Snakes from coastal islands of State of São Paulo, Southeastern Brazil

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

Oral microbiota of Brazilian captive snakes

The present work aimed to determine the oral microbiotic composition of snakes from Sao Jose do Rio Preto city, São Paulo State, Brazil. Ten snake species, comprising the families Boidae, Colubridae, Elapidae and Viperidae, were submitted to microbiological examination of their oral cavity, which indicated positivity for all buccal samples. Gram-negative bacilli, gram-negative cocci bacilli, gram-positive bacilli and gram-positive cocci were isolated from the snakes. Among isolated bacterium species, the occurrence of coagulase-negative staphylococci in the buccal cavity of Crotalus durissus (Viperiade), Eunectes murinus (Boidae), Mastigodryas bifossatus (Colubridae) and Bacillus subtilis, common to oral cavity of Bothrops alternatus (Viperidae) and Phalotris mertensi (Colubridae), was detected. It was observed higher diversity of isolated bacteria from the oral cavity of Micrurus frontalis (Elapidae) and Philodryas nattereri (Colubridae), as well as the prevalence of gram-positive baccillus and gram-positive cocci. The composition of the oral microbiota of the studied snakes, with or without inoculating fangs, is diverse and also related to the formation of abscesses at the bite site in the victims of the ophidian accidents, and to pathogenic processes in the snakes that host these microorganisms.

Snake assemblage in a disturbed grassland environment in Rio Grande do Sul State, southern Brazil: Population fluctuations of Liophis poecilogyrus and Pseudablabes agassizii

A mark-recapture study of a snake assemblage using pitfall traps with drift fences was carried out in a disturbed grassland environment (e.g. cattle breeding and cultivations), located in the Pampa Biome, in the central region of the Rio Grande do Sul State, southern Brazil. From February 2001 to January 2004 we caught 272 snakes belonging to 20 species from the following families: Elapidae (5%), Viperidae (10%), and Colubridae (85%). The assemblage had a unimodal seasonal pattern of activity, and the highest number of captures occurred between September and May. There was a positive and significant correlation between the number of captures and monthly minimum and maximum average temperatures. Recruitment was observed from January to April. During the study, the area was affected by human activities, which altered the community structure: Pseudablabes agassizii was negatively affected by habitat devastation while Liophis poecilogyrus took advantage of this. Our results reinforced the impression that Pseudablabes agassizii is a habitat specialist species. We extend the understanding of the susceptibility of this species to environmental destruction in open natural environments of South America, and propose its use as a potential bio-indicator of the Pampa biome. We also discuss the importance of conservation strategies for snakes in grasslands of southern Brazil. © Finnish Zoological and Botanical Publishing Board 2007.

Molecular phylogeny of advanced snakes (Serpentes, Caenophidia) with an emphasis on South American xenodontines: A revised classification and descriptions of new taxa

We present a molecular phylogenetic analysis of caenophidian (advanced) snakes using sequences from two mitochondrial genes (12S and 16S rRNA) and one nuclear (c-mos) gene (1681 total base pairs), and with 131 terminal taxa sampled from throughout all major caenophidian lineages but focussing on Neotropical xenodontines. Direct optimization parsimony analysis resulted in a well-resolved phylogenetic tree, which corroborates some clades identified in previous analyses and suggests new hypotheses for the composition and relationships of others. The major salient points of our analysis are: (1) placement of Acrochordus, Xenodermatids, and Pareatids as successive outgroups to all remaining caenophidians (including viperids, elapids, atractaspidids, and all other colubrid groups); (2) within the latter group, viperids and homalopsids are sucessive sister clades to all remaining snakes; (3) the following monophyletic clades within crown group caenophidians: Afro-Asian psammophiids (including Mimophis from Madagascar), Elapidae (including hydrophiines but excluding Homoroselaps), Pseudoxyrhophiinae, Colubrinae, Natricinae, Dipsadinae, and Xenodontinae. Homoroselaps is associated with atractaspidids. Our analysis suggests some taxonomic changes within xenodontines, including new taxonomy for Alsophis elegans, Liophis amarali, and further taxonomic changes within Xenodontini and the West Indian radiation of xenodontines. Based on our molecular analysis, we present a revised classification for caenophidians and provide morphological diagnoses for many of the included clades; we also highlight groups where much more work is needed. We name as new two higher taxonomic clades within Caenophidia, one new subfamily within Dipsadidae, and, within Xenodontinae five new tribes, six new genera and two resurrected genera. We synonymize Xenoxybelis and Pseudablabes with Philodryas; Erythrolamprus with Liophis; and Lystrophis and Waglerophis with Xenodon.

