Helix Pomatia Research Articles

SHELLS OF MOLLUSCS FROM MOTORONYN HILLFORT

The shells of 3 land molluscs’ species are found in the historical layer of VI—V century B.C. from the Motronyn hillfort of the Scythian age (Chyhyryn district of Cherkasy region, Ukraine): Helix pomatia, Austrotachea vindobonensis and Fruticicola fruticum. All these species are known to still occur in this area. Since it was disputable whether the H. pomatia was recently introduced in the Dnipro area, an evidence of its presence for at least 2400 years in this region is notable. Nature of damage to the shell of H. pomatia indicates that it was probably used for food by humans.

Comprehensive kinetic and substrate specificity analysis of an arylsulfatase from Helix pomatia using mass spectrometry.

Sulfatases hydrolyze sulfated metabolites to their corresponding alcohols and are present in all domains of life. These enzymes have found major application in metabolic investigation of drugs, doping control analysis and recently in metabolomics. Interest in sulfatases has increased due to a link between metabolic processes involving sulfated metabolites and pathophysiological conditions in humans. Herein, we present the first comprehensive substrate specificity and kinetic analysis of the most commonly used arylsulfatase extracted from the snail Helix pomatia. In the past, this enzyme has been used in the form of a crude mixture of enzymes, however, recently we have purified this sulfatase for a new application in metabolomics-driven discovery of sulfated metabolites. To evaluate the substrate specificity of this promiscuous sulfatase, we have synthesized a series of new sulfated metabolites of diverse structure and employed a mass spectrometric assay for kinetic substrate hydrolysis evaluation. Our analysis of the purified enzyme revealed that the sulfatase has a strong preference for metabolites with a bi- or tricyclic aromatic scaffold and to a lesser extent for monocyclic aromatic phenols. This metabolite library and mass spectrometric method can be applied for the characterization of other sulfatases from humans and gut microbiota to investigate their involvement in disease development.

SCIENTIFIC AND TECHNOLOGICAL SUBSTANTIATION OF PRODUCTION OF MIXED FODDERS FOR SNAILS

The article deals with the issues of modern snail breeding in Ukraine, problems and prospects for their cultivation. The main areas of use of snails are classified. With the proper cultivation of snails, the business is profitable, because meat and slime of snails are expensive, have a lot of useful qualities. The characteristic of the farms for the production of snails in Ukraine and the products of snails - snails, meat, caviar, mucus. The volumes and methods of growing snails in the world and Ukraine have been analyzed, the tendency of annual increase in livestock and consumption of snails, especially in Europe, is formed, which creates favorable market conditions that guarantee the constant demand for products of activity of snail farms in Ukraine. It was established that in 2018 volumes reached 750 thousand tons. The data of the main countries of producers, importers and consumers of snails in the world are presented. The advantages and disadvantages of the production of snails are determined in comparison with the cultivation of farm animals. The analysis and characterization of feed supplementation for Ahatinka snails and grain-calcium blend used in Ukraine is provided. Objects of research - Snails Akhaten and snails grape Helix pomatia, fodder for snails. Scientifically substantiated and calculated recipe of full-feed compound for snails using the software complex KormOptimaExpert, which includes grain, protein mineral components and biologically active substances. The step-by-step scheme of technology of production of mixed fodders for snails is indicated, indicating technological regimes of production, according to which experimental samples of mixed fodder are made. The physical, chemical and microbiological indices of full-fodder mixed fodders for the snails were studied. According to the organoleptic, physical properties and chemical composition, ready-mixed fodder for snails meets the requirements of the State Standard of Ukraine. The results of the zootechnical evaluation of the developed mixed fodder, according to which it was established that feed for snails during feeding gives a positive zootechnical effect, since the size of the shell at the end of the experiment relates to the commodity, the increase in the mass of snails increased by 13.6%, and the length by 30% at the coefficient the usefulness of a feed is equal to 1,2 ... 1,5.

