Kinetics of antibody production against Vibrio anguillarum antigens in turbot

Abstract

We evaluated the kinetics of antibody production by turbot ( Scophthalmus maximus L.) in response to intraperitoneal (i.p.) inoculation with Vibrio anguillarum (ATCC strain 19264) cells in the presence or absence of Freund's complete adjuvant. Antibody production was monitored by enzyme-linked immunosorbent assay (ELISA), using and anti-turbot Ig monoclonal antibody produced in our laboratory and three bacterial antigen preparations (whole cells, a whole-cell extract and O-antigen). Our results demonstrate: (a) that maximal antibody response can be achieved with a single inoculation in the presence of adjuvant, and is reached within 45 days; (b) that in the absence of adjuvant, an i.p. booster is required to achieve a maximal response; and (c) that the whole-cell extract (a cytosolic fraction obtained by sonication and subsequent ultracentrifugation at 100 000× g for 30 min) is the best antigen for determination by ELISA of anti- V. anguillarum antibodies in turbot.