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Studies on the extra- and intracellular acid-base status and its role on metabolic depression in the land snail Helix lucorum (L.) during estivation.

The aim of the present study was to examine the acid-base status of extra- and intracellular fluids and its possible role on the regulation of the metabolic rate of Helix lucorum during prolonged estivation. For this purpose, the rate of oxygen consumption for active and estivating snails was determined. The acid-base status was also examined in the hemolymph and tissues from active and estivating snails acclimated at 25 degrees C. In addition, the buffer values of hemolymph and tissues were determined in order to examine whether there is a change in the snails during estivation. The rate of oxygen consumption decreased significantly within the 1st 10 days of estivation from 122.51+/-10 microl.g(-1).h(-1) to 25.86+/-5.2 microl.g(-1).h(-1), indicating a marked decrease in metabolic rate. P(CO2)increased within the 1st 20 days of estivation from 13.52+/-0.68 mmHg to 25.09+/-2.05 mmHg, while the pH of hemolymph (pH(e)) decreased from 7.72+/-0.04 to 7.44+/-0.06. The level of bicarbonates decreased in the hemolymph of estivating snails, indicating a metabolic acidosis, which was moderate in extracellular fluids. In contrast to pH(e), the intracellular pH (pH(i)) was maintained in the tissues of estivating H. lucorum, indicating a regulation of pH(i) despite the developed hypercapnia. According to the results presented here, it seems that the timing of pH(e) changes does not correlate with the timing of metabolic rate reduction in estivating H. lucorum.

Acid-Base Equilibrium↗

Cold tolerance of Littorinidae from southern Africa: intertidal snails are not constrained to freeze tolerance.

All intertidal gastropods for which cold tolerance strategies have been assessed have been shown to be freeze tolerant. Thus, freeze tolerance is considered an adaptation to the intertidal environment. We investigated the cold tolerance strategies of three species of subtropical and temperate snails (Gastropoda: Littorinidae) to determine whether this group is phylogenetically constrained to freeze tolerance. We exposed 'dry' acclimated and 'wet' rehydrated snails to low temperatures to determine temperature of crystallisation (Tc), lower lethal temperature and LT(50) and to examine the relationship between ice formation and mortality. Tc was lowest in dry Afrolittorina knysnaensis (-13.6+/-0.4 degrees C), followed by dry Echinolittorina natalensis (-10.9+/-0.2 degrees C) and wet A. knysnaensis (-10.2+/-0.2 degrees C) . The Tc of both A. knysnaensis and E. natalensis increased with rehydration, whereas Tc of dry and wet Afrolittorina africana did not differ (-9.6+/-0.2 and -9.0+/-0.2 degrees C respectively). Wet snails of all species exhibited no or low survival of inoculative freezing, whereas dry individuals of A. knysnaensis could survive subzero temperatures above -8 degrees C when freezing was inoculated with ice . In the absence of external ice, Afrolittorina knysnaensis employs a freeze-avoidance strategy of cold tolerance, the first time this has been reported for an intertidal snail, indicating that there is no family-level phylogenetic constraint to freeze tolerance. Echinolittorina natalensis and A. africana both showed pre-freeze mortality and survival of some internal ice formation, but were not cold hardy in any strict sense.

Adaptation, Physiological↗

Interactions related to non-host snails in the host-finding process of Euparyphium albuferensis and Echinostoma friedi (Trematoda: Echinostomatidae) miracidia.

In order to determine whether the miracidia of Euparyphium albuferensis and Echinostoma friedi are sensitive to their host snail (HS) and capable of discriminating between HS and non-host snails (NHS), or whether these NHS can interfere and thus reduce the infection rates (decoy effect), a total of three experiments were conducted with HS, NHS and snail-conditioned water (SCW). Gyraulus chinensis is the HS for E. albuferensis miracidia, while Physa acuta, Radix peregra and Lymnaea fuscus are considered NHS. For E. friedi miracidia R. peregra, G. chinensis and L. fuscus are the HS, while P. acuta is the NHS. The NHS R. peregra produces the greatest decoy effect on Euparyphium albuferensis miracidia, while R. peregra, as the HS of Echinostoma friedi miracidia, is always subject to a NHS decoy effect. However, an increase in E. friedi miracidia infectivity is observed in its HS G. chinensis, in the presence of SCW of P. acuta. These experimental results might explain the low prevalence of snails infected with E. albuferensis miracidia in their natural habitat.

Animals↗

Nitric oxide synthesis is blocked in the enteral nervous system during dormant periods of the snail Helix lucorum.

