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PubMed2006

Use of the land snail Helix aspersa as sentinel organism for monitoring ecotoxicologic effects of urban pollution: an integrated approach.

Atmospheric pollution from vehicular traffic is a matter of growing interest, often leading to temporary restrictions in urban areas. Although guidelines indicate limits for several parameters, the real toxicologic impacts remain largely unexplored in field conditions. In this study our aim was to validate an ecotoxicologic approach to evaluate both bioaccumulation and toxicologic effects caused by airborne pollutants. Specimens of the land snail Helix aspersa were caged in five sites in the urban area of Ancona, Italy. After 4 weeks, trace metals (cadmium, chromium, copper, iron, manganese, nickel, lead, and zinc) and polycyclic aromatic hydrocarbons (PAHs) were measured and these data integrated with the analyses of molecular and biochemical responses. Such biomarkers reflected the induction of detoxification pathways or the onset of cellular toxicity caused by pollutants. Biomarkers that correlated with contaminant accumulation included levels of metallothioneins, activity of biotransformation enzymes (ethoxyresorufin O-deethylase, ethoxycoumarin O-deethylase), and peroxisomal proliferation. More general responses were investigated as oxidative stress variations, including efficiency of antioxidant defenses (catalase, glutathione reductase, glutathione S-transferases, glutathione peroxidases, and total glutathione) and total oxyradical scavenging capacity toward peroxyl and hydroxyl radicals, onset of cellular damages (i.e., lysosomal destabilization), and loss of DNA integrity. Results revealed a marked accumulation of metals and PAHs in digestive tissues of organisms maintained in more traffic-congested sites. The contemporary appearance of several alterations confirmed the cellular reactivity of these chemicals with toxicologic effects of potential concern for human health. The overall results of this exploratory study suggest the utility of H. aspersa as a sentinel organism for biomonitoring the biologic impact of atmospheric pollution in urban areas. Key words: atmospheric pollutants, bioindicators, biomarkers, DNA integrity, lysosomes, metallothioneins, oxidative stress, peroxisomes, polycyclic aromatic hydrocarbons, trace metals.

Air Pollutants↗
PubMed1996

Deterministic chaos in mathematical model of pacemaker activity in bursting neurons of snail, Helix pomatia.

Chaotic regimes in a mathematical model of pacemaker activity in the bursting neurons of a snail Helix pomatia, have been investigated. The model includes a slow-wave generating mechanism, a spike-generating mechanism, an inward Ca current, intracellular Ca ions, [Ca2+]in, their fast buffering and uptake by intracellular Ca stores, and a [Ca2+]in-inhibited Ca current. Chemosensitive voltage-activated conductance, gB*, responsible for termination of the spike burst, and chemosensitive sodium conductance, gNa*, responsible for the depolarization phase of the slow-wave, were used as control parameters. These conductances in intact snail bursting neuron are regulated by neuropeptides. Time courses of the membrane potential and [Ca2+]in were employed to analyse different regimes in the model. Histograms of interspike intervals, autocorrelograms, spectral characteristics, one-dimensional return maps, phase plane trajectories, positive Lyapunov exponent and especially cascades of period-doubling bifurcations demonstrate that approaches to chaos were generated. The bifurcation diagram as a function of gB* and the ([Ca2+]in-V) phase diagram of initial conditions reveal fractal features. It has been observed that a short-lasting depolarizing current of elevation of [Ca2+]in may evoke transformation of chaotic activity into a regular bursting one. These kinds of transitions do not require any changes in the parameters of the model. The results demonstrate that chaotic regimes of neuronal activity modulated by neuropeptides may play a relevant role in information processing and storage at the level of a single neuron.

Animals↗
PubMedDate not supplied

[The role of glucose in facilitating defensive reactions during feeding in the edible snail Helix lucorum].

The glucose role in a regulation of defense responses during feeding behaviour was studied in the snail Helix lucorum. It was found that 15-20 min after the beginning of the food intake the foot withdrawal response evoked by a train of electrical pulses was facilitated up to 20-30%. At the same time glucose concentration in the haemolymph increased from 0.5 +/- 0.1 mM in the starved molluscs to 0.9 +/- 0.5 mM in the snails at 20-25 min of eating. In the food satiated snails and 30-90 min after the food intake the glucose level in the haemolymph was 1.6 +/- 0.5 mM. It was shown that 0.5 ml glucose solution (5 mM) injected into the mantle cavity also facilitated the foot withdrawal defense response in the fastened snail, whereas injection of 2.5 mM or 10 mM glucose solutions were ineffective. These data support a supposition that glucose in one of the endogenous factors involved in regulation of defense behaviour during feeding in snails.

