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PubMed1983

Specificity and memory in increased defence reactions against bacteria in the pond snail Lymnaea stagnalis.

In Lymnaea stagnalis injections with dead Escherichia coli or Staphylococcus saprophyticus bacteria resulted in an enhanced clearance of both live S. saprophyticus and E. coli injected 4 days later. A non-specific activation of the internal defence system was concluded from these findings. The activation was dose-dependent: pre-injections with high doses resulted in a higher increase in the clearance capacity of the snails than pre-injections with low doses of bacteria. The state of increased activity of the defence system, induced with injection of dead E. coli, lasted at least 64 days. The heightened responses of the defence system were probably due to an activation of the blood cells (amoebocytes) since: 1) amoebocyte numbers increased faster in bacteria pretreated snails than in control animals; 2) ultrastructural observations revealed that the amoebocytes of bacteria pre-treated animals had a more ruffled outline than those of control snails; 3) amoebocytes from sensitized snails showed a higher phagocytic activity in vitro; 4) mitotic activity of amoebocytes increased after snails had been injected with bacteria.

Animals↗

PubMed1991

Isolation and functional characterization of snail hemocyte-modulating polypeptide from primary sporocysts of Schistosoma mansoni.

Infection with larval trematodes has been shown to inhibit several snail-host defences, including hemocyte phagocytosis, cytotoxicity, motility, and adherence. Certain plasma factors which mediate snail defence responses, and which may be produced by host hemocytes, also appear to be altered by these parasites. In this study we present protocols for the isolation of 2 proteins from larval Schistosoma mansoni excretory-secretory (ES) products and detail the effects of these components on Biomphalaria glabrata hemocyte protein synthetic/secretory (S/S) activity. Schistosome ES proteins, separated with a combination of membrane ultrafiltration, size exclusion, and ion exchange chromatography, were tested for their in vitro effect on cultured snail hemocytes, in the presence and absence of homologous plasma. A high-molecular-weight ultrafiltration fraction of parasite ES products (H30), in combination with plasma, was found to differentially affect susceptible (M-line) and resistant (10-R2) snail hemocytes. Secretion of metabolically labeled polypeptides by M-line cells was inhibited significantly while the S/S response of 10-R2 hemocyte polypeptides was not affected. In the absence of homologous plasma, little or no differential affect of ES polypeptides on hemocyte S/S activity was seen. Much of the inhibitory activity of H30 was attributable to a partially purified fraction, Peak I (PkI), of ES products. Evidence suggests that, in its native state, PkI is a high-molecular-weight protein aggregate comprising subunits of approximately 22-24 kDa. Thus, PkI, in the presence of homologous plasma components, is a potential mediator of schistosome-induced suppression of polypeptide synthesis or secretion in hemocytes of susceptible snails. In combination with other parasite and host factors, PkI may be involved in the host-parasite interaction which leads to the state of susceptibility or resistance found in our strains of B. glabrata.

Animals↗

PubMed1992

Studies on carbohydrates extracted from native and chemically treated Biomphalaria alexandrina snails.

1. Carbohydrates were extracted from total tissue extracts of Biomphalaria alexandrina snails and were analyzed to their monosaccharides using GLC. 2. The snails were chemically treated with thioxanthone derivatives (compounds I, II, III) and the change in the monosaccharide constituents of their carbohydrates was investigated. 3. The isolated monosaccharides from native and chemically pretreated snails were injected into mice and their protective effects were examined after infection of mice with cercariae of Schistosoma mansoni. 4. The results showed that the main monosaccharides in carbohydrates of snails were galactose, glucose, fucose and mannose and that chemical treatment caused a drop in the galactose content. 5. Moreover, monosaccharide fractions from snails treated with compound III were the most effective in inducing protection against Schistosoma infection in mice.

Animals↗

PubMed2006

Tissue distribution and subcellular localization of trace metals in the pond snail Lymnaea stagnalis with special reference to the role of lysosomal granules in metal sequestration.

