Diet of the colubrid snake, Thamnodynastes strigatus (Günther, 1858) from Paraná State, Brazil, with field notes on anuran predation

São apresentados dados da dieta de Thamnodynastes strigatus (Serpentes: Colubridae), a partir da dissecção de 44 espécimes. No tubo digestivo dos exemplares examinados foram encontrados anfíbios anuros (71,4% da amostra), roedores (14,3%), peixes (10,7%) e lagartos (3,6%). A maioria das espécies de anuros (Bufo sp., Leptodactylus sp., Physalaemus cuvieri e Scinax fuscovarius) encontradas no exame de conteúdo estomacal de T. strigatus, utiliza o solo ou o nível d'água como sítio de vocalização. Também são apresentados dados sobre a observação de eventos de predação na natureza.

When Basking Is Not an Option: Thermoregulation of a Viperid Snake Endemic to a Small Island in the South Atlantic of Brazil

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

Crystallization and preliminary diffraction data of BaP1, a haemorrhagic metalloproteinase from Bothrops asper snake venom

BaP1 is a metalloproteinase isolated from the venom of the Central American snake Bothrops asper (terciopelo). It is a 24 kDa protein consisting of a single chain which includes the metalloproteinase domain only, therefore being classified as a class P-I snake-venom metalloproteinase. BaP1 induces prominent local tissue damage, such as haemorrhage, myonecrosis, blistering, dermonecrosis and oedema. In order to elucidate its structure, BaP1 was crystallized by the hanging-drop vapour-diffusion technique in 0.1 M bicine pH 9.0, 10% PEG 20 000 and 2%(v/v) dioxane. Diffraction data were observed to a resolution of 2.7 Angstrom. Crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 38.22, b = 60.17, c = 86.09 Angstrom.

Thrombomodulin-independent activation of protein C and specificity of hemostatically active snake venom serine proteinases - Crystal structures of native and inhibited Agkistrodon contortrix contortrix protein C activator

Protein C activation initiated by the thrombin-thrombomodulin complex forms the major physiological anticoagulant pathway. Agkistrodon contortrix contortrix protein C activator, a glycosylated single-chain serine proteinase, activates protein C without relying on thrombomodulin. The crystal structures of native and inhibited Agkistrodon contortrix contortrix protein C activator determined at 1.65 and 1.54 angstrom resolutions, respectively, indicate the pivotal roles played by the positively charged belt and the strategic positioning of the three carbohydrate moieties surrounding the catalytic site in protein C recognition, binding, and activation. Structural changes in the benzamidine-inhibited enzyme suggest a probable function in allosteric regulation for the anion-binding site located in the C-terminal extension, which is fully conserved in snake venom serine proteinases, that preferentially binds Cl1- instead of SO42-.

Crotacetin, a novel snake venom C-type lectin homolog of convulxin, exhibits an unpredictable antimicrobial activity

Snake venom (sv) C-type lectins encompass a group of hemorrhagic toxins that are capable of interfering with blood stasis. A very well-studied svC-type lectin is the heterodimeric toxin, convulxin (CVX), from the venom of South American rattlesnake Crotalus durissus terrificus. CVX is able to activate platelets and induce their aggregation by acting via p62/GPVI collagen receptor. By using polymerase chain reaction homology screening, we have cloned several cDNA precursors of CVX subunit homologs. One of them, named crotacetin (CTC) beta-subunit, predicts a polypeptide with a topology very similar to the tridimensional conformations of other subunits of CVX-like snake toxins, as determined by computational analysis. Using gel permeation and reverse-phase high-performance liquid chromatography, CTC was purified from C. durissus venoms. CTC can be isolated from the venom of several C. durissus subspecies, but its quantitative predominance is in the venom of C. durissus cascavella. Functional analysis indicates that CTC induces platelet aggregation, and, importantly, exhibits an antimicrobial activity against Gram-positive and -negative bacteria, comparable with CVX.