Animals↗
PubMed1992

Channels activated by stretch in neurons of a helix snail.

Single-channel recordings from central neurons of the helix snail, Cepaea nemoralis, revealed two types of channels that could be activated by stretch (i.e., by the membrane deformation produced when suction is applied to the patch pipette). One, a K+ channel (58 pS in physiological solution), was evident in excised and cell-attached patches. Its conductance in symmetrical [K+] solutions indicated a channel of high K+ permeability (PK = 3.4 x 10(-13) cm/s). Though osmoregulation has been suggested as a function for such channels, comparisons among molluscs indicate osmotic milieu does not govern their expression; Cepaea is terrestrial, and stretch-activated K+ channels similar to those described here occur in aquatic and marine molluscs. The second type of channel, observed only in excised patches, was Cl- permeant; it had a large conductance (130 pS) and was inactive prior to patch excision. Membrane tension may not be the physiological activator of either the K+ or Cl- channel; the channels are designated as stretch-activated channels on the basis of their experimental behaviour during single-channel recording.

Animals↗
PubMed1996

Effectors of metabolic depression in an estivating pulmonate snail (Helix aspersa): whole animal and in vitro tissue studies.

We have examined metabolic depression in the land snail (Helix aspersa) during estivation, and have developed a tissue model of metabolic depression using an in vitro mantle preparation. The metabolic rate of H. aspersa is depressed by 84% in vivo within 4 weeks of onset of estivation, and this metabolic depression is accompanied by a decrease in haemolymph PO2 and pH, and an increase in haemolymph PCO2. The in vitro mantle preparation has a stable O2 consumption and energy charge, and an energy charge similar to that of mantle in vivo. The in vitro mantle is an O2-conforming tissue, with VO2 varying curvilinearly with PO2. Consequently, we have developed a mathematical method of calculating tissue VO2 at any PO2. These calculations show that under appropriate incubation conditions of pH and PO2, the mantle from estivating animals shows a stable in vitro metabolic depression of 48% compared to mantle from control snails. The extrinsic effects of PO2 and pH account for 70% of the total in vitro metabolic depression of mantle tissue; intrinsic effectors contribute a further 30%.

Adenosine Diphosphate↗
PubMedDate not supplied

[Mathematical model of the dual action of G protein on the Ca2+ current in identified neurons of the snail Helix aspersa].

INTRODUCTION: The effect of the dopamine on the calcium current of identified cells of the snail Helix aspersa consists on an initial decrease, followed by a subsequent increase when the drug is removed. It had been previously demonstrated that this effect is mediated by a G protein, supposing that the decrease of the current would be mediated by the G alpha subunit, while the increase would be produced by the G beta gamma subunit. OBJECTIVE: A mathematical model has been developed with the object to test if the hypothesis of the dual action of the G protein could explain the experimental results. MATERIAL AND METHODS: It has been recording by means of the 'patch-clamp' (whole cell) in identified cells of snail. On the other hand, it has been developed a mathematical model of the calcium current using a Hodgkin-Huxley model, and a simulation of the action of the G protein on this current. RESULTS: Adjusting the kinetic parameters of the channel by means of the experimental data, it has reproduced in a faithful way the behavior of the calcium current. The simulation of the action of dopamine reproduces the decrease and increase of the current by means of the serial action of the G protein's subunits. CONCLUSION: Although a mathematical model cannot demonstrate the logic necessity of the hypotheses on that is based, it is unequivocally demonstrated that the hypothesis is fully compatible with experimental results.

Animals↗
PubMed1990

Oligopeptides in the regulation of feeding and avoidance behaviour in the land snail (Helix lucorum).

