• Journal of fish pathology
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• v.36 no.2
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• pp.389-394
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• 2023

Infectious hematopoietic necrosis virus (IHNV) is s significant viral pathogen affecting cultured rainbow trout (Oncorhynchus mykiss) in Korea. In this study, five monoclonal antibodies (mAbs) (IHNV-1, 2, 3, 4, and 5) were produced using purified IHNV. Reactivities of these mAbs were analyzed by western blot (WB), enzyme-linked immunosorbent assay (ELISA), and indirect fluorescent antibody test (IFAT). These mAbs recognized glycoprotein (69 kDa, IHNV-1), nucleocapsid protein (39 kDa, IHNV-3, 4, and 5), or phosphoprotein (27 kDa, IHNV-2) of IHNV by WB analysis. ELISA results indicated that these five mAbs were specific to IHNV without showing any cross-reactivity against other fish viruses (hirame rhabdovirus, infectious pancreatic necrosis virus, and viral hemorrhagic septicemia virus). IFAT demonstrated specific fluorescence signals of IHNV-infected epithelioma papulosum cyprini (EPC) cells, whereas no reactivity of normal EPC cells was observed. These mAbs can be very useful for immuno-diagnosis of IHNV infection.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.3
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• pp.248-253
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• 2018

Freshwater farms are primarily located adjacent to rivers and lakes, facilitating the introduction and spread of pathogens into natural systems. Therefore, it is necessary to continuously monitor natural aquatic organisms, the breeding environment, and infection rates by pathogenic organisms. Fish and crustaceans were sampled 4 times in the Geum River estuary in 2016. The samples were analyzed for the presence of pathogens for reportable communicable diseases, including KHVD (koi herpesvirus disease), SVC (spring viraemia of carp), EUS (epizootic ulcerative syndrome) and WSD (white spot disease); parasite abundance was also examined. The dominant fish species were deep body bitterling Acanthorhodes macropterus (21.4%), followed by skygager Erythroculter erythropterus (12.7%). For crustaceans, Palaemon paucidens and Chinese mitten crab Eriocheir sinensis were dominant. Sixty fish and 36 crustacean species were examined for reportable communicable diseases. When using a specific primer set for each disease, PCR analysis did not detect any reportable communicable diseases in the samples. Some instances of Dactylogyrus, copepods, nematodes and metacercaria were detected. However, the PCR results indicated that the metacercaria were not Clonorchis sinensis.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.369-378
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• 2018

This study was conducted to evaluate effects of dietary microbial probiotics on the growth and disease resistance of olive flounder (Paralichthys olivaceus) in a recirculating aquaculture system (RAS), and the effects of the probiotic bioaugmentation on the microbial community structure and water quality. For the analysis, 80 juvenile fish (average weight, $25.7{\pm}7.6g$; average length, $15.2{\pm}1.7cm$) were fed a basal diet containing a commercial microbial product CES-AQ1 (CES; $1{\times}10^9\;CFU/kg$ diet) in an RAS for 8 weeks. Weight gain, the specific growth rate, feed efficiency, and protein efficiency ratio of the fish fed the CES diet in the RAS were 1.5~2.5 times higher than those of fish fed the basal diet alone, or the basal diet containing oxytetracycline (OTC), yeast plus bacterium, or Bacillus subtilis in a still water system. There was no significant difference in the pathogen challenge test between fish fed the OTC diet and fish fed the CES diet in the RAS, suggesting the CES-AQ1 probiotic used in the RAS as a potential replacement for antibiotics. The RAS biofilter maintained the highest microbial diversity and appeared to harbor microbial communities with ammonium oxidation, denitrification, and fish pathogen suppression functions. Ammonia, which is hazardous to fish, was significantly decreased to < 0.5 mg/L in 19 days, indicating the effectiveness of probiotic supplementation to maintain good water quality in RAS. These results suggest that the intestinal microbial communities of fish are stabilized by a probiotic-containing diet (CES) and that bioaugmentation with probiotics may be an eco-friendly and economical supplement for aquaculture of olive flounder, promoting both good water quality and fish health in an RAS.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.3
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• pp.443-450
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• 2020

No established method can be used to select effective antibiotics in antibiotic susceptibility tests for fish bacterial pathogens quickly and accurately. Here, we established the optimal conditions for determining the minimal inhibitory concentration (MIC) of major fish bacterial pathogens (Streptococcus spp., Edwardsiella tarda, Vibrio spp., Aeromonas spp., and Pseudomonas spp.) using the KRAQ1 and CAMPY2 panels. The MIC panel used 18 antibiotics of two types and we conducted experiments to establish the optimal culture medium and temperature for each species. The optimal conditions for incubating Streptococcus spp. were in cation-adjusted Mueller-Hinton broth with TES buffer (CAMHBT) at 28℃, using 5% lysed horse blood (LHB) as recommended by the Clinical Laboratory Standards Institute. For Vibrio spp., the optimal culture conditions were 28℃ in CAMHBT supplemented with 1% NaCl. The optimal conditions for culturing E. tarda, Aeromonas spp., and Pseudomonas spp. were in CAMHBT at 28℃.

