Laboratory Animals > Volume 25, Number 1 > Pp. 61-67

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

George S. Eyambe, Arthur J. Goven, L. C. Fitzpatrick, Barney J. Venables and Edwin L. Cooper

Environmental Effects Research Group, Department of Biological Sciences, University of North Texas, Denton, Texas 76203, USA; Environmental Effects Research Group, Department of Biological Sciences, University of North Texas, Denton, Texas 76203, USA; Environmental Effects Research Group, Department of Biological Sciences, University of North Texas, Denton, Texas 76203, USA; Environmental Effects Research Group, Department of Biological Sciences, University of North Texas, Denton, Texas 76203, USA; TRAC Laboratories, 113 Cedar St, Denton, Texas 76201, USA; Department of Anatomy and Cell Biology, School of Medicine, University of California Los Angeles, Los Angeles, California 90024, USA

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.

Key Words: EARTHWORMS • LUMBRICUS TERRESTRIS • IMMUNE RESPONSES • LEUKOCYTES • COELOMOCYTES • COELOMOCYTE COLLECTION TECHNIQUES

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