Evaluation of anticoagulants and hemocyte-maintaining solutions for the study of hemolymph components in the spiny lobster Panulirus argus (Latreille, 1804) (Decapoda: Achelata: Palinuridae)

Abstract

Functional studies on humoral or cellular responses in the hemolymph of crustaceans require the selection of suitable anticoagulant- and hemocyte-maintaining solutions. We studied the suitability of several anticoagulant- and hemocyte-maintaining solutions in the spiny lobster Panulirus argus (Latreille, 1804), with emphasis in the preservation of hemocyte number and viability. It was found that the modified Alsever solution was the ideal anticoagulant, while modified L-15 medium and Panulirus argus saline (PAS) were the best hemocyte-maintaining solutions. It is striking that whereas avoiding plasma clotting is relatively simple to achieve, avoiding lysis and aggregation of hemocytes could be challenging and variable among closely related crustaceans. The reasons are hardly known and might indicate different composition or sensitivity of both membrane-bound and soluble mediators in any of the three types of hemocytes identified among decapod crustaceans. Hemolymph volume average in P. argus was 10.5% of fresh body weight (more than 50 ml per adult individual), which makes this species an attractive model for functional studies of hemolymph components in crustaceans.