The present study was undertaken to elucidate the cellular mechanisms, which govern metal sequestration and detoxification in gastropods. For this purpose the pond snail, Lymnaea stagnalis was exposed to environmentally relevant concentrations of three species of metals (Al, Zn and Cd) for 30 days and the localization and fate of these metals were followed in different tissues of the snails. The measurement of relative distribution of metals between tissues revealed that the digestive gland and kidney account for most of the accumulated metals. Al and Cd (non-essential metals) were redistributed to the digestive gland, possibly because of the presence of specific binding entities in the digestive glands of the herein species. This study focuses on the role of intracellular metal-containing granules on metal sequestration. Three main types of granules were identified in the digestive gland cells namely small, green and yellow granules. The morphological examination and the progressive accumulation of elements within these granules revealed that they are developmental stages with the yellow granule being the mature one. The total number of these granules was found to be significantly increased upon exposure of the snails to Al only. This increase may be a response to the large amount of Al that is accumulated through feeding route of this grazing snail. X-ray microanalysis (XRMA) revealed that metals were localized in all three types of digestive gland granules. The increased amount of ligands (P and S) in the granules may give evidence for their role in metal sequestration. Levels of Al and P were positively correlated in the digestive gland granules. It is possible that aluminium is bound to phosphorus to render it insoluble and so to both immobilize it within the lysosome and to be excreted in a highly insoluble form. On the other hand, both Zn and Cd induced marked upregulation of S in mature (yellow) granules by 26- and 11-folds, respectively. The lysosomal codeposition of S and either Cd or Zn in the lysosomal granules in addition to the increase in RER cisternae may indicate that the exposure to these metals could induce metallothionein synthesis in the cells. The microscopical examinations in the present study revealed that metal detoxification from the digestive gland cells may occur via faeces or via basal exocytosis towards hemocytes dispersed by the connective tissue in the visceral mass. In the kidney, one type of granules, the excretory concretions, was identified in the nephrocytes. The significant increase in the number of these concretions in the snail L. stagnalis upon exposure to metals may give further evidence for their role in metal excretion.

Aluminum Compounds↗

PubMed2006

PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails.

Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and crushing. We developed a PCR assay for the sensitive and specific detection of F. hepatica in field-collected Lymnaea sp. snails. A primer pair was designed to amplify a 405 bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica. The PCR assay showed a limit of detection of 10 pg of genomic F. hepatica DNA. No cross-reactions were observed with samples from other related trematode species or from the snail hosts Lymnaea columella and Lymnaea viatrix. DNA sequencing of the amplicon showed 100% homology with F. hepatica, and 75-89% homology with other trematodes on regions that did not include the entire set of primers. Two samples from Argentina were analysed. For snails in sample 1 (n = 240), identified as L. columella, the infection rate was 17.5 and 51.3% by direct examination and PCR, respectively. For snails in sample 2 (n = 34), identified as L. viatrix, the infection rate was 2.9 and 61.8% by direct examination and PCR, respectively. Differences in infection rates between these diagnosis methods were significant for both samples. Our PCR technique showed to be effective for detecting specific F. hepatica infections of low intensity in the intermediate host, and hence it could be used to study the epidemiological situation in a given area, as well as to assess host suitability for the parasite.

Animals↗

PubMed2002

The symbiont Capsaspora owczarzaki, nov. gen. nov. sp., isolated from three strains of the pulmonate snail Biomphalaria glabrata is related to members of the Mesomycetozoea.