The effects of the oligopeptides pentagastrin, cholecystokinin octapeptide, substance P, and FMRF-amide on the feeding and avoidance behaviour of the land snail Helix lucorum were studied. Administration of 1 microM pentagastrin to fasted snails immediately before feeding led to a significant increase in the amount of food consumed in the first 20 min of the feeding period, although there was no change in the total amount of food consumed. Prefeeding injections of 0.05 microM pentagastrin or 0.05 microM or 1 microM solutions of cholecystokinin octapeptide, substance P, or FMRF-amide had no effect on feeding behaviour. Pentagastrin and cholecystokinin octapeptide (0.05 microM solutions) invoked a significant reversible decrease in the generalised avoidance reaction thresholds in fasted but not in prefed snails, whereas the same concentration of FMRF-amide reduced these thresholds only in prefed snails.

Animals↗
PubMed1997

The specificity of an alpha-(1-->2)-L-galactosyltransferase from albumen glands of the snail Helix pomatia.

The specificity of an L-galactosyltransferase (L-Gal-T) from albumen glands of the snail Helix pomatia has been studied. This enzyme transfers L-Gal from GDP-L-Gal to various disaccharides with beta-linked D-Gal in terminal non-reducing position, forming an alpha-(1-->2) linkage. The subterminal residue and the type of interglycosidie linkage proved to be of minor importance. However, the branched trisaccharide beta-D-Gal-(1-->3)-[beta-D-Gal-(1-->6)]-beta-D-Gal-(1-->O)Me is a very poor acceptor. The specificity of the L-Gal-T correlates well with the equimolar occurrence of L-Gal and the structural element-->2)-Gal- (1-->found in the storage polysaccharide of this snail. Since L-Fuc is also transferred from its GDP-activated form, the membrane preparations of the albumen glands can be used to synthesize fucosylated oligosaccharides.

Animals↗
PubMedDate not supplied

[The facilitation of defensive reactions during food consumption in the snail helix: the participation of glucose and gastrin/cholecystokinin-like peptide].

Molecular and cellular mechanisms of the interrelations between the feeding and defense behaviour were studied in a snail Helix lucorum. The dynamics of defense reactions was investigated in snails with different levels of feeding motivation. Defense reactions were suppressed in hungry snails, while 15-20 min after the beginning of food intake they were facilitated. The facilitation depended on a duration of starvation. Injection of 0.5 ml of 5 mM glucose solution (up to the glucose level in the haemolymph of a food satiated snail, 1.6-2.0 mM) or injections of 20-30 ng of synthetic analogues of the gastrointestinal peptides (pentagastrin of octapeptide cholecystokinin, CCK-8) facilitated the defense reaction in a hungry snail. Parameters of the facilitation were similar to those in the period of food intake. Activity of the command neurons of defense behaviour (L-PPL1) after the carrot juice application to the lip of a semi-intact preparation from a hungry snail was glucose-dependent. Similar glucose-dependent changes of L-PPL1 activity were found after CCK-8, but not FMRFamide application during the perfusion with 0.5 mM glucose. L-PPL1, but not L-PPa2-3 neurons were most sensitive to glucose and CCK-8 level changes in the Ringer solution. Adaptive significance of the behavioural phenomena as well as glucose and gastrin/CCK-like peptide participation in these processes are discussed.

Animals↗
PubMed2006

The potential role of CO2 in initiation and maintenance of estivation in the land snail Helix lucorum.

Elevated CO(2) levels are hypothesized to play a role in the initiation and maintenance of estivation in snails through disturbances of acid-base status. The aim of our study was to identify the ambient CO(2) threshold that induces disturbances in acid-base status in the air-breathing land snail Helix lucorum. Acid-base parameters were determined in the hemolymph of snails acclimated to 0.5%, 1%, 2%, 4%, and 8% CO(2) in air for 20 d. In addition, we evaluated the effects of long-term acclimation on metabolic rate and on levels of D-lactate dehydrogenase activity (D-LDH) and of D-lactate in snails after 20 d of exposure to increased CO(2) levels. Helix lucorum proved to be unable to compensate for a decrease in extracellular pH (pH(e)) when acclimated to levels higher than 1% CO(2) in air. The rate of oxygen consumption started to decrease when snails were acclimated to 0.5% CO(2) in air. However, there was no correlation between the drops in pH(e) and in metabolic rate. Long-term acclimation to elevated CO(2) levels induced an increase in the activity of D-LDH with a concomitant accumulation of D-lactate in tissues. This indicates that long-term acclimation to elevated ambient CO(2) levels could reduce the aerobic capacity of land snails and trigger expression of anaerobic pathways of ATP turnover. The threshold levels of ambient CO(2) that induce changes in acid-base status and elicit metabolic depression in adult land snails H. lucorum are higher than the future atmospheric levels that are expected to result from human use of fossil energy resources.

