Insecticidal activity of Leptodactylus knudseni and Phyllomedusa vaillantii crude skin secretions against the mosquitoes Anopheles darlingi and Aedes aegypti.

BACKGROUND: Mosquitoes are important vectors of several diseases, including malaria and dengue, and control measures are mostly performed using chemical insecticides. Unfortunately, mosquito resistance to commonly applied insecticides is widespread. Therefore, a prospection for new molecules with insecticidal activity based on Amazon biodiversity using the anurans Leptodactylus knudseni and Phyllomedusa vaillantii was performed against the mosquito species Anopheles darlingi and Aedes aegypti. METHODS: The granular secretion from anuran skin was obtained by manual stimulation, and lethal concentrations (LCs) for larvicidal and adulticidal tests were calculated using concentrations from 1-100 ppm. The skin secretions from the anuran species tested caused significant mortality within the first 24 hours on adults and larvae, but differed within the mosquito species. RESULTS: The skin secretions from the anuran species tested caused significant mortality within the first 24 hours on adults and larvae, but differed within the mosquito species. The calculated LC50 of L. knudseni skin secretions against An. darlingi was 0.15 and 0.2 ppm for adults and larvae, respectively, but much higher for Ae. aegypti, i.e., 19 and 38 ppm, respectively. Interestingly, the calculated LCs50 of P. vaillantii against both mosquito species in adults were similar, 1.8 and 2.1 ppm, respectively, but the LC50 for An. darlingi larvae was much lower (0.4 ppm) than for Ae aegypti (2.1 ppm). CONCLUSIONS: The present experiments indicate that skin secretions from L. knudseni and P. vaillantii contain bioactive molecules with potent insecticide activity. The isolation and characterization of skin secretions components will provide new insights for potential insecticidal molecules.

Antitumoral activity of snake venom proteins: new trends in cancer therapy.

For more than half a century, cytotoxic agents have been investigated as a possible treatment for cancer. Research on animal venoms has revealed their high toxicity on tissues and cell cultures, both normal and tumoral. Snake venoms show the highest cytotoxic potential, since ophidian accidents cause a large amount of tissue damage, suggesting a promising utilization of these venoms or their components as antitumoral agents. Over the last few years, we have studied the effects of snake venoms and their isolated enzymes on tumor cell cultures. Some in vivo assays showed antineoplastic activity against induced tumors in mice. In human beings, both the crude venom and isolated enzymes revealed antitumor activities in preliminary assays, with measurable clinical responses in the advanced treatment phase. These enzymes include metalloproteases (MP), disintegrins, L-amino acid oxidases (LAAOs), C-type lectins, and phospholipases A2 (PLA2s). Their mechanisms of action include direct toxic action (PLA2s), free radical generation (LAAOs), apoptosis induction (PLA2s, MP, and LAAOs), and antiangiogenesis (disintegrins and lectins). Higher cytotoxic and cytostatic activities upon tumor cells than normal cells suggest the possibility for clinical applications. Further studies should be conducted to ensure the efficacy and safety of different snake venom compounds for cancer drug development.

Purification and biochemical characterization of three myotoxins from Bothrops mattogrossensis snake venom with toxicity against Leishmania and tumor cells.

Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s), one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like), BmatTX-II (Lys49 PLA2-like), and BmatTX-III (Asp49 PLA2). The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T) and SK-BR-3 (breast adenocarcinoma) cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.