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The analytical and diagnostic performances of methods for detecting red sea bream iridovirus (RSIV), which infects marine fish, have not been evaluated. As disease management and transmission control depend on early and reliable pathogen detection, rapid virus detection techniques are crucial. Herein, we evaluated the diagnostic performance of a TaqMan-based real-time polymerase chain reaction (PCR) assay that detects RSIV rapidly and accurately. The assay amplified the RSIV, infectious spleen and kidney necrosis virus, and turbot reddish body iridovirus genotypes of Megalocytivirus and the detection limit was 10.96 copies/reaction. The assay’s performance remained uncompromised even in the presence of nine potential PCR inhibitors, including compounds commonly used in aquaculture. The variation of the cycle threshold values between assays performed by three technicians was evaluated using a plasmid DNA containing the major capsid protein gene sequence. The variation between replicates was low. The diagnostic sensitivity and specificity of the developed assay were evaluated using fish samples (n = 510) and were found to be 100% and 99.60%, respectively. Two technicians evaluated the reproducibility of the assay using fish samples (n = 90), finding a high correlation of 0.998 (p < 0.0001). Therefore, the newly developed real-time PCR assay detects RSIV both accurately and rapidly.

Red sea bream iridoviral disease (RSIVD) causes serious economic losses in the aquaculture industry. In this paper, we evaluated RSIV kinetics in rock bream under various rearing water temperatures and different RSIV inoculation concentrations. High viral copy numbers (approximately 103.7–106.7 RSIV genome copies/L/g) were observed during the period of active fish mortality after RSIV infection at all concentrations in the tanks (25 °C and 20 °C). In the group injected with 104 RSIV genome copies/fish, RSIV was not detected at 21–30 days post-infection (dpi) in the rearing seawater. In rock bream infected at 15 °C and subjected to increasing water temperature (1 °C/d until 25 °C) 3 days later, the virus replication rate and number of viral copies shed into the rearing seawater increased. With the decrease in temperature (1 °C/d) from 25 to 15 °C after the infection, the virus replicated rapidly and was released at high loads on the initial 3–5 dpi, whereas the number of viral copies in the fish and seawater decreased after 14 dpi. These results indicate that the number of viral copies shed into the rearing seawater varies depending on the RSIV infection level in rock bream.

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Background: Olive flounder, Paralichthys olivaceus, is an economically important aquaculture species in Korea. Olive flounders have been heavily damaged by streptococcal infections every year and are treated with antibiotics. However, antibiotic abuse is causing the emergence of resistant strains, and to overcome this, research has shown that new antibiotics must be applied. Tylosin is a relatively safe antibiotic and has good activity against Grampositive bacteria and mycoplasma. We studied the therapeutic effects and side effects of tylosin on Streptococcus parauberis-infected olive flounder. Methods: After artificial infection of olive flounder with S. parauberis SPOF18J3, an appropriate dose of tylosin was confirmed by intramuscular injection (I.M.) at 2.5, 5, 10, and 15 mg/kg, and oral administration at 10 and 20 mg/kg. After I.M. and oral administration dosing of tylosin, side effects were confirmed by serological analysis, histopathological analysis, and median lethal dose (LD 50) analysis at both an appropriate concentration and a high concentration. Statistical analysis was performed using one-way analysis of variance (ANOVA) and Tukey's test (p < 0.05). Results: The appropriate I.M. and oral administration concentration of tylosin administered to olive flounder infected with S. parauberis SPOF18J3 was found to be 10 mg/kg. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were showed not significantly different between the control group and the experimental groups. The histopathologic results showed mild inflammatory responses in muscle and tubular vacuolization and tubular atrophy appeared, but there were no significant differences between the groups. The LD 50 was confirmed to be 461 mg/kg. Conclusion: In this study, an effective treatment method was provided by verifying the treatment effects and side effects of tylosin in olive flounder infected with S. parauberis, which can be applied directly to aquaculture sites. In addition, these results may be used as a reference for evaluation required upon request to obtain approval for tylosin antibiotics as fishery antibiotics in Korea. After approval, it is possible that a fishery disease manager will be able to prescribe and sell the antibiotic tylosin.