Search Results (30)

The progressive commercial deployment of multi-walled carbon nanotubes (MWCNTs) raises concerns about their terrestrial ecotoxicity. We exposed adult Cornu aspersum (150 snails; five replicates of three animals per time-point) to 50 mg L⁻¹ MWCNT-dosed Lactuca sativa for 30 days and quantified five mechanistic biomarkers alongside survival. Hemocyte spread-cell area increased by 48% (from 243 ± 22 µm² at day 0 to 360 ± 18 µm² at day 14, p < 0.001). Lysosomal membrane stability (neutral red retention) fell twofold within 72 h and to 10 min by day 30 (controls ≈ 60 min), indicating early, persistent destabilization. Micronucleus frequency rose above the ecogenotoxic threshold of 5‰ after 7 days, peaking at 8.3 ± 0.7‰ on day 14 (p < 0.01). Hepatopancreas metallothionein concentrations doubled by day 3 (2.1 ± 0.3 vs. 1.0 ± 0.2 µg g⁻¹ ww in controls) and remained >150% of control throughout exposure, consistent with metal impurity mobilization. Acetylcholinesterase activity in cephalic tissue declined by 50% after 7 days and by 73% after 30 days, revealing sustained neurotoxicity. Despite these pronounced sub-individual disturbances, cumulative mortality reached only 19% at day 30, suggesting substantial, but finite, physiological compensation. Collectively, the data demonstrate that a 50 mg L⁻¹ dietary load of MWCNTs elicits rapid cytotoxic, genotoxic, and neurotoxic responses in C. aspersum that precede overt lethality, underscoring the utility of this gastropod and the chosen biomarker suite for monitoring nanotube contamination in agro-ecosystems and food-grade snail farming. Full article

The green synthesis of metal nanoparticles, mediated by extracts from various biological sources, leads to the formation of nanoparticles with unique characteristics and potential biomedical applications. In this study, a fraction of the mucus from the snail Cornu aspersum with MW > 20 kDa was used as a bioreducing and biostabilizing agent to obtain silver nanoparticles (AgNPs) in different pH media (pH 1.5, 3.5, and 7.0). The AgNPs were characterized using UV-visible spectroscopy, SEM, TEM, XRD, and FTIR. The synthesis at pH 1.5 and pH 3.5 in the presence of two reducing agents (i.e., the C. aspersum mucus fraction with MW > 20 kDa and ascorbic acid [AsA]) resulted in the formation of well-formed spherical nanoparticles (NPs) with larger sizes (20–80 nm) than the NPs obtained at pH 7.0 (20–60 nm) in the presence of only one reducing agent. Furthermore, the biosynthesized AgNPs significantly inhibited the growth of medically significant pathogens such as Gram-positive (Bacillus subtilis NBIMCC2353, Bacillus spizizenii ATCC 6633, Staphylococcus aureus ATCC 6538, and Listeria innocua NBIMCC8755) and Gram-negative (Escherichia coli ATCC8739, Salmonella enteitidis NBIMCC8691, Salmonella typhimurium ATCC 14028, Stenotrophomonas maltophilia ATCC 17666) bacteria compared to output mucus. Full article

(1) Background: Hibernation in Lutrasil-covered enclosures led to elevated mortality rates for Cornu aspersum on Romanian farms. This two-year study evaluated the feasibility of adapting indoor hibernation technology to the Italian outdoor snail farming (IOSF) system as a solution for overwintering mature C. aspersum snails. (2) Methods: Body weight, survival rates, and mortality factors during key hibernation phases (purging and overwintering) were monitored in three commercial snail farms using different hibernation scenarios. Labor efficiency was evaluated, comparing the use of micro shelters vs. hand-picking for snail collection. We analyzed post-purging/post-hibernation weight loss and mortality in hibernation spaces with and without proper thermal insulation. Mortality causes were also investigated. (3) Results: Using micro shelters significantly reduced labor time for snail collection. Weight loss during purging, but not during overwintering, were similar between groups. Post-hibernation survival ranged from 69% to 79% for comparable hibernation durations. Significantly lower survival was associated with significantly higher weight loss and using hibernation spaces with minimal thermal insulation. Predators exerted a very limited effect on post-hibernation survival, with most death appearing to be related to environmental causes. (4) Integrating indoor hibernation into the IOSF system is a promising strategy for the successful rearing of C. aspersum in colder climates. Full article

