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Interactions between intermediate snail hosts of the genus Bulinus and schistosomes of the Schistosoma haematobium group.

Within each of the four species groups of Bulinus there are species that act as intermediate hosts for one or more of the seven species of schistosomes in the Schistosoma haematobium group, which includes the important human pathogens S. haematobium and S. intercalatum. Bulinus species have an extensive distribution throughout much of Africa and some surrounding islands including Madagascar, parts of the Middle East and the Mediterranean region. Considerable variation in intermediate host specificity can be found and differences in compatibility between snail and parasite can be observed over small geographical areas. Molecular studies for detection of genetic variation and the discrimination of Bulinus species are reviewed and two novel assays, allele-specific amplification (ASA) and SNaPshot, are introduced and shown to be of value for detecting nucleotide changes in characterized genes such as cytochrome oxidase 1. The value and complexity of compatibility studies is illustrated by case studies of S. haematobium transmission. In Senegal, where B. globosus, B. umbilicatus, B. truncatus and B. senegalensis may act as intermediate hosts, distinct differences have been observed in the infectivity of different isolates of S. haematobium. In Zanzibar, molecular characterization studies to discriminate between B. globosus and B. nasutus have been essential to elucidate the roles of snails in transmission. B. globosus is an intermediate host on Unguja and Pemba. Further studies are required to establish the intermediate hosts in the coastal areas of East Africa. Biological factors central to the transmission of schistosomes, including cercarial emergence rhythms and interactions with other parasites and abiotic factors including temperature, rainfall, water velocity, desiccation and salinity are shown to impact on the intermediate host-parasite relationship.

Africa↗

Mixed strain schistosome infections of snails and the evolution of parasite virulence.

Mathematical models often propose that within-host competition between parasites can be a major factor in the evolution of increased parasite virulence. Kin selection predicts that as the coefficient of relatedness between infecting parasites decreases, the benefits of competition to individual genotypes increases. Thus where parasites can adjust their behaviour in response to current conditions, higher virulence is predicted in multiple genotype infections. There is limited experimental data, however, regarding the effects of mixed strain infections on host and parasite fitness. We investigated, for a snail-schistosome system, whether a conditional increase in replication rates occurred in mixed genotype infections and resulted in increased virulence. Four groups of Biomphalaria glabrata snails were exposed to 1 or 2 laboratory strains of Schistosoma mansoni. Mixed genotype infections were observed to be more virulent than single genotype infections, in terms of reductions in host reproductive success and survival. Parasite reproductive rate was also increased in mixed strain groups. Reduced host reproductive success was suggested to be directly due to the genetic heterogeneity of the parasitic infections resulting in increased host defence costs. Reduced host survival was consistent with an adaptive conditional parasite response.

Animals↗

Failure of Schistosoma mansoni to reinfect Biomphalaria glabrata snails: acquired humoral resistance or intra-specific larval antagonism?

Failure of snail reinfection by Schistosoma mansoni has been demonstrated in susceptible Biomphalaria glabrata infected with 1 miracidium and subsequently re-exposed to 1 or 5 homologous parasite larvae. The acquisition of 'resistance' to secondary parasite infection was time dependent since complete inhibition was observed at 2 weeks and longer following monomiracidial exposure. This phenomenon was still observed in snails challenged 8 weeks after primary infection. Histological observations revealed that sporocysts from the challenge infection were free of encapsulation, their development was stopped and they degenerated slowly in the absence of haemocytic reaction of the host. Under the hypothesis of an acquired homologous resistance mechanism, this strongly suggests that 1 or several unidentified humoral factors are responsible for the non-development of the sporocysts from the challenge infection. However, considering the time-dependent nature of the phenomenon, an intraspecific larval antagonism process between sporocysts resulting from the primary infection and those from the challenge infection may be involved.

Animals↗

Coordination of the activity of monoaminergic pedal neurons in freshwater snails.

