Defensive stinging by Parabuthus transvaalicus scorpions: risk assessment and venom metering

Venom is a metabolically expensive commodity that animals should use judiciously. The venom-metering hypothesis proposes that venomous animals control, or meter, the quantity of venom they deploy during predatory or defensive situations. We sought to clarify experimentally whether the buthid scorpion Parabuthus transvaalicus can regulate defensive venom expenditure based on perceived risk. Scorpions were tested under two threat conditions by inducing them to sting repeatedly a parafilm-covered cup. The high-threat condition involved five sting presentations at 5 s intervals, and the low-threat condition comprised five sting presentations at 5 min intervals. Venom metering appeared to be modulated at three levels: wet versus dry sting, composition of venom injected and volume of venom delivered. Scorpions delivered dry stings more often under the low-threat condition, but in both conditions were more likely to employ wet stings as the threat persisted. Venom appearance changed during successive stings from clear (potassium-rich ‘prevenom’), to opalescent, to milky (protein-rich ‘venom’), with biochemical analyses (protein assay and MALDI-TOF) confirming the compositional changes. However, progression through this sequence depended on quantity of venom expended, with milky secretion appearing only after the limited quantity of clear secretion was exhausted (i.e. composition and volume covaried). Scorpions injected approximately two-fold more venom per sting during the high-threat compared to the low-threat condition, with milky venom appearing more quickly within the sequence of five stings for the high-threat condition. Thus, these scorpions perceive risk and regulate venom expenditure during stinging according to level of threat, providing further support for the venom-metering hypothesis.