Seven rainbow trout cytokine genes (interleukin (IL)-2, IL-8, IL-15, IL-17, IL-1β, intracellular interferon (iIFN) 1a, and IFN-γ2) were evaluated for their adjuvant effects on a DNA vaccine, called pG, containing the glycoprotein gene of infectious hematopoietic necrosis virus (IHNV). Distinct DNA constructs in expression plasmid pcDNA3.1 encoding a cytokine gene were generated. Immunofluorescence assays in rainbow trout gonadal cells demonstrated successful protein expression from all these constructs. Subsequently, fish were immunized with pG alone or together with a cytokine expression plasmid. Results showed that each cytokine plasmids at an appropriate dose showed notable effects on immune gene expression. IL-17 and IFN-γ2 can enhance early specific IgM response. All cytokines, except IL-8, can benefit initial neutralizing antibody (NAb) titers. At 35 days post immunization (dpi), NAb titers of fish immunized with pG and IL-2, iIFN1a, or IFN-γ2 plasmids remained at high levels (1:160). NAb titers of fish immunized with pG alone decreased to 1:40. IL-8 or IL-1β can enhance antigen-specific proliferative T-cell responses at 14 dpi. At 28 dpi, coinjection of pG with IL-2, IL-8, IL-15, or IL-17 plasmids induced considerably stronger lymphocyte proliferation than that with injection of pG alone. All cytokine plasmids delivered with pG plasmid enhanced protection of trout against IHNV-mediated mortality. These results indicate that the type and dose of trout cytokine genes injected into fish affect quality of immune response to DNA vaccination.

Flavobacterium psychrophilum causes substantial economic losses in salmonid aquaculture worldwide. Although this bacterium is also believed to be a disease source in China, published reports of its presence do not yet exist.

Yersinia ruckeri is the causative agent of enteric redmouth disease or yersiniosis, which affects salmonids and several other species of fish. However, there are no reports on the characteristics and pathogenicity of Y. ruckeri isolated from farm-raised Amur Sturgeon Acipenser schrencki. Here, we isolated and characterized Y. ruckeri strain H01 from the diseased Amur Sturgeon in China. The phenotypic and genotypic characteristics of Y. ruckeri were observed, and its virulence was tested by examining experimentally infected sturgeons. Examination of the flagellar morphology of Y. ruckeri by transmission electron microscopy showed five to eight peritrichous flagella located on the cell body. Actively dividing cells with an obvious cell membrane were approximately 0.64 μm in diameter and between 1.7 and 2.5 μm in length. The dose that was lethal to 50% of the test fish after intraperitoneal injection was determined to be 7.2×10⁶ CFU, and Y. ruckeri could be reisolated from the liver and kidneys of infected sturgeon. Antimicrobial susceptibility tests showed that H01 was susceptible to 10 antimicrobial agents. Part of the 16S rRNA sequences (563 base pair) was amplified and sequenced to study the genotypic characterization in Y. ruckeri (GenBank accession number JQ657818). The phylogenetic tree revealed H01 was clustered together with Y. ruckeri strains. Together, this study describes the isolation, characterization, and phenotypic–genotypic analysis of a Y. ruckeri strain isolated from farm-raised Amur Sturgeon. The results discovered may provide some theoretical basis for the prevention and control of yersiniosis in Amur Sturgeon. Received July 23, 2012; accepted September 5, 2012