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Biochemical studies on the terrestrial snail, Eubania vermiculata (Müller) treated with some pesticides.

The in vivo effects of methomyl, thiodicarb and metaldehyde on total soluble proteins, total lipids and glycogen content, in addition, the activity of glutamic oxaloacetic transaminase, (GOT), (GPT) glutamic pyruvic transaminase and catalase (CAT) enzymes of terrestrial E. vermiculata snails was studied. The experimental snails were treated with low concentration of 0.2% brain bait w/w of the pesticides for a period of 1,3,5,7 and 10 days. The results showed that methomyl and thiodicarb lead to significant reduction in total soluble proteins, lipids, and glycogen content, while significant increases in the activity of all enzymes tested were noted. Metaldehyde treatment showed no significant effect on total soluble proteins, lipids and GOT level, whereas a significant increase in GPT and CAT enzymes was observed. Also, metaldehyde resulted a significant reduction in glycogen content of snails.

Acetaldehyde↗

Effects of prolonged acclimation to cold on the extra--and intracellular acid-base status in the land snail Helix lucorum (L.).

The aim of this study was to examine the effect of prolonged acclimation to cold on the acid-base status of extra- and intracellular fluids in the land snail Helix lucorum. For this purpose, acid-base parameters in the hemolymph and tissues were determined. In addition, the buffer values of hemolymph and tissues were determined in order to examine whether they change in the snails during acclimation to cold. According to the results presented, there is an inverse pH-temperature relationship in the hemolymph within the first day of acclimation, which is consistent with alphastat regulation. The Pco2 decreased, and pH in the hemolymph (pH(e)) increased by 0.32 U within the first day of acclimation to cold, which corresponds to a change of 0.013 U degrees C(-1). After the first day of acclimation, Pco2 increased in the hemolymph, resulting in a significant drop in pH(e) by 90 d of acclimation to cold. Acclimation of snails to low temperatures did not change the buffer value of the hemolymph. Also, intracellular pH (pH(i)) and intracellular buffer values remained stable during acclimation to cold for prolonged periods. The latter results in conjunction with those obtained by the in vitro determination of the passive component of intracellular fluids indicate an active regulation of pH(i) in H. lucorum during acclimation to cold.

Acclimatization↗

Seasonal and artificially elevated temperatures influence bioenergetic allocation patterns in the common pond snail, Physella virgata.

Allocation of organic carbon (OC) to primary energetic pathways was estimated under seasonal and artificially elevated ambient temperatures for a field population of a freshwater pulmonate snail, Physella virgata. Allocation to respiration increased with temperature. Snails allocated most assimilated OC to reproduction within their natural temperature range (15 degrees -35 degrees C), where assimilation efficiencies remained relatively stable at 25%-35%. However, in artificially heated waters exceeding 35 degrees C, declining assimilation rates and increasing respiratory demands inhibited allocation to reproduction and growth. At the species' 40 degrees C upper thermal limit, assimilation efficiencies fell below 10%, while average consumption levels more than doubled relative to snails unaffected by the thermal effluent. Ambient temperature substantially influenced OC allocation over P. virgata's natural temperature range and negatively affected growth and reproduction at temperatures approaching or exceeding maximum natural levels.

Animals↗

dCtBP mediates transcriptional repression by Knirps, Krüppel and Snail in the Drosophila embryo.

The pre-cellular Drosophila embryo contains 10 well characterized sequence-specific transcriptional repressors, which represent a broad spectrum of DNA-binding proteins. Previous studies have shown that two of the repressors, Hairy and Dorsal, recruit a common co-repressor protein, Groucho. Here we present evidence that three different repressors, Knirps, Krüppel and Snail, recruit a different co-repressor, dCtBP. Mutant embryos containing diminished levels of maternal dCtBP products exhibit both segmentation and dorsoventral patterning defects, which can be attributed to loss of Krüppel, Knirps and Snail activity. In contrast, the Dorsal and Hairy repressors retain at least some activity in dCtBP mutant embryos. dCtBP interacts with Krüppel, Knirps and Snail through a related sequence motif, PXDLSXK/H. This motif is essential for the repression activity of these proteins in transgenic embryos. We propose that dCtBP represents a major form of transcriptional repression in development, and that the Groucho and dCtBP co-repressors mediate separate pathways of repression.

Alcohol Oxidoreductases↗

Bioaccumulation of 95mTc in fish and snails.

Concentration factors for technetium recommended in radiological assessment models for freshwater biota are default values based on the behavior of iodine in the environment. In this study a small experimental freshwater pond was spiked with 95mTc to obtain data for calculating concentration factors for fish and snails. A model using the pond data was developed to calculate steady-state body burdens for freshwater biota. The concentration factors based on the calculated body burden for carp (Caprinus carpio), mosquitofish (Gambusia affinis), and snails (Helisoma sp.) were 11, 75 and 121, respectively. The concentration factor for carp was less than the recommended value of 15 listed in the USNRC Regulatory Guide 1.109 for calculating radiation dose to man; however, the concentration factors for mosquitofish and snails exceeded the recommended values by 5 and 24 times, respectively.

