Identification and analysis of venom gland-specific genes from the coastal taipan (Oxyuranus scutellatus) and related species

Australian terrestrial elapid snakes contain amongst the most potently toxic venoms known. However, despite the well-documented clinical effects of snake bite, little research has focussed on individual venom components at the molecular level. To further characterise the components of Australian elapid venoms, a complementary (cDNA) microarray was produced from the venom gland of the coastal taipan (Oxyuranus scutellatus) and subsequently screened for venom gland-specific transcripts. A number of putative toxin genes were identified, including neurotoxins, phospholipases, a pseudechetoxin-like gene, a venom natriuretic peptide and a nerve growth factor together with other genes involved in cellular maintenance. Venom gland-specific components also included a calglandulin-like protein implicated in the secretion of toxins from the gland into the venom. These toxin transcripts were subsequently identified in seven other related snake species, producing a detailed comparative analysis at the cDNA and protein levels. This study represents the most detailed description to date of the cloning and characterisation of different genes associated with envenomation from Australian snakes.

Novel natriuretic peptides from the venom of the inland taipan (Oxyuranus microlepidotus): isolation, chemical and biological characterisation

Three natriuretic-like peptides (TNP-a, TNP-b, and TNP-c) were isolated from the venom of Oxyuranus microlepidotus (inland taipan) and were also present in the venoms of Oxyuranus scutellatus canni (New Guinea taipan) and Oxyuranus scutellatus scutellatus (coastal taipan). They were isolated by HPLC, characterised by mass spectrometry and Edman analysis, and consist of 35-39 amino acid residues. These molecules differ from ANP/BNP through replacement of invariant residues within the 17-membered ring structure and by inclusion of proline residues in the C-terminal tail. TNP-c was equipotent to ANP in specific GC-A assays or aortic ring assays whereas TNP-a and TNP-b were either inactive (GC-A over-expressing cells and endothelium-denuded aortic rings) or weakly active (endothelium-in tact aortic rings). TNP-a and TNP-b were also unable to competitively inhibit the binding of TNP-c in endothelium-denuded aortae (GC-A) or endothelium-in tact aortae (NPR-C). Thus, these naturally occurring isoforms provide a new platform for further investigation of structure-function relationships of natriuretic peptides. (C) 2004 Elsevier Inc. All rights reserved.

Comparison of the in vitro neuromuscular activity of venom from three Australian snakes (Hoplocephalus stephensi, Austrelaps superbus and Notechis scutatus): Efficacy of tiger snake antivenom

1. Tiger snake antivenom, raised against Notechis scutatus venom, is indicated not only for the treatment of envenomation by this snake, but also that of the copperhead (Austrelaps superbus ) and Stephen's banded snake (Hoplocephalus stephensi ). The present study compared the neuromuscular pharmacology of venom from these snakes and the in vitro efficacy of tiger snake antivenom. 2. In chick biventer cervicis muscle and mouse phrenic nerve diaphragm preparations, all venoms (3-10 mug/mL) produced inhibition of indirect twitches. In the biventer muscle, venoms (10 mug/mL) inhibited responses to acetylcholine (1 mmol/L) and carbachol (20 mumol/L), but not KCl (40 mmol/L). The prior (10 min) administration of 1 unit/mL antivenom markedly attenuated the neurotoxic effects of A. superbus and N. scutatus venoms (10 mug/mL), but was less effective against H. stephensi venom (10 mug/mL); 5 units/mL antivenom attenuated the neurotoxic activity of all venoms. 3. Administration of 5 units/mL antivenom at t(90) partially reversed, over a period of 3 h, the inhibition of twitches produced by N. scutatus (10 mug/mL; 41% recovery), A. superbus (10 mug/mL; 25% recovery) and H. stephensi (10 mug/mL; 50% recovery) venoms. All venoms (10-100 mug/mL) also displayed signs of in vitro myotoxicity. 4. The results of the present study indicate that all three venoms contain neurotoxic activity that is effectively attenuated by tiger snake antivenom.

Comparative analysis of prothrombin activators from the venom of Australian elapids

A key component of the venom of many Australian snakes belonging to the elapid family is a toxin that is structurally and functionally similar to that of the mammalian prothrombinase complex. In mammals, this complex is responsible for the cleavage of prothrombin to thrombin and is composed of factor Xa in association with its cofactors calcium, phospholipids, and factor Va. The snake prothrombin activators have been classified on the basis of their requirement for cofactors for activity. The two major subgroups described in Australian elapid snakes, groups C and D, are differentiated by their requirement for mammalian coagulation factor Va. In this study, we describe the cloning, characterization, and comparative analysis of the factor X- and factor V-like components of the prothrombin activators from the venom glands of snakes possessing either group C or D prothrombin activators. The overall domain arrangement in these proteins was highly conserved between all elapids and with the corresponding mammalian clotting factors. The deduced protein sequence for the factor X-like protease precursor, identified in elapids containing either group C or D prothrombin activators, demonstrated a remarkable degree of relatedness to each other (80%-97%). The factor V-like component of the prothrombin activator, present only in snakes containing group C complexes, also showed a very high degree of homology (96%-98%). Expression of both the factor X- and factor V-like proteins determined by immunoblotting provided an additional means of separating these two groups at the molecular level. The molecular phylogenetic analysis described here represents a new approach for distinguishing group C and D snake prothrombin activators and correlates well with previous classifications.