Background/Objectives: The treatment of diabetic foot ulcers (DFU) is a challenging medical problem of extreme clinical and social importance, as a consequence of the emerging antibiotic resistance and decreased quality of life of diabetic patients due to impaired wound healing. One of the current trends in world science is the search for biologically active substances derived from living organisms. Biologically active peptides from snail mucus attract considerable scientific interest because of their pleiotropic pharmacological properties. The aim of our study was to evaluate the activity of a combination between a snail mucus protein fraction (MW > 20 kDa) obtained from the garden snail Cornu aspersum and the clinically applied antibacterial chemotherapeutic ciprofloxacin on pathogenic bacterial strains isolated from DFU. Results: The test bacterial strains were characterized as multidrug resistant. The combination between ciprofloxacin and the snail mucus fraction of interest led to additive or synergistic effects depending on the test strain. The mucus fraction exerted a well-pronounced wound-healing effect and no cytotoxicity on normal human fibroblasts and keratinocytes. Methods: The snail mucus was obtained by a patented technology (BG Utility model 2097/2015) and its electrophoretic profile was presented by SDS-PAGE. The bacterial strains were identified and tested for antimicrobial susceptibility (BD Phoenix M50 and Kirby–Bauer assay). The in vitro cytotoxicity of the mucus was evaluated by ISO 10995-5. The antimicrobial activity and combination effects were tested through ISO 20776/1 and the Checkerboard assay. Conclusions: The obtained results are promising and open new horizons for the development of novel combination treatment schemas for healing of infected DFU. Full article

Fungal infections are a significant global public health challenge because of their widespread occurrence, morbidity, and profound social and economic consequences. Antifungal resistance is also an increasing concern, posing a substantial risk to public health. There is a growing interest in searching for new antifungal drugs isolated from natural sources. This study aimed to evaluate the antifungal activity of novel mollusk fractions against fungal strains resistant to nystatin and amphotericin B. In addition, the role of oxidative stress in the mechanism of damage was determined. The mucus from the garden snail Cornu aspersum (MCa/1-20) and the hemolymph fraction from the marine snail Rapana venosa (HLRv/3-100) were obtained and characterized via 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometric -analyses. The results demonstrate that the spores and biomass of both mollusk fractions have a significant fungicidal effect against Penicillium griseofulvum, and Aspergillus niger. Compared to the control group, the release of intracellular proteins and reducing sugars was significantly increased in the treated groups. The data showed increased levels of oxidative stress biomarkers (lipid peroxidation and oxidatively damaged proteins) and a downregulated antioxidant enzyme defense, corresponding to increased antifungal activity. To our knowledge, this is the first study evaluating oxidative stress as a factor in mollusk fractions’ antifungal activity. Full article

Many biologically active compounds have been identified in the mucus of the garden snail Cornu aspersum, which are effective in the treatment of several diseases such as cancer, ulcers, wounds, etc. The incorporation of these compounds into the green synthesis of copper nanoparticles (CuONPs-Muc) was demonstrated in our previous study. Based on the synergistic effect of two reducing agents—C. aspersum snail mucus and ascorbic acid (AsA)—on CuSO₄.5H₂O, which also act as stabilizers of the resulting compound, a new method for the “green” synthesis of CuONPs-Muc is presented. Using two reducing agents has several advantages, such as forming spherical nanoparticles with a diameter of about 150 nm and reducing the formation time of CuONPs-Muc to 3 h. Analyses by ultraviolet–visible spectroscopy (UV-Vis) and Fourier transform infrared spectroscopy (FT-IR) show the formation of CuONPs-Muc, composed of a mixture of copper and copper oxide. This was confirmed by scanning electron microscopy combined with energy-dispersive spectroscopy (SEM/EDS) and X-ray diffraction (XRD). Another important advantage of CuONPs obtained by the new method with two reducing agents is the stronger inhibitory effect on the bacterial growth of some Gram-positive and Gram-negative bacterial strains, compared to CuONPs-Muc prepared with only one reducing agent, i.e., a fraction of mucus with an MW > 20 kDa. Full article

