BRACHYURAN CRABS (DECAPODA, BRACHYURA) ASSOCIATED WITH THE GREEN SPONGE AMPHIMEDON VIRIDIS (DEMOSPONGIAE) FROM ITAGUA BEACH, SOUTH-EASTERN COAST of BRAZIL

This study provides information on the community structure of brachyuran crabs associated with the green sponge, Amphimedon viridis Duchassaing & Michelotti, 1864, from a beach on the south-eastern coast of Brazil. Monthly collections from May 2002 through August 2003 were carried out on the rocky bottoms off Itagua Beach (23 degrees 27'07 '' S 45 degrees 02'49 '' W). Crabs on the green sponge banks were hand-caught during snorkelling sessions, and 638 crabs belonging to 31 species, 19 genera, and 10 families were obtained. Biological parameters, including sponge volume, number of individuals, density, richness, diversity, evenness, and dominance changed considerably over time. Species recruitment and foraging habit are some of the factors that drive the richness, dominance, and diversity patterns of this community. The green sponge banks are important sites for recruitment, refuge, and feeding of many species of brachyurans. Therefore, they are also important for the conservation of this group on the south-eastern coast of Brazil.

Structure determination of a tetrahydro-beta-carboline of arthropod origin: A novel alkaloid-toxin subclass from the web of spider Nephila clavipes

The orb-web spiders are polyphagous animals in which the web plays a very important role in the capture of preys; oily droplets usually cover the capture-web of the spider Nephila clavipes and seem to be of great importance for prey capture. The knowledge of the chemical composition of these droplets is necessary to understand the function of this adhesive material in web mechanics and prey capture. A novel subclass of spider toxins, tetrahydro-beta-carboline, was identified among the weaponry of compounds present inside of oily droplets. This type of alkaloid is not common among the natural compounds of spider toxins. Apparently, when the prey arthropods get caught by the spider web, their bodies are covered with many adhesive oily droplets, which disrupt delivering the tetrahydro-beta-carboline to the direct contact with the prey integument. Toxicity assays demonstrated a potent lethal effect of the alkaloid toxin to the spider preys; topical applications of the teirahydro-beta-carboline at first caused clear signs of neurotoxicity, followed by the death of preys. The structure of the major component, a tetrahydro-beta-carboline, among the alkaloid toxins was elucidated by means of UV spectrophotometry, ESI mass spectrometry, H-1-NMR spectroscopy, and high-resolution mass spectrometry. The structure of the natural toxin was determined as 1-(2-guanidinoethyl)-1,2,3,4-tetrahydro-6-hydroxymethyl)-beta-carboline; the investigation of the pharmacological properties and neurotoxic actions of this compound may be used in the future as reference for the development of new drugs to be applied at level of pest control in agriculture.

Multiple bradykinin-related peptlides from the capture web of the spider Nephila clavipes (Araneae, Tetragnatidae)

Three bradykinin-related peptides (nephilakinins-I to -III) and bradykinin itself were isolated from the aqueous washing extract of the capture web of the spider Nephila clavipes by gel permeation chromatography on a Sephacryl S-100 column, followed by chromatography in a Hi-Trap Sephadex-G25 Superfine column. The novel peptides occur-red in low concentrations and were sequenced through ESI-MS/MS analysis: nephilakinin-I (G-P-N-P-G-F-S-P-F-R-NH2), nephilakinin-Il (E-A-P-P-G-F-S-P-F-R-NH2) and nephilakinin-III (P-S-P-P-G-F-S-P-F-R-NH2)- Synthetic peptides replicated the novel bradykinin-related peptides, which were submitted to biological characterizations. Nephilakinins were shown to cause constriction on isolated rat ileum preparations and relaxation on rat duodenum muscle preparations at amounts higher than bradykinin; apparently these peptides constitute B-2-type agonists of ileal and duodenal smooth muscles. All peptides including the bradykinin were moderately lethal to honeybees. These bradykinin peptides may be related to the predation of insects by the webs of N. clauipes. (c) 2005 Elsevier B.V. All rights reserved.

