Passive immunisation of goldfish with the serum of those surviving a Cyprinid herpesvirus 2 infection after high temperature water treatment

Herpesviral haematopoietic necrosis of goldfish caused by cyprinid herpesvirus 2 (CyHV-2) can be controlled by raising water temperature to a virus non-permissive temperature of 34℃. Consequently, the goldfish can survive and acquire resistance to the disease; the underlying mechanism of acquired resistance, however, remains unclear. In this study, we investigated serological changes in the surviving goldfish, with a focus on their humoral immunity, and examined whether sera of the surviving goldfish conferred passive immunity to naive goldfish. Levels of the anti-CyHV-2 antibodies in 8 of the 9 survivors measured via ELISA were higher than those in control fish. Neutralising antibodies were detected in the sera of 2 survivors, but no direct correlation was observed between ELISA optical density value and neutralising antibody titer. Passive immunisation tests showed that recipients injected with the serum containing neutralising antibodies showed higher survival rates than the control group. The sera from 6 other survivors showed no effect on the recipient\u27s mortality regardless of anti-CyHV-2 antibody levels. These results suggest that neutralising antibodies can contribute to acquired immunity in survivors, and other protective factors, including cell-mediated immunity, may work in the survivors that show no detectable neutralising antibodies

Molecular characterization and validation of commercially available methods for haptoglobin measurement in bottlenose dolphin

AbstractHaptoglobin (Hp) is a positive acute-phase protein and a valuable marker of inflammation in both human and veterinary medicine. The aim of this study was to validate the molecular characterization of Hp in dolphins and to validate commercially available Hp measurement methods such as Hp-ELISA (originally designed for pigs) and Hp–hemoglobin (Hb) binding assay. The dolphin Hp (dHp) amino acid sequence appeared most similar to pig Hp by sequence homology and phylogenetic clustering. Amino acid sequence analysis revealed that dHp comprises the Hp1 form of α1 and β chains. The anti-pig Hp antibody cross-reacted with both recombinant dHp, expressed by Escherichia coli, and dHp from serum. The intra- and inter-assay levels of imprecision of pig Hp-ELISA and the Hp–Hb binding assay were found to be tolerable for the determination of Hp in dolphin, and there was no significant discrepancy between the two determination methods. The ability of the assay to differentiate between healthy and inflammation groups was investigated, and a significant increase in Hp concentration was detected in inflammatory conditions. Thus, Hp is a useful inflammation marker for dolphin, and the Hp concentration in dolphin serum samples can be reliably measured using commercially available pig Hp-ELISA and Hp–Hb binding assay