Animals↗

Reconsolidation of a context long-term memory in the terrestrial snail requires protein synthesis.

We investigated the influence of the protein synthesis blocker anisomycin on contextual memory in the terrestrial snail Helix. Prior to the training session, the behavioral responses in two contexts were similar. Two days after a session of electric shocks (5 d) in one context only, the context conditioning was observed as the significant difference of behavioral response amplitudes in two contexts. On the day following testing of context learning, a session of "reminding" was performed, immediately after which the snails were injected with anisomycin or vehicle. Testing of long-term context memory has shown that only anisomycin injections impaired the context conditioning. In control series, the snails were injected after the training session with anisomycin/saline without reminding, and no impairment of the long-term context memory was observed, while injection of anisomycin during the training session completely abolished the long-term memory. No effects of anisomycin on the short-term memory were observed. Surprisingly, injection of anisomycin after the reminding combined with reinforcing stimuli elicited no effect on the context memory. Differences between single-trial and multisession learning are discussed.

Animals↗

Family-level covariation between parasite resistance and mating system in a hermaphroditic freshwater snail.

Genetic compatibility, nonspecific defenses, and environmental effects determine parasite resistance. Host mating system (selfing vs. outcrossing) should be important for parasite resistance because it determines the segregation of alleles at the resistance loci and because inbreeding depression may hamper immune defenses. Individuals of a mixed mating hermaphroditic freshwater snail, Lymnaea ovata, are commonly infected by a digenetic trematode parasite, Echinoparyphium recurvatum. We examined covariation between quantitative resistance to novel parasites and mating system by exposing snail families from four populations that differed by their inbreeding coefficients. We found that resistance was unrelated to inbreeding coefficient of the population, suggesting that the more inbred populations did not carry higher susceptibility load than the less inbred populations. Most of the variation in resistance was expressed among the families within the populations. In the population with the lowest inbreeding coefficient, resistance increased with outcrossing rate of the family, as predicted if selfing had led to inbreeding depression. In the other three populations with higher inbreeding coefficients, resistance was unrelated to outcrossing rate. The results suggest that in populations with higher inbreeding some of the genetic load has been purged, uncoupling the predicted relationship between outcrossing rate and resistance. Snail families also displayed crossing reaction norms for resistance when tested in two environments that presented low and high immune challenge, suggesting that genotype-by-environment interactions are important for parasite resistance.

Animals↗

Mitochondrial haplotypes and the New Zealand origin of clonal European Potamopyrgus, an invasive aquatic snail.

The small aquatic snail Potamopyrgus antipodarum is an important invading species in Europe, Australia and North America. European populations are generally believed to derive from accidental introductions from New Zealand, probably dating back to the mid-19th century. We have employed mitochondrial DNA sequences to test the proposed New Zealand origin of European Potamopyrgus, and to learn more about its genealogical history. Using a 481-bp region of the 16S ribosomal RNA gene, we identified 17 distinct haplotypes among 65 snails from New Zealand. In marked contrast, only two haplotypes were found across all European samples, which cover a large geographical area. Importantly, these two haplotypes are shared with snails from the North Island of New Zealand. Due to sampling limitations we cannot rule out a South Island origin for one of the haplotypes, but our results clearly demonstrate the New Zealand origin of European populations. The marked divergence among the two European haplotypes implies the successful colonization by two distinct mitochondrial lineages, which is consistent with previous data based on nuclear markers.

Animals↗

Molecular cloning of the antibacterial protein of the giant African snail, Achatina fulica Férussac.

An expression cDNA library was constructed with poly(A)-rich RNA extracted from the collar of the giant African snail, Achatina fulica Férussac. A 1.9-kbp cDNA clone encoding a precursor of antibacterial glycoprotein of the snail, achacin, was isolated from the cDNA expression library. The cDNA sequence contains an open reading frame with 1593-nucleotide residues. The deduced amino acid sequence of this achacin precursor starts with a 29-residue leader peptide followed by a 502-residue mature peptide (56 kDa) with four possible N-glycosylation sites, Asn-Xaa-Ser or Asn-Xaa-Thr. The Northern-blot analysis proved that the achacin precursor was specifically expressed in the tissue of snail collar and processed to mature achacin. cDNA inserts encoding achacin precursor were subcloned into expression plasmids. Three kinds of expressed polypeptides were cross-reacted with rabbit antiserum raised against achacin. The largest polypeptide (M(r) 63,000) should be the achacin precursor.

Amino Acid Sequence↗

Purification and primary structure of snail metallothionein. Similarity of the N-terminal sequence with histones H4 and H2A.