Structural characterization of a new acylpolyaminetoxin from the venom of Brazilian garden spider Nephilengys cruentata

Mario Sergio Palma, Yasuhiro Itagaki, Tsuyoshi Fujita, Hideo Naoki and Terumi Nakajima. Structural characterization of a new acylpolpaminetoxin from the venom of Brazilian garden spider Nephilengys: cruentata. Toxicon 36, 455-493, 1998.-The use of mass spectrometry, in which high-energy CID and charge remote fragmentation both of protonated and sodium-attached molecular ions was applied, afforded the structural elucidation of a new acylgolyaminetoxin with M-W= 801 da from the venom of the Brazilian garden spider Nephilengys cruentata. In spite of having the same M-W of the NPTX-2, previously described in the venom of the Joro spider Nephila clavata, neither toxins are isomers. In order to differentiate them by using the most usual nomenclature, the new toxin was named NPTX-801C and the NPTX-2 was renamed to NPTX-801E. Both toxins have as common structure the 4-hydroxyindole-3-acetyl-asparaginyl-cadaveryl moiety in their molecules and their structure may be represented in a simplified way: NPTX-801E is HO-indole-Asn-Cad-Pta-Orn-Arg and NPTX-801C is HO-indole-Asn-Cad-Gly-Put-Pta-Pta. (C) 1998 Elsevier B.V. Ltd. All rights reserved.

Structure determination of hydroxytrypargine: A new tetrahydro-beta-carboline toxin from the venom of the spider Parawixia bistriata

A new, highly active tetrahydro-p-carboline toxin from the spider Parawixia bistriata, the most-common species of social spider occurring in Brazil, was isolated. The new toxin was identified as 1,2,3,4-tetrahydro-6-hydroxy-beta-carboline (= N-[3-(2,3,4,9-tetrahydro-6-hydroxy-1H-pyrido[3,4-b]indol-1-yl)propyl]guanidine; 3). This type of alkaloid, not common among spider toxins, was found to be the most-potent constituent of the spider's chemical weaponry to kill prey. When P bistriata catch arthropods in their web, they apparently attack their prey in groups of many individuals injecting their venoms. In vivo toxicity assays with 3 demonstrated a potent lethal effect to honeybees, giving rise to clear neurotoxic effects (paralysis) before death. The compound's toxicity (LD50 value) was determined to be ca. 8 ng/g of honeybee. The investigation of the pharmacological properties and neurotoxic actions of 3 may be used in the future for the development of new drugs to be applied for pest control in agriculture.

The venomous secrets of the web droplets from the viscid spiral of the orb-weaver spider Nephila clavipes (Araneae, Tetragnatidae)

The capture web of N. clavipes presents viscous droplets, which play important roles in web mechanics and prey capture. By using scanning and transmission electron microscopy, it was demonstrated that the web droplets are constituted of different chemical environments, provided by the existence both of an aqueous and a lipid layer, which, in turn, present a suspension of tenths of vesicles containing polypeptides and/or tipids. GC/EI-MS Analysis of the contents of these vesicles led to the identification of some saturated fatty acids, such as decanoic acid, undecanoic acid, dodecanoic acid, tetradecanoic acid, octadecanoic acid, and icosanoic acid, while other components were unsaturated fatty acids, such as (Z)-tetradec-9-enoic acid, (Z)-octadec-9-enoic acid, and (Z)-icosa-11-enoic acid; and polyunsaturated fatty acids like (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid, and (11Z,14Z)-icosa-11,14-dienoic acid. Toxic proteins such as calcium-activated proteinase and metalloproteinase jararhagin-like precursor were also identified by using a proteomic approach, indicating the possible involvement of these enzymes in the pre-digestion of spiders' preys web-captured. Apparently, the mixture of fatty acids are relatively toxic to insects by topical application (LD50 64.3 +/- 7.6 ng mg(-1) honeybee), while the proteins alone present no topical effect; however, when injected into the prey-insects, these proteins presented a moderate toxicity (LD50 40.3 +/- 4.8 ng mg(-1) honeybee); the mixture of fatty acids and proteins is very toxic to the preys captured by the web droplets of the viscid spiral of Nephila clavipes when topically applied on them (LD50 14.3 +/- 1.8ng mg(-1) honeybee).

