• The Journal of the Korea institute of electronic communication sciences
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• v.16 no.4
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• pp.661-674
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• 2021

Edwardsiellosis is difficult to treat in cultured olive flounder, Paralichthys olivaceus. It is present in the fish for a long period during all growth stages, and it often leads to mass mortalites. In this paper, the clinical patterns of Edwardsiellosis were analyzed by dividing the data into the whole-water temperature, low-water temperature, low-high water temperature, high-water temperature, and high-low water temperature groups based on various clinical signs of diseased cultured olive flounder using a decision tree technique. In the clinical sign patterns in the decision trees analyzed in the experiment, clinical signs in the liver, such as liver nodules, liver hemorrhages, and liver degeneration, were selected as the criteria for determining Edwardsiellosis. The selected clinical signs were known as the major clinical signs of Edwardsiellosis, and through consultation with fishery disease experts, the analysis confirmed that the clinical signs of Edwardsiellosis were successfully found in this study.

• Journal of fish pathology
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• v.34 no.2
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• pp.225-231
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• 2021

In biofloc culture for olive flounder, Paralichthys olivaceus, the possibility of antibiotics treatment was investigated against edwardsiellosis. After inducing edwardsiellosis by immersion in Edwardsiella tarda 1.2 × 10⁵/mL suspension, the survival trends on various biofloc water management and some physiological changes were observed. For biofloc water management, six types of water treatments were carried out, which were no exchange without antibiotics as negative control, the exchange to stored biofloc water, the exchange to stored biofloc water with 20% flow-through, the exchange to fresh biofloc water, half fresh biofloc water and half sea water, and the complete flow-through. There was no significant physicochemical change on water qualities in any type. The exchange to fresh biofloc water was shown the highest survival ratio as 72.3%, and in case of stored biofloc water with 20% flow-through, the survival ratio was also significantly high as 62%. Plasma glucose, cholesterol, total protein, calcium, and magnesium were analyzed as physiological index. Mostly, there was no significant change, but plasma cholesterol showed an initial decrease in low survival group, and an initial increase with high survival group. Consequently, antibiotic treatment against a bacterial disease during biofloc culture is possible as long as the biofloc water management follow along properly.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.5
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• pp.620-628
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• 1996

To avoid the self-agglutination of Staphylococcus aureus sensitized with rabbit antibody in the absence of antigen, we determined the optimum concentration of rabbit antibody for sensitization. It was analyzed by using three different kinds of S. aureus strains at various concentraions of antibody. The optimized coagglutination test using the S. aureus sensitized with rabbit antibody was applied to the diagnosis of edwardsiellosis in field and in laboratory. The presence of E. tarda as low as $10\;{\mu}g/ml$ was detected by this method. Moreover, it showed good coagglutination results against several different forms of antigens such as FKC, EDTA or heat extracted antigen of E. tarda. E. tarda strains, isolated from the flounders suffering from edwardsiellosis in fields, showed some cross-reactions to the E. tarda 219 analyzed by both agglutination and coagglutination test with rabbit anti-E, tarda 219 antibody. The degree of cross-reactions analyzed was enough to apply the coagglutination test for the diagnosis of edwardsiellosis in field. Thus, even 1,000 fold diluted tissue homogenate of infected flounder naturally contained enough E. tarda as an antigen to show good coagglutination with S. aueus sensitized with rabbit anti-E, tarda 219 antibody. The successful application of this method to the homogenate and heat extract of tissues from naturally or artificially infected flounder or tilapia preyed that coagglutination test was a simple and rapid reliable dignostic technique suitable for using in laboratory and field without any special equipments.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 1996.06a
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• pp.177.2-184
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• 1996

• Journal of Microbiology and Biotechnology
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• v.33 no.5
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• pp.668-679
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• 2023

Edwardsiella tarda is one of the most significant fish pathogens, causes edwardsiellosis in a variety of freshwater fish species, and its antibiotic resistance against multiple drugs has made it a health risk worldwide. In this study, we aimed to investigate the antibiotic resistance (ABR) genes of E. tarda and establish its antibiotic susceptibility. Thus, 540 fish (299 Oreochromis niloticus, 138 O. mossambicus, and 103 O. aureus) were collected randomly from twelve fish farms in three districts of Punjab in Pakistan. E. tarda was recovered from 147 fish showing symptoms of exophthalmia, hemorrhages, skin depigmentation, ascites, and bacteria-filled nodules in enlarged liver and kidney. Antimicrobial susceptibility testing proved chloramphenicol, ciprofloxacin, and streptomycin effective, but amoxicillin, erythromycin, and flumequine ineffective in controlling edwardsiellosis. Maximum occurrence of qnrA, blaTEM, and sul3 genes of E. tarda was detected in 45% in the liver, 58%, and 42% respectively in the intestine; 46.5%, 67.2%, and 55.9% respectively in O. niloticus; 24%, 36%, and 23% respectively in summer with respect to fish organs, species, and season, respectively. Motility, H₂S, indole, methyl red, and glucose tests gave positive results. Overall, E. tarda infected 27.2% of fish, which ultimately caused 7.69% mortality. The Chi-squared test of independence showed a significant difference in the occurrence of ABR genes of E. tarda with respect to sampling sites. In conclusion, the misuse of antibacterial agents has led to the emergence of ABR genes in E. tarda, which in association with high temperatures cause multiple abnormalities in infected fish and ultimately resulting in massive mortality.

