Aspectos da ecologia reprodutiva de Trachycephalus cunauaru (Anura: Hylidae) na Amazônia meridional / Aspects of the reproductive ecology of Trachycephalus cunauaru (Anura: Hylidae) in the southern Amazon

Trachycephalus cunauaru é um hilídeo amazônico que utiliza fitotelmatas para se reproduzir. Existem relativamente poucas informações sobre a espécie, principalmente devido à dificuldade de acesso aos seus sítios reprodutivos. Nesse trabalho, reunimos dados de ecologia e história natural de T. cunauaru em uma área da Amazônia meridional, no estado de Mato Grosso, Brasil. Além de fitotelmatas naturais, utilizamos baldes instalados a uma altura de 10 m como fitotelmatas artificiais. Comparamos características físicas e químicas, bem como a presença e ausência de girinos, entre fitotelmatas naturais e artificiais. Também coletamos dados sobre o comportamento reprodutivo da espécie por meio de armadilhas fotográficas. Registramos uma densidade de 14,1 sítios reprodutivos por km2 . Os parâmetros ambientais diferiram significativamente entre fitotelmatas artificiais e naturais. Em sítios artificiais a presença de girinos esteve diretamente relacionada à árvores com maior diâmetro. Registramos, pela primeira vez, oofagia para a espécie e observamos que machos podem utilizar mais de um fitotelmata. Também registramos a presença de cobras nos sítios reprodutivos. Constatamos que sítios artificiais e armadilhas fotográficas representam uma alternativa satisfatória para registros comportamentais para T. cunauaru e, possivelmente, para outras espécies com hábitos similares. (AU)

Marinobufagin, a molecule from poisonous frogs, causes biochemical, morphological and cell cycle changes in human neoplasms and vegetal cells.

Skin toad secretion present physiologically active molecules to protect them against microorganisms, predators and infections. This work detailed the antiproliferative action of marinobufagin on tumor and normal lines, investigate its mechanism on HL-60 leukemia cells and its toxic effects on Allium cepa meristematic cells. Initially, cytotoxic action was assessed by colorimetric assays. Next, HL-60 cells were analyzed by morphological and flow cytometry techniques and growing A. cepa roots were examined after 72 h exposure. Marinobufagin presented high antiproliferative action against all human tumor lines [IC50 values ranging from 0.15 (leukemia) to 7.35 (larynx) µM] and it failed against human erythrocytes and murine lines. Human normal peripheral blood mononuclear cells (PBMC) were up to 72.5-fold less sensitive [IC50: 10.88 µM] to marinobufagin than HL-60 line, but DNA strand breaks were no detected. Leukemia treaded cells exhibited cell viability reduction, DNA fragmentation, phosphatidylserine externalization, binucleation, nuclear condensation and cytoplasmic vacuoles. Marinobufagin also reduced the growth of A. cepa roots (EC50: 7.5 µM) and mitotic index, caused cell cycle arrest and chromosomal alterations (micronuclei, delays and C-metaphases) in meristematic cells. So, to find out partially targeted natural molecules on human leukemia cells, like marinobufagin, is an amazing and stimulating way to continue the battle against cancer.

Antiplasmodial and Cytotoxic Activities of Toad Venoms from Southern Amazon, Brazil.

The drug-resistance of malaria parasites is the main problem in the disease control. The huge Brazilian biodiversity promotes the search for new compounds, where the animal kingdom is proving to be a promising source of bioactive compounds. The main objective of this study was to evaluate the antiplasmodial and cytotoxic activity of the compounds obtained from the toad venoms of Brazilian Amazon. Toad venoms were collected from the secretion of Rhinella marina and Rhaebo guttatus in Mato Grosso State, Brazil. The powder was extracted at room temperature, yielding 2 extracts (RG and RM) and a substance ('1') identified as a bufadienolide, named telocinobufagin. Growth inhibition, intraerythrocytic development, and parasite morphology were evaluated in culture by microscopic observations of Giemsa-stained thin blood films. Cytotoxicity was determined against HepG2 and BGM cells by MTT and neutral red assays. The 2 extracts and the pure substance ('1') tested were active against chloroquine-resistant Plasmodium falciparum strain, demonstrating lower IC50 values. In cytotoxic tests, the 2 extracts and substance '1' showed pronounced lethal effects on chloroquine-resistant P. faciparum strain and low cytotoxic effect, highlighting toad parotoid gland secretions as a promising source of novel lead antiplasmodial compounds.

