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Nocardiosis caused by Nocardia seriolae is one of the major threats in the aquaculture of Seriola species (yellowtail; S. quinqueradiata, amberjack; S. dumerili and kingfish; S. lalandi) in Japan. Here, we report the complete nucleotide genome sequence of N. seriolae UTF1, isolated from a cultured yellowtail. The genome is a circular chromosome of 8,121,733 bp with a G+C content of 68.1% that encodes 7,697 predicted proteins. In the N. seriolae UTF1 predicted genes, we found orthologs of virulence factors of pathogenic mycobacteria and human clinical Nocardia isolates involved in host cell invasion, modulation of phagocyte function and survival inside the macrophages. The virulence factor candidates provide an essential basis for understanding their pathogenic mechanisms at the molecular level by the fish nocardiosis research community in future studies. We also found many potential antibiotic resistance genes on the N. seriolae UTF1 chromosome. Comparative analysis with the four existing complete genomes, N. farcinica IFM 10152, N. brasiliensis HUJEG-1 and N. cyriacigeorgica GUH-2 and N. nova SH22a, revealed that 2,745 orthologous genes were present in all five Nocardia genomes (core genes) and 1,982 genes were unique to N. seriolae UTF1. In particular, the N. seriolae UTF1 genome contains a greater number of mobile elements and genes of unknown function that comprise the differences in structure and gene content from the other Nocardia genomes. In addition, a lot of the N. seriolae UTF1-specific genes were assigned to the ABC transport system. Because of limited resources in ocean environments, these N. seriolae UTF1 specific ABC transporters might facilitate adaptation strategies essential for marine environment survival. Thus, the availability of the complete N. seriolae UTF1 genome sequence will provide a valuable resource for comparative genomic studies of N. seriolae isolates, as well as provide new insights into the ecological and functional diversity of the genus Nocardia.

Immunoglobulins (Igs), also termed antibodies, orchestrate host-acquired immune responses against foreign antigens, including invasive pathogens. In fish, IgM, which is present predominantly circulating in the blood, is particularly important for humoral systemic immunity and protecting the host from pathogens. The efficacy of inactivated vaccines, a major type of vaccine commonly used worldwide in fish, is directly linked to the serum antibody level; however, the timing of the appearance of systemic IgM circulating in the blood has not been determined in fish. In the present study, we examined the dynamics of serum IgM levels in juvenile Japanese amberjack Seriola quinqueradiata, using a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) that we developed for IgM. We found that serum IgM concentration in young fish up to 72 days post-hatching (d.p.h.) (mean ± standard error of the mean [SEM]; body weight: 5.73 ± 0.38 g, standard length [S.L.]: 72.2 ± 1.94 mm) was sustained at a low level, but that the level significantly increased from 79 d.p.h. onward, reaching a mean of 84.76 ± 9.23 μg/mL at 85 d.p.h. (body weight: 14.05 ± 0.92 g, S.L.: 101.1 ± 2.07 mm). These results suggest that systemic immunity mediated by IgM is only partially matured in the early growth stage of juveniles. The present findings could help establish effective vaccination programs for infectious diseases in young fish.

Largemouth bass Micropterus salmoides were immunized with four different N. seriolae strains—two α-glucosidase-positive (961113, KU040801) and two α-glucosidase-negative (94260, OTTS) strains—along with Freund’s incomplete adjuvant. After primary immunization (week 0), a booster was administered at weeks 4 and 8. Nonspecific immune responses to multiple immunizations with the different N. seriolae strains were determined based on serum lysozyme activity and nitroblue tetrazolium (NBT)-positive cells in peripheral blood. The serum lysozyme activity and NBT-positive cells in peripheral blood were not significantly increased even after the two booster immunizations. Specific antibody responses against N. seriolae cells were investigated by enzyme-linked immunosorbent assay. At 4 weeks after immunization, all groups immunized with N. seriolae antigens showed significant increases in their specific antibody levels. The sera from fish immunized with different N. seriolae strains exhibited reactivity with N. seriolae sonicated antigens of 28, 30, 36 and 84 kDa by western blot analysis. After two boosters, fish were challenged with live N. seriolae to assess the vaccine’s efficacy; however, multiple injections of the N. seriolae strains did not reduce mortality, irrespective of the bacterin.