An efficient and versatile synthesis of acylpolyamine spider toxins

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

Biology and behaviour of the neotropical ant-mimicking spider Aphantochilus rogersi (Araneae: Aphantochilidae): nesting, maternal care and ontogeny of ant-hunting techniques

Aphantochilus rogersi is an ant-mimicking spider that preys exclusively on cephalotine ants. The spiders oviposit in close proximity to nests of the model ant Zacryptocerus pusillus, and emergent spiderlings tend to remain in the vicinity of natal egg sacs. Females of A. rogersi actively defend their egg sacs against approaching workers of Z. pusillus, but the latter may sometimes destroy the eggs. Feeding specialization on these ants is confirmed by more than 300 observations of young and adult A. rogersi carrying ant corpses in the held. Although A. rogersi possesses several behavioural traits which may reduce the risk of being injured by ants during subjugation, field and laboratory observations showed that social defence by Z. pusillus may cause mutilation to the spiders. Tests in captivity revealed an ontogenetic change in the prey-capture techniques employed by A. rogersi. Early-instar spiderlings can apparently only seize the ant's petiole tightly if they are able to approach the ant from the front. As the ant is paralysed, the spiderling positions itself vertically in relation to the substratum. Larger spiders, on the other hand, attack ants most frequently from behind, and seem better equipped to seize the ant's petiole firmly with their larger chelicerae. Owing to their greater strength, late-instar spiders are able to Lift the struggling ant aloft. The selection of a suitable oviposition site, the mother's ability to defend herself and the eggs from nearby ants, and the capacity to capture and subdue ants safely from emergence to maturity, are regarded as crucial traits inherent in the mimetic and feeding specialization by A. rogersi.

Isolation and sequence determination of peptides in the venom of the spider wasp (Cyphononyx dorsalis) guided by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry

Micro-scale (sub-pmol) isolation and sequence determination of three peptides from the venom of the solitary spider wasp Cyphononyx dorsalis is described. We isolated two novel peptides Cd-125 and Cd-146 and a known peptide Thr(6)-bradykinin from only two venom sacs of solitary spider wasp Cyphononyx dorsalis without bioassay-guided fractionation. but instead guided by MALDI-TOF MS. The MALDI-TOF MS analysis of each fraction showed the purity and molecular weight of the components, which led to the isolation of the peptides virtually without loss of sample amount. The sequences of the novel peptides Cd-125 (Asp-Thr-Ala-Arg-Leu-Lys-Trp-His) and Cd-146 (Ser-Glu-Thr-Gly-Asn-Thr-Val-Thr-Val-Lys-Gly-Phe-Ser-Pro-Leu-Arg) were determined by Edman degradation together with mass spectrometry. and finally corroborated by solid-phase synthesis. The known peptide Thr(6)-bradykinin (Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg) was identified by comparison with the synthetic authentic specimen. This is the first example for any kinins to be found in Pompilidae wasp venoms. The procedure reported here can be applicable to studies on many other components of solitary wasp venoms with limited sample availability. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Structure determination of an organometallic 1-(diazenylaryl)ethanol: A novel toxin subclass from the web of the spider Nephila clavipes

A novel chemical subclass of toxin, [1-(3-diazenylphenyl) ethanol]iron, was identified among the compounds present in the web of the spider Nephila clavipes. This type of compound is not common among natural products, mainly in spider-venom toxins; it was shown to be a potent paralytic and/or lethal toxin applied by the spider over its web to ensure prey capture only by topical application. The structure was elucidated by means of ESI mass spectrometry, H-1-NMR spectroscopy, high-resolution (HR) mass spectrometry, and ICP spectrometry. The structure of [1-( 3-diazenylphenyl)ethanol] iron and the study of its insecticidal action may be used as a starting point for the development of new drugs for pest control in agriculture.

Fecundity of the crab Callinectes ornatus Ordway, 1863 (Decapoda, Brachyura, Portunidae) from the Ubatuba region, São Paulo, Brazil

The objective of the present study was to characterize the fecundity of Callinectes ornatus from the northern coast of São Paulo, Brazil. To study the fecundity and its relationships to size, brood weight, and egg size, the ovigerous females were collected at 2-month intervals for two consecutive years (January, 1991 to November, 1992) in the Ubatuba region using a fishing boat equipped with an otter-trawl type of net. After collecting, the animals were screened, bagged, labeled, and stored frozen. Only data from 38 females carrying early stage eggs were considered. In the laboratory, the specimens were thawed at room temperature, the pleopod structure with the egg mass was removed and the eggs were carefully removed from the pleopods. The samples were then fixed in 10% formalin and stored in 70% ethanol, until the time for processing. Processing followed the method of Hines (1982, 1988). Following the frequency determination of carapace width and fecundity, mean egg number, mean volume and mean dry weight of brooded egg mass were determined for each class obtained. Data were analyzed by regressions of log-log transformations for allometric plots (Y = aX(b)) of reproductive variables versus carapace width. Carapace width was one of the main factors for the determination of fecundity, which ranged 171 570 +/- 94 634 eggs, with females of the same size class presenting a wide amplitude of variation. This supports the hypothesis that portunid females present staggered spawning, possibly presenting more than one period of reproduction within one year. The present species showed lower fecundity than the remaining portunids studied, but they did show a rapid and efficient embryonic development, presumably capable of assuring reproductive success.