Alzheimer’s disease (AD) is the most widespread neurodegenerative disorder. Recently, it was found that mucus extract from Cornu aspersum has beneficial effects on memory and cognitive processes in a rat scopolamine model of AD. The present study elucidated the mechanisms of action of standardized mucus snail extract (SE) enriched with a fraction above 20 kDa on Alzheimer-type dementia in rats. Using proteomic analysis on two-dimensional polyacrylamide gel electrophoresis (2D–PAGE) on rat cortex extracts, we compared protein expression in both groups: the first group was treated intraperitoneally with scopolamine (Sco, 2 mg/kg, 11 days) and the second (Sco + SE) group was treated intraperitoneally with Sco (Sco, 2 mg/kg) and protected by SE (0.5 mL/100 g bw) applied daily orally for 11 days. Brain cortex was separated and the expressions of various proteins related to memory and cognitive functions were identified. We found that the expression of Ubiquitin carboxyl-terminal hydrolase isozyme L1, Calbindin, Vacuolar ATP synthase catalytic subunit A, Tropomyosin beta chain, 14-3-3 zeta/delta, Kinesin-1 heavy chain, and Stathmin-4 significantly differs in SE-protected rats as compared to dement animals treated only by Sco, and these brain proteins might be potential therapeutic targets for Alzheimer’s-type dementia treatment. Full article

Heliciculture farms are susceptible to significant biotic issues that can impact snail breeding, among them, the entomofauna predation of snails. Predatory insects can cause damage to snail shells during predation, and sometimes, the specific type of damage may be characteristic of certain insect families or species. Under laboratory conditions, we analysed the predatory activity of the species Silpha tristis Illiger, 1798 (Coleoptera: Silphidae), Ocypus olens (Müller, 1764) (Coleoptera: Staphylinidae), Carabus (Macrothorax) morbillosus constantinus Kraatz, 1899 (Coleoptera: Carabidae), and Lampyris sardiniae Geisthardt, 1987 (Coleoptera: Lampyridae) against the gastropod Cornu aspersum (Müller, 1774) reared on snail farms located in the Sardinian region. The adult and larval stages of each species were tested, except for L. sardiniae, as only the larval stage preys upon snails. This study showed that among all of the species considered, only two insects were able to damage the shell of C. aspersum: C. morbillosus constantinus and O.olens. This may prove valuable in C. aspersum breeding for the recognition of specific lesions, even in the absence of the predator. Full article

The discovery and investigation of new natural compounds with antimicrobial activity are new potential strategies to reduce the spread of antimicrobial resistance. The presented study reveals, for the first time, the promising antibacterial potential of two fractions from Cornu aspersum mucus with an MW < 20 kDa and an MW > 20 kDa against five bacterial pathogens—Bacillus cereus 1085, Propionibacterium acnes 1897, Salmonella enterica 8691, Enterococcus faecalis 3915, and Enterococcus faecium 8754. Using de novo sequencing, 16 novel peptides with potential antibacterial activity were identified in a fraction with an MW < 20 kDa. Some bioactive compounds in a mucus fraction with an MW > 20 kDa were determined via a proteomic analysis on 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) and bioinformatics. High homology with proteins and glycoproteins was found, with potential antibacterial activity in mucus proteins named aspernin, hemocyanins, H-lectins, and L-amino acid oxidase-like protein, as well as mucins (mucin-5AC, mucin-5B, mucin-2, and mucin-17). We hypothesize that the synergy between the bioactive components determined in the composition of the fraction > 20 kDa are responsible for the high antibacterial activity against the tested pathogens in concentrations between 32 and 128 µg/mL, which is comparable to vancomycin, but without cytotoxic effects on model eukaryotic cells of Saccharomyces cerevisiae. Additionally, a positive effect, by reducing the levels of intracellular oxidative damage and increasing antioxidant capacity, on S. cerevisiae cells was found for both mucus extract fractions of C. aspersum. These findings may serve as a basis for further studies to develop a new antibacterial agent preventing the development of antibiotic resistance. Full article