Amino acid sequence and crystal structure of BaP1, a metalloproteinase from Bothrops asper snake venom that exerts multiple tissue-damaging activities

BaP1 is a 22.7-kD P-I-type zinc-dependent metalloproteinase isolated from the venom of the snake Bothrops asper, a medically relevant species in Central America. This enzyme exerts multiple tissue-damaging activities, including hemorrhage, myonecrosis, dermonecrosis, blistering, and edema. BaP1 is a single chain of 202 amino acids that shows highest sequence identity with metalloproteinases isolated front the venoms of snakes of the subfamily Crotalinae. It has six Cys residues involved in three disulfide bridges (Cys 117-Cys 197, Cys 159-Cys 181, Cys 157-Cys 164). It has the consensus sequence H(142)E(143)XXH(146)XXGXXH(152), as well as the sequence C164I165M166, which characterize the metzincin superfamily of metalloproteinases. The active-site cleft separates a major subdomain (residues 1-152), comprising four a-helices and a five-stranded beta-sheet, from the minor subdomain, which is formed by a single a-helix and several loops. The catalytic zinc ion is coordinated by the N-epsilon2 nitrogen atoms of His 142, His 146, and His 152, in addition to a solvent water molecule, which in turn is bound to Glu 143. Several conserved residues contribute to the formation of the hydrophobic pocket, and Met 166 serves as a hydrophobic base for the active-site groups. Sequence and structural comparisons of hemorrhagic and nonhemorrhagic P-I metalloproteinases from snake venoms revealed differences in several regions. In particular, the loop comprising residues 153 to 176 has marked structural differences between metalloproteinases with very different hemorrhagic activities. Because this region lies in close proximity to the active-site microenvironment, it may influence the interaction of these enzymes with physiologically relevant substrates in the extracellular matrix.

Antimicrobial activity of an L-amino acid oxidase isolated from Bothrops leucurus snake venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Pseudechis australis Venomics: Adaptation for a Defense against Microbial Pathogens and Recruitment of Body Transferrin

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

Enzymatic toxins from snake venom: structural characterization and mechanism of catalysis

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

Snake Venomic of Crotalus durissus terrificus-Correlation with Pharmacological Activities

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

Snake venomics of the Siamese Russell's viper (Daboia russelli siamensis) - Relation to pharmacological activities

The venom proteome of Daboia russelli siamensis, a snake of medical importance in several Asian countries, was analysed by 2-D electrophoresis, subsequent MS/MS and enzymatic assays. The proteome comprises toxins from six protein families: serine proteinases, metalloproteinases, phospholipases A(2), L-amino acid oxidases, vascular endothelial growth factors and C-type lectin-like proteins. The venom toxin composition correlates with the clinical manifestation of the Russell's viper bite and explains pathological effects of the venom such as coagulopathy, oedema, hypotensive, necrotic and tissue damaging effects. The vast majority of toxins are potentially involved in coagulopathy and neurotoxic effects. The predominant venom components are proteinases capable of activating blood coagulation factors and promoting a rapid clotting of the blood, and neurotoxic phospholipase A(2)s. The analysis of the venom protein composition provides a catalogue of secreted toxins. The proteome of D. r. siamensis exhibits a lower level of toxin diversity than the proteomes of other viperid snakes. In comparison to the venoms of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis, the venom from D. r. siamensis showed quantitative differences in the proteolytic, phospholipase A2, L-amino acid oxidase and alkaline phosphatase activities. (c) 2009 Published by Elsevier B.V.

Purification, crystallization and preliminary X-ray diffraction analysis of crotamine, a myotoxic polypeptide from the Brazilian snake Crotalus durissus terrificus

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

Penetration time of Salmonella Heidelberg through shells of white and brown commercial eggs

This study aimed at determining the minimum time required for the penetration of Salmonella Heidelberg inside the eggs after contact with contaminated material. Recently-collected brown and white eggs from laying hens between 45-50 weeks of age, reared in a commercial poultry house, were artificially contaminated by contact with wood shavings moistened with liquid inoculum of Salmonella Heidelberg in stationary-growth phase (10³-10(4) CFU g-1). According to type (white or brown), eggs were distributed into three different groups, with four replicates each: negative control group (no artificial contamination), positive control group (analyzed externally immediately after contamination and internally after the maximum storage period of the test group) and test group. Eggs were stored at controlled environmental temperature varying from 25ºC to 30ºC. In the test group, eggs contents (yolk and albumen) were pooled and analyzed after 1:00, 1:30, 2:00, 2:30, 3:00, 3:30, and 4:00 hours after contamination for the presence of Salmonella Heidelberg in 25g of this pool. The experimental unit consisted of five eggs in each test. The analysis protocol included pre-enrichment, selective enrichment, plating on selective agar, and biochemical and serological tests. The results obtained were submitted to logistic regression, which indicated that the presence of Salmonella Heidelberg was verified after 2:16 h and 2:44 h of contact with white and brown eggs, respectively.

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.