RELATIVE GROWTH OF THE SPECKLED SWIMMING CRAB ARENAEUS-CRIBRARIUS (LAMARCK, 1818) (BRACHYURA, PORTUNIDAE), NEAR UBATUBA, STATE OF SAO-PAULO, BRAZIL

Arenaeus cribrarius is a mainly tropical crab that occurs in the Western Atlantic Ocean, Brazil being its type-locality. The species ranges from Vinevard Sound, Massachusetts, USA to La Paloma, Uruguay. Information about this species is scarce. The relative growth of A. cribrarius was analyzed, based on some morphometric relations, where the carapace width, excluding lateral spines (CW), was used as an independent variable. A total of 403 specimens (189 males and 214 females), was collected in Ubatuba, State of São Paulo, Brazil, with otter-trawls. The animals were sexed and sorted to maturation phase (juvenile or adult). Some measurements were made: carapace (length and width excluding lateral spines), abdomen (greatest width of the fifth somite in females and the sixth in males) and major chela (greatest length, width and height, dactylus length). This study was made by the application of the power function (y=a.x(b)) which was fitted to the data and the pattern of growth established for each parameter by the ''b''-value (constant of allometry), as positive allometry (b>1), negative allometry (b<1) or isometry (b=1). The morphometric relations of the carapace showed a tendency to isometry. In females, the abdominal width grew in positive allometry, higher in juveniles (b=1.33) than in adults (b=1.18). In this case, an overlap and discontinuity was noticed between the phases over a carapace width range of 55 to 70 mm, where the puberty molt occurs. The majority of relationships showed that the major chela of the males grew in positive allometry, however, the greatest allometric difference between the phases was observed towards the propodus length with 1.09 as juvenile and 1.26 as adult ''b''values. In the males, this variable showed an inflection between the CW range of 45' to 55 mm, where the transition to the maturation phase occurs. The relative growth of this species is similar to those of previously studied species. This indicates, that the propodus length and the abdominal width are the morphometric variables most appropriate to estimate the size of the beginning of the sexual maturity for males and females of this species, respectively.

Isolation and chemical characterization of PwTx-II: A novel alkaloid toxin from the venom of the spider Parawixia bistriata (Araneidae, Araneae)

Brazil has many species of spiders belonging to Araneidae family however, very little is known about the composition, chemical structure and mechanisms of action of the main venom components of these spiders. The main objective of this work was to isolate and to perform the chemical characterization of a novel beta-carboline toxin from the venom of the spider Parawixia bistriata, a typical species of the Brazilian 'cerrado'. The toxin was purified by RP-HPLC and structurally elucidated by using a combination of different spectroscopic techniques (UV, ESI-MS/MS and H-1 NMR), which permitted the assignment of the molecular structure of a novel spider venom toxin, identified as 1-4-guanidinobutoxy-6-hydroxy-1,2,3,4-tetrahydro-beta-carboline, and referred to here as PwTx-II. This compound is toxic to insects (LD50 = 12 +/- 3 eta g/mg honeybee), neurotoxic, convulsive and lethal to rats (LD50 = 9.75 mg/kg of male Wistar rat). (c) 2005 Elsevier Ltd. All rights reserved.

Biometric relationship analysis of the wet weight by carapace width to the swimming crab Arenaeus-cribrarius (Lamarck, 1818) (Crustacea, Brachyura, Portunidae)

The biometric relationship between the weigth and the carapace width in the swimming crab A. cribrarius was compared with the results from other portunid crabs studied previously. During November/1988 to October/1989, a total of 403 specimens (189 males and 214 females) were collected with otter-trawl nets in north coast of the São Paulo State, Brazil. The animals were measured (carapace width excluding lateral spines = LC! and weighed (wet weight = PE). The empiric points of this relation were fit according to the power function (Y = a.X(b)) for each sex, maturation phases and total of individuals. The relation PE x LC indicates that the mole's growth changes during the ontogenesys from isometric (in juvenile phase) to allometric positive (in adult phase). For the females the growth is isometric in the two phases. The weight grows in a higher proportion than the carapace width variable (allometric positive growth). The data can be grouped in a single equation (PE = 7.85.10(-5).LC(3.14)) for the convertion between the variables there was a greater similarity between the equations obtained far each sex. In spite of this, the males present the fattening grade value (''a'') slightly higher than that of the females, possibly because of the greater size reached in its devellopment. The mean weight of the males is greater than the females one (p < 0.01). In the range 80 proves 90mm the males were more abundant, probably due to the females terminal ecdysis is near this size. The females only have the mean weight greater than the males in the 60 proves 70mm range (p < 0.01) when the puberty molt occurs and they present morphological changes in their reproductive system.