In the freshwater pulmonate pond snail and flateye snail, contact of the pneumostoma (the respiratory organ) with the water surface and tactile stimulation of the mantle induced phase-coordinated excitation and inhibition of activation of symmetrical pedal neurons RPeD1 (the GDC--giant dopamine cell) and LPeD1 (the GSC--giant serotonin cell). GDC and GSC are not primary sensory neurons and the mechanism by which their actions are coordinated is polysynaptic. Injections of depolarizing current into the GDC or GSC induced spike discharges in one of the columellar nerves. It is suggested that coordination the activities of the GDC and GSC is functionally important for controlling the right and left halves of the columellar complex of muscles during respiration.

Animals↗

Development of the nitric oxide/cGMP system in the embryonic and juvenile pond snail, Lymnaea stagnalis L. A comparative in situ hybridization, histochemical and immunohistochemical study.

Recent studies have indicated that nitric oxide (NO)-induced cGMP synthesis is involved in different steps of neurogenesis in invertebrates. The development of putative NO synthetising elements was described earlier in the embryonic and juvenile pond snail, Lymnaea stagnalis, applying NADPH-diaphorase histochemistry (Serfozo et al., 1998). In the present study, we examined the distribution of NO synthase (NOS) during Lymnaea development by in situ hybridization for Lymnaea-NOS mRNA, histochemical, and immunohistochemical techniques for the NOS and NO-stimulated cGMP. Peripheral fibers projecting to the CNS and terminating in the ganglionic neuropils showed NOS immunoreactivity from 85% of embryonic development. At the same time, a fine dot-like, immunostaining indicated the presence of cGMP in the neuropil area. In the CNS, Lymnaea-NOS mRNA positive, as well as NOS and cGMP immunoreactive perikarya were detected first during postembryonic development; their number significantly increased from P3 juvenile stage. Some of the cell groups in the CNS containing NOS immunoreactive material also displayed Lymnaea-NOS mRNA hybridization signal and were cGMP-positive. However, in the subesophageal ganglia, the distribution of Lymnaea-NOS mRNA positive cell groups did not correspond to that of the NOS immunoreactive cells. Neurons revealing transient NOS and cGMP immunoreactivity, respectively, could also be detected in this part of the CNS. In most of the ganglia the number of Lymnaea-NOS mRNA containing and cGMP immunopositive neurons, respectively, exceeded that of the NOS immunoreactive cells from P4 juvenile stage. The localization of NADPH-diaphorase reaction also correlated well with that of the NOS immunoreactivity in the developing CNS. At the periphery, colocalization of Lymnaea-NOS mRNA signal, NOS and cGMP immunoreactivities were observed in the epithelial cells of the esophagus and mantle after hatching. The findings suggest the functional maturity of the NO/cGMP signal transduction pathway at both central and peripheral levels during the development of the snail, Lymnaea stagnalis. The differences in the localization of Lymnaea-NOS mRNA labeling and NOS immunoreactivity in the CNS and PNS can be explained by the existence of different NOS isoforms, posttranslational regulation of NOS, and/or some non-specific antibody labeling.

Animals↗

Primary structure and origin of schistosomin, an anti-gonadotropic neuropeptide of the pond snail Lymnaea stagnalis.

In the pond snail Lymnaea stagnalis infected with the schistosome parasite Trichobilharzia ocellata, a peptide called schistosomin is released from the central nervous system, which counteracts the bioactivity of a number of gonadotropic hormones. This leads to inhibition of the reproductive activities of the infected snail. In order to determine the structure of schistosomin, the neuropeptide was purified from the central nervous system using gel-permeation chromatography and reverse-phase h.p.l.c. The complete primary structure of the peptide was determined by N-terminal sequencing and peptide mapping. Schistosomin is a single-chain molecule of 79 amino acids with a molecular mass of 8738 Da. The peptide contains eight cysteine residues which may give rise to four intramolecular disulphide bridges that fold the peptide into a stable globular structure. A database search did not reveal any known peptides that show significant sequence similarity to schistosomin. By means of immunocytochemistry, the peptide was shown to be localized in the growth-controlling neurosecretory light green cells, which are located in the cerebral ganglia of the central nervous system of Lymnaea. In addition to schistosomin, these neurons are known to produce various insulin-related peptides.