While investigating the resistance of some strains of Biomphalaria glabrata to infection with Schistosoma mansoni, a unicellular eukaryotic symbiont was noted in the snail haemolymph. It was similar in appearance to Nuclearia sp. reported from B. glabrata. Sequences comprising the 18S, ITS1, 5.8S, ITS2 and the beginning of the 28S rDNA gene regions were obtained from symbionts isolated from three strains of B. glabrata, and compared with the same sequences obtained from a culture of Nuclearia sp. 18S rDNA sequences were identical for all four isolates. 18S rDNA sequences were used in a phylogenetic analysis to produce minimum evolution, maximum parsimony, maximum likelihood and Bayesian trees. All four analyses indicated that the B. glabrata symbiont is not closely related to Nuclearia but instead to the Mesomycetozoea, a recently recognised clade of symbiotic eukaryotes. Based on phylogenetic analysis, life history and morphological differences, the symbiont is described as a new genus and species, Capsaspora owczarzaki. Distinguishing characters are the presence of life cycle stage(s) that occur within snail haemolymph; ability to kill and ingest digenetic trematode larvae; ability to undergo asexual fission to produce daughter cells; absence of flagella, a mucous sheath and membranes containing chitin, elastin, or collagen; and presence of long unbranching pseudopodia and a penetration process. Using both polymerase chain reaction (PCR) and culturing techniques, the S. mansoni-resistant Salvador and 13-16-R1 strains were found to be significantly more likely to harbour the symbiont than the susceptible M line strain. Small but consistent sequence differences were noted among symbiont isolates from different snail strains, raising the possibility that the symbiont has diverged in different snail lineages. This suggests further that the symbiont is not restricted to albino lab-reared snails. A role, if any, of the symbiont in resistance awaits further study.

Animals↗

PubMed2000

Measurement of ingestion rate of Hydrobia ulvae (Pennant) on intertidal epipelic microalgae: the effect of mud snail density.

The individual mean ingestion rate of Hydrobia ulvae was measured experimentally in controlled microcosms, in the dark to avoid primary production during measurement and at constant temperature. The experimental design was based on the addition of prelabelled epipelic microalgae to microcosms in a constant proportion with unlabelled diatoms, and in such a way that algal food availability was not a limiting factor within the range of tested densities (0.3 to 4.1 snails cm(-2)). Results show that the individual mean ingestion rate decreased significantly from 26.6+/-1.1 ng Chl a snail(-1) h(-1) to 22.4+/-1.0 ng Chl a snail(-1) h(-1) between 0.7 and 3 snails cm(-2). We hypothesize that this sharp decrease (the threshold density was between 1.4 and 2.5 snails cm(-2)) may account for a density-dependent effect. We have tested this hypothesis by using a simple random walk model including basic behavioural processes such as a break in feeding activity when two individuals contact each other. The model represents quantitatively well the threshold effect, suggesting that behavioural processes have to be taken into account for estimating a global feeding activity of H. ulvae populations.

Journal Article↗

PubMed2003

Effect of orthosilicic acid on the accumulation of trace metals by the pond snail Lymnaea stagnalis.

Silicon (Si) has a marked affinity for aluminium (Al(III)), but not other trace metals such as cadmium (Cd(II)) and zinc (Zn(II)). Exogenous orthosilicic acid (Si(OH)(4)) ameliorates the toxicity of Al(III) to the pond snail Lymnaea stagnalis, but its mechanism of action is unclear. Here, studies were conducted to ascertain whether interaction between orthosilicic acid and Al(III) occurs in the water column to prevent Al(III) uptake, or in the tissues to reduce the toxicity of accumulated metal. Silicon did not reduce the accumulation of Al(III) by the digestive gland (the main "sink" for trace metals in L. stagnalis) following exposure of the snail for 30 days to 500 microg l(-1) added Al(III) and 13-fold molar excess of orthosilicic acid. However, Si concentrations correlated well with Al(III) levels in the digestive gland (R(2)=0.77), giving a ratio of 2.5:1 (Al(III):Si). Exposure to Zn(II) or Cd(II) and 13-fold molar excess of orthosilicic acid did not prevent uptake of these metals, or result in a correlation between metal and Si concentrations of the snail digestive gland. These data show that aquated orthosilicic acid does not prevent Al(III) accumulation by L. stagnalis. However, following exposure, the ratio of Al(III) to Si in the digestive gland is suggestive of the early formation of hydroxyaluminosilicates, probably proto-imogolites (2-3:1 Al(III):Si). Whether hydroxyaluminates are formed ex vivo in the water column and taken up by snails into the digestive gland, or formed in situ within the digestive gland remains to be established. Either way, orthosilicic acid clearly prevents the in vivo toxicity of Al(III) rather than reducing its uptake. Silicon appears to have an important role in the handling Al(III) by the pond snail which may also have wider relevance in understanding the role of Si in ameliorating Al(III) toxicity.

