E.L.A. Malpezzi

Characterization of peptides in sea anemone venom collected by a novel procedure

Peptide neurotoxins were isolated from the venom obtained by electrical stimulation of the sea anemone Bunodosoma caissarum. This technique allows almost pure venom to be collected, and the animals to survive. Three neurotoxins (assayed on crustacean nerves) were isolated by gel filtration and reversed-phase high performance liquid chromatography. Hemolysins were also detected in the venom. The amino acid sequence of a major neurotoxin BcIII was determined. BcIII has 48 amino acid residues with six half-cystine residues. This sequence has homology with the type 1 long sea anemone neurotoxins. Two minor toxins (BcI and II) have similar amino acid composition and amino-terminal sequences to BcIII.

Chemical and pharmacological characterization of halitoxin from Amphimedon viridis (Porifera) from the southeastern Brazilian coast

The halitoxin complex from the marine sponge Amphimedon viridis, collected in the São Sebastião channel (southeastern Brazilian coast), was isolated by gel-filtration chromatography on Sephadex LH-20. Spectroscopic data (1H, 13C and 2D-NMR) of halitoxin from A. viridis indicated that it has the same two alkylpyridine monomers composition of the previously isolated halitoxin from Amphimedon compressa (as Haliclona rubens). Nevertheless, analysis by High Performance Gel Permeation Chromatography indicated that the halitoxin complex of A. viridis has a lower molecular weight (500, 2000 and 5000 Da; the major component corresponding to the fraction of 2000 Da MW) when compared to the previously isolated halitoxin complex from A. compressa. Some pharmacological properties of the halitoxin complex from A. viridis were evaluated in terms of lethality, antimitosis, hemolysis and neurotoxicity. The possible chemotaxonomic value of alkyl pyridine alkaloids is discussed.

Hemolytic activity in extracts of the diatom Nitzschia

The few reports about diatom toxins are related to central nervous system toxicity, induced by domoic acid. In the present work Nitzschia sp. (Bacillariophyceae) was studied. The cells were cultured in f/2 medium, under 4000 lux and 14/10 hr light/dark cycle. After massive growth (5 x 10(6) cells/ml) the diatom cells were filtered, and an extract was prepared and partitioned in two fractions (polar and apolar). After cell harvesting by filtration, the diatom cells were shaken in artificial sea water to extract the water-soluble extracellular matrix (mucilage). An extract was prepared with the washed cells (free of mucilage), and polar and apolar fractions were obtained. Hemolytic assays were performed using 4.0 and 0.5% erythrocyte suspensions. Both the diatom polar and apolar fractions showed hemolytic activity. The membrane phospholipid sphingomyelin was tested as an acceptor for the hemolysins in the polar and apolar fractions. The mucilage did not exhibit hemolytic activity.

Occurrence of toxins, other than paralysing type, in the skin of tetraodontiformes fish

Puffer fish (Tetraodontidae and Diodontidae) possess paralysing toxins (tetrodotoxin and analogues) that are secreted upon stimulation. In a previous work it was demonstrated that mucous secretion from the puffer fish Sphoeroides spengleri, when mixed in sea water passing through the orobranchial cavity of groupers, induced cardiorespiratory alterations. In the present study, skin secretions from Ciclichthys spinosus, S. spengleri and Diodon hystrix were tested on crustacean nerves, sea urchin eggs and mouse erythrocyte suspensions to verify neurotoxic and cytotoxic activities. Ciclichthys spinosus and D. hystrix secretions induced transient depolarizations with 0.16 mg and blocked crustacean nerve conduction after prolonged exposure. Both secretions had cytotoxic effects; when applied to sea urchin eggs they caused cleavage inhibition and anomalies in a dose-dependent manner (ED50 +/- S. E. M. = 2.59 +/- 0.08 mg/ml for C. spinosus and 1.23 +/- 0.07 mg/ml for D. hystrix); moreover, hemolysis occurred with an ED50 = 0.76 mg/ml of 0.5% mouse erythrocyte suspensions to C. spinosus and 0.59 mg/ml to D. hystrix. These secretions were not lethal in acute toxicity tests, even at 335 mg/ml. The neurotoxic components were thermolabile while the hemolytic activity was resistant to boiling. Tests with the secretion from S. spengleri did not show cytotoxic effects but promptly blocked action potentials of crustacean nerves and were lethal for mice in acute toxicity rests. When applied to groupers, the C. spinosus secretion caused cardiorespiratory alterations. These results suggest the presence of neurotoxins (other than tetrodotoxin) and cytotoxins in skin of diodontid puffer fish.

A case of envenoming by Portuguese man-of-war from the Brazilian coast

This is an individual case report on envenoming caused by the cnidarian Portuguese man-of-war. The reported local reactions and the clinical symptoms are similar to those already known for Portuguese man-of-war envenoming. As far as we know it is the first clinical case documented for the Brazilian shores.

This histamine-like effects of Phallusia nigra extract: evidences for direct activity at H1 receptors

The methanol extract (mol. wt lower than 3,000 Da) of the sea squirt Phallusia nigra has stimulatory activity on guinea-pig ileum preparations. This effect was inhibited by cyproheptadine and mepyramine, but not be atropine. Mepyramine antagonized competitively the extract activity with a pA2 of 10.09 +/- 1.12, suggesting a direct activity on H1 histamine receptors. The extract was also assayed on guinea-pig right atria, however, only a mild increase in spontaneous contractions was observed compared to histamine, showing that the extract was a rather poor activator of cardiac H2 receptors. Histamine was not detected upon TLC analysis of the extract by comparison with an authentic standard.