Several biologically active compounds involved in the green synthesis of silver and gold nanoparticles have been isolated from snail mucus and characterized. This paper presents a successful method for the application of snail mucus from Cornu aspersum as a bioreducing agent of copper sulfate and as a biostabilizer of the copper oxide nanoparticles (CuONPs-Muc) obtained. The synthesis at room temperature and neutral pH yielded nanoparticles with a spherical shape and an average diameter of 150 nm. The structure and properties of CuONPs-Muc were characterized using various methods and techniques, such as ultraviolet–visible spectroscopy (UV–vis), high-performance liquid chromatography (HPLC), one-dimensional polyacrylamide gel electrophoresis (1D-PAGE), up-conversion infrared spectroscopy Fourier transform (FTIR), scanning electron microscopy combined with energy dispersive spectroscopy (SEM/EDS), Raman spectroscopy and imaging, thermogravimetric analysis (TG-DSC), etc. Mucus proteins with molecular weights of 30.691 kDa and 26.549 kDa were identified, which are involved in the biogenic production of CuONPs-Muc. The macromolecular shell of proteins formed around the copper ions contributes to a higher efficiency of the synthesized CuONPs-Muc in inhibiting the bacterial growth of several Gram-positive (Bacillus subtilis NBIMCC2353, Bacillus spizizenii ATCC 6633, Staphylococcus aureus ATCC 6538, Listeria innocua NBIMCC8755) and Gram-negative (Escherichia coli ATCC8739, Salmonella enteitidis NBIMCC8691, Salmonella typhimurium ATCC 14028, Stenotrophomonas maltophilia ATCC 17666) bacteria compared to baseline mucus. The bioorganic synthesis of snail mucus presented here provides CuONPs-Muc with a highly pronounced antimicrobial effect. These results will expand knowledge in the field of natural nanomaterials and their role in emerging dosage forms. Full article

A large variety of microorganisms ingested with food constitute animals’ intestinal microflora, enhancing and maintaining the homeostasis of the host. Rations enriched with probiotics are a method recommended to prevent undesirable conditions. To date, research has been limited to farmed animals and reared fish, creating a knowledge gap concerning the effect of probiotics on the growth rate, physiological responses, and energy metabolism of invertebrates such as the land snail Cornu aspersum. Herein, juvenile snails (26.23 ± 0.5 mm shell diameter and 8.23 ± 1.02 g body weight) were fed L. plantarum probiotic-enriched rations in two different proportions (1.25 mg and 2.5 mg), and their growth rate was monitored for three months. Additionally, the RNA/DNA and Bax/Bcl-2 ratios, HSP gene expression and protein levels, and ND2 expression, were measured in the hepatopancreas, digestive tract, and mantle. Although the snails’ growth rate was not affected, the RNA/DNA ratio presented an increase in various tissues, indicating an intense physiological response. Also, probiotic administration demonstrated low levels of the Bax/Bcl-2 ratio. HSP levels were higher in the presence of probiotics, probably signaling an attempt by the animal to face potentially stressful situations. Finally, ND2 expression levels in the hepatopancreas indicate intense metabolic and antioxidant activity. Full article

