An efficient and versatile synthesis of acylpolyamine spider toxins

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

SEASONAL DYNAMICS OF MACROALGAL COMMUNITIES IN THE PRETO RIVER BASIN, SAO-PAULO, SOUTHEASTERN BRAZIL

Macroalgal seasonality was analyzed in nine stream segments of the Preto River basin: five of these were sampled monthly for one year and four bimonthly during two years. Seasonal variation of macroalgae was correlated with stream variables (temperature, current velocity, specific conductance, turbidity, surface light, pH and oxygen). Multiple linear regression was applied to evaluate the macroalgal seasonal variability determined by a combination of temperature, current velocity and turbidity, the most influential environmental parameters on macroalgal seasonality. Most species revealed a clear seasonal trend, such that late fall through early spring was the period with highest macroalgal abundance in most sites. Temperature, current velocity and turbidity account for 24 to 83 % of macroalgal species number variation and 24 to 79 % of macroalgal abundance in the basin. The results suggest that the seasonal flow determined by the precipitation regime associated with turbidity and temperature were the major factors influencing the seasonal dynamics of macroalgal communities. The highest values of macroalgal species number and abundance were found under cooler temperatures, lower current velocities and lower values of turbidity.

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.

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.

Macrobenthic intertidal communities from wave to tide dominated beach environments. A case study in two Brazilian beaches

Modification showed by intertidal macrofaunal communities between two nearby sites that change from a wave to a tide dominated beach environment, was analyzed on the present study. At each site, eleven intertidal sampling stations were distributed along a transect, from the drift line to the spring low tide water level. Four macrofaunal samples one meter long-shore spaced were collected at, each station with an iron core of 0.05 m(2) surface area, taken to a depth of 20 cm. Major,differences on sediments between sites were the offshore decrease of mean particle size diameter and increase of kurtosis and water content at the tide dominated site. KIDS ordination showed major similarities between the lowest stations of this site, that represents the dissipative low tide-terrace portion of the beach. Two lower station of the wave dominated site presented similarities with this group. The other stations of the tide-dominated site, that represents the reflective high tide beach portion, grouped distant from the former. (canonical Correspondence Analysis (CCA) showed a similar spatial distribution of stations, suggesting the importance of environmental factors on the explanation of species distribution patterns. Sediment water content and water table depth, with the highest inertia value, seems to be the principal physical factor. Increase on water content affects the macrofaunal distribution by the expansion of typical infralitoral species, as was the case of Bathyporeiapus ruffoi, on the dissipative low tide terrace beach portion. A disrupted distribution with the lost of a gradate zonation along a physical gradient is one of the major modification presented by macrobenthic communities on the transition from a wave to a tide dominated beach environment.

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.

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.

Relative growth of spider crab Acanthonyx scutiformis (Dana, 1851) (Crustacea, Decapoda, Majidae)

The dimensions of chelar propodus and abdomen were utilized for the morphometric analysis about the relative growth studies of Acanthonyx scutiformis (Dana, 1851). A total of 297 specimens (114 males and 183 females), was collected in Ubatuba, State of São Paulo, Brazil. The animals were sexed and soarted to maturation phase (immature and mature). The following measurements were made: carapace (length and width), abdomen width and chelar propodus (length and height). In this study was applicated the power function (y= a.x(b)), in which the growth was considered positive allometric with isometric with b=1, and negative allometric with b<1. The alometry becomes isometric, when the results concerning immature and mature females and males are analysed. In the Brachyura male chelipeds are utilized in territorial defense, fight, display and courtship and female abdomens are utilized as a chamber that protect and incubate the eggs.

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)

Changes in coral reef communities and an associated reef fish species, Cephalopholis cruentata (Lacepede), after 30 years on Curacao (Netherlands Antilles)

Using the same methodology and identical sites, we repeat a study dating from 1973 and quantify cover of hard coral species, soft corals, sponges, hard substratum and soft substratum, and density of a commercially important reef fish species, the graysby Cephalopholis cruentata, along a depth-gradient of 3-36 m oil the coral reefs of Curacao. The objective was to determine the multi-decade change in benthic coral reef cover and structural complexity, and their effect oil densities of an associated reef fish species. Total hard coral cover decreased on average from 52% in 1973 to 22% in 2003, representing a relative decline of 58%. During this time span, the cover of hard substratum increased considerably (from 11 to 58%), as did that of soft corals (from 0.1 to 2.2%), whereas the cover of sponges showed no significant change. Relative decline of hard coral cover and of reef complexity was greatest in shallow waters (near the coast), which is indicative of a combination of anthropogenic influences from shore and recent storm damage. Cover of main reef builder coral species (Agaricia spp., Siderastrea siderea, Montastrea annularis) decreased more than that of other species, and resulted in a significant decrease in reef complexity. Although density of C. cruentata was highly correlated to cover of Montastrea and Agaricia in 1973, the loss of coral cover did not show any effect on the total density of C. cruentata in 2003. However, C. cruentata showed a clear shift in density distribution from shallow water in 1973 to deep water in 2003. It call be concluded that the reefs of Curacao have degraded considerably in the last three decades, but that this has had no major effect on the population size of one commercially important coral-associated fish species.

The role of spatial and temporal descriptors for neotropical tadpole communities in southern Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biofilm microbial communities of denture stomatitis

Introduction: Denture stomatitis is a common lesion that affects denture wearers. Its multifactorial etiology seems to depend on a complex and poorly characterized biofilm. The purpose of this study was to assess the composition of the microbial biofilm obtained from complete denture wearers with and without denture stomatitis using culture-independent methods.Methods: Samples were collected from healthy denture wearers and from patients with denture stomatitis. Libraries comprising about 600 cloned 16S ribosomal DNA (rDNA) bacterial sequences and 192 cloned eukaryotic internal transcribed spacer (ITS) region sequences, obtained by polymerase chain reactions, were analyzed.Results: The partial 16S rDNA sequences revealed a total of 82 bacterial species identified in healthy subjects and patients with denture stomatitis. Twenty-seven bacterial species were detected in both biofilms, 29 species were exclusively present in patients with denture stomatitis, and 26 were found only in healthy subjects. Analysis of the ITS region revealed the presence of Candida sp. in both biofilms.Conclusion: The results revealed the extent of the microbial flora, suggesting the existence of distinct biofilms in healthy subjects and in patients with denture stomatitis.

Impact of sewage sludge on the soil bacterial communities by DNA microarray analysis

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