Amino Acid Sequence↗

Red blood with blue-blood ancestry: intriguing structure of a snail hemoglobin.

The phylogenetic enigma of snail hemoglobin, its isolated occurrence in a single gastropod family, the Planorbidae, and the lack of sequence data, stimulated the present study. We present here the complete cDNA and predicted amino acid sequence of two hemoglobin polypeptides from the planorbid Biomphalaria glabrata (intermediate host snail for the human parasite Schistosoma mansoni). Both isoforms contain 13 different, cysteine-free globin domains, plus a small N-terminal nonglobin "plug" domain with three cysteines for subunit dimerization (total M(r) approximately 238 kDa). We also identified the native hemoglobin molecule and present here a preliminary 3D reconstruction from electron microscopical images (3 nm resolution); it suggests a 3 x 2-mer quaternary structure (M(r) approximately 1.43 MDa). Moreover, we identified a previously undescribed rosette-like hemolymph protein that has been mistaken for hemoglobin. We also detected expression of an incomplete hemocyanin as trace component. The combined data show that B. glabrata hemoglobin evolved from pulmonate myoglobin, possibly to replace a less-efficient hemocyanin, and reveals a surprisingly simple evolutionary mechanism to create a high molecular mass respiratory protein from 78 similar globin domains.

Amino Acid Sequence↗

Snail shape and growth rates: Evidence for plastic shell allometry in Littorina littorea.

The periwinkle Littorina littorea exhibits morphological variation among southern New England populations that appear to be genetically continuous. In dense populations, individuals have relatively elongate shells in comparison to individuals in sparse populations, which have rounder, globose shells. We experimentally demonstrate that this shell variation is a function of snail growth rate. Rapidly growing snails develop thin, globose shells that accommodate more body mass than thicker, more elongate shells. The implications of these results are discussed in relation to interpreting morphological variation in extant gastropods and in the molluscan fossil record.

Journal Article↗

Intense natural selection caused a rapid morphological transition in a living marine snail.

Shell shape and shell thickness of the intertidal snail Littorina obtusata changed markedly between 1871 and 1984 in northern New England. Shells collected prior to 1900 were high-spired with thin walls, whereas shells collected in 1982-84 were low-spired with thick walls. An intertidal crab (Carcinus maenas) which preys on L. obtusata expanded its range into northern New England around 1900. This suggests that the change in snail shell form was a response to predation by Carcinus. Field and laboratory experiments demonstrated that the high-spired form of L. obtusata, which can still be found in some Maine localities, is more vulnerable to predation by Carcinus than is the low-spired form of L. obtusata. Electrophoretic comparisons of high- and low-spired populations of L. obtusata confirmed that these populations represent different morphological forms of L. obtusata rather than different species [Nei's D (unbiased measure of genetic distance) = 0.003]. These data demonstrate that classical Darwinian selection can produce a rapid morphological transition without speciation.

Journal Article↗

Metacercarial utilization of a naturally infected single species (Lymnaea peregra) snail community by Echinoparyphium recurvatum.

Utilization of a single-species molluscan community of Lymnaea peregra by metacercariae of Echinoparyphium recurvatum over a summer (July-September) period in south-east England showed an increase in the mean number of cysts per host with host size and time of exposure. Aggregation resulting from host and habitat-related factors increased with host size and time of exposure. Encystment within the host was restricted to the peripheral organs in smaller juvenile snails but as snails increased in size, metacercariae were distributed throughout the tissues.

Animals↗

Prevalence and intensity of schistosomiasis in two rural areas in Zimbabwe and their relationship to village location and snail infection rates.