During dormancy of terrestrial snails, the whole neuromodulation of the nervous system is deeply modified. In this work we studied the adaptation of a previously described, putatively nitric oxide (NO) forming enteral network to the long-term resting periods of the snail Helix lucorum. The standard NADPH diaphorase (NADPHd) technique, which is an accepted method for histochemical NO synthase (NOS) detection, labeled the same enteric neurons of the midintestine in active or hibernated snails. Quantification of the NO-derived nitrite by the Griess reaction established that the nitrite formation is confined to the NADPHd-reactive network containing the midintestinal segment. In active snails, the nitrite formation could be enhanced by the NOS substrate L-arginine (10 microM-1 mM), but decreased by the known NOS inhibitors 1 mM N(omega)-nitro-L-arginine (NOARG) and 10 mM aminoguanidine (AG). Application of 1 mM L-arginine and 1 mM NOARG decreased the amplitude of the midintestinal muscle contractile activity, but did not affect the rectal motility. In dormancy, the nitrite formation was reduced in the NADPHd-reactive midintestinal network. Application of l-arginine could not provoke nitrite production and did not influence the midintestinal motility. Our findings indicate that NO is involved in the neural transmission to intestinal muscles of gastropods, but enteric release of NO is blocked during dormancy. The decreased NO synthesis is possibly due to an as yet undefined mechanism, by which the L-arginine/NO conversion ability of NOS could temporarily be inhibited in the long-term resting period of H. lucorum.

Animals↗

Regulation of tentacle length in snails by odor concentration.

The upper tentacle of the snail, bearing the olfactory organ, produces complex movements when the snail explores a new environment. Tentacle trajectories were reconstructed in the presence and absence of odors using two simultaneous video recordings. Reconstructions showed that in the absence of odor, snails constantly scanned the surrounding space with the extended tentacles. Presentation of an odor elicited rapid flexion, independent of the odor concentration, accompanied by concentration-dependent tentacle contractions. Activation of identified motoneuron MtC3 is known to elicit tentacle contraction. Recordings made in semi-intact preparations showed that the dynamics and duration of the spike activity of MtC3 produced in response to odors correlated with the degree of tentacle contraction in response to odors. These data suggest that the central motoneuron MtC3, which triggers tentacle contraction, is involved in controlling the margins of the scanning field. Slow contraction or extension of the tentacle, associated with the level of MtC3 activity, may operate to tune the snail's investigative behavior to the conditions of the sensory environment.

Action Potentials↗

Biological control of the snail hosts of Schistosoma mansoni in the Caribbean area using Thiara spp.

Field observations and experiments using thiarid snails as competitors of Biomphalaria spp., potential intermediate hosts of Schistosoma mansoni in the Caribbean area, are reviewed. The parthenogenetic snails, Thiara granifera and T. (= Melanoides) tuberculata, were introduced to the Neotropical area in recent decades. In numerous islands and countries, these oriental species have demonstrated their capacity to colonize rapidly and densely many types of habitats while at the same time reducing and even eliminating populations of Biomphalaria spp. The results of field experiments, carried out in several Caribbean islands, have shown the efficiency as well as the limitations of T. tuberculata as a competitor of B. glabrata and B. straminea. In St. Lucia, B. glabrata was apparently eliminated from marshes and streams, 6 to 22 months after the introduction of the competitor. In Martinique, T. tuberculata was introduced into two groups of water-cress beds which constituted the last transmission sites of schistosomiasis on the island. In just less than three years after the introduction of the competitor, both B. glabrata and B. straminea have been eliminated from the transmission sites. In Guadeloupe, several introductions have been carried out in different types of habitat such as permanent ponds, canals, streams and temporary marshes. The findings of all field experiments have indicated that thiarid snails as competitors of pulmonates are favoured by the presence of permanent and stable habitats, preferably shallow, with emergent plants and well oxygenated. On the other hand, the competitor snails are at a disadvantage in waterbodies which are temporary, extremely deep, poorly oxygenated or with a dense mat of floating aquatic vegetation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Schistosoma mansoni: male-biased sex ratios in snails and mice.