A cadmium-binding metallothionein has been purified from metal-exposed Roman snails (Helix pomatia) using gel-permeation, ion-exchange and reverse-phase high-performance liquid chromatography. The S-methylated protein was digested with trypsin and the endoproteinases Asp-N, Glu-C and Arg-C. While most of the resulting peptides could be sequenced by Edman degradation, the intact protein, as well as the N-terminal peptide, proved to be blocked. Analysis by mass spectrometry showed that the N-terminal amino acid was an acetylated serine residue. Snail metallothionein, which is suggested to be involved in the detoxification of cadmium, contains 66 amino acid residues, 18 of which are cysteine residues arranged in seven Cys-Xaa-Cys motifs. The calculated molecular mass of the protein is 6.62 kDa. The primary structure of snail metallothionein reveals a clear relationship with molluscan and vertebrate metallothioneins, but lower similarity with metallothioneins of other invertebrate species. The N-terminal region of the isolated protein proved to be unique among the metallothionein sequences determined so far, showing high degrees of similarity with the N-terminal sequences of histones H2A and H4 which may be important for regulatory functions.

Amino Acid Sequence↗

Identification of trematode cercariae carrying Neorickettsia risticii in freshwater stream snails.

We provide evidence of Neorickettsia (Ehrlichia) risticii Holland, the agent of Potomac horse fever, in trematode larval stages found in aquatic snails and insects collected from a stream in Korea, using nested polymerase chain reaction (PCR) and sequence analysis of the 16S rRNA gene fragment amplified from several cercaria species. It was observed that of 423 (13.1%) of 3,219 snails infected with cercariae, 77 (20.8% of the 371) were infected with N. risticii. Five families of trematode cercariae, Schistosomatidae, Echimostomatidae, Heterophyidae, Microphallidae, and Acanthocopidae were identified morphologically within Semisulcospira libertina, Radix auricularia coreana, and S. gottschei snails. Echinostoma cinetorchis, E. hortense, and Metagonimus sp. were identified based on both the cercarial morphology as well as by phylogenetic analysis of the amplified 18S rRNA gene sequences. Adult aquatic insects were also collected from the same sites and were sorted into five species, Ischnura asiatica in Coenagrionidae and Calopteryx japonica, Sympetrum darwinianum, Symptrum eroticum, and Symptrum parvulum in Calopterygoidae. One thousand and two hundred eighty five metacercariae (classified into groups A through F) were isolated from 310 adult aquatic insects, and the average number of metacercariae per aquatic insect was 4.1. However, there was no amplification of N. risticii from these metacercariae.

Anaplasmataceae Infections↗

Reduced competition and altered feeding behavior among marine snails after a mass extinction.

Extinction may alter competitive interactions among surviving species, affecting their subsequent recovery and evolution, but these processes remain poorly understood. Analysis of predation traces produced by shell-drilling muricid snails on bivalve prey reveals that species interactions were substantially different before and after a Plio-Pleistocene mass extinction in the western Atlantic. Muricids edge- and wall-drilled their prey in the Pliocene, but Pleistocene and Recent snails attacked prey only through the shell wall. Experiments with living animals suggest that intense competition induces muricid snails to attack shell edges. Pliocene predators, therefore, probably competed for resources more intensely than their post-extinction counterparts.

Animals↗

Larval trematodes: double infections in common mud-flat snail.

Larvae of the trematode Zoogonus lasius are involved in most double infections of Nassarius obsoleta. The two most common trematode parasites of this snail do not occur together in double infections. Double infections were found in 14 of 340 infected snails in a total sample of 5025 snails.

Animals↗

Vertical distribution of an estuarine snail altered by a parasite.

Estuarine snails Ilyanassa obsoleta bearing larvae of the trematode Gynaecotyla adunca behave singularly in comparison with conspecifics lacking this parasite. Following high tides, and especially at night, infected snails were found stranded high on beaches and sandbars. Semiterrestrial crustaceans living well up on the shore serve as the next host, and the modified (induced) snail behavior is apparently a parasite adaptation facilitating cercarial transmission to these crustaceans. The altered behavior is unusual because of its apparent enhancement of host-to-host transmission by cercariae rather than predation, the process commonly recognized as being enhanced by parasitic modification of host behavior.

Animals↗

Isolation of salmonellae and other potential pathogens from the freshwater aquarium snail Ampullaria.

The freshwater aquarium snail (Ampullaria spp.) was demonstrated to carry as many as 10(8) viable mesophilic bacteria per g of meat plus shell. Some 16 genera of bacteria were identified, with gram negatives predominating. Enrichment culture techniques enabled the isolation of salmonellae from 24 to 42 lots of 200 g each. The salmonellae comprised eight different serotypes, including Salmonella newport, Salmonella saint-paul, and Salmonella infantis. This association of salmonellae with snails may contribute to cases of human salmonellosis, since other aquarium species have already been shown to contribute to many such cases. The snails were also found to commonly harbor Pseudomonas aeruginosa and, occasionally, Edwardsiella tarda.

