• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.48 no.5
• /
• pp.725-730
• /
• 2015

We observed yellow catfish Pelteobagrus fulvidraco fingerlings cultured in land ponds in Korea swimming in a corkscrew spiral pattern while hanging head-up and tail-down at the water surface, before eventually dying. Externally, these fish displayed “hole in the head” disease, pale gills, and hemorrhages in the base of the pectoral and caudal fins; internally they had liver hemorrhages and kidney discoloration. The bacterium Edwardsiella ictaluri (YCK-01 and YCL-01) was identified in the kidneys and livers of diseased fish via phenotypic characteristics and PCR analysis using the ictaluri-specific primers IVS (an intervening sequence) and IRS (the inter-ribosomal spacer). Infectivity challenges by intraperitoneal and immersion routes showed that a representative bacterial strain (YCK) exhibited strong virulence to yellow catfish, with an LD₅₀ of 3.2×10⁴ CFU/fish and 2.5×10⁶ CFU/mL, respectively. This is the first report of E. ictaluri infection in yellow catfish from Korea.

• Journal of fish pathology
• /
• v.34 no.1
• /
• pp.31-38
• /
• 2021

The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H₂S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.2
• /
• pp.167-179
• /
• 2023

The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD₅₀). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 10³ CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 10⁷ CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 10⁷ CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.

• Journal of fish pathology
• /
• v.33 no.2
• /
• pp.111-118
• /
• 2020

The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.

• Journal of fish pathology
• /
• v.7 no.2
• /
• pp.105-112
• /
• 1994

A new bacterial infection occurred among the cultured Korean catfish, Silurus asotus, in Chunbuk prefecture, Korea, 1993. This disease produced about 30% mortality in the fish for 4 months. The diseased fish was swimming listless at the water surface with head up and tail down, sometimes spinnig in circles. The most outstanding clinical sign was ulceration on the skull and at the base of the pectoral fins. The causative organism was isolated from the brain, kidney, spleen and liver of diseased fish, and identified as Edwardsiella icaluri by the biochemical and biophysical characteristics. After intraperitoneal innoculaton of the isolate, the pathogenicity was proved positive for Korean catfish, S. asotus, and channel catfish, Ictalurus punctatus, but negative for carp, Cyprinus carpio.

• Journal of fish pathology
• /
• v.9 no.1
• /
• pp.79-85
• /
• 1996

The effect of $\beta$-glucan administration on the resistance of Korean catfish(Silurus asotus) to experimental Edwardsiella ictaluri and Aeromonas hydrophila infection was evaluated. Fish were either intraperitoneally received $\beta$-glucan($200\sim1000{\mu}g$/100g body weight) dissolved in physiological saline once or twice at an intervals of 3 days, or placed in $\beta$-glucan bath($100{\mu}g/m\ell$) prepared with filtered tap water for 30 or 60 min. Bacterial challenge was performed by intraperitoneal injection of 0.1 ml of bacteria suspension($2{\times}10^7 CFU/m\ell$) 3 days after $\beta$-glucan administration. The $\beta$-glucan injected fish showed an significantly enhanced resistance against experimental infections. The resistance was much higher in the twice-injected fish than in the once-injected fish. But glucan bath did not affect survival rate after the challenge. The protective effect in the $\beta$-glucan injected fish was higher to A. hydrophila than to E. ictaluri. These results indicate that $\beta$-glucan injection can increase the resistance of Korean catfish against experimental E. ictaluri and A. hydrophila infection and that twice injection of $\beta$-glucan at an 3 day-intervals is more effective to the resistance.

• Proceedings of the Korean Society of Fisheries Technology Conference
• /
• 1996.06a
• /
• pp.155-160
• /
• 1996

• Journal of fish pathology
• /
• v.7 no.2
• /
• pp.113-117
• /
• 1994

The antimicrobial activity of Artemisia princeps var. orientalis essential oil against a partial fish pathogenic bacteria was examined. The growth of Aeromonas hydrophila, Aeromonas salomonicida, Aeromonas sorbia, Edwardsiella tarda and Streptococcus sp. (yellowtail) were inhibited at concentrations of 1,000 to 2,000 ppm. The A. salmonicida was inhibited at 1,000 ppm, A. hydrophila, A. sorbia, E, tarda and Streptococcus sp. (yellowtail) at 1,500 ppm, but Vibrio anguillarum, Vibrio ordalii, Edwardsiella ictaluri and Streptococcus sp. (SF 1) were grown on 100-2,000 ppm.

• Journal of fish pathology
• /
• v.10 no.2
• /
• pp.75-86
• /
• 1997

Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.