The complete mitogenome of the Roman snail Helix pomatia Linnaeus 1758 (Stylommatophora: Helicidae)

This study reports the complete mitochondrial sequence of the Roman snail, one of the largest terrestrial snails of Europe. Two specimens of Helix pomatia were sequenced, which had a sequence length of 14,070 and 14,072 base pairs. The mitogenome has the common metazoan makeup (13 protein-coding genes, two ribosomal RNAs, and 21 transfer RNAs) and the ‘typical’ Helicid gene order (Cytochrome Oxidase subunit III and tRNA-T translocated compared to other Helicoidea). All protein coding genes could be annotated with proper start and stop codons (no completion of stop codons by polyadenylation). The specimens have a sequence similarity of 99.0% (146 differences) and the average base composition is 29.7% A, 15.0% C, 17.9% G, and 37.4% T. Phylogenetic anlyses with available Helicid mitogenomes show Helicinae as monophyly and H. pomatia as the sister-group of (Cepaea nemoralis, Theba pisana, and Cornu aspersum).

Seasonally dependent morphological variations of circulating hemocytes in Helix pomatia

Seasonally dependent morphological variations of circulating hemocytes in Helix pomatia

Вміст жирних кислот у м'ясі різних видів равликів

Вміст жирних кислот у м'ясі різних видів равликів

Fatty Acid Profile in Fat Obtained from Edible Part of Land Snails Harvested in Poland.

IntroductionThe objective was to determine the content of fatty acids in edible snail fat by snail species, collection site, and processing stage.Material and MethodsThe research material comprised 180 edible fat samples from the three genera of edible snails collected in Poland: free-living Helix pomatia (HP) and two cultivated Cornu subspecies: C. aspersa maxima (CAM) and C. aspersum aspersum (CAA). All snails came from the Greater Poland and Lower Silesian Provinces: HP from their natural habitat and CAM and CAA from heliciculture farms. The studies focused on the raw meat, cooked meat, and frozen meat processing stages. Fatty acid (FA) profiles were determined by the gas chromatography method.ResultsHelix pomatia fat showed a higher saturated fatty acid (SFA) content, whereas the fat of Cornu genus snails had a higher unsaturated fatty acid (UFA) component, i.e. monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA). Thermal processing of snail meat increased all the determined SFA and decreased all the PUFA values, and increased the content of C18:1, C20:1, and C22:1 acids in the MUFA group. The material collection site had limited impact on FA content as differences were noted only in levels of C18:1, C18:2 n6, and C20:5. The differences pertained only to the fat of farmed snails of the Cornu genus.ConclusionDue to the high content of UFA and a favourable ratio of n6:n3 acids and PUFA:SFA, snail fat can be regarded as nutritionally valuable.

POSSIBILITIES OF LECTIN HELIX POMATIA AGGLUTININ (HPA) USE IN RABBITS INSEMINATION

The aim of this work was to analyse the effect of incubation HPA (lectin Helix pomatia agglutinin) with ID (insemination dose) on reproduction parameters (conception ratio, number of live and dead born of pups and birth weight of pups) of rabbits. The experiments were carried out under conditions of 2 rabbit breeds (production breed P and experimental breed E). Fresh ID, in sperm concentrations 25 mil. / 0.5 ml ID, were divided into control ID (without HPA) and experimental ID (with HPA). HPA was added to the experimental ID in concentrations 1µg HPA / 1x106 spermatozoa (30 - 60 minutes of incubation before AI – artificial insemination). Conception ratios obtained from 2 different breeds (P, E) showed minor improvement ( + 2.67 % P; + 7.36 % E) between the monitored groups of females, however without statistically significant differences (P χ2 = 0.03; E χ2 = 0.11). Number of live born pups in litter obtained from breeds P and E showed minor improvement (+ 0.37 pcs P (p>0.05); + 0.95 pcs E (p≤0.05)) compared to the control groups. The weight of live pups among the monitored groups was comparable. It can be concluded from the present study that incubation of ID with lectin HPA had been positively affected by the observed reproductive parameters. The statistically significant difference was found only in the number of live-born pups in the litter. The price of lectin is too high for breeding in common breeding practice.