Acclimatization↗
PubMed1991

DARPP-32 like protein in specific snail (Helix aspersa) neurones.

Monoclonal antibodies to DARPP-32 recognise an antigen which is present in specific neurones in the snail (Helix aspersa). Consecutive sections 10-microns-thick processed for the localisation of DARPP-32 and tyrosine-hydroxylase immunoreactivity did not show a coexistence in any neuronal structures. DARPP-32 positive cells were, however, often morphologically closely associated with tyrosine-hydroxylase positive cells, implying a functional relationship consistent with the proposed role of DARPP-32. Immunochemical analysis of the DARPP-32 immunoreactive material in the snail nervous system shows that the substance has a molecular weight of 28 kDa and therefore different from the DARPP-32 protein found in the rat brain.

Animals↗
PubMed1985

Horseshoe crab (Limulus polyphemus) lectin but not garden snail (Helix pomatia) lectin elicits insulin-like activities in vitro.

Lectins from the horseshoe crab (Limulus polyphemus) and the garden snail (Helix pomatia) were tested for insulinomimetic activities in isolated rat epididymal adipocytes. The sialic acid binding horseshoe crab lectin suppressed epinephrine-induced lipolysis and augmented lipogenesis from D-[3-3H]-glucose while the N-acetylgalactosamine binding snail lectin was inactive. The results suggest that the insulin receptor on rat adipocytes contains sialic acid in its carbohydrate moiety but does not possess non-reducing alpha-D-galactopyranosyl or 2-acetamido-2-deoxy-alpha-D-galactopyranosyl end groups.

Adipose Tissue↗
PubMed2001

The garden snail (Helix aspersa) as a bioindicator of organophosphorus exposure: effects of dimethoate on survival, growth, and acetylcholinesterase activity.

The garden snail (Helix aspersa) is currently used as bioindicator of metallic pollution. Our objective was to extend its use to organic chemicals by studying the effects and tissue concentrations of the organophosphorus pesticide dimethoate following dietary uptake. After exposure for four weeks to increasing doses of pesticide in the diet, the median lethal concentration (LC50) was determined to be 3,700 microg/g food. Clinical signs indicated a no-observed-effect concentration of 100 microg/g and a lowest-observed-effect concentration of 250 microg/g. The growth parameters were decreased with increasing exposure to the pesticide. The median effective concentration (EC50), which was evaluated based on both shell diameter and dry weight inhibitions, was 665 and 424 microg/g, respectively, and the EC10 was 180 and 145 microg/g, respectively. Accumulation in the viscera was related to the amount of dimethoate in the food. The bioconcentration factors were low (>6 x 10(-3)). Acetylcholinesterase (AChE) activity was strongly decreased (80% from 250 microg/g). In conclusion, we demonstrated that the species H. aspersa could be a useful sentinel organism for organophosphorus contamination surveys. Among the effects measured, the inhibition of AChE activities and clinical signs were the most sensitive, followed by the growth parameters. These results confirm the suitability of the garden snail for development of sublethal toxicity tests using primary consumers and aboveground organisms.

Acetylcholinesterase↗
PubMed1997

Molecular and phenotypic features of aeromonads isolated from snails (Helix aspersa) affected with a new summer disease.

Forty-four aeromonads were isolated during the summer of 1994 from snails (Helix aspersa) affected or not by a newly described disease. Snta 1 rRNA gene restriction patterns (ribotyping) and nine selected conventional phenotypic tests were performed to retrospectively determine relationships among these isolates regarding the disease. Results from ribotyping were found highly discriminant in strain differentiation and suggest the possible existence of a group of Aeromonas associated with the disease. Biochemical and molecular characterizations indicated that most of these disease-associated aeromonads could belong to the phenospecies A. hydrophila. Phenospecies of aeromonads from healthy snails were distinct from those isolated from diseased snails and could be identified as A. caviae or remained undetermined. To the authors' knowledge, this is the first report of a probable pathogenic aeromonad found in snails, extending the host-pathogen spectrum of Aeromonas.

