Effect of handling, confinement and crowding in HSP70 production in Pachygrapsus marmoratus, a model species for climate change experiments

The aim of this study was to investigate the effect of handling, confinement and crowding on HSP70 production in an intertidal crab. HSP70 has been widely used as a biochemical indicator of thermal stress; however studies on the effect of experimental conditions and handling on HSP70 expression are very scarce. The response of marbled crab. Pachygrapsus marrnoratus, collected in the Portuguese coast, was investigated through an experiment in captivity, where 42 juveniles (10-19 mm of carapace width) were confined in an experimental container (60 x 42 x 10 cm) and exposed to three treatments: 1) increasing temperatures and periodic handling, 2) only to periodic handling and 3) no handling, for 12 h. The objective of this work was to investigate whether the HSP70 production measured in an increasing temperature experiment is a response to temperature alone or whether manipulation, confinement and crowding during the experiment also increase HSP70 production. Three individuals were sampled after t = 0, t = 4, t = 6, t = 8, t = 10 and t = 12 h, for the quantification of HSP70 production in the hemolymph, in the three trials. It was concluded that while temperature has a significant effect on HSP70 production, periodic handling does not, nor does confinement to the experimental container or crowding. It can thus be concluded that experimental conditions and handling are not parasitic variables in experiments with this species considering increasing temperatures. P. marmoratus can thus be used as a model species in climate warming experiments involving handling, confinement and crowding. (C) 2012 Elsevier B.V. All rights reserved.