A, Silver catfish were intraperitoneally injected with BSA alone (BSA+PBS, 200 [micro]g/fish) or BSA adjuvanted with aluminum hydroxide (AIOH), Freund's complete adjuvant (FCA), Freund's incomplete adjuvant
(FIA) and Montanide; B, bSa alone or combined with CpG 1668, CpG 2102, CpG 2133 and CpG 2143; C, BSA alone or combined with Montanide, and [beta]-glucan (0.02; 0.06 and 0.1%).
Subsequently, we gave the rats two booster injections using the same conjugate in Freund's incomplete adjuvant
. After 2 months of immunization, we checked the antibody titer, using rabbit anti-rat IgG-coated plates and europium-labeled EG with time-resolved fluorescence detection.