Metamorphosis in balanomorphan, pedunculated, and parasitic barnacles:a video-based analysis

Cypris metamorphosis was followed using video microscopy in four species of cirripeds representing the suspension-feeding pedunculated and sessile Thoracica and the parasitic Rhizocephala. Cirripede metamorphosis involves one or more highly complex molts that mark the change from a free cypris larva to an attached suspension feeder (Thoracica) or an endoparasite (Rhizocephala). The cyprids and juveniles are so different in morphology that they are functionally incompatible. The drastic reorganization of the body implicated in the process can therefore only commence after the cyprid has irreversibly cemented itself to a substratum. In both Megabalanus rosa and Lepas, the settled cyprid first passes through a quiescent period of tissue reorganization, in which the body is raised into a position vertical to the substratum. In Lepas, this is followed by extension of the peduncle. In both Lepas and M. rosa, the juvenile must free itself from the cypris cuticle by an active process before it can extend the cirri for suspension feeding. In M. rosa, the juvenile performs intensely pulsating movements that result in shedding of the cypris carapace ∼8 h after settlement. Lepas sp. sheds the cypris cuticle ∼2 days after settlement due to contractile movements of the peduncle. In Lepas anserifera, the juvenile actively breaks through the cypris carapace, which can thereafter remain for several days without impeding cirral feeding. Formation of the shell plates begins after 1-2 days under the cyprid carapace in Lepas. In M. rosa, the free juvenile retains its very thin cuticle and flexible shape for some time, and shell plates do not appear until sometime after shedding of the cypris cuticles. In Sacculina carcini, the cypris settles at the base of a seta on the host crab and remains quiescent and aligned at an angle of ∼60° to the crab’s cuticle. The metamorphosis involves two molts, resulting in the formation of an elongated kentrogon stage with a hollow injection stylet. Due to the orientation of the cyprid, the stylet points directly towards the base of the crab’s seta. Approximately 60 h after settlement the stylet penetrates down one of the cyprid antennules and into the crab. Almost immediately afterwards the unsegmented vermigon stage, preformed in the kentrogon, passes down through the hollow stylet and into the crab’s hemocoel in a process lasting only 30 s. In S. carcini, the carapace can remain around the metamorphosing individual without impeding the process

Chitin is a functional component of the larval adhesive of barnacles

Barnacles are the only sessile crustaceans, and their larva, the cyprid, is supremely adapted for attachment to surfaces. Barnacles have a universal requirement for strong adhesion at the point of larval attachment. Selective pressure on the cyprid adhesive has been intense and led to evolution of a tenacious and versatile natural glue. Here we provide evidence that carbohydrate polymers in the form of chitin provide stability to the cyprid adhesive of Balanus amphitrite. Chitin was identified surrounding lipid-rich vesicles in the cyprid cement glands. The functional role of chitin was demonstrated via removal of freshly attached cyprids from surfaces using a chitinase. Proteomic analysis identified a single cement gland-specific protein via its association with chitin and localized this protein to the same vesicles. The role of chitin in cyprid adhesion raises intriguing questions about the evolution of barnacle adhesion, as well as providing a new target for antifouling technologies