Taxocenoses de serpentes em grupos fitofisionômicos de cerrado no Parque Nacional de Sete Cidades, Piracuruca, Piauí, Brasil

Este trabalho teve como objetivo principal inventariar a fauna de serpentes do Parque Nacional de Sete Cidades, Piraracuruca, Piauí, Brasil, enfocando os seguintes aspectos: composição, riqueza e abundância de espécies nos seus diversos habitats, padrões de atividade diária, dieta, reprodução e comparação de composição com outras áreas estudadas por outros autores. Foram realizadas seis expedições ao Parque, entre setembro de 2005 e agosto de 2006, que totalizaram 120 dias de trabalho de campo. Para a amostragem de campo, foram utilizados três métodos: procura limitada por tempo, armadilhas de interceptação e queda e encontros ocasionais. Foram registradas 87 serpentes, distribuídas em quatro famílias (Boidae, Colubridae, Elapidae, Viperidae), 18 gêneros e 24 espécies. A espécie dominante foi Thamnodynastes sp. A (13,1%), seguida de Oxyrhopus trigeminus e Micrurus ibiboboca (10, 3%). Houve um predomínio de espécies terrestres e com períodos de atividade diurna. Como em outras taxocenoses de áreas abertas os colubrídeos mostraram-se dominantes. A fitofisionomia Cerrado Típico apresentou maior diversidade de espécies, sendo as menores diversidades registradas no Campo Limpo e Cerrado Rupestre. O método que apresentou melhor desempenho foi procura limitada por tempo, contudo, a utilização dos métodos de coleta isolados não se mostrou eficiente para inventariar a fauna de serpentes, sendo necessário o uso conjunto destes métodos para uma melhor amostragem da área. Através da análise de ACOP e da análise de agrupamento, foi possível observar que apesar de haver uma semelhança florística e fisionomicamente com Cerrado, a composição de espécies mostrou maior similaridade faunística com taxocenoses de áreas transição Cerrado/Caatinga e Caatinga.

Filogenia molecular da família Dipsadidae (serpentes : Colubroidea)

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Pseudechis guttatus venom proteome: Insights into evolution and toxin clustering

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversity, natural history, and geographic distribution of snakes in the Caatinga, Northeastern Brazil

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

Levantamento das espécies de serpentes de Botucatu

Throughout the analysis of the collections JJ, CEVAP, IBSP, ZUEC and UEL, a study on the ophiofauna of Botucatu was made. We present a preliminary list of the species of snakes here found, with small comments on their natural history. The number of specimens studied was 943. We registered, for Botucatu, 51 species of snakes, distributed in 31 genera and 6 families. The families here found were Anomalepididae (1 spp.), Boidae (1 spp.), Colubridae (7 spp.), Dipsadidae (32 spp.), Elapidae (3 spp.) and Viperidae (7 spp.). The analysis of the relative abundance shows that Dipsadidae was the most abundant family, with n=425 (44,83%), followed by Viperidae, with n=388 (40,93%), Boidae, n=62 (6,54%), Colubridae, n=57 (6,01%), Elapidae, n=15 (1,6%) and, at last, Anomalepididae, with n=1 (0,1%). The five more representative species were Crotalus durissus (n=135, 14,31%), Oxyrhopus guibei (n=123, 12,8%) Bothropoides jararaca (N=121, 12,6%), Bothropoides neuwiedi (N=95, 9,88%) and Sibynomorphus mikani (N=65, 6,76%). A higher number of individuals collected was registered for the months of january to april that, together, sum up to almost 50% of the total. The months of june to september registered a lower number of individuals. The spacial distribution analysis shows that a higher number of snakes was found on the country areas of Botucatu (n=270, 41,54%), specially on pastures. Due to the lack of studies of Botucatu’s ophiofauna, this list is probably underestimated. It is mandatory that future studies approaching this group and its ecological components on this region are made, using appropriate sampling methodologies, in order to form an accurate list of the species of snakes of Botucatu

Biological characterization of the Amazon coral Micrurus spixii snake venom: isolation of a new neurotoxic phospholipase A2

