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.

To be or not to be . . . a flower? A test of possible cues influencing hunting site selection in subadult females of the crab spider Epicadus heterogaster (Guerin 1812) (Araneae: Thomisidae)

Females of some Thomisidae species are known to use visual and olfactory stimuli to select high quality hunting sites. However, because studies about foraging behavior in this family are concentrated on a few species, the comprehension of the process related to hunting behavior evolution in crab spiders may be biased. In this study we investigated the hunting site selection of a previously unstudied crab spider, Epicadus heterogaster. We performed three experiments to evaluate the hypothesis that subadult females are able to use visual and olfactory stimuli to select hunting sites. In the first experiment, females did not preferentially select flower paper models that matched their body coloration. However, after choosing a model that had the same body color as the spider, they remained on it for longer periods than on models with different colors. In the second experiment, females did not discriminate between flower paper models, natural flower models and crumpled paper models. Females did also not discriminate among different olfactory stimuli in the third experiment. It is possible that subadult females of E. heterogaster need to establish and experience a given hunting site before evaluating its quality. However, it remains to be investigated if they use UV cues to select a foraging area before experiencing it.

Using visual cues of microhabitat traits to find home: the case study of a bromeliad-living jumping spider (Salticidae)

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

Bromeliad architectural complexity and vertical distribution predict spider abundance and richness

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

Fine-scale microhabitat selection in a bromeliad-dwelling jumping spider (Salticidae)

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

Spatial variation in the strength of mutualism between a jumping spider and a terrestrial bromeliad: Evidence from the stable isotope N-15

Psecas chapoda, a neotropical jumping spider strictly associated with the terrestrial bromeliad Bromelia balansae in cerrados and semi-deciduous forests in South America, effectively contributes to plant nutrition and growth. In this study, our goal was to investigate if spider density caused spatial variations in the strength of this spider-plant mutualism. We found a positive significant relationship between spider density and delta N-15 values for bromeliad leaves in different forest fragments. Open grassland Bromeliads were associated with spiders and had higher delta N-15 values compared to forest bromeliads. Although forest bromeliads had no association with spiders their total N concentrations were higher. These results suggest that bromeliad nutrition is likely more litter-based in forests and more spider-based in open grasslands. This study is one of the few to show nutrient provisioning and conditionality in a spider-plant system. (c) 2008 Elsevier Masson SAS. All rights reserved.

Effects of habitat fragmentation on the spider community (Arachnida: Araneae) in three Atlantic forest remnants in southeastern Brazil.

This work was developed in three remnants of Atlantic forest in southeastern Brazil. We aimed to assess edge effects in the spider community in a well conserved fragment and to study the variation of spider diversity among fragments of different sizes. The spider families with the highest richness were Theridiidae (38 sp), Araneidae (31 sp) and Salticidae (25 sp). The control area showed the highest diversity (D=0.98) and exclusive species (58.9%). We concluded that spider richness is higher in the large and best preserved fragment. In addition, we found that species richness and abundance increased towards the interior.

Photosynthetic response of soybean to twospotted spider mite (Acari: Tetranychydae) injury

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

On the development of the brown spider Loxosceles gaucho Gertsch (Araneae, Sicariidae): the nympho-imaginal period

Observations of the developmental biology of Loxosceles gaucho Gertsch, 1967, the brown spider of southeastern Brazil are scarce. The present study reports the method of individualized rearing of 18 populations of L. gaucho, kept in laboratory under varied diet conditions, and characterizes their nympho-imaginal period as well as factors related to their reproduction. Females built the first egg sac about 20 days after copulation and nymphs hatched 40 days after the laying date. Average offspring was 61.3 spiderlings and females usually built three to four successive egg sacs in a period of five to seven months. First nymphs initiated their predatory activity between the 5th and 8th days after hatching and the majority reached adulthood within six moults (range of five to eight) in approximately 15 to 17 months (male) and 15.5 to 18 months (female). The average sex ratio equaled 1.0:1.7 (male:female). The wide individual variability of this species intermoult intervals is herewith expressed by the intermoult rate, which was fairly uniform for both intra and interpopulations and gives a relevant aspect for a general idea of the life cycles of spiders.

