Preparatory Immunodiffusion for Production of Specific Anti-Adult Southern Pine Beetle Serum

Abstract

Preparatory immunodiffusion was used to produce an antiserum specific for the southern pine beetle, Dendroctonus frontalis (Zimm.), and the black turpentine beetle, D. terebrans (Olivier). Precipitin bands were excised from two-dimensional electroimmunodiffusion agar gel plates of crude antigen and antiserum, extracted in saline, emulsified with Freund's incomplete adjuvant, and injected subcutaneously near the inguinal lymph nodes of rabbits. The specific antiserum did not cross react with Ips avulsus Eichhoff, I. calligraphus Germar, or I. grandicollis (Eichhoff) when tested at high antigen concentrations by agar gel double diffusion, Crowle's template micro-double diffusion, two-dimensional electroimmunodiffusion, counter immunoelectrophoresis, or cellulose acetate membrane electrophoresis with immunofixation. The antiserum is genus specific in all tests, and species specific when run in the agar gel double diffusion test against a standard Dendroctonus antigen (7.0 mg protein/ ml).