HelixComplex snail mucus exhibits pro-survival, proliferative and pro-migration effects on mammalian fibroblasts.

Snail mucus is a mixture of active substances commonly thought to have healthy properties for the treatment of skin disorders. Although snail mucus is an ingredient of several cosmetic and para-pharmaceutic products, a comprehensive characterization of chemical composition and biological effects is still missing. Crude purified extracts from Helix aspersa muller mucus (HelixComplex) were prepared and, after chemical characterization, tested on in vitro experimental models. Differently from what expected, HelixComplex was characterized by the presence of small amounts of glycolic acid and allantoin. By using different in vitro assays on fibroblast cultures, we found that HelixComplex lacked of cytotoxicity, protected cells from apoptosis (p < 0.05) and, importantly, was able to significantly induce cell proliferation and migration through direct and indirect mechanisms. These effects were associated to morphological changes, cytoskeleton re-organization and release of cytokines. In conclusion, our findings suggest that snail mucus biological effects are attributable to cell proliferation and migration, and pave the way for further investigating snail mucus potential as therapeutic agent.

Distinct Bacterial Microbiomes in Sexual and Asexual Potamopyrgus antipodarum, a New Zealand Freshwater Snail.

Different reproductive strategies and the transition to asexuality can be associated with microbial symbionts. Whether such a link exists within mollusks has never been evaluated. We took the first steps towards addressing this possibility by performing pyrosequencing of bacterial 16S rRNA genes associated with Potamopyrgus antipodarum, a New Zealand freshwater snail. A diverse set of 60 tissue collections from P. antipodarum that were genetically and geographically distinct and either obligately sexual or asexual were included, which allowed us to evaluate whether reproductive mode was associated with a particular bacterial community. 2624 unique operational taxonomic units (OTU, 97% DNA similarity) were detected, which were distributed across ~30 phyla. While alpha diversity metrics varied little among individual samples, significant differences in bacterial community composition and structure were detected between sexual and asexual snails, as well as among snails from different lakes and genetic backgrounds. The mean dissimilarity of the bacterial communities between the sexual and asexual P. antipodarum was 90%, largely driven by the presence of Rickettsiales in sexual snails and Rhodobacter in asexual snails. Our study suggests that there might be a link between reproductive mode and the bacterial microbiome of P. antipodarum, though a causal connection requires additional study.

Microbiological quality of raw and processed wild and cultured edible snails.

BACKGROUND: An increasing interest in snail farming in Greece and other European countries has been observed. Despite the fact that edible snails have been involved with problems of Salmonella spp. contamination, there are to our knowledge only limited studies regarding microbiological safety and hygiene of such products. Enumeration of microbial populations and presence/absence of Salmonella spp. in snail meat and intestines of wild Cornu aspersum, Helix lucorum and cultured Cornu aspersum snails from indoor/outdoor type farms was conducted. Furthermore, snail-processing steps were simulated in the laboratory and the population reduction in snail meat was determined. RESULTS: Microbial populations were higher in intestines than snail meat in almost all cases. Escherichia coli/coliforms and Enterococcus spp. populations were lower in the intestines and snail meat of cultured C. aspersum. Salmonella spp. were detected in the intestines and snail meat of wild snails only. The high levels of bacterial populations were considerably reduced after the appropriate processing. CONCLUSION: The lower populations of E. coli/coliforms, Enterococcus spp. and especially the absence of Salmonella spp. in cultured snails show that the controlled conditions decrease the possibility of pathogen presence and contribute to food safety and public health.

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.

[Conjugal plasmid transfer in Bacillus subtilis in soil microcosms]. / Kon"iugativnyi perenos plazmid u Bacillus subtilis v usloviiakh pochvennykh mikrokosmov.

Conjugal transfer of the small plasmid pUB110 between Bacillus subtilis strains was studied under conditions of microcosms with sterile and nonsterile soil. Plasmid transfer proved to be possible after soil inoculation with vegetative partner cells or with their spores. Plasmid transfer occurred at temperatures of 30 degrees C and 22-23 degrees C.

The effect of antibiotic treatment on the supercooling ability of the land snail Helix aspersa (Gastropoda: Pulmonata).