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ophioluxin, a convulxin-like C-type lectin from Ophiophagus hannah (King cobra) is a powerful platelet activator via glycoprotein VI

Ophioluxin, a potent platelet agonist, was purified from the venom of Ophiophagus hannah (King cobra). Under nonreducing conditions it has a mass of 85 kDa, similar to convulxin, and on reduction gives two subunits with masses of 16 and 17 kDa, slightly larger than those of convulxin. The N-terminal sequences of both subunits are very similar to those of convulxin and other C-type lectins. Ophioluxin induces a pattern of tyrosine-phosphorylated proteins in platelets like that caused by convulxin, when using appropriate concentrations based on aggregation response, because it is about 2-4 times more powerful as agonist than the latter. Ophioluxin and convulxin induce [Ca(2+)](i) elevation both in platelets and in Dami megakaryocytic cells, and each of these C-type lectins desensitizes responses to the other. Convulxin agglutinates fixed platelets at 2 microg/ml, whereas ophioluxin does not, even at 80 microg/ml. Ophioluxin resembles convulxin more than echicetin or alboaggregin B because polyclonal anti-ophioluxin antibodies recognize both ophioluxin and convulxin, but not echicetin, and platelets adhere to and spread on ophioluxin- or convulxin-precoated surfaces in the same way that is clearly different from their behavior on an alboaggregin B surface. Immobilized ophioluxin was used to isolate the glycoprotein VI-Fcgamma complex from resting platelets, which also contained Fyn, Lyn, Syk, LAT, and SLP76. Ophioluxin is the first multiheterodimeric, convulxin-like snake C-type lectin, as well as the first platelet agonist, to be described from the Elapidae snake family.

Novel microbicidal agents derived from Naja atra cardiotoxin 1 (CTX-1)

During my PhD course, I focused my research on antimicrobial peptides (AMPs), in particular on the aspects of their computational design and development. This work led to the development of a new family of AMPs that I designed, starting from the amino acid sequence of a snake venom toxin, the cardiotoxin 1 (CTX-1) of Naja atra. Naja atra atra cardiotoxin 1, produced by Chinese cobra snakes belonging to Elapidae family, is included in the three-finger toxin family and exerts high cytotoxicity and antimicrobial activity too. This toxin family is characterized by specific folding of three beta-sheet loops (“fingers”) extending from the central core and by four conserved disulfide bridges. Using as template the first loop of this toxin, different sequences of 20 amino acids linear cationic peptides have been designed in order to avoid toxic effects but to maintain and strengthen the antimicrobial activity. As a result, the sequence NCP-0 (Naja Cardiotoxin Peptide-0) was designed as ancestor and subsequently other 4 variant sequences of NCP0 were developed. These variant sequences have shown microbicidal activity towards a panel of reference strains of Gram-positive and Gram-negative bacteria, fungi and an enveloped virus. In particular, the sequence designed as NCP-3 (Naja Cardiotoxin Peptide-3) and its variants NCP-3a and NCP-3b have shown the best antimicrobial activity together with low cytotoxicity against eukaryotic cells and low hemolytic activity. Bactericidal activity has been demonstrated by minimum bactericidal concentration (MBC) assay at values below 10 μg/ml for Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii ( clinical isolates), Moraxella catharralis ATCC 25238, MRSA ATCC 43400, while towards Staphylococcus aureus ATCC 25923, Enterococcus hirae ATCC 10541 and Streptococcus agalactiae ATCC 13813 the bactericidal activity was demonstrated even below 1.6 μg/ml concentration. This potent antimicrobial activity was confirmed even for unicellular fungi Candida albicans, Candida glabrata and Malassezia pachydermatis (MBC 32.26-6.4 μg/ml), and also against the fast-growing mycobacteria Mycobacterium smegmatis DSMZ 43756 and Mycobacterium fortuitum DSMZ 46621 (MBC 100 μg/ml). Moreover, NCP-3 has shown a virucidal activity on the enveloped virus Bovine Herpesvirus 1 (BoHV1) belonging to herpesviridae family. The bactericidal activity is maintained in a high salt concentration (125 and 250 mM NaCl) medium and PB +20% Mueller Hinton Medium for E. coli, MRSA and Pseudomonas aeruginosa reference strains. Considering these in vitro obtained data, we propose NCP-3 and its variants NCP-3a and NCP-3b as promising antimicrobial candidates. For this reason, the whole novel AMPs family has been protected by a national patent (n°102015000015951).