Animals↗

Analysis of 16S rRNA and 51-kilodalton antigen gene and transmission in mice of Ehrlichia risticii in virgulate trematodes from Elimia livescens snails in Ohio.

Operculate snails (the family Pleuroceridae: Elimia livescens) were collected between June and October 1998 from a river in central Ohio where repeated cases of Potomac horse fever (PHF) have occurred. Of collected snails, consistently 50 to 80% carried a combination of cercariae and sporocysts of digenetic virgulate trematodes. The trematodes obtained from each snail were pooled and tested for Ehrlichia risticii, the agent of PHF, by nested PCR using primers specific to the 16S rRNA gene. Out of a total of 209 trematode pools, 50 pools were found to be positive by PCR. The DNA sequence of the 16S rRNA gene identified in one trematode pool was identical to that of the type strain of E. risticii, and the sequence of the gene identified in another pool differed from that of the type strain by 1 nucleotide. Comparison of the deduced amino acid sequence of the partial 51-kDa antigen gene from various sources revealed that Maryland, Ohio (except Ohio 081), and Kentucky strains are in a cluster distinct from the sequences obtained from sources in California and Oregon. Ohio 081 was shown previously by antigenic composition analysis to be distinct from other groups. However, all sequences examined were not segregated according to their sources: horse blood or infected trematodes. E. risticii was found to be transmittable from trematodes to mice and was subsequently passaged from infected mice to additional mice, as determined by PCR analysis. Our findings suggest the evolution of E. risticii in the natural reservoir in separate geographic regions and persistent infection of trematode populations with E. risticii during summer and early fall. The study also suggests that the mouse can be used to isolate E. risticii from the infected trematode.

Amino Acid Sequence↗

Pulsed-field gel electrophoresis profiles of aeromonads isolated from healthy and diseased Helix aspersa from French snail farms.

The XbaI digestion patterns of chromosomal DNA of 42 aeromonads isolated from French breeding snails during a new epizootic disease, which rapidly progressed to death during the summer of 1994, were analyzed by pulsed-field gel electrophoresis. Biochemical identification to species level was also performed. Interestingly, we found that 76% of the aeromonads isolated from diseased snails clustered into a unique pulsotype (P1) whatever their geographic origin, and were assessed to belong to Aeromonas hydrophila. Other strains belonged to Aeromonas caviae or remained unspecified. Our results provide retrospective supplementary epidemiological evidence for implication of A. hydrophila strains in the snail summer disease.

Aeromonas↗

Cyclooxygenase-2-dependent regulation of E-cadherin: prostaglandin E(2) induces transcriptional repressors ZEB1 and snail in non-small cell lung cancer.

Elevated tumor cyclooxygenase-2 (COX-2) expression is associated with tumor invasion, metastasis, and poor prognosis in non-small cell lung cancer (NSCLC). Here, we report that COX-2-dependent pathways contribute to the modulation of E-cadherin expression in NSCLC. First, whereas genetically modified COX-2-sense (COX-2-S) NSCLC cells expressed low E-cadherin and showed diminished capacity for cellular aggregation, genetic or pharmacologic inhibition of tumor COX-2 led to increased E-cadherin expression and resulted in augmented homotypic cellular aggregation among NSCLC cells in vitro. An inverse relationship between COX-2 and E-cadherin was shown in situ by double immunohistochemical staining of human lung adenocarcinoma tissue sections. Second, treatment of NSCLC cells with exogenous prostaglandin E(2) (PGE(2)) significantly decreased the expression of E-cadherin, whereas treatment of COX-2-S cells with celecoxib (1 mumol/L) led to increased E-cadherin expression. Third, the transcriptional suppressors of E-cadherin, ZEB1 and Snail, were up-regulated in COX-2-S cells or PGE(2)-treated NSCLC cells but decreased in COX-2-antisense cells. PGE(2) exposure led to enhanced ZEB1 and Snail binding at the chromatin level as determined by chromatin immunoprecipitation assays. Small interfering RNA-mediated knockdown of ZEB1 or Snail interrupted the capacity of PGE(2) to down-regulate E-cadherin. Fourth, an inverse relationship between E-cadherin and ZEB1 and a direct relationship between COX-2 and ZEB1 were shown by immunohistochemical staining of human lung adenocarcinoma tissue sections. These findings indicate that PGE(2), in autocrine or paracrine fashion, modulates transcriptional repressors of E-cadherin and thereby regulates COX-2-dependent E-cadherin expression in NSCLC. Thus, blocking PGE(2) production or activity may contribute to both prevention and treatment of NSCLC.

Cadherins↗