Influence of heat treatment on amino acid composition of snails meat

Amino acids are organic compounds that are the structural component of the protein. That is, protein is the main building material of the tissues of the body. It is needed for muscle mass growth and is indispensable in fat burning – all about amino acids, of which protein is formed. The main source of amino acids are food that is rich in protein. However, based on the content of one or another amino acid proteins contained in food, can be divided into complete and inferior. Complete proteins contain all the essential amino acids. These products include, primarily, products of animal origin: meat, poultry, fish, eggs, dairy products. In order to plant sources of full protein is soybean. Inferior proteins – in their composition there is at least one irreplaceable acid. Accordingly, in their “quality” defective proteins can be very different. The source of defective proteins is mainly products of planting: legumes, cereals, nuts and seeds. The purpose of our work was to determine the content of amino acids in raw and cooked meat of the food species Helix pomatia, Helix aspersa maxima and Helix aspersa muller and to make an analysis on the effect of heat treatment on the amino acid composition of snail meat. This article presents the results of the content of amino acids in meat of food snails using electrophoresis “Capel-105/105M”. Three types of snails have identified the presence and amount of amino acids. In general, the meat of snails contains amino acids – arginine, histidine, serine, alanine, glycine, tyrosine and proline, and irreplaceable – lysine, phenylalanine, leucine + isoleucine, methionine, valine, threonine.

Identification of Nanomolar Lectin Ligands by a Glycodendrimer Microarray.

Carbohydrate–protein interactions play key roles in a wide variety of biological processes. These interactions are usually weak, with dissociation constants in the low millimolar to high micromolar range. Nature uses multivalency to reach high avidities via the glycoside cluster effect. Capitalizing on this effect, numerous synthetic multivalent glycoconjugates have been described and used as ligands for carbohydrate-binding proteins. However, valency is only one of the several parameters governing the binding mechanisms that are different for every biological receptor, making it almost impossible to predict. In this context, ligand optimization requires the screening of a large number of structures with different valencies, rigidities/flexibilities, and architectures. In this article, we describe a screening platform based on a glycodendrimer array and its use to determine the key parameters for high-affinity ligands of lectin. Several glycoclusters and glycodendrimers displaying varying numbers of α-N-acetylgalactosamine residues were covalently attached on glass slides, and their bindings were studied with the fluorophore-functionalized Helix pomatia agglutinin (HPA) used as a lectin model. This technique requires minimal quantities of glycoconjugate compared to those for other techniques and affords useful information on the binding strength. Building of the glycodendrimer array and quantification of the interactions with HPA are described.

Hemocyanin genes as indicators of habitat shifts in Panpulmonata?

Hemocyanin is the primary respiratory protein for the majority of the Mollusca and therefore directly interfaces with the physiological requirements of each species and the environments to which they are adapted. Hemocyanin is therefore likely to have been evolutionarily imprinted by significant habitat shifts. In the gastropod clade Panpulmonata (>30,000 species) major realm transitions have occurred multiple times independently and may have contributed to the diversification of this group. Yet, little is known about the adaptive changes linked to these habitat shifts. In order to gain deeper insight into the evolution of panpulmonate hemocyanins and to infer possible impacts associated with those scenarios, we have assembled and analysed hemocyanin isoforms from 4 panpulmonate species: (i) Helix pomatia, (ii) Cantareus aspersus (both Helicidae, Stylommatophora), (iii) Arion vulgaris (Arionidae, Stylommatophora) and (iv) Lymnaea stagnalis (Lymnaeidae, Hygrophila). Additionally, we describe a new hemocyanin isoform within the genome of the euopisthobranch Aplysia californica. Using these newly acquired hemocyanin data, we performed a phylogenetic analysis that reveals independent duplication events of hemocyanin within lineages that correlate with significant habitat shifts.

Systematic Evaluation and Validation of Benzodiazepines Confirmation Assay Using LC-MS-MS.