Aluminum↗

PubMed1998

The origins, fate, and ecological significance of free amino compounds released by freshwater pulmonate snails.

The mass-specific accumulation rates (MSAR) of both total (TFAC) and individual free amino compounds (FAC) in conditioned media were measured by HPLC, using the orthophthaldialdehyde (OPA) methods, in the following cases: (a) laboratory-reared freshwater snails (B. glabrata) with chemosterilized shells; (b) Biomphalaria glabrata with non-chemosterilized shells; (c) B. glabrata faeces; (d) isolated shells of B. glabrata; and (e) 10 other species of freshwater gastropods from the Lewes Brooks, East Sussex, U.K. The MSAR values for B. glabrata show that 95% of the TFAC's (predominantly ethanolamine, phosphoserine, and the amino acids leucine, isoleucine, valine, aspartic acid, and glycine/threonine) originated from the snails themselves as the faeces and shells contributed only 5.0 and 0%, respectively. In contrast, epizootic organisms on the shells of all 10 snail species from the Lewes Brooks released significant amounts of FAC with the two smallest species (Planorbis vortex and Planorbis contortus) having the highest MSAR values. The MSAR for isolated B. glabrata mucus was 42.45 micromol x g(-1)h(-1). As 500 mg snails can release 16.67 mg of mucus daily, this could potentially result in the daily loss of 707.5 micromol of FAC. The cost/benefits of mucus secretion and the various anatomical, physiological, biochemical, and ecological mechanisms which allow freshwater snails to recover FAC's lost as a result of a high rate of urine production in their hypotonic environment, are discussed.

Amines↗

PubMed1986

Development of Trichobilharzia australis Blair & Islam, 1983 in the snail, Lymnaea lessoni Deshayes and in an experimental definitive host, the Muscovy duck.

The development of Trichobilharzia australis Blair & Islam, 1983 in the intermediate host, Lymnaea lessoni Deshayes and in experimental definitive hosts, Muscovy ducks is described. 24 hours after entry into the snail, miracidia had lost their cilia, epidermal plates and lateral processes and became young mother sporocysts. They were located in the tissues of the head-foot organ of the snail and were oval in cross section but still retained the shape of a miracidium, and measured 0.058-0.066 X 0.041-0.049 mm. From the seventh day onward young mother sporocysts were tubular, thin-walled, irregular in shape and were in the tissues of the lung and kidney of the snail. On the 24th day mature mother sporocysts and young daughter sporocysts were found in the digestive gland. Between the 24th and 29th day mature daughter sporocysts with fully developed cercariae ready to emerge, or already emerged, could be seen in the digestive gland of the snail. Cercariae emerged from the snail from the 29th to 46th day after exposure at 25 degrees +/- 1 degree C. The prepatent period of T. australis in the Muscovy ducks was 22 to 42 days after the beginning of exposure to cercariae.

Animals↗

PubMed1987

Transient non-susceptibility to Schistosoma mansoni associated with atrial amoebocytic accumulations in the snail host Biomphalaria glabrata.

In some Biomphalaria glabrata-Schistosoma mansoni combinations snails are susceptible to infection as juveniles, but have variable susceptibility as adults. These snails become non-susceptible at the onset of egg-laying and typically revert to susceptibility in old age. Certain stocks of B. glabrata have the capacity to form amoebocytic accumulations in the atrium, and this ability is under genetic control. The atrial amoebocytic accumulations are transitory, typically appearing at onset of egg-laying and disappearing after a few months. A snail stock which has genetic tendencies for both adult variable susceptibility and atrial amoebocytic accumulations was studied. An association between the time of occurrence of adult non-susceptibility and atrial accumulation is revealed as snails never became infected with S. mansoni when amoebocytic accumulations were present. Developing parasites, however, were not necessarily encapsulated and destroyed by amoebocytes. Some sporocysts were able to delay development until the amoebocytic accumulations disappeared. The timing of atrial amoebocytic accumulations and resulting transient non-susceptibility in this host-parasite combination could influence snail population dynamics.

