Taxonomia e filogenia das serpentes do gênero Typhlops Oppel, 1811 (Squamata: Typhlopidae) ocorrentes na Mata Atlântica brasileira

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Atividade antiofídica do decocto das folhas de jatropha gossypiifolia l. frente o veneno de bothrops jararaca

Antiophidic activity from decoct of Jatropha gossypiifolia L. leaves against Bothrops jararaca venom. Snakebites are a serious worldwide public health problem. In Latin America, about 90 % of accidents are attributed to snakes from Bothrops genus. Currently, the main available treatment is the antivenom serum therapy, which has some disadvantages such as inability to neutralize local effects, risk of immunological reactions, high cost and difficult access in some regions. In this context, the search for alternative therapies to treat snakebites is relevant. Jatropha gossypiifolia L., a medicinal plant popularly known in Brazil as “pinhão-roxo”, is very used in folk medicine as antiophidic. So, the aim of this study is to evaluate the antiophidic properties of this species against enzymatic and biological activities from Bothrops jararaca snake venom. The aqueous leaf extract of J. gossypiifolia was prepared by decoction. The inhibition studies were performed in vitro, by pre-incubation of a fixed amount of venom with different amounts of extract from J. gossypiifolia for 60 min at 37 °C, and in vivo, through oral or intraperitoneal treatment of animals, in different doses, 60 min before venom injection. The proteolytic activity upon azocasein was efficiently inhibited, indicating inhibitory action upon metalloproteinases (SVMPs) and/or serine proteases (SVSPs). The extract inhibited the fibrinogenolytic activity, which was also confirmed by zymography, where it was possible to observe that the extract preferentially inhibits fibrinogenolytic enzymes of 26 and 28 kDa. The coagulant activity upon fibrinogen and plasma were significantly inhibited, suggesting an inhibitory action upon thrombin-like enzymes (SVTLEs), as well as upon clotting factor activators toxins. The extract prolonged the activated partial thromboplastin time (aPTT), suggesting an inhibitory action toward not only to SVTLEs, but also against endogenous thrombin. The defibrinogenating activity in vivo was efficiently inhibited by the extract on oral route, confirming the previous results. The local hemorrhagic activity was also significantly inhibited by oral route, indicating an inhibitory action upon SVMPs. The phospholipase activity in vitro was not inhibited. Nevertheless, the edematogenic and myotoxic activities were efficiently inhibited, by oral and intraperitoneal route, which may indicate an inhibitory effect of the extract upon Lys49 phospholipase (PLA2) and/ or SVMPs, or also an anti-inflammatory action against endogenous chemical mediators. Regarding the possible action mechanism, was observed that the extract did not presented proteolytic activity, however, presented protein precipitating action. In addition, the extract showed significant antioxidant activity in different models, which could justify, at least partially, the antiophidic activity presented. The metal chelating action presented by extract could be correlated with SVMPs inhibition, once these enzymes are metal-dependent. The phytochemical analysis revealed the presence of sugars, alkaloids, flavonoids, tannins, terpenes and/or steroids and proteins, from which the flavonoids could be pointed as major compounds, based on chromatographic profile obtained by thin layer chromatography (TLC). In conclusion, the results demonstrate that the J. gossypiifolia leaves decoct present potential antiophidic activity, including action upon snakebite local effects, suggesting that this species may be used as a new source of bioactive molecules against bothropic venom.

Análise fitoquímica e avaliação das atividades anti-inflamatória, antipeçonhenta e citotóxica de extratos aquosos de Aspidosperma pyrifolium e Ipomoea asarifolia

