Caso sospechoso de envenenamiento por araña reclusa (loxosceles) y revisión de la literatura

El Loxoscelismo en una enfermedad causada por la picadura de una araña del género Loxosceles, que se puede manifestar con compromiso cutáneo y/o sistémico e incluso llevar a la muerte. La distribución geográfica de la araña es mundial, con mayor incidencia en países como Perú, Chile y Brasil, y en algunos lugares de Norte América. En Colombia, se ha identificado este tipo de arácnido, sin embargo no se ha reportado ningún caso, se cree que probablemente exista subregistro. El cuadro clínico puede ir desde pacientes asintomáticos hasta dermonecrosis severa, con gran riesgo de secuelas e incapacidad funcional. Cuando hay compromiso sistémico, cursa con falla renal, hemólisis, coagulación intravascular diseminada y alto riesgo de muerte. El diagnóstico es clínico y epidemiológico, pues no hay pruebas serológicas confirmatorias disponibles. A pesar de que se han descrito múltiples opciones terapéuticas, no hay un consenso que permita hacer recomendaciones, por la escasa evidencia científica que existe al respecto. Este artículo presenta el caso de un varón de 11 años, con antecedente de picadura por animal desconocido, con evolución clínica sugestiva de loxoscelismo cutáneo y sistémico, que recibió manejo con suero antiarácnido, oxígeno hiperbárico, inhibidor de polimorfonucleares y reconstrucción quirúrgica. Se revisa la literatura y la situación de la enfermedad en Colombia, con el fin de llamar la atención sobre una enfermedad poco conocida y favorecer la sospecha diagnóstica y el enfoque adecuado.

Loxosceles chapadensis (Araneae: Sicariidae): a new recluse spider species of the gaucho group from Brazil

A new species of the medically important recluse spider genus Loxosceles Heinecken & Lowe 1832 is described from the State of Bahia, Brazil. The species occurs between rocks and crevices, as well as in and around man-made structures. The new species belongs to the gaucho group, as evidenced by the spermathecal shape and color pattern. The presence of a long male palpal tibia is unusual in the gaucho group; thus, the inclusion of the new species in this group is discussed.

Necrotic skin lesion in a dog attributed to Loxosceles (brown spider) bite: a case report

Envenomations caused by Loxosceles (brown spider) have been reported throughout the world. Clinical signs associated to bites of these spiders involve dermonecrotic lesions and intense local inflammatory response, besides systemic manifestations such as intravascular hemolysis, thrombocytopenia, disseminated intravascular coagulation and acute renal failure. The present study aimed to report and to describe dermonecrotic lesions probably caused by a Loxosceles envenomation in a four year-old poodle female dog, treated at the Dermatology Service of the Veterinary Hospital of the Veterinary Medicine and Animal Husbandry School, São Paulo State University, Botucatu, Brazil. Initially, the animal presented two skin lesions with blackish aspect that evolved into ulcerative crusts. The owner reported the presence of a brown spider near the place where the animal spent most of the time. Histological examination of lesions revealed necrosis of the epidermis extending to adnexa and panniculi, which is compatible with Loxosceles bite reaction. The animal was treated with systemic antibiotic and local curatives. Lesions healed by second intention in two months.

Effect of architectural angularity on refugia selection by the brown spider, Loxosceles gaucho

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.

SMase II, a new sphingomyelinase D from Loxosceles laeta venom gland: Molecular cloning, expression, function and structural analysis

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

Crystallization and preliminary X-ray diffraction analysis of a class II phospholipase D from Loxosceles intermedia venom

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

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.

Kinetic and mechanistic characterization of the Sphingomyelinases D from Loxosceles intermedia spider venom

Envenomation by arachnids of the genus Loxosceles leads to local dermonecrosis and serious systemic toxicity mainly induced by sphingomyelinases D (SMase D). These enzymes catalyze the hydrolysis of sphingomyelin resulting in the formation of ceramide-phosphate and choline as well as the cleavage of lysophosphatidyl choline generating the lipid mediator lysophosphatidic acid. We have, previously, cloned and expressed two functional SMase D isoforms, named P1 and P2, from Loxosceles intertnedia venom and comparative protein sequence analysis revealed that they are highly homologous to SMase I from Loxosceles laeta which folds to form an (alpha/beta)(8) barrel. In order to further characterize these proteins, pH dependence kinetic experiments and chemical modification of the two active SMases D isoforms were performed. We show here that the amino acids involved in catalysis and in the metal ion binding sites are strictly conserved in the SMase D isoforms from L. intermedia. However, the kinetic studies indicate that SMase P1 hydrolyzes sphingomyelin less efficiently than P2, which can be attributed to a substitution at position 203 (Pro-Leu) and local amino acid substitutions in the hydrophobic channel that could probably play a role in the substrate recognition and binding. (c) 2005 Elsevier Ltd. All rights reserved.

Sistematização da Assistência de Enfermagem: acidente por Loxosceles gaucho.