Animals↗

PubMed1989

A comparative evaluation of snail sampling and cercariometry to detect Schistosoma mansoni transmission in a large-scale, longitudinal field-study in Machakos, Kenya.

In an operational Schistosoma mansoni field-study in an area about 20 km 2 (population approximately 8000), transmission detection by simple snail sampling was compared with cercariometry. Between 1985 and 1987, 62 field sites were sampled at fortnightly intervals. Of a total of 2758 field observations, 89.8% gave full snail data; 64.4% full cercarial data; and 61.7% complete data for both methods. The complete data sets showed significant but not strong correlations between Biomphalaria pfeifferi (total and infected with S. mansoni or other trematodes) and cercarial (S. mansoni and non-human) recoveries. Non-human (but not S. mansoni) cercarial recovery decreased with deteriorating cercariometry filter quality. Both snail and cercarial recoveries diminished significantly with increasing water flows at the time of collection. Many samples yielded infected snails or cercariae, but not both, and neither method detected significantly more transmission sites. The method of choice for detecting transmission in a large-scale field-study depends on logistical and financial considerations. Relatively simple snail sampling allows quick, cheap and widespread data collection adequate for most purposes but more complicated cercariometry is still valuable for specific, small-scale studies.

Animals↗

PubMed2001

Coevolution and compatibility in the snail-schistosome system.

In stark contrast to the huge body of theoretical work on the importance of hosts and parasites as selective agents acting on each other, until recently, little systematic empirical investigation of this issue has been attempted. Research on snail-schistosome interactions have, therefore, the potential for making an important contribution to the study of coevolution or reciprocal adaptation. This may be particularly pertinent since snail-schistosomes represent an indirectly transmitted macroparasite system, so often overlooked amongst both theoretical and empirical studies. Here we review ideas and experiments on snail-schistosome interactions, with particular emphasis on those that may have relevance to the potential coevolution between host resistance and parasite infectivity and virulence. We commence with an introduction and definition of the general concepts, before going into detail of some specific studies to illustrate these: evidence of snail-schistosome coevolutionary process in the field; evidence of coevolutionary processes in the laboratory; a general assessment of the applicability of coevolutionary models in snail-schistosome interactions; and finishing with a section on conclusions and areas for further study.

Animals↗

PubMed2000

Evaluation of environmental methods to control snails in an irrigation system in Central Morocco.

The Moroccan Ministry of Public Health has launched a programme to eliminate schistosomiasis. One of the components in this process is the control of Bulinus truncatus, the intermediate host snail of Schistosoma haematobium. We evaluated three environmentally safe measures to control B. truncatus in siphon boxes, the main breeding sites for these snails in the Tessaout Amont irrigation system. The first method involved covering the siphon boxes to exclude light and reduce algal growth, the second consisted of increasing the frequency of emptying and cleaning the siphon boxes, and the third method increased water velocity to hinder the establishment of the intermediate hosts. The results showed that covering had a pronounced effect on snail and egg mass density, was accepted by the local community and prevented water contact. Cleaning the siphons three times during the irrigation season led to a reduction in snail density although it was not statistically significant and recolonization was rapid. Increasing water velocity by reducing the dimensions of siphon boxes delayed recolonization, but such a control measure can be applied only in specific situations where it does not pose hydraulic problems. The three interventions were selectively effective against B. truncatus, whereas other snails such as Physa acuta and Lymnaea peregra were hardly affected. Covering, the most promising control measure, could be useful in the Moroccan schistosomiasis eradication programme. However, further investigations are needed to assess its impact on water quality.

Animals↗

PubMed1999

Influence of low temperatures on the cercarial shedding of Paramphistomum daubneyi from the snail Lymnaea truncatula.