Envenomation caused by venomous animals, mainly scorpions and snakes, are a serious matter of public health. Tityus serrulatus is considered the most venomous scorpion in South America because of the high level of toxicity of its venom. It is responsible for causing serious accidents, mainly with kids. The species Bothrops jararaca is a serpent that has in its venom a complex mixture of enzyme, peptides and other molecules. The toxins of the venom of B. jararaca induce local and systemic inflammatory responses. The treatment chosen to serious cases of envenomation is the intravenous administration of the specific antivenom. However, the treatment is not always accessible to those residents in rural areas, so that they use medicinal plant extracts as the treatment. In this context, aqueous extracts, fractions and isolated compounds of Aspidosperma pyrifolium (pereiro) and Ipomoea asarifolia (salsa, salsa-brava), used in popular medicine, were studied in this research to evaluate the anti-inflammatory activity in the peritonitis models induced by carrageenan and peritonitis induced by the venom of the T. serrulatus (VTs), and in the local oedema model and inflammatory infiltrate induced by the venom of the B. jararaca, administrated intravenously. The results of the assays of cytotoxicity, using the MTT, showed that the aqueous extracts from the plant species presented low toxicity to the cells that came from the fibroblast of the mouse embryo (3T3).The chemical analysis of the extracts by High Performance Liquid Chromatography revealed the presence of the rutin flavonoid, in A. pyrifoliu, and rutin, clorogenic acid and caffeic acid, in I. asarifolia. Concerning the pharmacological evaluation, the results showed that the pre-treatment using aqueous extracts and fractions reduced the total leukocyte migration to the abdominal cavity in the peritonitis model caused by the carrageenan and in the peritonitis model induced by the T. serulatus venom. Yet, these groups presented anti-oedematous activity, in the local oedema model caused by the venom of the B. jararaca, and reduced the inflammatory infiltrate to the muscle. The serum (anti-arachnid and anti-bothropic) specific to each venom acted inhibiting the inflammatory action of the venoms and were used as control. The compounds identified in the extracts were also tested and, similar to the plant extracts, showed meaningful anti-inflammatory effects, in the tested doses. Thus, these results are indicating the potential anti-inflammatory activity of the plants studied. This is the first research that evaluated the possible biological effects of the A. pyrifolium and I. asarifolia, showing the biological potential that these species have.

Inibição dos efeitos locais induzidos pelas peçonhas das serpentes Bothrops erythromelas e Bothrops jararaca pelo extrato aquoso das folhas de Jatropha mollissima (Pohl) Bail

Snakebites are a serious public health problem in tropical and subtropical countries and Bothrops genus is responsible for the accidents in Brazil and throughout Latin America (90% of cases). The local effects (pain, edema, hemorrhage and myonecrosis) and systemic (cardiovascular alterations, shock and blood clotting disorders) caused by the venom of Bothrops are due to the numerous protein and non-protein components, which are part of the constitution of the poison. The only form of therapy is scientifically validated antivenom serum therapy which, however, is not effective with respect to local effects produced, risk of immunological reactions, high cost and difficult access in some regions. Thus, the search for new alternatives to serum therapy becomes important, and in this context, many medicinal plants have been highlighted by the popular use as antiophidic. Among these plants, we can mention the species Jatropha mollissima (Euphorbiaceae) which has popular use in traditional medicine as antiophidic, anti-inflammatory, antimicrobial and antipyretic. Therefore, this study aims to evaluate the neutralizing potential of local effects induced by the venom of Bothrops erythromelas and Bothrops jararaca with the aqueous extract of the leaves of J. mollissima. The leaf extracts were prepared by decoction, fractionated (by liquid-liquid partition) and characterized by thin layer chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Antiophidic activity of the extract was evaluated in model of paw edema, peritonitis, bleeding and myotoxicity induced by venoms of B. jararaca and B. erythromelas. In all models, the extract was evaluated by intraperitoneal route at the doses of 50, 100 and 200 mg/kg, administered 30 minutes prior to injection of the venom (pretreatment protocol). Stains suggestive of the presence of flavonoids: apigenin, luteolin, orientin, isoorientin, vitexin and vitexin-2-O-rhamnoside were detected in the extract by co-CCD. By means of HPLC were identified isoorientin, orientin, vitexin and isovitexin. All tested doses of J. mollissima extract reduced the paw edema induced by the venom with intensity similar to dexamethasone. The aqueous extract of J. mollissima leaves on all evaluated doses, inhibited cell migration induced by B. jararaca and B. erythromelas promoting inhibition of recruitment of mononuclear cells and the polymorphonuclear cells. Local bleeding induced by B. jararaca venom was significantly inhibited by the extract. Both venoms were inhibited by the extract in myotoxic activity. These results indicate that the aqueous extract of J. mollissima leaves have snakebite potential, particularly with respect to local effects, which may justify the use of this plant in traditional medicine and complementary therapy as anti-venom serum.