The land snail Helix aspersa is a partially freezing tolerant species whose supercooling ability is limited to ca. -3 to -5 degrees C. One hundred adult snails were subjected to the following two experimental conditions: (i) a starved group, provided with water; (ii) an antibiotic-treated group that was provided with a solution containing a mixture of two antibiotics. The antibiotic group exhibited a T(c) significantly lower than the starved group (-3.94 +/- 1.32 degrees C, n = 40 and -3.07 +/- 0.99, n = 30, t test, p < 0.005). This study showed that bacteria of the gut are likely to elevate animal supercooling points. It is also the first report in which a possible ice-nucleating activity of the gut microflora in a land snail has been suggested by the action of antibiotics on the T(c).

Characterization of yellow-pigmented and motile enterococci isolated from intestines of the garden snail Helix aspersa.

AIMS: Enterococci associated with garden snails (Helix aspersa) were studied in order to obtain reliable species identification and characterization. METHODS AND RESULTS: Twelve yellow-pigmented and motile enterococci, isolated from the intestines of garden snails, were phenotypically close to Enterococcus casseliflavus, but they showed certain unusual biochemical characteristics. tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all strains studied into two groups, in full agreement with biochemical test results. 16S rDNA sequencing, DNA base composition analysis and DNA-DNA hybridization results showed unambiguously that the enterococci studied belonged to the species Ent. casseliflavus. The representative strains of described ecovars were deposited in the Czech Collection of Microorganisms (CCM) as Ent. casseliflavus CCM 4868, 4869, 4870 and 4871. CONCLUSIONS: Enterococcus casseliflavus associated with garden snails can be subdivided into groups. SIGNIFICANCE AND IMPACT OF THE STUDY: Enterococcus casseliflavus differs from other enterococcal species in that it is typically associated with plants, soil, water and invertebrate animals. The different groups that can be found in these widely occurring bacteria are possibly source-specific ecovars, as exemplified by the Ent. casseliflavus inhabiting the intestines of snails.

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.

Molluscan immunobiology: isolation of an Aeromonas formicans which escapes the internal defense system of Helix pomatia.

Bacterial infections are surprisingly rare in wild populations of gastropod molluscs. A strain of Aeromonas formicans, which has now been isolated, kills Helix pomatia, a land snail, when injected at doses greater than 10(6) viable cells g-1 snail weight. Koch's postulates were fulfilled with this organism. Initial clearance of Aeromonas from the circulation was followed within hours by a bacteremia; since neither culture supernatants, killed cells nor cell extracts killed Helix, there is evidently no specific lethal toxin; death appeared to be due to a general septicemia.

Thermal susceptibility of Salmonella in the Moroccan food snail, Helix aspersa.

Thirty samples of 10-15 Helix aspersa food snails were examined for Salmonella by a surface rinsing method and by analysis of whole snails rinsed with 70% ethanol. Thirteen samples were positive by the rinsing method and 6 were positive by whole snail analysis, with this difference being significant (P less than 0.01). Although Salmonella contamination in H. aspersa appeared to be predominantly surface, the pathogen was also found within the snail meat. The ability of surface and subsurface Salmonella organisms to survive in cooked snails was determined in 90 samples. Thermocouple readings indicated that an internal temperature of at least 200 degrees F (93 degrees C) was reached within the snail meat during cooking by a typical recipe. This temperature was sufficient to kill both surface and subsurface Salmonella in 29 samples positive for the pathogen prior to heating. A variety of serotypes representing several somatic groups was isolated.

The moroccan food snail, Helix aspersa, as a source of Salmonella.

A total of 270 samples, nine lots of 30 samples each, of imported Moroccan food snails was examined for the presence of Salmonella. Eighty-four samples (an overall incidence of 31.11%) and all nine lots contained Salmonella. No significant difference (P greater than 0.25) in the number of positive samples was observed by using either selenite cystine both or tetrathionate broth when the samples had been pre-enriched in lactose broth. When used as direct selective enrichments with samples not pre-enriched in lactose broth, tetrathionate broth was significantly (P less than 0.05) more productive than selenite cystine broth. The overall detection of Salmonella-positive samples by direct enrichment was significantly greater (P less than 0.001) than by pre-enrichment. A variety of uncommon serotypes occurrence and incidence, and the concomitant human health potential, of Salmonella in one species of live, imported food snails.