Lymnaeids in nature are subjected to temperature fluctuations that may have an influence on the shedding of cercariae. Thus, experimental infections of Lymnaea truncatula by Paramphistomum daubneyi were performed to determine whether a sudden fall in temperature--daily dipping of infected snails into spring water at 14-15 degrees C, or at 6-8 degrees C--followed by its increase at 20 degrees C in the subsequent hour, had an influence on the characteristics of snail infection and cercarial production. The immersion of infected snails in cold water during a short period delayed the first cercarial shedding, at day 66 on average, in the 6-8 degrees C group vs at day 57 in the 14-15 degrees C group, or at day 49 in the 20 degrees C group. The percentage of cercaria-shedding snails was greater in the 6-8 degrees C group than in the 14-15 degrees C and the 20 degrees C ones: 41.8% vs 17.3% and 7.1%, respectively. The total number of cercariae given by each infected snail was also higher in the 6-8 degrees C group. A fall in the temperature of water during daily change, followed by its increase at 20 degrees C within the subsequent hour, stimulated the cercarial shedding of P. daubneyi.

Animals↗

PubMed1992

Dorsal-ventral patterning in Drosophila: DNA binding of snail protein to the single-minded gene.

The Drosophila snail gene is required for proper mesodermal development. Genetic studies suggest that it functions by repressing adjacent ectodermal gene expression including that of the single-minded (sim) gene. The snail gene encodes a protein with a zinc-finger motif, and here we report that the snail gene product is a sequence-specific DNA binding protein. The snail protein recognizes a 14-base-pair consensus sequence that is found nine times in a 2.8-kilobase sim regulatory region. These results provide evidence for the direct control of sim transcription by snail.

Adenovirus Early Proteins↗

PubMed2003

Cadmium toxicity and snail-digenean interactions in a population of Lymnaea spp.

The toxicity of cadmium to a population of Lymnaea peregra and L. stagnalis naturally infected with a range of digeneans and collected from a number of sites in the lower Thames Valley, UK was investigated. Lymnaeid snails were exposed to 100 microg l-1 cadmium and the effects on host survival and emergence of cercariae recorded. Overwintered L. peregra, but not L. stagnalis, showed significantly reduced survival compared to seasonally infected snails, i.e. snails which have acquired an infection during the spring or summer. A significant increase in survival with increasing snail size was demonstrated for L. stagnalis and for seasonally infected L. peregra only. Only L. stagnalis infected with Diplostomum spathaceum and L. peregra infected individually with D. spathaceum, Sanguinicola inermis, Echinoparyphium recurvatum and Notocotylus attenuatus demonstrated a significantly reduced survival compared to laboratory-bred controls. The exposure of L. stagnalis to cadmium resulted in a significant reduction in the emergence of D. spathaceum over a 5-day period but cadmium-exposed L. peregra showed no difference in the emergence of E. recurvatum cercariae over a 3-day exposure period. The mechanisms and importance of metal toxicity to snail-digenean interactions are discussed.

Animals↗

PubMed2005

Storage and incubation of Echinostoma revolutum eggs recovered from wild Branta canadensis, and their infectivity to Lymnaea tomentosa snails.

Echinostoma revolutum eggs recovered from naturally infected wild Canada geese (Branta canadensis) were cold stored (4-6 degrees C) for up to 72 weeks. Successful hatching followed incubation for from 6 to 8 days at an optimum temperature of between 25 and 30 degrees C. A partial life cycle from adult worm to metacercarial encystment in Lymnaea tomentosa snails was completed in the laboratory. Snails were infected both by free miracidia and by ingestment of unhatched embryonated eggs. Infection was equally successful in environmental temperature ranges from 10 to 25 degrees C, and at challenge levels of 2, 5 or 10 embryonated eggs per snail. Exposure to 10 eggs was lethal. Ingestion by snails of embryonated eggs with successful infection at 10 degrees C suggests that embryonated eggs may be used to infect wild snails when the environmental water temperature has reached 10 degrees C.

Animals↗