Ultrastructure of the excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis (Lepidoptera : Pyralidae)

The excretory duct in the silk gland of the sugarcane borer Diatraea saccharalis consists of two morphologically distinct regions, recognized by scanning and transmission electron microscopy. The thin posterior region, adjacent to the glandular region, presents a regular surface. Secretory vesicles containing either electron-dense or fibrillar cuticular-like materials are observed in their apical cytoplasm; the same cuticular materials were detected as extracellular deposits among the microvilli. The short anterior region, near the common duct, exhibits surface protrusions; there are no secretory vesicles in their apical cytoplasm. These results show that only the duct cells at the posterior region are involved in the secretion of the cuticular intima elements. Desmosome-like structures were visualized linking together adjacent microvillar membranes only in the cells of anterior duct region, with unknown function. The transition between the duct and the glandular region is abrupt; the cells of the glandular and posterior duct regions present large amounts of microtubules. Nerve fibers can be observed between the duct cells in their two regions, suggesting that control of silk secretion may occur in the excretory duct via neurotransmitter liberation. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Mantispa minuta Fabricius (Neuroptera : Mantispidae) on egg sacs of the orb-weaver spider Parawixia bistriata (Araneidae) in forests of Eucalyptus grandis in Brazil

This is the first report of the interaction between the two entomophagous agents Mantispa minuta Fabricius (Mantispidae) and Parawixia bistriata Rengger, an orb-weaver spider (Araneidae). From egg sacs kept in the laboratory we observed the emergence of adult neuropterans followed by the emergence of a high number of spiderlings. This association illustrates the interactions among the invertebrate predators coexisting in Eucalyptus plantations managed without chemical treatment, in the State of São Paulo, Brazil.

Kinetic and mechanistic characterization of the Sphingomyelinases D from Loxosceles intermedia spider venom

Envenomation by arachnids of the genus Loxosceles leads to local dermonecrosis and serious systemic toxicity mainly induced by sphingomyelinases D (SMase D). These enzymes catalyze the hydrolysis of sphingomyelin resulting in the formation of ceramide-phosphate and choline as well as the cleavage of lysophosphatidyl choline generating the lipid mediator lysophosphatidic acid. We have, previously, cloned and expressed two functional SMase D isoforms, named P1 and P2, from Loxosceles intertnedia venom and comparative protein sequence analysis revealed that they are highly homologous to SMase I from Loxosceles laeta which folds to form an (alpha/beta)(8) barrel. In order to further characterize these proteins, pH dependence kinetic experiments and chemical modification of the two active SMases D isoforms were performed. We show here that the amino acids involved in catalysis and in the metal ion binding sites are strictly conserved in the SMase D isoforms from L. intermedia. However, the kinetic studies indicate that SMase P1 hydrolyzes sphingomyelin less efficiently than P2, which can be attributed to a substitution at position 203 (Pro-Leu) and local amino acid substitutions in the hydrophobic channel that could probably play a role in the substrate recognition and binding. (c) 2005 Elsevier Ltd. All rights reserved.

An efficient and versatile synthesis of acylpolyamine spider toxins

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

LARVAL DEVELOPMENT OF THE SPIDER CRAB ACANTHONYX-PETIVERII EDWARDS,H.MILNE, 1834 (DECAPODA, MAJIDAE) IN THE LABORATORY

The larval development of Acanthonyx petiverii H. M. Edwards, 1834, was studied in the laboratory through eggs hatched from ovigerous females collected in Ubatuba, state of São Paulo, Brazil. The rearings were carried out in a climatic room with constant temperature (25 degrees +/- 1 degrees C) and salinity (34,5 parts per thousand). The larvae were maintained individually and the food consisted of Artemia nauplii. The larval development of A. petiverii consists of two zoeal stages and a megalopa. All the larval stages were drawn and described in detail. Tables include those presenting morphological characters that allow the identification of zoeae and megalopa of A. petiverii. A comparative study was realized with previously studied majid species that occur in southern and southeastern Brazil.