Complex organization of a cryptic satellite DNA in the genome of the marine invertebrate Rapana thomasiana Grosse (Gastropoda).

Isopicnic centrifugation in Cs2SO4-Ag+ gradients at pH 7.0 reveals that the genome of the marine snail Rapana thomasiana Grosse (Gastropoda) contains an AT-rich satellite fraction comprising 5% of the DNA. Restriction enzyme analysis shows that the satellite DNA is composed of a number of related subsets arranged in tandem arrays. They have evolved from the segmental amplification of an 1460 bp long monomer unit with a complex inner organization. Most probably, the present basic repeat originates from an ancestral 400-500 bp long sequence in which some insertions and/or deletions have occurred.

Purification and molecular properties of two cadmium-binding glycoproteins from the hepatopancreas of a whelk, Buccinum tenuissimum.

Two cytoplasmic cadmium-binding glycoproteins from the hepatopancreas of a whelk, Buccinum tenuissimum, were purified to homogeneity by Sephadex G-75 gel filtration and two DEAE-Sephadex A-25 chromatographies in the presence of 2-mercaptoethanol. These two cadmium-binding glycoproteins, termed FIIA and FIIB, each showed a single band after electrophoresis on 10% polyacrylamide gel at pH 8.9. The molecular weight of FIIA was estimated as 8000 and that of FIIB as 13000 by SDS-polyacrylamide gel electrophoresis. Based on molecular weights of 6500 and 11900 for the protein moiety of FIIA and FIIB, respectively, the total number of amino acid residues was 52 in the former and 94 in the latter. Three and two cysteine residues in FIIA and FIIB, respectively, were titratable with p-chloromercuribenzoate. FIIB also contained two more half-cystine residues. The sugar contents of FIIA and FIIB were about 20.5% and 8.7% by weight, respectively, consisting of galactose, mannose, fucose and amino sugar. The purified glycoproteins FIIA and FIIB contained about 0.6% and 1.0% cadmium by weight, respectively, and both showed strong metal-binding capacity, especially for cadmium, copper and mercury. The apparent cadmium dissociation constants for FIIA and FIIB after treatment with 2-mercaptoethanol were 7.3 X 10(-6) and 9.1 X 10(-7) M, respectively. Cadmium contents at saturation were nearly 6 and 8 gatom per mole for FIIA and FIIB, respectively.

Immunocytochemical evidence of vertebrate bioactive peptide-like molecules in the immuno cell types of the freshwater snail Planorbarius corneus (L.) (Gastropoda, Pulmonata).

An immunocytochemical investigation was carried out on round and spreading hemocytes of Planorbarius corneus by using 20 antisera to vertebrate bioactive peptides. The immunotests showed the presence of alpha 1-antichymotrypsin-bombesin-, calcitonin-, CCK-8 (INC)-, CCK-39-, gastrin-, glucagon-, Met-enkephalin-, neurotensin-, oxytocin-, somatostatin-, substance P-, VIP-, and vasopressin-immunoreactive molecules in the spreading hemocytes. The round hemocytes were only positive to anti-bombesin, anticalcitonin, anti-CCK-8 (INC), anti-CCK-39, anti-neurotensin, anti-oxytocin, anti-substance P and anti-vasopressin antibodies. No immunostaining was observed with anti-CCK-8 (Peninsula), anti-insulin, anti-prolactin, anti-thyroglobulin and anti-thyroxin (T4) antibodies. As probably in vertebrates, these bioactive peptides may modulate immuno cell function.

Detection of snail hemocyanin in the whole hemolymph by serologically specific electron microscopy.

This study demonstrates the applicability of the serologically specific electron microscopy (SSEM) technique in the detection of hemocyanin molecules in the whole hemolymph of the snail, Megalobulimulus ovatus. The results are positive and easily reproducible. The SSEM might be useful as a technique for taxonomic studies of snails as well as to study structural aspects of their hemocyanin molecules.

Snailine: a possible diagnostic reagent from a common marine snail, the southern periwinkle, Littorina angulifera.

A substance which strongly precipitates human serum proteins is released into the environment by the southern periwinkle, Littorina angulifera. Two-dimensional crossed immunoelectrophoresis revealed that the snail precipitin reacts with a number of human serum proteins, including 11 not detected by mammalian antiserum. The precipitin is readily collected in saline solution, which then is referred to as snailine, a potentially useful diagnostic reagent. Precipitins released into the environment by marine species may represent a little-known external immunologic system.

Structural studies on glycolipids of shellfish. V. Gala-6 series glycosphingolipids of the marine snail, Chlorostoma argyrostoma turbinatum.

