Abstract. Chemical and visual defenses are used by many organisms to avoid being approached or eaten by predators. An example is inking molluscs-including gastropods such as sea hares and cephalopods such as squid, cuttlefish, and octopus-which release a colored ink upon approach or attack. Previous work showed that ink can protect molluscs through a combination of chemical, visual, and other effects. In this study, we examined the effects of ink from longfin inshore squid, Doryteuthis pealeii, on the behavior of two species of predatory fishes, summer flounder, Paralichthys dentatus, and sea catfish, Ariopsis felis. Using a cloud assay, we found that ink from longfin inshore squid affected the approach phase of predation by summer flounder, primarily through its visual effects. Using a food assay, we found that the ink affected the consummatory and ingestive phase of predation of both sea catfish and summer flounder, through the ink's chemical properties. Fractionation of ink showed that most of its deterrent chemical activity is associated with melanin granules, suggesting that either compounds adhering to these granules or melanin itself are the most biologically active. This work provides the basis for a comparative approach to identify deterrent molecules from inking cephalopods and to examine neural mechanisms whereby these chemicals affect behavior of fish, using the sea catfish as a chemosensory model.

SUMMARYAntipredator defenses are ubiquitous and diverse. Ink secretion of sea hares (Aplysia) is an antipredator defense acting through the chemical senses of predators by different mechanisms. The most common mechanism is ink acting as an unpalatable repellent. Less common is ink secretion acting as a decoy (phagomimic) that misdirects predatorsʼ attacks. In this study, we tested another possible mechanism -sensory inactivation -in which ink inactivates the predatorʼs reception of food odors associated with would-be prey. We tested this hypothesis using spiny lobsters, Panulirus argus, as model predators. Ink secretion is composed of two glandular products, one being opaline, a viscous substance containing concentrations of hundreds of millimolar of total free amino acids. Opaline sticks to antennules, mouthparts and other chemosensory appendages of lobsters, physically blocking access of food odors to the predatorʼs chemosensors, or over-stimulating (short term) and adapting (long term) the chemosensors. We tested the sensory inactivation hypotheses by treating the antennules with opaline and mimics of its physical and/or chemical properties. We compared the effects of these treatments on responses to a food odor for chemoreceptor neurons in isolated antennules, as a measure of effect on chemosensory input, and for antennular motor responses of intact lobsters, as a measure of effect on chemically driven motor behavior. Our results indicate that opaline reduces the output of chemosensors by physically blocking reception of and response to food odors, and this has an impact on motor responses of lobsters. This is the first experimental demonstration of inactivation of peripheral sensors as an antipredatory defense. Supplementary material available online at