The pathway and the lifestyle of known enterococcus species are too complicated. The aim of the present study is to trace the path of pathogenicity of enterococci isolated from seven habitats (Cornu aspersum intestine; Bulgarian yoghurt; goat and cow feta cheese—mature and young, respectively; Arabian street food—doner kebab; cow milk; and human breast milk) by comparing their pathogenic potential. In total, 72 enterococcal strains were isolated and identified by MALDI-TOF, sequencing, and PCR. Hemolytic and gelatinase activity were biochemically determined. PCR was carried out for detection of virulence factors (cylB, esp, gls24, nucl, psaA, agg, gelE, and ace) and antibiotic resistance (erm, ermB, blaZ, vanA, aphA, mefA, gyrA, cat_pIP501, and aac6′-aph2″). Phenotypic antibiotic resistance was assigned according to EUCAST. Eleven representatives of the genus Enterococcus were identified: E. mundtii, E. casseliflavus, E. gilvus, E. pseudoavium, E. pallens, E. malodoratus, E. devriesei, E. gallinarum, E. durans, E. faecium, and E. faecalis. Twenty-two strains expressed α-hemolysis. Thirteen strains had the cylB gene. Only two strains expressed α-hemolysis and possessed the cylB gene simultaneously. Positive amplification for gelE was found in 35% of the isolates, but phenotypic gelatinase activity was observed only in three strains. All isolates showed varying antibiotic resistance. Only E. faecalis BM15 showed multiple resistance (AMP-HLSR-RP). Correlation between genotypic and phenotypic macrolide resistance was revealed for two E. faecalis strains. Full article

Edible snails are an attractive protein source due to their high growth rate, cost-efficiency, and nutritional value. Calcium is crucial for snail growth, reproduction, and shell formation, while magnesium plays a role in enzyme function and muscle tone. This study aimed to optimise calcium and magnesium levels in Cornu aspersum diets to optimise the production and technological characteristics of the derived animal products. Snails were fed specific diets in controlled conditions with varying calcium and magnesium levels (44.3, 66.1, 88.7, 103.5 Ca g/kg feed and 3.3, 5.6, 7.2 Mg g/kg feed) for four months. Their growth, shell characteristics, and meat composition were evaluated. As calcium in the feed increased, carcass and shell weights were higher. Also, the crushing force of the shells was higher with increasing amount of calcium in the feed. In the group with 10.35% calcium and 0.72% magnesium, snail growth significantly slowed down after three months, with lower mortality. It is suggested that a shortened fattening cycle by 3–4 weeks compared to the magnesium-free diet is possible. However, based on meat, shell, mortality, and feed intake analysis, a 0.56% magnesium concentration in the feed seems to give better results, as magnesium content at 0.72% might be toxic to snails. Further investigation is to confirm the possibility of neutralising the negative effects of magnesium in the diet through increasing calcium and phosphorus intake. Full article

Cornu aspersum, the land snail, is recognized as a suitable intermediate host of Aelurostrongylus abstrusus; however, there is little information both on larval development as well as on the intermediate host’s immune system reaction to the parasite. The aim of the study was to assess the histological reaction of C. aspersum’s immune system against A. abstrusus. Sixty-five snails were provided by a snail farm. Five of them were digested to assess the absence of natural parasitic infections. The remaining sixty were divided into five groups. Three groups of snails were infected with A. abstrusus using by-contact or injection methods; one group was injected only with saline solution and one group was left untreated as the control. The snails of group A were sacrificed and digested on study days 2, 10, and 18; snails of the other groups were collected and examined for histopathological analysis on study days 2, 10, and 18. On study day 2, in the infected snails, several free L1s were observed along with the absence of immune system reactions. On day 10, the L2s elicited an intense reaction in the internal layer of the muscular foot. On day 18, all L3s partially encapsulated by the snail’s immune system were observed in the outermost part of the muscular foot, which is near and among the goblet cells. This last finding suggests that L3s could be shed with the snail’s mucus and spread in the environment, representing an alternative route of transmission for this feline lungworm. Full article