Experience report carried out at a university hospital involving a patient victimized by an accident with a spider of the Loxosceles gaucho genus. This type of accident can be classified as mild, moderate and severe, depending on the period of time elapsed between the occurrence of the accident and the moment of care provision. We aimed at applying nursing care systematization in a comprehensive and humanized manner. The following nursing diagnoses were established: acute pain, damaged skin integrity, risks for infection, constipation and low self-esteem. The therapeutic relationship favored student/patient interactions and enabled the recognition of the needs that deserved nursing interventions. The nursing process was a valuable instrument and provided important elements for the patient's daily development and planning adjustment by prioritizing care quality.

Crystallization and preliminary X-ray diffraction analysis of a novel sphingomyelinase D from Loxosceles gaucho venom

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Disputa por fêmeas entre machos da aranha marrom Loxosceles gaucho (Araneae: Sicariidae)

O comportamento agonístico de machos de Loxosceles gaucho (“aranha-marrom”), sem assimetrias de dominância de território e experiência prévia, minimizando-se a diferença de tamanho, foi avaliado em presença do recurso fêmea. Reavaliamos a importância da diferença de tamanho e pudemos sugerir o papel relevante da reação do animal a ambiente novo (primeiro a movimentar-se) na disputa por fêmea reprodutiva. Vimos que a competição por fêmeas se resolve por disputas pouco intensas, ou até mesmo sem qualquer interação observável entre os machos. Os dados mostram uma disputa que prioriza a economia de energia e impõe menor risco de danos físicos aos animais, característica possivelmente associada ao hábito gregário de L. gaucho.

Sphingomyelinase D/Ceramide 1-Phosphate in Cell Survival and Inflammation

Sphingolipids are major constituents of biological membranes of eukaryotic cells. Many studies have shown that sphingomyelin (SM) is a major phospholipid in cell bilayers and is mainly localized to the plasma membrane of cells, where it serves both as a building block for cell architecture and as a precursor of bioactive sphingolipids. In particular, upregulation of (C-type) sphingomyelinases will produce ceramide, which regulates many physiological functions including apoptosis, senescence, or cell differentiation. Interestingly, the venom of some arthropodes including spiders of the genus Loxosceles, or the toxins of some bacteria such as Corynebacterium tuberculosis, or Vibrio damsela possess high levels of D-type sphingomyelinase (SMase D). This enzyme catalyzes the hydrolysis of SM to yield ceramide 1-phosphate (C1P), which promotes cell growth and survival and is a potent pro-inflammatory agent in different cell types. In particular, C1P stimulates cytosolic phospholipase A2 leading to arachidonic acid release and the subsequent formation of eicosanoids, actions that are all associated to the promotion of inflammation. In addition, C1P potently stimulates macrophage migration, which has also been associated to inflammatory responses. Interestingly, this action required the interaction of C1P with a specific plasma membrane receptor, whereas accumulation of intracellular C1P failed to stimulate chemotaxis. The C1P receptor is coupled to Gi proteins and activates of the PI3K/Akt and MEK/ERK1-2 pathways upon ligation with C1P. The proposed review will address novel aspects on the control of inflammatory responses by C1P and will highlight the molecular mechanisms whereby C1P exerts these actions.

Lonomia obliqua (Lepidoptera, Saturniidae) caterpillar bristle extract induces direct lysis by cleaving erythrocyte membrane glycoproteins

Lonomia obliqua caterpillar bristle extract induces hemolysis in vitro on washed human and rat erythrocytes, in either the absence or presence of exogenous lecithin. In the former condition, phospholipases A(2) are key enzymes involved in hemolysis. However, the mechanism whereby this extract causes direct hemolysis is not known. Thus, the aim of this study was to investigate the hemolytic mechanism of the crude extract of the caterpillar L obliqua on human erythrocytes in the absence of lecithin. The extract significantly increased the erythrocyte osmotic fragility and promoted the removal of glycophorins A and C, and band 3 from the erythrocyte membrane. The use of Ca(2+) and Mg(2+) ions significantly potentiated glycoprotein removal, remarkably of erythrocyte band 3. The composition of fatty acids was analyzed by HPLC in both L obliqua caterpillar bristle extract and human erythrocyte membranes incubated with the extract. The levels of unsaturated fatty acids were remarkably augmented in erythrocytes incubated with the extract than in control erythrocytes, modifying thereby the saturated/unsaturated fatty acid ratio. Altogether, evidence is provided here that the interplay of at least three mechanisms of action accounts for the direct activity of the bristle extract on erythrocyte membrane, leading to hemolysis: the removal of glycoproteins and band 3; the insertion of fatty acids; and the action of phospholipases. Such mechanisms might affect erythrocyte flexibility and deformability, which may induce hemolysis by increasing erythrocyte fragility. However, whether the direct hemolytic activity of L obliqua caterpillar is the major cause of intravascular hemolysis during envenomation still needs further investigation. (C) 2010 Elsevier Ltd. All rights reserved.

Brown Recluse Spider Venom: Proteomic Analysis and Proposal of a Putative Mechanism of Action

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