SEROLOGICAL COMPARISONS AMONG FOUR CLASSES OF MOLLUSCA 1 CHARLES A. LEONE AND CARLON W. PRYOR 2 Department of Zoology, University of Kansas, Laivrcnce Kansas Serological comparisons of the proteins of organisms are a means of studying the results of biochemical evolution. With the precipitin test the chemical similar-ities of the proteins of animals can be measured quantitatively and the degrees of relationship thereby estimated. The purpose of this paper is to report on some serological comparisons of the proteins from the four Classes Amphineura, Gastro-poda, Pelecypoda, and Cephalopoda in the Phylum Mollusca. MATERIALS AND METHODS Antigens Table I lists the species of Mollusca from which antigens were obtained. Sera from Octopus vulgaris and Sepia officinalis were collected at the Stazione Zoologica di Napoli, Naples, Italy in the summer of 1948. Serum from Busycon caricum was obtained at the laboratory of the United States Fish and Wildlife Service, Beaufort, North Carolina, in the summer of 1951. The other antigens were col-lected at the Friday Harbor Marine Laboratory, University of Washington, Friday Harbor, Washington, in the summer of 1952. 'Merthiolate' in a final concentration of 0.01 per cent (1 : 10,000) was added as a preservative at the time of collection. Antisera The antisera used in this study were produced in healthy, adult rabbits. Table I shows the schedule of injections followed for the production of these antisera. Mul-tiple series of injections were given to all rabbits except the one which produced the antiserum against the proteins of Tonicella lincata. This rabbit received a single series of injections ; seven injections were given on alternate days 1.0, 1.5, 2.0, 2.0, 5.0, 5.0, and 5.0 ml. The fifth and sixth were intraperitoneal and the others were intravenous. Each rabbit was given a trial bleeding from the median artery of the ear eight days after the last injection of a series. At the end of the injection sched-ule those rabbits with good levels of antibody were completely exsanguinated by intracardial puncture after a fast of 18 hours to obtain sera free from dissolved lipids. Whole blood, obtained either by bleeding from the median artery of the ear, or by intracardial puncture, was placed in lusteroid centrifuge tubes, permitted to clot, rimmed, and held at 2 C. for 1 to 10 hours. Expressed serum was 1 These studies were aided by a contract between the Office of Naval Research, Department of the Navy and the University of Kansas, NR 163-012. -Public Health Service Predoctorate Research Fellow of the National Microbiological Institute. 411
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