Inibição dos efeitos locais induzidos pelas peçonhas das serpentes Bothrops erythromelas e Bothrops jararaca pelo extrato aquoso das folhas de Jatropha mollissima (Pohl) Bail

Snakebites are a serious public health problem in tropical and subtropical countries and Bothrops genus is responsible for the accidents in Brazil and throughout Latin America (90% of cases). The local effects (pain, edema, hemorrhage and myonecrosis) and systemic (cardiovascular alterations, shock and blood clotting disorders) caused by the venom of Bothrops are due to the numerous protein and non-protein components, which are part of the constitution of the poison. The only form of therapy is scientifically validated antivenom serum therapy which, however, is not effective with respect to local effects produced, risk of immunological reactions, high cost and difficult access in some regions. Thus, the search for new alternatives to serum therapy becomes important, and in this context, many medicinal plants have been highlighted by the popular use as antiophidic. Among these plants, we can mention the species Jatropha mollissima (Euphorbiaceae) which has popular use in traditional medicine as antiophidic, anti-inflammatory, antimicrobial and antipyretic. Therefore, this study aims to evaluate the neutralizing potential of local effects induced by the venom of Bothrops erythromelas and Bothrops jararaca with the aqueous extract of the leaves of J. mollissima. The leaf extracts were prepared by decoction, fractionated (by liquid-liquid partition) and characterized by thin layer chromatography (TLC) and High Performance Liquid Chromatography (HPLC). Antiophidic activity of the extract was evaluated in model of paw edema, peritonitis, bleeding and myotoxicity induced by venoms of B. jararaca and B. erythromelas. In all models, the extract was evaluated by intraperitoneal route at the doses of 50, 100 and 200 mg/kg, administered 30 minutes prior to injection of the venom (pretreatment protocol). Stains suggestive of the presence of flavonoids: apigenin, luteolin, orientin, isoorientin, vitexin and vitexin-2-O-rhamnoside were detected in the extract by co-CCD. By means of HPLC were identified isoorientin, orientin, vitexin and isovitexin. All tested doses of J. mollissima extract reduced the paw edema induced by the venom with intensity similar to dexamethasone. The aqueous extract of J. mollissima leaves on all evaluated doses, inhibited cell migration induced by B. jararaca and B. erythromelas promoting inhibition of recruitment of mononuclear cells and the polymorphonuclear cells. Local bleeding induced by B. jararaca venom was significantly inhibited by the extract. Both venoms were inhibited by the extract in myotoxic activity. These results indicate that the aqueous extract of J. mollissima leaves have snakebite potential, particularly with respect to local effects, which may justify the use of this plant in traditional medicine and complementary therapy as anti-venom serum.

Herpetofauna associada à bromélia rupícola, encholirium spectabile, no semiárido brasileiro: revisão da literatura, ecologia das espécies e comportamento de psychosaura agmosticha (squamata: mabuyidae)

