non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immuno-toxicity studies

We have compared two methods for collecting earthworm leukocytes (coelomocytes) with respect to cell yield, viability and behaviour in immunoassays. Non-invasive extrusion was more efficient than puncturing the coelomic cavity. Extrusion does not produce trauma to earthworms maintained under long term laboratory conditions. Neither technique modified immune assays, as determined by erythrocyte and secretory rosette formation, and phagocytosis, since all 3 were functionally equivalent. After an initial extrusion, sequential leukocyte collections by extrusion are possible at intervals of 6 weeks without affecting total and differential cell counts and rosette formation.