Benzodiazepines is a large drug group used extensively to treat sleep disorders and anxiety; these drugs are frequently associated with various crimes such as murder and drug-facilitated sexual assault. With growing use and misuse of these compounds, confirmatory assays are increasingly required in clinical laboratories. In this study, 4 β-glucuronidase enzymes were systematically evaluated for their hydrolysis efficiencies. Additionally, the matrix effects and extraction recoveries of three protein precipitation plates were systematically evaluated. The recombinant IMCSzyme β-glucuronidase enzyme showed higher hydrolysis efficiency than did β-glucuronidase from abalone, Patella vulgata and Helix pomatia. Lower ion suppression and more favorable overall recovery were observed in the Supelco protein precipitation plate than in the two other plates. Analytes were separated on a superficially porous particle column (ultra biphenyl) within 4.5 min and characterized through electrospray ionization-tandem mass spectrometry in the positive ion multiple-reaction monitoring mode. The accuracy, carryover, extraction recovery, detection limit, matrix effect, quantification limit and precision of the method were validated. Sixty opioid-positive urine samples were analyzed; a high incidence of benzodiazepines was detected in these samples. The proposed technique is robust and suitable for efficiently monitoring the numerous benzodiazepines that occur in urine samples.

Seasonal distributions of heavy metal concentrations in different snail (Helix pomatia) tissues from an urban environment in Serbia.

The aim of this study was to evaluate the differences between concentrations of heavy metals (cadmium, copper, zinc, iron, and manganese) in terrestrial gastropods, Helix pomatia, collected in four different environment locations in Pančevo city, Serbia. Metal concentrations in the foot and in the digestive gland were measured. Heavy metals cadmium, copper, zinc, iron, and manganese were measured in triplicate by atomic absorption spectrophotometry. Also, this study investigated correlations between the trace element content in the soft tissues and different seasons (summer and fall). The results showed that heavy metal concentrations of the measured elements in both snail tissues were significantly higher in polluted sites when compared to the background levels in the city. In digestive gland tissues, the concentration of heavy metals was higher compared with that in foot tissues. The bioaccumulation (heavy metal concentrations in soft tissues) of heavy metals (Cd, Cu, Zn, Fe, and Mn) in snails, Helix pomatia, has been assessed and related to seasons and sites. Heavy metal accumulation in the soft tissues of snails could provide a powerful monitoring tool for the assessment of environmental heavy metal pollution.

LECTIN-HISTOCHEMICAL REGULARITIES OF DIFFERENTIATION OF THE HUMAN PAROTID GLAND EPITHELIAL GERMS

Glycopolymer compounds make up structural and functional basis of cells and tissues of a living organism. The necessity of anatomical-lectinohistochemical examination of the parotid gland in early prenatal period of ontogenesis is substantiated, as the evidences concerning its topography are fragmentary and not systematized, and certain aspects of its ontogenesis are disputable. 35 human embryos and pre-fetuses aged from 21 days to 12 weeks of intrauterine development were examined. Glycopolymers were found by means of treatment of serial sections with lectins conjugated with horseradish peroxidase. The dynamics of prenatal morphogenesis of the embryos and pre-fetuses of the 4th-12th week of embryogenesis expression of glycopolymers – lectinreceptors on the surface of cells, in the cytoplasm and on the basal membrane of the parotid gland epithelial germs and the human oral cavity with its derivatives by re-distribution of glycopolymers are similar, which can be the evidence of ectodermal source of the parotid gland epithelial germ. Immersion of the epithelial cells of the buccal-alveolar pockets into the lower adjacent mesenchyme with the formation of primary parotid gland germs in embryos with 11,0-12,5 mm of PCL and their transformation into the epithelial taeniae is associated with accumulation of sialic glycopolymers (N-acetyl neuraminic acid), N-acetyl-D-glucosamine – specific to Wheat germ agglutinin (WGA) and Sambucus nigra agglutinin (SNA); N-acetyl-2-desoxy-2-amino-D-glucopyranose, screened by sialic acid of ß-D-galactose and α-L-fucose – specific to Helix pomatia (edible snail) agglutinin (HPA), Rhizoctonia solani agglutinin (RCA) and Laburnum anagyroides (Golden Rain shrub) agglutinin (LABA) respectively. These glycopolymers are present during the first 12 weeks both on the cytolemma of the cells of the parotid gland epithelial germ and in their cytoplasm. The results of lectin-histochemical examination of the early prenatal ontogenesis of the parotid gland can form the basis for the work of laboratories dealing with screening of morphological material in order to assess the degree of maturation and prognosis of fetus viability and diagnostics of deviations from normal development.