Nigriventrine: A low molecular mass neuroactive compound from the venom of the spider Phoneutria nigriventer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Monoamine oxidase inhibitory activities of indolylalkaloid toxins from the venom of the colonial spider Parawixia bistriata: Functional characterization of PwTX-I

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influence of Prey Size on the Capture of Social Wasps (Hymenoptera: Vespidae) by the Orb-Web Spider Nephilengys cruentata

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Brown Recluse Spider Venom: Proteomic Analysis and Proposal of a Putative Mechanism of Action

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Extreme degree of chromosome number variability in species of the spider genus Scytodes (Araneae, Haplogynae, Scytodidae)

Genus Scytodes includes most species of the spider family Scytodidae. Until now, 187 species of the genus have been described. In spite of this great diversity, only three Scytodes species were karyotyped so far. The present paper provides for the first time karyotype analysis of two synanthropic species, Scytodes fusca and Scytodes itapevi. Furthermore, new data on karyotype of Scytodes globula are also provided using conventional and differential cytogenetical procedures. The diploid number in the genus Scytodes varied considerably, namely from 2n = 13 to 2n = 31. The diploid number found in S. globula (2n male = 13) is the lowest in haplogyne spiders with monocentric chromosomes. Except S. globula, this number has been found only in one haplogyne spider with monocentric chromosomes, namely Ochyrocera sp. (Ochyroceratidae). on the contrary, the diploid number of S. fusca (2n male = 31) is one of the highest diploid numbers recorded in haplogyne spiders. The degree of intrageneric variation found in the genus Scytodes is the highest recorded in araneomorph spiders with monocentric chromosomes so far. Some karyotype characteristics (diploid number, chromosome morphology, total chromosome length, and distribution of constitutive heterochromatin) allowed us to postulate a close relationship between S. globula and S. itapevi. According to the karyotype data, S. fusca is not closely related to these two species. This conclusion corroborates a recent taxonomic work that grouped S. globula, S. itapevi, and other four Scytodes species in the 'globula group'.

Effects of Spider Venom Toxin PWTX-I (6-Hydroxytrypargine) on the Central Nervous System of Rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The chelation of metal ions by the acylpolyamine toxins from the web-spider Nephilengys cruentata: effects in the intoxication/detoxification of preys

Orb-web-spiders present a series of different strategies for prey capture, involving the use of different types of silk for web building, the use of adhesive traps in the webs, the secretion of toxic compounds to the spider's preys in the adhesive coating of the capture web and the biosynthesis of a wide range of structurally related acylpolyamine toxins in their venoms. The polyamine toxins usually block neuromuscular junctions and/or the central nervous system (CNS) of Arthropods, targeting specially the ionotropic glutamate receptors; this way these toxins are used are as chemical weapons to kill / paralyze the spider's prey. Polyamine toxins contain many azamethylene groups involved with the chelation of metal ions, which in turn can interact with the glutamate receptors, affecting the toxicity of these toxins. It was demonstrated that the chelation of Ni+2, Fe+2, Pb+2, Ca+2 and Mg+2 ions by the desalted crude venom of Nephilengys cruentata and by the synthetic toxin JSTX-3, did not cause any significant change in the toxicity of the acylpolyamine toxins to the model-prey insect (honeybees). However, it was also reported that the chelation of Zn+2 ions by the acylpolyamines potentiated the lethal / paralytic action of these toxins to the honeybees, while the chelation of Cu+2 ions caused the inverse effect. Atomic absorption spectrometry and Plasma-ICP analysis both of N.cruentata venom and honeybee's hemolymph revealed that the spider's venom concentrates Zn+2 ions, while the honeybee's hemolymph concentrates Cu+2 ions. These results are suggesting that the natural accumulation of Zn+2 ions in N. cruentata venom favors the prey catching and/or its maintenance in the web, while the natural accumulation of Cu+2 ions in prey's hemolymph minimizes the efficiency of the acylpolyamine toxins as killing/paralyzing tool.

Predation on Leptodactylus marmoratus (Anura: Leptodactylidae) by the spider Ctenus medius (Araneae: Ctenidae) in the Atlantic Forest, southeast Brazil.

We report herein a case of predation by the ctenid spider Ctenus medius on a leptodactylid frog Leptodactylus marmoratus, observed in a remnant of the Atlantic Forest, municipality of Sao Vicente, São Paulo state, southeast Brazil. This is the first record of predation by C. medius upon L. marmoratus. Nevertheless, due to the high abundance of both groups, we suggest that the interaction between spiders and amphibians could be very common on the floor of the Atlantic Forest.

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.