Adult sex ratios of Schistosoma mansoni, in mice, were shown to be biased toward males (3:1) despite the finding that sex ratios of miracidia were 50:50. The adult male bias was caused by greater male infectivity of miracidia for snails and cercariae for mice. A significantly higher percentage of male miracidia developed to cercarial production in unimiracidial infections (57 male, 34 female), and a significantly higher percentage of male cercariae developed to adulthood in mice (143 male, 79 female worms resulted from 900 male and 900 female cercariae). No significant differences were found between male and female parasites for longevity of miracidia (both sexes, 10 hr) and cercariae (males 21.3 +/- 5.75 hr, females 25.0 +/- 7.02 hr), prepatent periods in snail hosts (male 34 +/- 2.92 days, females 33 +/- 2.36 days), longevity of snail infections (males 96.6 +/- 25.15 days, females 115.2 +/- 82.43 days), and the numbers of cercariae produced per snail lifetime (males 30,751.44 +/- 18,064.33, females 34,083.00 +/- 33,732.82). Present results provide a better understanding of the life cycle of S. mansoni, are of theoretical significance for theories of biased sex ratios (which at present cannot account for the male-biased ratio of S. mansoni), and also suggest that schistosomiasis transmission models assuming a 50:50 sex ratio at all stages of the life cycle should be reassessed.

Animals↗

Novelty-induced opioid analgesia in the terrestrial snail, Cepaea nemoralis.

Exposure for 1-15 min to the surface of either a non-functional (22 degrees C) hot-plate or a polypropylene tube increased the thermal (38.5 degrees C) nociceptive thresholds of the terrestrial snail, Cepaea nemoralis. This "analgesic response," which was evident 10-15 sec after exposure to the new stimuli and lasted for 15-30 min, was blocked by the opiate antagonist naloxone. The analgesic response was not observed in snails previously made familiar with the test situation. Snails exposed to a functional (36.5 degrees C) stressful hot-plate surface to which they were aversive also displayed a naloxone-reversible analgesia. However, the level and duration of analgesia was markedly greater than that observed following exposure to the novel non-physically stressful stimuli and was not inhibited by prior familiarization with the stimuli. These observations demonstrate that novelty per se can activate endogenous opioid system(s) and induce an analgesic response in a snail. They also suggest a phylogenetic continuity in responses to novel stimuli and environmental conditions.

Animals↗

Environmental specificity of tolerance to morphine-induced analgesia in a terrestrial snail: generalization of the behavioral model of tolerance.

Terrestrial snails, Cepaea nemoralis, develop tolerance to morphine-induced analgesia, such that after 7-9 days of treatment with morphine (10 mg/kg) their response latencies to an aversive thermal stimuli (38.5 degrees C) are not significantly different from those of untreated control animals. In Experiment A snails were rendered tolerant to morphine using either of two pre-injection cues (light and dark background brightness or color) and then assessed for morphine-induced alterations in thermal nociceptive responses in both environments. In Experiment B snails were made tolerant to morphine in the presence of one of two different thermal cues (a stressful temperature of 35 degrees C that is normally avoided or an ambient temperature of 22 degrees C) and then tested for morphine-induced alterations in nociceptive responses in both environments. In the two experiments tolerance to morphine-induced analgesia was displayed when snails were exposed to the pre-injection environmental cue normally associated with the administration of morphine, but not when exposed to the alternative pre-injection cue. These results demonstrate that various environmental factors (background colors or brightness as well as temperature cues and potentially thermal stress), can function as environmental specific cues for the development of tolerance to morphine-induced analgesia in molluscs, in a manner consistent with a behavioral mechanism of tolerance. Thus, these results suggest that environmental specificity of tolerance involving either classical (Pavlovian) conditioning or habituation may be a general phenomenon having an early evolutionary development and broad phylogenetic continuity.

Analgesia↗

Comparative metabolism of fenitrothion in aquatic organisms. II. Metabolism in the freshwater snails, Cipangopaludina japonica and Physa acuta.

When freshwater snails, Cipangopaludina japonica and Physa acuta, were exposed to 0.1 ppm [14C]fenitrothion in a dynamic flow system, the concentrations of fenitrothion and 14C in the body reached equilibrium on Day 1 of exposure. The maximum bioaccumulation ratios of fenitrothion were 18 and 53 in C. japonica and P. acuta, respectively. These snails metabolized the compound primarily by demethylation, hydrolysis, and reduction. The liberated phenol moiety was found to be conjugated with sulfate in C. japonica and mainly with glucose in P. acuta. When the snails were transferred to a freshwater stream, fenitrothion and its metabolites were rapidly excreted, and the half-life of the parent compound was less than 0.5 day in both snails. Fenitrothion and its decomposition products were mainly distributed in liver of P. acuta, as evidenced by whole-body radioautography.

