2001, ideas for a snail genome project were discussed at the American Society of Parasitologists meeting (New Mexico) and a snail genome consortium was subsequently established (the first consortium meeting was held in 2005). A proposal for sequencing the snail genome was submitted to the National Human Genome Research Institute, and Biomphalaria glabrata was prioritized as a non-mammalian sequencing target in 2004. The sequencing of the genome of this medically important snail is now underway.

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Detection of Fasciola gigantica infection in snails by polymerase chain reaction.

Polymerase chain reaction (PCR) was used to detect Fasciola gigantica infection in the snail intermediate host. Fasciola specific primers amplified a 124 bp fragment in PCR when the genomic DNA isolated from F. gigantica infected Lymnaea auricularia snails was used as template. In addition to the 124 bp amplicon, a ladder of DNA fragments representing amplification of the 124 bp repetitive sequences was observed. Genomic DNA of the parasite was used as a positive control, which also gave an amplification of the 124 bp fragment. DNA isolated from non-infected snails was used as a negative control and no amplification of this sequence was observed. This technique is highly specific and sensitive and possesses fairly good prospects of its utility as an epidemiological tool for ascertaining the infectivity status in ubiquitous snail populations.

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The control of the schistosome-transmitting snail Biomphalaria glabrata by the plant Molluscicide Euphorbia splendens var. hislopii (syn milli Des. Moul): a longitudinal field study in an endemic area in Brazil.

Under laboratory conditions, latex from Euphorbia splendens has shown promise as a plant molluscicide for control of Biomphalaria species, intermediate hosts for Schistosoma mansoni. The purpose of this study was to evaluate its efficiency under field conditions. Application of filtered latex at 12 ppm to one stream in an endemic rural area in Minas Gerais state, Brazil, in September 1995, did result in a reduction in snail density as compared to an untreated stream but the snail population recovered quickly. However, two applications with a two-week interval of unfiltered E. splendens latex at 5 ppm in November 1996 in the same stream resulted in complete disappearance of B. glabrata and snails did not reappear until the 14th month after the applications. In the control stream, without treatment, the snails were found during all months. Laboratory studies confirmed that unfiltered latex is a more potent molluscicide than filtered latex. Considering the advantages of the latex such as its low toxicity to other aquatic animals and its photobiodegradability, as well as the simple method of application, this natural product is promising as an effective molluscicide.

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Phylogeography and genetic divergence of some lymnaeid snails, intermediate hosts of human and animal fascioliasis with special reference to lymnaeids from the Bolivian Altiplano.

A population genetic study using starch gel electrophoresis was performed on populations of several species of lymnaeid snails acting as intermediate hosts for Fasciola hepatica (Trematoda, Plathyhelminth). Lymnaea viatrix was collected in 16 sites from the Bolivian Northern Altiplano. L. cubensis were obtained in one site from Venezuela, one site from Guadeloupe, three sites from Cuba and one site from the Dominican Republic. L. truncatula were collected in one site from France, one from Portugal and one from Morocco. Multilocus enzyme electrophoresis (MEE) were determined for 282 snails at 18 loci. A complete monomorphism was encountered at each geographic site. However, among these 18 loci, 13 are polymorphic and low and high levels of genetic divergence were observed between samples. Two genotypic groups can be differentiated by their multilocus genotypes. The western genotypic group associates together samples from Venezuela, Guadeloupe, Cuba and Dominican Republic (L. cubensis) while samples from France, Portugal and Morocco (L. truncatula) belong to the eastern genotypic group. Surprisingly, the Northern Bolivian Altiplano populations (L. viatrix) do not present any genetic divergence with the Portuguese sample. Therefore, the Bolivian snails belong entirely to the eastern genetic group. Within each group slight genetic divergences were observed. These results strongly support the European origin of the lymnaeid snails from the Northern Bolivian Altiplano.

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Training in a novel environment improves the appetitive learning performance of the snail, Lymnaea stagnalis.

The effect of novelty, an environmental background variable affecting feeding and appetitive learning performance, was examined in a behavioral study of the pond snail Lymnaea stagnalis. Transfer of snails into a novel aquatic environment (clean water) evoked exploratory behavior which manifested itself in an increased number of spontaneous rasping movements of the mouth over the second to fifth minute after exposure to the novel environment. The intensity of this behavior was much weaker in a familiar environment (used water from the home tank). Similarly, sucrose-induced feeding rates were highest when the snails were given the sucrose stimulus in a novel environment. The effectiveness of appetitive conditioning using tactile stimulus paired with food (Kemenes & Benjamin, 1989a) improved when the snails were subjected to conditioning in a novel environment. Satiety, an internal variable, suppressed the stimulating effects of the novel environment on the spontaneous, unconditioned, and conditioned feeding alike. After training in the novel environment, the conditioned response was retained for up to 12 days and thus provided a robust behavioral paradigm for the extension of the analysis to the neurophysiological mechanisms of factors affecting appetitive learning in molluscs.

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Snail control strategies for reduction of schistosomiasis transmission.

As intermediate hosts, molluscs play a major role in the transmission of schistosomes; they are the sites of an intense multiplication of parasites. Thus, snail control strategies are considered a priority for the reduction of schistosomiasis transmission. Here, Vinca Lardans and Colette Dissous review the efficacy of environmental management and the use of molluscicides and biological agents to control snail populations. They then describe the development of diagnostic tests, based on the detection of parasite antigens or specific parasite DNA sequences in snail tissues, to detect the early infection of snails. Finally, they discuss progress in studying the molecular basis of susceptibility and resistance phenotypes, and the possible application of the genetic manipulation of molluscs.

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Dissociation of food-finding and tentacle-lowering, following food-attraction conditioning in the snail, Helix aspersa.

The assumption that tentacle-lowering and food-finding may be used interchangeably as measures of food-attraction conditioning was examined in the snail, Helix aspersa. A brief pairing of an odor with the opportunity to feed (food-attraction conditioning) resulted in increased tendency to orient to that odor (food-finding), when tested the following day. In addition, conditioned snails exhibited increased levels of tentacle-lowering. A more detailed analysis revealed that a subset of conditioned snails exhibited successful food-finding in the absence of tentacle-lowering, and that another subset of conditioned snails exhibited increased levels of tentacle-lowering in the absence of successful food-finding. These results suggest that caution should be observed when comparing results across these two response systems.

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