A series of glycosphingolipids and phosphonoglycosphingolipid containing only galactose as the sugar component were isolated from the marine snail, Chlorostoma argyrostoma turbinatum. The structures of these lipids were studied by methylation analysis, hydrogen fluoride degradation, proton magnetic resonance spectroscopy and fast atom bombardment mass spectrometry, and characterized as follows: the glycosphingolipids galactosyl beta(1-1)ceramide, galactosyl beta(1-6)galactosyl beta(1-1)ceramide, galactosyl beta(1-6)galactosyl beta(1-6)galactosyl beta(1-1)ceramide and galactosyl beta(1-6)galactosyl beta(1-6)galactosyl beta(1-6)galactosyl beta(1-1)ceramide, and phosphonoglycosphingolipid N-methylaminoethylphosphonyl galactosyl(1-1)ceramide. The main molecular species of the ceramide moiety were hexadecanoyl-octadecasphingenine and hydroxyhexadecanoyl-octadecasphingadienine in all of these sphingolipids.

Differential hydrogen ion titrations of the histidine residues in Helix pomatia haemocyanin.

The properties of the histidine residues in Helix pomatia haemocyanin have been studied by differential hydrogen ion titrations. In oxy-and deoxyhaemocyanin 31 X 10(-5) histidine residues per g protein are titrated in contrast to 35 X 10(-5) residues in apohaemocyanin. The difference corresponds to a stoichiometry of one histidine residue per copper atom bound. Even in apohaemocyanin about 6 X 10(-5) histidine residues per g protein are not titrated in their normal pH region. In the presence of sufficient calcium to displace the dissociation completely out of the titration region, the titration curve of apohaemocyanin could be linarized according to the model of Linderstrom--Lang. In oxy-and deoxyhaemocyanin, however, a distinct deviation from linearity was found under the same conditions. In the absence of calcium the effect of the dissociation adds up to this deviation. The electrostatic interaction factors were determined for the protein at 0.1 M KC1 and for the dissociation products: halves and tenths at 1.0 M KC1. The electrostatic interaction factor for the wholes and the halves are much smaller than the values calculated from the Linderstrom--Lang equation, using the radius of the equivalent sphere either obtained from electron microscopy or from the partial specific volume. This probably due to solvent penetration. For the tenths at 1.0 M KC1, this effect is small.

Isolation of eburnetoxin, a vasoactive substance from the Conus eburneus venom.

Eburnetoxin, a powerful vasoactive protein has been isolated from the venom of the marine snail Conus eburneus, monitored by the contractile effect to the rabbit aorta. The molecular weight was estimated to be 28,000 by gel permeation chromatography and slab gel electrophoresis. The purified protein was electrophoretically homogeneous. The toxin at concentrations above 3 X 10(-7) g/ml elicited a marked contractile response of aorta, which was inhibited by verapamil (10(-6) M). The minimum lethal dose in the fish Rhodeus ocellatus smithi was 1 microgram/g body weight.

Characterization of aminoalkylphosphonyl cerebrosides in muscle tissue of Turbo cornutus.

From muscle tissues of the marine snail (Turbo cornutus) aminoalkylphosphonyl cerebrosides, which had been shown to be present in visceral parts were isolated. Their structure was determined by degradative methods and by characterization of components by gas chromatography-mass spectrometry. The aminoalkylphosphonyl cerebroside fraction consisted of a major portion of 1-O-[6'-O-(N-methylaminoethylphosphonyl) galactosyl] ceramide and a minor portion of a novel lipid, 1-O-[6'-O-(aminoethylphosphonyl)galactosyl] ceramide. The fatty acids of the fraction were mainly palmitic (53.3%) and 2-hydroxy palmitic acid (14.6%). The long chain bases were mainly dihydroxy C22: 2(36.6%), C18:1 (14.6%) and trihydroxy bases were also found as minor components.

Methylation studies of the polysaccharides resulting from sequential Smith-degradations of the galactan from the snail Strophocheilus oblongus.

When the galactan from the albumen glands of the snail Strophocheilus oblongus was submitted to three Smith-degradation, the degraded polysaccharide, isolated in 6% yield, was much more linear. Methylation analysis showed that the Smith-degraded polysaccharide gave an increased percentage of 2,4,6-tri, decreased percentages of 2,3,4,6-tetra-and 2,4-di, and a large variation in the amount of 2,3,4-tri-O-methyl-D-galactose. The sugars in the polysaccharide which result in the formation of 2,3,4,6-tetra- and 2,3,4-tri-O-methyl-D-galactose are destoryed in subsequent degradation procedures. The above observations suggest that the degradation by periodate oxidation proceeds via non-reducing end-groups and though some internal residues that are exposed as the degradation proceeds. As a result of the overall process, new non-reducing end-groups are formed and new (1 leads to 6)-linked D-galactose residues are then exposed. The isolation of glycosides of low molecular weight supports the suggestion that the molecule, in the main, is sequentially degraded from the external layers.

Potential for employing the distribution of anomalous non-methylene-interrupted dienoic fatty acids in several marine invertebrates as part of food web studies.