Surveys for schistosomiasis of 2498 people of various ages from 22 villages in northeast Zimbabwe are reported. A high prevalence of Schistosoma haematobium (53.1%) and a medium prevalence of S. mansoni (17.5%) were found. Schistosoma haematobium prevalence among males was significantly higher than among females. Age prevalence curves for both schistosome species showed a typical pattern with the peak in the ten to 20 years age group, which is also the age group with the highest potential of contaminating water with excreta. Snail surveys at 12 local sites showed the presence of Bulinus globosus and Biomphalaria pfeifferi. Both species were found shedding cercariae, and snail infection rates showed a focal distribution which could be related to the prevalence of infection in the human population.

Adolescent↗

Elimination of the cyanobacterial hepatotoxin microcystin from the freshwater pulmonate snail Lymnaea stagnalis jugularis (say).

It has been suggested that little to no microcystin (MC), a cyanobacterial hepatotoxin, accumulates within freshwater pulmonate snails because the toxin is associated primarily with undigested gut contents that are eliminated from the animal via egestion. To test this, Lymnaea stagnalis exposed to MC-containing cyanobacteria were placed into toxin-free environments and sampled over short (24 h at 21 degrees C) and long (30 d at 22 and 10 degrees C) time periods. Within 8 h after being removed from exposure to microcystin-containing phytoplankton, the gizzard and cecal string fractions of the feces were eliminated, accounting for 57% of the initial MC concentration. However, detectable concentrations remained beyond 24 h, likely in association with the digestive-gland contents, which can be retained up to 100 h. Long-term MC loss was biphasic at two ambient temperatures. The greatest change (fast phase) occurred over the first 3 d after exposure. By 6 d, the cumulative MC loss from L. stagnalis was 80 and 95% at 10 and 22 degrees C, respectively. Toxin loss over this period was attributed to egestion of indigestible cells/colonies from gizzard and cecum, as well as elimination of unassimilated MC-laden fragments and vacuolate excretion of residues from the digestive gland. The fast-phase depuration rate constant was significantly higher at 22 than at 10 degrees C, indicating an influence of ambient temperature on the rate of toxin loss from pulmonate snails. Depuration continued at slower rates until 30 d, when most (97.5 and 99.5% at 10 and 22 degrees C, respectively) of the initial MC was eliminated.

Animals↗

The genetics of a small autosomal region of Drosophila melanogaster containing the structural gene for alcohol dehydrogenase. VII. Characterization of the region around the snail and cactus loci.

The genetic interval 35C to 36A on chromosome arm 2L of Drosophila melanogaster has been saturated for mutations with visible or lethal phenotypes. 38 loci have been characterized, including several maternal-effect lethals (vasa, Bic-C, chiffon, cactus and cornichon) and several early embryonic lethals, including snail and fizzy. About 130 deletions have been used to order these loci. Complex interactions between mutant alleles have been uncovered in the immediate genetic environs of the snail gene, as has further evidence for an interaction between this region and that including the nearby genes no-ocelli and elbow.

Alcohol Dehydrogenase↗

Microsatellites and the genetics of highly selfing populations in the freshwater snail Bulinus truncatus.

Hermaphrodite tropical freshwater snails provide a good opportunity to study the effects of mating system and genetic drift on population genetic structure because they are self-fertile and they occupy transient patchily distributed habitats (ponds). Up to now the lack of detectable allozyme polymorphism prevented any intrapopulation studies. In this paper, we examine the consequences of selfing and bottlenecks on genetic polymorphism using microsatellite markers in 14 natural populations (under a hierarchical sampling design) of the hermaphrodite freshwater snail Bulinus truncatus. These population genetics data allowed us to discuss the currently available mutation models for microsatellite sequences. Microsatellite markers revealed an unexpectedly high levels of genetic variation with < or = 41 alleles for one locus and gene diversity of 0.20-0.75 among populations. The values of any estimator of Fis indicate high selfing rates in all populations. Linkage disequilibria observed at all loci for some populations may also indicate high levels of inbreeding. The large extent of genetic differentiation measured by Fst, Rst or by a test for homogeneity between genic distributions is explained by both selfing and bottlenecks. Despite a limited gene flow, migration events could be detected when comparing different populations within ponds.