Ocorrência de Gonipterus scutellatus Gyllenhal (Coleoptera: Curculionidae) em plantações de eucalipto no Estado do Espírirto Santo

O gorgulho do eucalipto Gonipterus scutellatus Gyllenhal foi encontrado no Estado do Espírito Santo atacando plantações clonais de Eucalyptus urophylla x E. grandis (híbrido “urograndis”). Essa espécie de gorgulho é conhecida no Brasil desde 1979, tendo sua distribuição geográfica restrita aos estados da região Sul e São Paulo. A introdução do inseto no Estado do Espírito Santo foi provavelmente acidental e sua ocorrência tem se limitado à região central do Estado. Nos levantamentos de campo não foi detectado o parasitóide de ovos Anaphes nitens (Hymenoptera: Mymaridae), que é o principal inimigo natural do gorgulho do eucalipto.

Biologia de Gonipterus scutellatus (Coleoptera: Curcu- lionidae) em Eucalyptus spp. em diferentes temperaturas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Revision of the genus Physoclypeus Hendel, 1907 (Diptera, Lauxaniidae), with description of seven new species

The genus Physoclypeus Hendel, 1907 has its distribution restricted to the Neotropical region. In this study, its species have been redescribed, three new combinations have been proposed, three lectotypes have been designated, seven new species have been described, and an identification key to the species is presented. An updated list of species of Physoclypeus is presented as: P. annulatus Hendel, 1925; P. coquilletti (Hendel, 1908); P. farinosus (Hendel, 1925); P. flavus (Wiedemann, 1830); P. hendeli sp. nov. (Type locality, Jamaica, N. Irish Town); P. lineatus (Williston, 1896) new comb.; P. montanus (Becker, 1919) new comb.; P. plaumanni sp. nov. (Type locality, Brazil, Santa Catarina); P. risaraldensis sp. nov. (Type locality, Colombia, Risaralda); P. saltensis sp. nov. (Type locality, Argentina, Salta); P. scutellatus (Curran, 1926) new comb.; P. unimaculatus sp. nov. (Type locality, Mexico, Vera Cruz); P. vitattus sp. nov. (Type locality, Brazil, Santa Catarina) and P. zebrinus sp. nov. (Type locality, Costa Rica, Limón).

Sobre o gênero Lophyraspis Stål e descrição de novas espécies (Hemiptera, Aetalionidae, Biturritiinae)

O gênero Lophyraspis e as seguintes espécies, juntamente com algumas mudanças nomenclaturais, são tratados: Lophyraspis Stål, 1869 = Gerridius Fowler, 1896 syn. rest.; Lophyraspis muscaria (Fabricius, 1803) = Gerridius scutellatus Fowler, 1896 syn. nov. = Gerridius abbreviatus Baker, 1907 syn. nov.; Lophyraspis pygmaea (Fabricius, 1803); Lophyraspis scutellata (Fabricius, 1803) = L. cristata Stål, 1869 syn. nov. = L. armata Haviland, 1925 syn. nov.; Lophyraspis spinosa (Funkhouser, 1930) comb. nov. (anteriormente em Mina Walker, 1858); Lophyraspis fenestrata sp. nov. (Brasil, Amazonas); Lophyraspis diminuta sp. nov. (Brasil, Mato Grosso).

Revision of the genus Physoclypeus Hendel, 1907 (Diptera, Lauxaniidae), with description of seven new species

The genus Physoclypeus Hendel, 1907 has its distribution restricted to the Neotropical region. In this study, its species have been redescribed, three new combinations have been proposed, three lectotypes have been designated, seven new species have been described, and an identification key to the species is presented. An updated list of species of Physoclypeus is presented as: P. annulatus Hendel, 1925; P. coquilletti (Hendel, 1908); P. farinosus (Hendel, 1925); P. flavus (Wiedemann, 1830); P. hendeli sp. nov. (Type locality, Jamaica, N. Irish Town); P. lineatus (Williston, 1896) new comb.; P. montanus (Becker, 1919) new comb.; P. plaumanni sp. nov. (Type locality, Brazil, Santa Catarina); P. risaraldensis sp. nov. (Type locality, Colombia, Risaralda); P. saltensis sp. nov. (Type locality, Argentina, Salta); P. scutellatus (Curran, 1926) new comb.; P. unimaculatus sp. nov. (Type locality, Mexico, Vera Cruz); P. vitattus sp. nov. (Type locality, Brazil, Santa Catarina) and P. zebrinus sp. nov. (Type locality, Costa Rica, Limón).