A group of homologous, nonmethylene-interrupted dienoic fatty acids (NMID) recently reported in oysters has been found in several other shellfish species and also in other marine phyla. The distribution of most other "normal" fatty acids among several species of shellfish is basically similar but mantle lipids from two other molluscan species, both squid, differ radically. The squid mantle fatty acids do not include NMID, suggesting that in molluscs NMID may accumula-e primarily in filter-feeders or herbovores. The distribution of these anomalous fatty acid components in higher species suggests that they reflect invertebrates in the diet and are biochemically inert.

Natural infection of land-snails by protostrongylids on a pasture grazed by sheep in the Rabat area of Morocco.

The output by sheep of first-stage protostrongylid larvae, and the population density and level of infection of terrestrial molluscs were assessed monthly during a 1-year study. Limax maximus was the most infected mollusc. Juveniles were less infected than adults, for Cernuella virgata and Theba pisana; the reverse was observed for Otala lactea. Banded individuals had a lower intensity of infection than the non-banded ones for C. virgata and T. pisana. Sixty-one percent of protostrongylid larvae in molluscs on the pastures studied were found in C. virgata, and 25% in O. lactea. The infectivity of the pasture for sheep was related to rainfall.

Histamine release from rat mast cells induced by the lectin from the snail, Cepaea nemoralis L. - a counterpart to the triggering action of polycations?

One of the two lectins, occurring in the albumen gland of the snail, Cepaea nemoralis L., is shown to be a histamine releaser from isolated rat mast cells as well as in vivo. The release reaction is induced in the same dosage range as the agglutination of human red blood cells occurs. Both the reactions are inhibitable by the addition of N-acetyl-neuraminic acid. The releasing activity of the lectin seems to be a counterpart to the triggering action of polycations, because its specificity is directed against this sugar.

Agglutination of bacteria by a sialic acid-specific lectin of the snail Cepaea hortensis.

Aqueous extracts from lyophilized albumen glands of the garden snail (Cepaea hortensis) exhibit, after separation from a second lectin, a specificity against sialic acid-containing polymers. In agglutination studies with bacteria, the lectin interacts with all strains of group B streptococci (Streptococcus agalactiae) containing type-specific polysaccharides. In contrast, group B streptococci containing only protein type antigens as well as all strains of other Lancefield streptococcal groups and several other bacterial strains did not react.

Isolation of nematode inhibitor from hemolymph of the snail, Helix aspersa.

Hemolymph plasma of the snail Helix aspersa which inhibits maturation and reproduction of its mantle cavity-inhabiting nematode, Rhabditis maupasi, was separated biochemically for the active proteinaceous component. Isolation of the active inhibitor was performed using ion-exchange chromatography in combination with subsequent gel filtration. The isolated peaks were bio-assayed in vitro on nematode larvae. The fractions harboring inhibitory protein suppressed larval growth and adult reproduction in vitro. The isolated fraction was purified by gel filtration and characterized on the basis of a single band on starch zone electrophoresis and positive reaction only with folin-phenol reagent.

Comparison of carbon and nitrogen content of infected and uninfected snails, Succinea ovalis, and the trematode Leucochloridium variae.

In June, 6.7% of adult Succinea ovalis collected near Urbana, Ohio, were infected with the trematode, Leucochloridium variae. The effects of parasitism were assessed as total organic carbon (equivalent to calorific values) and as total nitrogen. The parasite represents 23.8% of total (parasite + snail tissue) dry tissue weight, 21.4% of total carbon and 19.8% of total nitrogen of infected snails. The higher C : N ratio for parasite tissue indicates a higher proportion of nonproteinaceous compounds (e.g., fats and/or carbohydrates) as compared to host tissue. There is less snail tissue in parasited S. ovalis. The C : N ratios for parasitized and nonparasitized snail tissue suggest identical percentage compositions of proteins, fats, and carbohydrates.

A relationship between avian carcasses and living invertebrates in the epizootiology of avian botulism.

A survey of the sources of Clostridium botulinum type C toxin possibly utilized as food by aquatic birds in an epizootic area of avian botulism in northern Utah showed that living aquatic and terrestrial invertebrates normally found in close association with dead, decomposing birds commonly carried the toxin. Of 461 samples associated with 21 species of avian carcasses, 198 were toxin-positive. Invertebrate species not normally scavengers of vertebrate tissues were less commonly and less highly toxic, particularly when captured 30 cm or more from a carcass; six of 237 samples of such aquatic invertebrates low-level toxin. Of the species tested, blow fly larvae (Calliphoridae) were the most consistently and highly toxic, although others, particularly adult and larval stages of several species of beetles (Coleoptera), contained toxin at levels probably significant in the epizootiology of the disease. An estimated 0.05 to 0.25 g of the most toxic fly larvae or 15 g of the most toxic beetles tested carried a mediam lethal dose for an adult mallard duck. Examination of stomach contents of aquatic birds dead of botulism showed that some had consumed invertebrates.