Bromeliads are an important microhabitat for the herpetofauna, for being widely used as refuge from predators and their leaf architecture allows humidity maintenance and relatively constant temperature inside, setting a favorable environment for amphibians and reptiles, especially in areas under hydric stress. However, studies addressing this relationship are still incipient and more concentrated in fitotelmatas bromeliad. For non-fitotelmatas rupicolous bromeliads of the gender Encholirium, which develops into rocky outcrops and contains species of semi-arid regions such as the Caatinga, animal-plant relationships are almost unknown. In this context, this study aimed to know the herpetological fauna inhabitant of macambiras bromeliads, Encholirium spectabile, analyzing occupation and use of these bromeliads by different taxa, and the behavioral ecology of the lizard Psychosaura agmosticha, seeking to identify factors associated with this strict relationship in Caatinga. An extensive review of the world literature on the subject “lizards in bromeliads” subsidized this study from the ecological perspective of this association. The field work was carried out at Fazenda Tanques, municipality of Santa Maria / RN, mesoregion of Agreste Potiguar. The observations and/or data collection in daytime and in the evening was conducted monthly during three consecutive days, from January 2011 to August 2012, totaling 450 hour.man of sampling effort. Sixteen species were registered: six lizards (Mabuyidae, Tropiduridae, Gekkonidae and Phyllodactylidae Families), six snakes (Boidae and Dipsadidae Families) and four of amphibians of Hylidae Family. The effect of the forest edge on the distribution of species along the outcrop was significant, with most species found in outcrop edges. Significant difference was found between some pairs of species concerning use of bromeliads, and almost total niche overlap in the use of microhabitat. 62.5% of the species are nocturnal and use these plants for sheltering, breeding and feeding. Regarding the relations between 4 the lizard Psychosaura agmosticha and macambiras bromeliads, behaviors of thermoregulation and foraging in the dry and wet seasons were recorded. Activity periods were concentrated between 7 and 10 am and between 3 and 5 pm in both seasons, showing a clear bimodal pattern. The species basically used the green leaves and there were no significant differences between males and females in the use of bromeliads. Positive associations were found between body temperature and temperatures of bromeliads and air. This species spent 1.95% ± 3.8 of the time moving (PTM) and moved on average 0:36 ± 2.1 seconds per minute (MPM), with significant differences between the wet and dry to PTM, and between the average time of stop and average duration of movements, being considered a sedentary forager. Psychosaura agmosticha, in the study area, is bromelicolous and uses macambiras primarily for thermoregulation and foraging. The results of this study elevate the rupicolous bromeliads Encholirium spectabile as key elements for the maintenance of amphibians and reptiles associated with it, and a clear advantageous association for the conservation of the groups involved.

Efeito anticonvulsivante de frações isoladas da peçonha da formiga Dinoponera quadriceps (Formicidae: Ponerinae)

A epilepsia é uma patologia crônica do sistema nervoso central que afeta cerca de 65 milhões de indivíduos no mundo. Aproximadamente 30% desses indivíduos desenvolvem crises convulsivas que persistem apesar do tratamento monitorado com drogas antiepiléptica s. Assim, há uma evidente necessidade do desenvolvimento de novos fármacos antiepilépticos e as peçonhas podem ser uma excelente fonte de modelos. Nesse contexto, enquanto já vários estudos sobre peçonhas de serpentes, escorpiões e aranhas, pouco se sabe s obre as peçonhas de formigas. Estudos prévios do nosso laboratório demonstraram que a peçonha desnaturada da formiga Dinoponera quadríceps protegeu camundongos de crises convulsivas e morte induzidas por bicuculina (BIC). Nesse contexto, o objetivo desse t rabalho foi investigar o potencial anticonvulsivante de frações isoladas da peçonha de D. quadríceps em crises convulsivas induzidas pela BIC, bem como uma análise dos efeitos dessas frações no comportamento natural dos camundongos no campo aberto. Os anim ais foram divididos em grupos, os quais receberam injeções (1 mg/ml i.c.v.) de seis frações distintas e tiveram seu comportamento geral observado no campo aberto durante 3 0 min. No segundo experimento, o s animais receberam as mesmas frações 20 min antes da administração de bicuculina (10 mg/ml). Em seguida, foi analisado o comportamento motor convulsivo desses animais durante 30 minutos no campo aberto. No primeiro experimento, não foram observadas alterações comportamentais. Já no segundo experimento, a ad ministração prévia de DqTx1, DqTx3, DqTx4 e DqTx6 aumentou a latência para o desenvolvimento de crises tônico - clônicas. Além disso, todas as frações, exceto DqTx5, aumentaram a latência para a morte dos animais. Ainda, os melhores resultados foram obtidos com a fração DqTx6, que protegeu 62,5% dos animais testados contra o desenvolvimento de crises tônico - clônicas e 100% dos animais contra a morte.