• Journal of fish pathology
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• v.12 no.1
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• pp.24-31
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• 1999

Optimization and standardization of solid phase enzyme immunoassay were done for the diagnosis of edwardsiellosis in fish. The analyzed degree of immobilized antibody on surface of solid phase with peroxidase saturation method showed the optimized result by using partially purified $50{\mu}g/ml$ of rabbit anti-E. tarda Edk-2 antibody in sodium bicarbonate buffer for overnight incubation to cover the surface of polystyrene beads. Optimized immunoreaction was observed in the treatment of $50{\mu}g/ml$ of biotin conjugated antibody followed extravidin-peroxidase diluted 1 : 2,000 in PBS. The detectable concentrations of the this method were $1{\times}10^5$ cells/ml and $1{\times}10^5$ cells/ml expressed as the source of antigen amount for EDTA extraction and heat extraction, respectively. High cross-reaction of solid phase ELISA with the prepared rabbit and-E. tarda Edk-2 was observed against E. tarda strains isolated from flounder suffering from edwardsiellosis in aquatic farms of Korea. It suggested that the potential of this solid phase of ELISA technique is very powerful for the application to different strains of E. tarda isolated in farms of many different areas.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1273-1280
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• 2019

Edwardsiella piscicida is the causative agent of edwardsiellosis, which has caused enormous economic losses worldwide. In our previous research, an attenuated live vaccine known as WED and based on the virulent strain E. piscicida EIB202 can effectively protect turbots against edwardsiellosis via intraperitoneal injection, while vaccination by immersion exhibits a weaker effect. During the development of the immersion vaccine, we surprisingly found the counts of ${\Delta}pEIB202/EIB202$ colonized on zebrafish were 100 times lower than those of EIB202. However, pEIB202 carries 53 predicted ORFs and has several copies in E. piscicida EIB202, impeding the study of its function. Thus, the replication region is located at a 1,980 bp fragment (from 18,837 to 20,816 bp), containing a transcriptional repressor and a replication protein. Moreover, the minimal replication plasmid, named pRep-q77, has low copies in both E. coli and E. piscicida, but is more stable in E. piscicida than in E. coli. This work lays a foundation for further examination of the function of the virulence plasmid pEIB202.

• Journal of Life Science
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• v.22 no.6
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• pp.788-793
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• 2012

High concentration of antibiotics has been used to treat the outbreak of edwardsiellosis caused by Edwardsiella tarda in aquaculture. However, not all of the bacteria have been killed with high concentrations of antibiotics treatment by the formation of persister cells with a dormant state. The main objective of this study was to kill persister cell using antibiotics with the addition of natural plant compounds. Antibiotics used in this study consist of 100 mg/ml florfenicol and 100 mg/ml amoxicillin. Ten natural plant compounds with persister cell inhibitor activity to E. coli were obtained from Protein Engineering and Systems Biology Lab. of Sungkyunkwan University. The persister cell inhibition activities of those natural plant compounds were evaluated in test tube. Concentrations of the antibiotics were in the ranges of 25~200 ${\mu}g/ml$. The persister cell formation was observed after 16 hours of culture. Persister cells were killed by antibiotics with natural plant compounds. Among ten natural plant compounds, Gynostemma pentaphyllum, Mallotus japonicus, and Orixa japonica showed persister cell formation inhibition activities. The optimal concentrations of G. pentaphyllum, M. japonicus, and O. japonica for the inhibitor of persister cell formation were 100 ${\mu}g/ml$, 100 ${\mu}g/ml$, and 200 ${\mu}g/ml$, respectively. In vivo study was carried out to evaluate the effect of the antibiotics with natural plant compounds using aquacultural fish, olive flounder, as test animals. G. pentaphyllum, M. japonicus, and O. japonica of 30 ${\mu}g/ml$, 10 ${\mu}g/ml$, and 10 ${\mu}g/ml$ with antibiotics reduced cumulative mortalities, showing the effectiveness of persister cell inhibition.

• Journal of Fisheries and Marine Sciences Education
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• v.27 no.3
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• pp.672-681
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• 2015

Edwardsiellosis has become a serious diseases problem in cultured eels for many years. This study was performed to investigate possibility of vaccination against edwardsiellosis caused by Edwardsiella tarda. We conducted a immersion and/or oral vaccination using formalin-killed E. tarda in eel Anguilla japonica. Three groups of fish ($26.8{\pm}1.2g$, $7.1{\pm}0.7g$ and $2.2{\pm}0.4g$) were used in this study. The protection (relative percentage survival, RPS) and serum antibody response (agglutination titer) were evaluated in the vaccinated fish. No correlation between agglutination titer and survival rate was observed in vaccinated fish. However, there was a satisfactory protective (RPS>50%) in vaccinated fish. Immersion (10 mg/mL, 1 hr) and immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of $26.8{\pm}1.2g$, immersion (10 mg/mL, 1 hr) plus oral (10 mg/g, 10 days) of $7.1{\pm}0.7g$ showed RPS of 62.6%, 52.2% and 56.8%, respectively.

• Journal of fish pathology
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• v.17 no.3
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• pp.171-177
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• 2004

This study was performed to investigate the effects of prolonged immersion vaccination in a dilute vaccine on the farmed olive flounder, Pararlichthys olivaceus. Juvenile olive flounders were immersed in the several diluted ($10^0$, $10^{-1}$, $10^{-2}$ and $10^{-3}$) commercial edwardsiellosis bacterins (5㎎/ℓ) for 2min (short immersion, SI) and 24 h (prolonged immersion, PI). After immunization, fish were analyzed to investigate on non-specific (serum lysozyme and bactericidal activity), specific immune responses (antibody agglutinin titer) and resistances against Edwardsiella tarda infection. The PI of the commercial vaccine diluted with $10^{-1}$ and $10^{-2}$ showed better results over the untreated control in terms of serum lysozyme, bactericidal activities and resistance against E. tarda infection. These results suggest that the prolonged treatments of the diluted vaccine might be a new approach improving the vaccination potential.