This study was undertaken to develop an experimental protocol using insects as biological models to assay venom toxicity of the following spiders Loxosceles gaucho, Phoneutria nigriventer, Nephilengys cruentata and Tityus serrulatus scorpion. Three different insect species were bioassayed: Apis mellifera (Hymenoptera), Grillus assimilis (Orthoptera), and Diatraea saccharalis (Lepidoptera). Venoms were injected into the hemocele of insects with a microsyringe at concentrations that caused dose/weight-dependent effects; doses causing either paralysis (ED50) or death (LD50) were recorded for each venom and insect test-species. T. serrulatus and L. gaucho venoms were lethal to all tested species, while P. nigriventer venom caused paralysis and death, and N. cruentata venom caused only paralysis at the doses assayed. A comparison between the insect test species described above revealed that A. mellifera was highly sensitive to all venoms tested; even a tiny amount of N. cruentata non-lethal venom caused a change in the walking pattern leading to transient paralysis. D. saccharalis larvae were very resistant to all four venoms
Field bioassays were used to demonstrate that aggressive behavior of Polybia paulista (Ihering) workers is elicited by alarm pheromones present in the venom reservoirs of nest defenders and that the brood care pheromone (pupal odor) produced by the young inside the nest also plays an important defensive role. Pupal odor was extracted from the surface of pupa bodies with methanol. When bioassayed alone, the pupal odor elicited only attractiveness of workers towards the odor source, but no stinging attacks were observed. However, in the presence of alarm pheromones, the brood care pheromone potentiated the effect caused by the pupal odors, increasing the number of stinging attacks during an action of colony defense. Thus, the presence of pupae within the nest evidently not only releases brood care but also enhances the aggressiveness of workers in P. paulista colonies
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