MOLECULAR DIAGNOSIS of LEISHMANIA AMAZONENSIS IN A CAPTIVE SPIDER MONKEY IN BAURU, São Paulo, BRAZIL

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

Population biology and habitat utilization of the stone crab Menippe nodifrons in the Ubatuba region, São Paulo, Brazil

The aim of the current study was to evaluate a population of Menippe nodiffons in a reef of sabellariid worms, Phragmatopoma lapidosa, with regard to recruitment, population structure and sex ratio. Sampling was carried out each other month from September 1994 up to and including July 1995, on the rocky shores of the Tenorio beach, São Paulo, Brazil. It resulted in 183 individuals, whose average carapace width in this biotope was 9.1 +/- 5.6 m. The animals were grouped in nine size classes and the frequency distribution showed that 80% occupied the three first classes. This means that the polychaete worm colonies are of great importance for the establishment of individuals in the first juvenile stages.

Plant glandular trichomes mediate protective mutualism in a spider-plant system

1. Although several species of Peucetia (Oxyopidae) live strictly in association with plants bearing glandular trichomes worldwide, to date little is known about whether these associations are mutualistic.2. In this study we manipulated the presence of Peucetia flava on the glandular plant Rhynchanthera dichotoma in the rainy and post-rain season, to test the strength of its effects on leaf, bud, and flower damage and plant reproductive output. In addition, we ran independent field experiments to examine whether these sticky structures improve spider fidelity to plants.3. Peucetia suppressed some species of foliar phytophages, but not others. Although spiders have reduced levels of leaf herbivory, this phenomenon was temporally conditional, i.e. occurred only in the post-rain but not in the rainy season. Floral herbivory was also reduced in the presence of spiders, but these predators did not affect plant fitness components.4. Plants that had their glandular trichomes removed retained fewer insects than those bearing such structures. Spiders remained longer on plants with glandular trichomes than on plants in which these structures had been removed. Isotopic analyses showed that spiders that fed on live and dead labelled flies adhered to the glandular hairs in similar proportions.5. Spiders incurred no costs to the plants, but can potentially increase individual plant fitness by reducing damage to reproductive tissues. Temporal conditionality probably occurred because plant productivity exceeded herbivore consumption, thus dampening top-down effects. Specialisation to live on glandular plants may have favoured scavenging behaviour in Peucetia, possibly an adaptation to periods of food scarcity.

Population biology and reproduction of the hermit crab Clibanarius vittatus (Decapoda: Anomura) in an estuarine region of southern Brazil

The population dynamics and reproduction of the hermit crab Clibanarius vittatus were evaluated on Pescadores Beach, located on the estuarine channel of Sao Vicente (São Paulo), Brazil. The hermit crabs were captured by hand during low tide,from May 2001 to April 2003. A total of 2554 hermit crabs were captured, of which 701 were males, 1741 non-ovigerous females, 48 ovigerous females and 64 intersex individuals. The size-frequency distribution of the males was represented by a platykurtic bell-shaped curve, which differed from the leptokurtic bell-shaped curve of the females. The smaller and intermediate classes were composed mainly of females (modal size 6.5-7.5 mm carapace shield length (CSL)), and the larger classes only by males (modal size 9.5-10.5 mm CSL). The overall sex-ratio was skewed toward females (0.39:1/M:F), differing significantly from the expected 1:1. A seasonal reproductive pattern was recorded for C. vittatus in this location, with more intensive reproductive activity in the warmer months. The absence of juveniles suggests that their recruitment area is different than the area inhabited by adults, possibly another area with more protection and specialized or different resources for young.

Spider assemblages in widely-separated patches of cerrado in São Paulo State, Brazil.

Brazilian cerrado is a biologically-rich, poorly understood, yet rapidly disappearing habitat. Composition of the spider assemblages from areas of cerrado from three separate sites in the State of São Paulo, Brazil were sampled by beating the canopies and adjacent shrubs of three Myrcia (Myrtaceae; "myrtle") tree species. These produced a total of 859 spiders 'belonging to 21 families and 75 species. The most undisturbed and densest cerrado habitat had the largest number and greatest diversity of spider species, encompassing stalkers, ambushers, space web-weavers, and foliage runners. The other two areas were dominated by foliage runners. Spider distribution in this natural and complex habitat was evaluated by classifying the samples into 12 habitat/microhabitat groups according to local of the patch, tree species, and microhabitat (target tree or adjacent shrub). Correspondence analysis was used for ordination of species and groups based on their abundance. Environmental factors such as patches type (p=0.027) and plant species (p=0.046) had significant effects in explaining the ordination. Canonical correspondence analysis was applied for relating the patterns in species richness and/or abundance to the significant environmental factors. A comparison of the results showed that the family composition among the patches is rather similar, and there is a tendency of spiders species overlap an interregional level (patches effect, p=0.027). However, the most similar spider assemblages living on woody vegetation occurred in Myrcia venulosa and Myrcia guianensis at São Carlos and Pirassununga, demonstrating an interregional similarity (plant species effect, p=0.046) that indicates an association between spiders and particular vegetation.