Optimisation and standardisation of functional immune assays for striped catfish (Pangasianodon hypophthalmus) to compare their immune response to live and heat killed Aeromonas hydrophila as models of infection and vaccination

Abstract

Aquaculture production of Pangasianodon hypophthalmus is growing rapidly in South East Asia, especially in Vietnam. As it is a relatively new aquaculture species there are few reports evaluating its immune response to pathogens. Thus, functional assays for P. hypophthalmus were optimised to evaluate both innate and adaptive immune responses, and were then used to examine immune response following stimulation with live and heat-killed Aeromonas hydrophila. These were used as models of infection and vaccination, respectively.Four treatment groups were used, including a control group, a group injected intraperitonally (IP) with adjuvant only, a group injected with heat-killed A. hydrophila (1 × 109 cfu ml−1 mixed with adjuvant), and a group injected with a subclinical dose of live A. hydrophila. Samples were collected at 0, 1, 3, 7, 14 and 21 days post-injection (d.p.i.) to assess their immune response. The results indicated that challenge with live or dead bacteria stimulated the immune response in P. hypophthalmus significantly above the levels observed in control groups with respect to specific antibody titre, plasma lysozyme and peroxidase activity, and phagocytosis by head kidney macrophages at 7 or/and 14 d.p.i. At 21 d.p.i., total and specific antibody (IgM) levels and plasma lysozyme activity in fish injected with either live or dead A. hydrophila were significantly different to the control groups. Differential immune responses were observed between fish injected with either live or dead bacteria, with live A. hydrophila significantly stimulating an increase in WBC counts and plasma peroxidase activity at 3 d.p.i., with the greatest increase in WBC counts noted at 21 d.p.i. and in phagocytosis at 14 d.p.i. By 21 d.p.i. only the macrophages from fish injected with dead A. hydrophila showed significantly stimulation in their respiratory burst activity. This study provides basic information on the immune response in pangasius catfish that can be useful in the health control of this species.