Animals↗

The interactions of courtship, feeding, and locomotion in the behavioral hierarchy of the snail Helix aspersa.

Starvation both increased the number of snails that exhibited feeding (i.e., increased feeding proclivity) and decreased the latencies of response to a food stimulus (i.e., increased food arousal). Feeding inhibited locomotion, although starvation itself had no effect. Sexual arousal (the intensity of courtship) increased locomotion, but only in the absence of a mating partner. Sexual proclivity (the likelihood of a snail to court with conspecifics) had no effect on locomotion. In general, proclivity and arousal had different effects on behavior. Food deprivation did not alter the preference of sexually aroused snails for sexual stimuli over food stimuli Both starved and fed courting pairs responded to a food stimulus only during periods of low sexual arousal, although when not sexually aroused, starved snails usually increased the amount of time they spent in contact with a food stimulus. These results suggest that courtship can suppress feeding and that the expression of feeding behavior is dependent upon the occurrence of low levels of sexual arousal (time sharing).

Animals↗

Immunocytochemistry of peptidergic systems in the pond snail Lymnaea stagnalis.

Evidence suggests that there exists in the animal kingdom a family of biologically active peptides whose members are related to the molluscan cardio-active tetrapeptide FMRFamide (Phe-Met-Arg-Phe-NH2). Immunocytochemical and ultrastructural studies indicate that several family-members occur in the pond snail Lymnaea stagnalis. Monoclonal antibodies were raised to whole brain homogenates of the pond snail. Selection of antibody producing hybridomas was carried out by staining sections of the central nervous system of the snail with the supernatants of the hybridomas. Certain antibodies stain selectively known (neuro)endocrine centres of the snail, others are directed against particular groups of neurons. It is argued that these antibodies were raised against biologically active peptides and/or their precursors. The antibodies may be used for the isolation of these peptides.

Amino Acid Sequence↗

The impairment of mobility and development in freshwater snails (Physa fontinalis and Lymnaea stagnalis) caused by herbicides.

1. The pulmonate freshwater snails Physa and Lymnaea and the earthworms Eisenia and Lumbricus can take up and concentrate a carbamate herbicide (chlorpropham). 2. The mobility of freshwater snails was diminished in the presence of the herbicides chlorpropham (carbamate), cycloate (thiocarbamate), pentanochlor (amide) and chloroxurone (urea derivate). 3. The egg-assemblies of Lymnaea stagnalis turned out to be suitable objects for testing the influences of herbicides upon embryonic development. 4. In the presence of chlorpropham, chloroxurone, cycloate, propanil, simazine and terbutryne, all applied in concentrations lower than in practical use, the period of egg-maturing was delayed and the total number of dead embryos increased. 5. Developing snail eggs were very sensitive towards triazine herbicides (simazine, terbutryne) reflecting in an ED50-value lower than 10(-7) M. 6. Similar herbicidal-induced effects suggested that the developing stages of snail embryos may be suitable models for the ecologically more important but experimentally less accessible earthworms.

Animals↗

Cold hardiness abilities vary with the size of the land snail Cornu aspersum.

The land snail Cornu aspersum (syn. Helix aspersa) living in Brittany (France) can be considered partially freezing tolerant as it possesses a low ability to supercool and a limited capacity to bear freezing of its body tissues. The absence of a marked cold hardiness strategy permits the emphasis of the role of parameters such as individual size or water mass (W(M)) contained by the organism. Adult snails (shell diameter 30-32 mm) had a supercooling ability, about 1-1.5 degrees C lower than that of immatures (shell diameter 12-20 mm) and survived longer to an exposure to -5 degrees C, with an Lt(50) comprised between 6.0 and 9.8 h against 2.6 to 4.2 h for immature snails. This better ability to bear freezing was explained by the faster dynamic of body ice formation observed in small individuals, which attained ice lethal quantity more rapidly. At the species level, large snails will then tend to be more tolerant to freezing and small ones to be freezing avoidant, a statement also observable at the phylum level.

Acclimatization↗

Toxicity of anti-fouling biocides to encysted metacercariae of Echinoparyphium recurvatum (Digenea: Echinostomatidae) and their snail hosts.