Assay for galactose-deficient IgA1 enables mechanistic studies with primary cells from IgA nephropathy patients.

Aims:IgA nephropathy, the most common primary glomerulonephritis worldwide, is characterized by glomerular deposition of galactose-deficient IgA1 and elevated serum levels of this IgA1 glycoform. Current ELISA methods lack sensitivity to assess galactose deficiency using small amounts of IgA1, which limits studies in primary cells due to modest IgA1 production in isolated peripheral-blood lymphocytes.Methods:Lectin from Helix pomatia was conjugated to biotin or acridinium ester and used in ELISA to detect galactose deficiency of IgA1 using small amounts of IgA1.Results:Lectin conjugated to acridinium had an approximately log-fold increased sensitivity compared with biotin-labeled lectin.Conclusions:The new method of using lectin from Helix pomatia conjugated to acridinium increased assay sensitivity, allowing future mechanistic studies with cultured primary cells.

Concentration levels of cadmium and lead in the raw and processed meat of Helix pomatia snails.

This work reports on cadmium and lead contaminations in the edible snail Helix pomatia harvested in Poland. One hundred and 24 samples of Helix pomatia meat collected from seven provinces (voivodeships) of Poland were analyzed for their trace metal levels by graphite furnace atomic absorption spectrometry (GFAAS). The research was conducted in 2 stages. The 1st stage analyzed snail meat prior to any further technological treatment (raw meat). In the 2nd stage, the trace element levels were measured in meat subjected to technological treatment (processed meat). The trace element contents in raw meat samples ranged from 0.06 mg kg-1 to 0.22 mg kg-1 for Cd and from 0.06 mg kg-1 to 0.18 mg kg-1 for Pb. The analyses revealed an increase in the cad- mium content from 0.12 mg kg-1 to 0.18 mg kg-1 in thermally treated snail meat and no changes in lead concentration during the two-stage heat treatment. Regulation (EC) 1881/2006 does not specify the Cd and Pb residue limits in meat of terrestrial edible snails. The limits are set for in- vertebrate aquatic organisms meat (i.e. shellfish, mollusc, cephalopod) and range from 0.5 mg/kg to 1.5 mg/kg of tissue fresh weight for Pb and from 0.5 mg kg -1 to 1 mg kg-1 for Cd (EU Commis- sion 2006). The results demonstrate that the land snail Helix pomatia has a tendency to bioaccu- mulate trace elements, and the cooking process is likely to affect (increase) the Cd content in the snail meat.

Binding of aberrant glycoproteins recognizable by Helix pomatia agglutinin in adrenal cancers.

BackgroundAberrant glycosylation is a hallmark of cancer cells and plays an important role in oncogenesis and cancer progression including metastasis. This study aimed to assess alteration in cellular glycosylation, detected by lectin Helix pomatia agglutinin (HPA) binding, in adrenal cancers and to determine whether such altered glycosylation has prognostic significance.MethodsHPA binding lectin histochemistry was performed on archival paraffin wax‐embedded specimens of adrenocortical cancers excised from patients attending two tertiary referral centres. Benign tumours were used as controls. Demographic, histological and survival data were collected and compared between patients with HPA‐positive and HPA‐negative tumours.ResultsThirty‐two patients were treated for adrenal cancer between 2000 and 2016; their median age was 49 (range 23–79) years. Fifteen patients had functioning tumours (14 adrenal Cushing's tumours and 1 Conn's tumour). Mean(s.d.) tumour size was 127·71(49·70) mm. None of 10 control tumours expressed HPA‐binding glycoproteins. Invasion was associated with HPA‐binding glycoproteins (P = 0·018). Local recurrence or metastatic disease did not significantly differ between HPA‐positive and HPA‐negative adrenocortical cancers. Overall survival was significantly longer in patients with HPA‐negative tumours (median survival not reached versus 22 months in patients with HPA‐positive tumours; P = 0·002).ConclusionAltered cellular glycosylation detected by lectin HPA is associated with poor survival in patients with adrenocortical cancer.