Predation of bat (Molossus molossus: Molossidae) by the centipede Scolopendra viridicornis (Scolopendridae) in Southern Amazonia / Predação de morcego (Molossus molossus: Molossidae) pela centopeia Scolopendra viridicornis (Scolopendridae) no sul da Amazônia

Centipedes are opportunistic carnivore predators, and large species can feed on a wide variety of vertebrates, including bats. The aim of this study was to report the third record of bat predation by centipedes worldwide, the first record in the Amazon region, while covering aspects of foraging, capture and handling of prey. We observed the occurence in a fortuitous encounter at Cristalino State Park, located in the Amazon region of the state of Mato Grosso, Brazil. The attack took place in a small wooden structure, at about three meters from the floor, and was observed for 20 minutes. During the observation, the centipede stung the neck and abdominal region of the bat several times, grabbing the prey with its 15 pairs of front legs while hanging from the ceiling with its hind legs. This type of observation suggests that vertebrates can be important preys for invertebrates such as giant centipedes, both for its nutritional composition and for the amount of energy available in a single prey.

Centopeias são predadores carnívoros oportunistas e grandes espécies podem se alimentar de uma grande variedade de vertebrados, inclusive de morcegos. O objetivo desse trabalho é reportar o terceiro registro de predação de morcego por centopeia, e o primeiro para a região Amazônica, abrangendo aspectos do forrageamento, captura e manipulação da presa. O registro foi realizado de forma fortuita, no Parque Estadual do Cristalino, região de Floresta Amazônica, no estado de Mato Grosso, Brasil. O ataque ocorreu em uma estrutura de madeira a cerca de três metros de altura e foi observado por 20 minutos. Durante a observação, a centopeia picou várias vezes o pescoço e região abdominal do morcego, agarrando a presa com seus 15 pares de pernas anteriores e se pendurando ao teto com as pernas posteriores. Esse tipo de observação sugere que vertebrados podem ser importantes presas para invertebrados, como centopeias gigantes, tanto por sua composição nutricional quanto pela quantidade de energia disponível em uma única presa.

Antiproliferative activity of Rhinella marina and Rhaebo guttatus venom extracts from Southern Amazon.

The venom of amphibians is a fascinating source of active substances. In view of their medical importance and aiming to explore the amazing Brazilian biodiversity, we conducted bioprospecting of antiproliferative activity in extracts of Rhinella marina and Rhaebo guttatus toads occurring in the Southern Amazon of Mato Grosso, Brazil. LC-MS and HPLC analysis of the venom extracts of R. marina revealed four bufadienolides (telocinobufagin, marinobufagin, bufalin and resibufogenin. R. guttatus venom extracts contained only marinobufagin. First, R. marina and R. guttatus venom extracts were evaluated for cytotoxicity against tumor cell lines by the MTT assay. All extracts revealed cytotoxicity, where R. marina extracts were comparable to doxorubicin (IC50 values ranging from 0.01 to 0.23 µg/mL). Only extracts of R. guttatus toad venom caused membrane disruption of human erythrocytes. The extracts were investigated for selective activity by determining their effect on stimulated human peripheral blood mononuclear cells (PBMC) with the Alamar Blue™ assay. The extracts were up to 80-fold more selective against leukemia cells when compared to dividing leukocytes. Aiming to confirm these antiproliferative effects, BrdU incorporation into DNA was measured in HL-60 treated cells with R. marina venom extracts. These extracts decreased BrdU incorporation at both concentrations tested. In summary, nine extracts of R. marina and R. guttatus venom showed pronounced lethal and discriminating effects on tumor lines, especially those from R. marina, highlighting toad parotoid gland secretions as a promising source for novel lead anticancer chemicals.