The kinetics of antibody production in mucus and serum of European eel (Anguilla anguilla L.) after vaccination against Vibrio vulnificus: development of a new method for antibody quantification in skin mucus.

Vibrio vulnificus serovar E, a bacterial pathogen for eels cultured in intensive systems, is transmitted through water and enters into new hosts mainly via gills. The main objective of this work was to study the kinetics of antibody production to V. vulnificus in serum and mucus and their relationship with protection after vaccination. To quantify local mucus antibodies, a new "in situ" dot blot immunoassay using image analysis has been developed. This assay was applied to measure antibody production in the skin zone next to the gills. We found that (i) the immune response in mucus was faster (peak at days 3-4) and shorter in duration (titres significantly elevated up to day 5 and 11 for skin zone next to the gills and for general cutaneous mucus, respectively) than in serum (peak at day 7; titres significantly elevated for more than 25 days); (ii) the exposure of vaccinated eels with basal levels of local antibodies to sub-lethal dose of the pathogen stimulated a more lasting secreted antibody production (for more than 14 days); (iii) protection and antibody levels in serum were clearly correlated, and (iv) immunised eels with basal levels of serum antibodies and maximal levels of local antibodies were partially protected.