Content of inorganic elements in snails meat

Currently, there are several classifications of inorganic elements, although each has its own advantages and disadvantages. The most studied group is macroelements, because they have a significant spread in the body and serve as a function of structural elements. The trace elements perform their functions, being part of the biologically active substances, which makes it difficult to study them, since the same element, depending on the form, can have a toxic effect or be vital. The macroelements include: K, Ca, Mg, Na, S, P, Cl. The trace elements are distributed as follows: Al, B, Br, V, I, Li, Si, As, Rb, Se, F, Sr and heavy metals, namely Zn, Cr, Ge, Fe, Cd, Co, Cu, Mn, Mo, Sn, Ni, Pb. Macroelements are contained in our body in a significant amount (more than 0.01% of the body weight, in other words, their content in the body of an adult is measured in grams and even in kilograms). The trace elements are involved in all processes of life and are the catalysts of biochemical reactions. Their daily intake is less than 200 mg, and they are contained in the body in small doses (less than 0.001% of body weight). The article presents the results of our own research on the content of inorganic elements in the meat of snails of various species Helix pomatia, Helix aspersa maxima and Helix aspersa muller. It is established that fresh meat of Helix aspersa maxima snails contains the least amount of heavy metals and the highest amount of selenium. Of the 12 inorganic elements that we studied in meat of snails from trace elements, Bromus is most often kept by Helix aspersa muller snails and is 1.91 ± 0.23 mg, Selenium and Manganese from Helix aspersa maxima snails are 0.32 ± 0.01mg and 29.29 ± 0.27 respectively. In the case of heavy metals in the meat of Helix aspersa maxima, there was no evidence of flatulence at all, and the most commonly found zinc in this specimen was 27.64 ± 0.22 mg in accordance. The calcium is the most enriched meat of Snails Helix pomatia and is 12571.25 ± 304.51 mg. As for the vital calcium macroelement, most of all it enriched the meat of the grape snail Helix pomatia. Based on the foregoing, it can be concluded that the fresh meat of Helix aspersa maxima snails contains the least amount of heavy metals and can be successfully used in the relevant industries, and especially in the food industry.

Application of β-glucuronidase-immobilised silica gel formulation to microfluidic platform for biotransformation of β-glucuronides.

To improve the efficiency of reactions of β-glucuronidase (GUS)-assisted glucuronic acid (GluA) removal within a microfluidic system. β-glucuronidase from Helix pomatia was immobilised and characterised in silica-based sol-gel monoliths. Efficiency of the GUS-doped silica monoliths was tested for hydrolysis of p-Nitrophenyl-β-D-glucuronide (pNP-GluA) in both ml-scaled medium via batch reactions and microfluidic environment via continuous-flow reactions. In the microfluidic platform, within a duration of 150min of continuous operation (flow rate: 1 µL/min), the obtained highest pNP yield was almost 50% higher than that of the corresponding batchwise reaction. However, increased flow rates (3, 5, and 10 µL/min) resulted in lower conversion yields compared to 1 µL/min. The microfluidic platform demonstrated continuous hydrolytic activity for 7days with considerable reaction yields while using a small amount of the enzyme. These results revealed that usage of the microreactors has considerable potential to efficiently obtain bioactive GluA-free aglycons from various plant-derived β-glucuronides for pharmaceutical applications.

Estimation of the Genetic Structure of Helix pomatia L. Populations (Mollusca, Pulmonata) by Capillary Electrophoresis of ISSR DNA Fragments

Using the capillary electrophoresis of ISSR DNA fragments, the genetic structure of the adventitious populations of Helix pomatia L. (Mollusca, Pulmonata) inhabiting the territory of Eastern Europe was studied. A significant decrease in genetic heterogeneity (He = 0.095 ± 0.002) and population differentiation level (Fst = 0.074) was revealed in comparison with similar data obtained using the ISSR method with electrophoretic detection in agarose gel and allozymes in polyacrylamide gel. Similar to other methods, an effect of spatial isolation (RM =–0.476) was found on the basis of regression analysis between genetic and geographical distances. The effective population size, calculated with the help of M. Slatkin’s model, turned out to be the largest of the previously calculated values (Ne = 41.8), which indicates the high viability of the studied groups of Roman snails in newly developed territories.