The toxicity of the anti-fouling biocides tributyltin (TBTO), copper, and Irgarol 1051 (irgarol) at a nominal concentration of 10 microg/l over a 30 day period were investigated against the viability of metacercarial cysts of the digenean parasite Echinoparyphium recurvatum resident within the body of two common freshwater snails, Lymnaea peregra and Physa fontinalis. Reduced parasite viability was found under most exposures in both snail species. However a greater effect of toxicant exposure was found in cysts within P. fontinalis compared to those in L. peregra. This was associated with an increased mortality of the host snail. Among all tested biocides, TBTO exposures induced the highest mortality to both the parasite and their hosts. These results suggest that parasite viability is interlinked with survival of the host snail. The mechanisms of differing toxicity between host species and its relevance to successful parasite transmission to the next host are discussed.

Animals↗

Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in a freshwater snail (Bellamya aeruginosa) from a large shallow, eutrophic lake of the subtropical China.

Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in a freshwater snail (Bellamya aeruginosa) were studied monthly in a large shallow, eutrophic lake of the subtropical China during June-November, 2003. Microcystins (MCs) were quantitatively determined by High-Performance Liquid Chromatography (HPLC) with a qualitative analysis by a Finnigan LC-MS system. On the average of the study period, hepatopancreas was the highest in MC contents (mean 4.14 and range 1.06-7.42 microg g(-1)DW), followed by digestive tracts (mean 1.69 and range 0.8-4.54 microg g(-1)DW) and gonad (mean 0.715 and range 0-2.62 microg g(-1)DW), whereas foot was the least (mean 0.01 and range 0-0.06 microg g(-1)DW). There was a positive correlation in MC contents between digestive tracts and hepatopancreas. A constantly higher MC content in hepatopancreas than in digestive tracts indicates a substantial bioaccumulation of MCs in the hepatopancreas of the snail. The average ratio of MC-LR/MC-RR showed a steady increase from digestive tracts (0.44) to hepatopancreas (0.63) and to gonad (0.96), suggesting that MC-LR was more resistant to degradation in the snail. Since most MCs were present in the hepatopancreas, digestive tracts and gonad with only a very small amount in the edible foot, the risk to human health may not be significant if these toxic parts are removed prior to snail consumption. However, the possible transference of toxins along food chains should not be a negligible concern.

Animals↗

Fasciola hepatica: identification of molecular markers for resistant and susceptible Pseudosuccinea columella snail hosts.

Protein electrophoresis, RAPD-PCR and nuclear rDNA ITS sequencing were performed to search for genetic differences between Pseudosuccinea columella snails susceptible and resistant to Fasciola hepatica infection. Of the 21 enzymatic loci analyzed in both populations, none of them exhibited neither within- or between-group variation. Such an absence of enzyme polymorphism support the hypothesis of selfing as the "prevalent" mating system for this hermaphroditic species. Conversely, the RAPD profiles displayed clear differences between susceptible and resistant isolates for 17 of the 26 primers tested while no within-group variation was detected. rDNA ITS sequence analysis from snails of each isolates showed only two bases that differed between groups accounting for a 0.17% of variation confirming that susceptible and resistant snails belong to the same species. This is the first time that a genetic variation using RAPD markers is demonstrated between susceptible and resistant lymnaeid snails vis-a-vis of F. hepatica infection in absence of experimental selection.

Animals↗

Chronic toxicity of tributyltin on development and reproduction of the hermaphroditic snail Physa fontinalis: Influence of population density.

Tributyltin (TBT) is toxic to aquatic organisms and occurs widely in sediments and surface waters of American and European rivers and lakes. This study investigated TBT effects on development and population growth rate (r) of the common, hermaphroditic European freshwater snail Physa fontinalis. Egg ropes of similar age (1-3 days old) were exposed to a control (solvent only) and nominal concentrations of 0.01, 1.0 and 10 microg TBT l(-1) in triplicate. Hatching and mortality were recorded during 0-40 days of exposure. At day 40, 18 juveniles were randomly selected from each concentration (i.e., six from each test vessel) and individually exposed to the same concentration of TBT in 50 ml beakers. A cohort of 20 juveniles was allowed to continue developing in the original test vessels, so that individual and grouped results could be compared. Mortality and reproduction were recorded at 48-h intervals throughout the study period (110 days). Abnormal embryonic development was observed at 1 and 10 microg TBT l(-1). Although 50% of eggs hatched at 10 microg TBT l(-1), all these hatchlings failed to survive. Survivorship of hatchlings was significantly reduced by TBT at 1 microgl(-1). In general, there was a delay in egg production in isolated snails when compared with the grouped snails. Survival, fecundity and population growth rate (r) were reduced in both individual and grouped P. fontinalis at 1.0 microg TBT l(-1). Only a decline in r was observed in snails exposed individually to 0.01 microg TBT l(-1).

Animals↗