Aeromonas↗
PubMed1990

Single and dual infections of the land snail Helix aspersa with Muellerius capillaris and Alloionema appendiculatum (Nematoda).

Helix aspersa, a land snail, has been experimentally infected with 2 species of nematodes Muellerius capillaris and Alloionema appendiculatum. A significant reduction of infection with M. capillaris was recorded in snails preinfected with A. appendiculatum, whereas snails preinfected with M. capillaris were as susceptible to A. appendiculatum.

Analysis of Variance↗
PubMed1972

The antagonistic actions of calcium and magnesium on the superfused ventricle of the snail Helix pomatia.

1. The isolated, superfused half-ventricle of the snail (Helix pomatia) maintains a degree of tonic activity even when not beating, since the membrane is depolarized beyond the tension threshold. Beating may be initiated by lateral stretch of the ventricle and by 5-hydroxytryptamine.2. Ca and Mn each have a hyperpolarizing action, while K and to a lesser extent Na cause depolarization: the tonic activity of the ventricle is affected accordingly.3. An increase in the extracellular concentration of Mg also causes the tension to fall, but without change in membrane potential. It can also initiate beating.4. Contractures induced by 30 mM-K are steadily maintained, but start with a more-or-less distinct twitch-like contraction. A contracture induced by a concentration of K above about 50 mM is poorly sustained and is followed by a further brief contracture when the K concentration is reduced to normal.5. Relations between contracture tension and the concentrations of Ca and Mg accord with the hypothesis that the two ions compete for attachment to a binding site on the cell surface and that tension is proportional to the amount of bound Ca. On this hypothesis, the apparent dissociation constant of the Mg complex is 11.4 mM and that of the Ca complex 0.15 mM or less.6. This effect of Mg is like that of Na on frog ventricle and some of the differences in the behaviour of snail and frog ventricles are abolished by appropriate adjustment of the extracellular concentrations of these ions.

Animals↗
PubMed1992

Effects of adenine nucleosides and nucleotides on the isolated heart of the snail Helix aspersa and the slug Arion ater.

1. Adenine nucleosides and nucleotides were examined for pharmacological activity in hearts isolated from the snail Helix aspersa and the slug Arion ater. 2. Adenosine, AMP, ADP and ATP (above 100 microM) produced either an excitation or an inhibition in the isolated hearts of the snail and slug. 3. 2-Chloroadenosine, alpha, beta-methylene ATP and 2-methylthio ATP were inactive at concentrations up to 1 mM. 4. Responses were not blocked by any commonly accepted vertebrate purinoceptor antagonists, indicating that these purinoceptors are dissimilar to vertebrate purinoceptors and cannot be classified according to accepted purinoceptor classifications. 5. Electrical field stimulation of the snail heart produced frequency-dependent responses: 1-4 Hz produced predominantly excitation, 8-32 Hz predominantly inhibition. These responses were unaffected by the purines up 3 mM.

Adenine Nucleotides↗
PubMed2000

Hemocytic lysosome response in the snail Helix aspersa after exposure to the fungicide copper oxychloride.

This study investigated the use of lysosomal responses of hemocytes of the common garden snail, Helix aspersa, as biomarker of stress due to exposure to the fungicide copper oxychloride. The neutral red retention (NRR) time assay was employed for this purpose. Two groups of snails were exposed to 80 microg g(-1) and 240 microg g(-1) copper oxychloride in their food, respectively, for a period of 6 weeks. They were compared with a control group to which no copper oxychloride was added. The two groups exposed to the fungicide exhibited significantly higher (p < 0.001) whole body copper concentrations (200.85 +/- 53.5 and 272.24 +/- 67.15 microg g(-1) dry mass, respectively), and significantly shorter (p < 0.001) NRR times (10.22 +/- 3.53 and 2.67 +/- 2.83 min, respectively), after 6 weeks, compared to the control group (67.85 +/- 31.08 microg g(-1) dry mass and 24.44 +/- 8.35 min). In both exposure groups NRR times became progressively shorter as body copper concentrations increased over time. Thus, both exposure concentration and exposure time of copper oxychloride were shown to be important factors influencing lysosomal responses (and therefore NRR times) of H. aspersa hemocytes. It was concluded that these responses in this species, as measured by the NRR time assay, could be considered a useful cellular biomarker of stress resulting from exposure to copper oxychloride.

Animals↗