Animals↗

OOCYTE DEGENERATION AND ALTERED OVIPOSITORY ACTIVITY INDUCED BY PARAQUAT IN THE FRESHWATER SNAIL PHYSA FONTINALIS (GASTROPODA: PULMONATA).

The freshwater snail Physa fontinalis was used as a bioindicator to study the effects of the herbicide Paraquat (PQ) in laboratory assays. The test solutions used, 0.125, 0.25, and 0.5 mg/l PQ, were in the range of the concentrations recommended for aquatic weed control. The study was carried out in two stages to determine the influence of PQ on the ovipository activity of Physa fontinalis, and the histological effects on these snails. Specimens exposed to PQ continued to be reproductively active, but the number of egg masses and eggs laid decreased significantly. Mortality was almost the same in all the experimental lots, but was significantly related to the production of egg masses only in the controls. The histological analysis showed a clear trend among PQ concentrations and degenerating oocytes, but no visible effects on the male sex-line were observed. By interfering with fertility, PQ has an action that may go well beyond its lethal effect on individuals, suggesting that this herbicide should be strictly regulated in weed control programmes. Moreover, since PQ was observed to interfere with the reproductive process, its endocrine disrupting action must not be excluded.

Journal Article↗

Effects of target tissue on growth of snail neurones in collagen gel culture.

The aim of this study was to assess the effect of potential target tissue on regenerating neurones of the snail Lymnaea stagnalis using the three-dimensional collagen gel culture system. Mammalian type I collagen supported the regenerative outgrowth of snail neurones, and the neurofilament antibody SMI31 specifically labelled regenerating processes both within the gel and those growing over the surface of the ganglia. Using these techniques we tested the effect of co-culturing ganglia with either additional nervous tissue, previously shown to produce trophic substances, or buccal muscle on both the amount and direction of outgrowth. We conclude that, under the conditions used, neither target tissue provided trophic or tropic support in collagen gel cultures.

Animals↗

Predator avoidance and immune defence: costs and trade-offs in snails.

Organisms are often confronted by both predators and pathogens. Defending against such widely divergent enemies requires more than one type of defence. Multiple defences, however, raise the possibility of trade-offs among defences. We tested for such trade-offs by manipulating the level of predator-avoidance behaviour and immune function in the freshwater snail Lymnaea stagnalis (Gastropoda: Pulmonata). Our results show that predator avoidance and immune function had clear costs in terms of reproduction and survival. Further, we show that increased levels of predator-avoidance behaviour reduced the snails' ability to defend against potential pathogens. Predator-avoidance behaviour may thus have the additional indirect cost of reduced immunocompetence and increased susceptibility to pathogens. Our results suggest that ecological factors (e.g. predator density) may considerably modify the expression and costs of immune defences.

Animals↗

A systems approach to the cellular analysis of associative learning in the pond snail Lymnaea.

We show that appetitive and aversive conditioning can be analyzed at the cellular level in the well-described neural circuitries underlying rhythmic feeding and respiration in the pond snail, Lymnaea stagnalis. To relate electrical changes directly to behavior, the snails were first trained and the neural changes recorded at multiple sites in reduced preparations made from the same animals. Changes in neural activity following conditioning could be recorded at the level of motoneurons, central pattern generator interneurons and modulatory neurons. Of significant interest was recent work showing that neural correlates of long-term memory could be recorded in the feeding network following single-trial appetitive chemical conditioning. Available information on the synaptic connectivity and transmitter content of identified neurons within the Lymnaea circuits will allow further work on the synaptic and molecular mechanisms of learning and memory.

Animals↗