• Korean Journal of Veterinary Research
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• v.56 no.2
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• pp.125-127
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• 2016

Gradual mortality of look down fish (Selene vomer) was observed in a private aquarium in Seoul, showing abnormal swimming behavior and lethargy. A bacterial pathogen from kidney was cultured, identified, and confirmed as Vibrio harveyi using Vitek System 2 and 16S rRNA gene sequencing. A predominant bacterial strain, SNUVh-LW2 was proved to be most closely related to isolates from China by phylogenetic analysis with minimum evolution method. Also, tetracycline was considered as the most sensitive antibiotic agent via antibiotic susceptibility test. The group of fish was treated according to the diagnostic result and no more mortality was observed.

• Journal of fish pathology
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• v.29 no.1
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• pp.13-23
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• 2016

Lectins play significant roles in the innate immune responses through binding to pathogen-associated molecular patterns (PAMPs) on the surfaces of microorganisms. In the present study, tissue distribution and expression analysis of olive flounder lectins were performed after viral hemorrhagic septicemia virus (VHSV) challenge. Fish egg lectin and serum lectin were found to be predominantly expressed in the gills and liver, these results indicate that the transcript expression of olive flounder lectins is concentrated in immune-related tissues. Following a VHSV challenge, an overall increase in the transcript levels of the genes was observed and the expression patterns were distinctly divided into early and later responses during VHSV infection. In conclusion, olive flounder lectins are specifically expressed in immune-related organs and induced in both the immediate and long-lasting immune responses to VHSV in the olive flounder. These results indicate that lectins may be play important roles in the host defense mechanism and involved in the innate and adaptive immune response to viruses in fish.

• Journal of Aquaculture
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• v.21 no.1
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• pp.13-18
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• 2008

Marine birnavirus(MABV) are well known fish pathogens in Asian countries such as Korea, Japan, and China. Prevalence of viral disease, geological distribution and host or reservoir of viruses were investigated from wild marine fishes in southern coast of Korea in 2003 and 2005. RT-PCR results showed that MABV were detected in 17 fishes(10.6%) from 160 fishes. Comparative analysis of the nucleotide sequences and the deduced amino acids of MABV genome from wild fishes were similar to reference strains of MABV and distinguished with IPNV strains.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1324-1334
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• 2019

Fish mycobacteriosis is a common bacterial disease in many species of freshwater and marine fish and has caused severe loss of fish production. Mycobacterium marinum has been the most prevalent pathogen observed in several outbreaks of mycobacteriosis of farmed sturgeons in China. However, the immune responses and pathology of sturgeons in mycobacterial infection are rarely studied. Therefore, we used the Illumina RNA-seq method to analyze the transcriptome profile of Acipenser schrenckii challenged with Mycobacterium marinum. To begin, 168,220 non-redundant contigs were acquired from the infection and control groups, and among these, 33,225 contigs have acquired annotations. A total of 4,043 differently expressed (DE) contigs between the two groups were identified, and among these, 2479 were up-regulated and 1564 were down-regulated in the infected fish. A total of 1,340 DE contigs with acquired annotations in KEGG were enriched for 124 pathways including the TNF signaling pathway, and the Toll-like receptor signaling pathway. The roles of DE genes involved in significant pathways and other processes were discussed. The 2,209 DE contigs that have yet to acquire proper annotation may represent candidate genes associated with infection in sturgeons and are expected to serve as immunogenetic resources for further study. To our best knowledge, this is the first transcriptome study on sturgeons under bacterial infection.

• Journal of fish pathology
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• v.34 no.1
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• pp.105-110
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• 2021

We experienced mortality of dark-banded rockfish (Sebastes inermis) maintained in the rearing facility located in Gangneung, after transportation. Moribund fish showed various symptoms such as exophthalmia, skin ulcers, tail rots, gill rots, discoloration of liver with petechiae, yellowish fluid in intestines and ascites. Two different colonies were dominantly appeared after spreading the lesions on the agar plates and incubation. One isolate (SI_1) showed swarming movement on TSA, and formed yellow colonies on TCBS agar. The other (SI_2) showed no swarming motility and green colonies on TCBS agar. Both of them were Gram-negative. All of these results are similar with those of Genus Vibrio. They were identified as V. harveyi and V. gigantis by PCR with subsequent sequencing of 3 different genes (16 rDNA, recA, rpoA). V. harveyi is well-known as a serious pathogen of marine fish and invertebrates, while V. gigantis is known to be often isolated from marine invertebrates, but the pathogenicity is still unknown. We suspect V. harveyi as the cause of the mortality of dark-banded rockfish, but challenge experiments with these 2 Vibrio species are thought to be necessary to make a clear conclusion.

• Journal of fish pathology
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• v.35 no.2
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• pp.157-165
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• 2022

Single-cycle viruses generated by reverse genetic technology are replication-incompetent viruses due to the elimination of gene(s) essential for viral replication, which provides a way to overcome the safety problem in attenuated viruses. Infectious hematopoietic necrosis virus (IHNV) is a major pathogen causing severe damage in cultured salmonid species. In the present study, we generated a single-cycle IHNV lacking the transmembrane and cytoplasmic domain in the G gene (rIHNV-GΔTM) and evaluated the prophylactic potential of rIHNV-GΔTM in rainbow trout (Oncorhynchus mykiss). To produce rIHNV-GΔTM, IHNV G protein-expressing Epithelioma papulosum cyprini (EPC) cells were established. However, as the efficiency of rIHNV-GΔTM production in EPC cell clones was not high, fish were immunized with a low-tittered single-cycle virus (1.5 × 10² PFU/fish). Despite the low dose, the single-cycle IHNV induced significant protection in rainbow trout against IHNV infection, suggesting high immunogenicity of rIHNV-GΔTM. No significant difference in serum ELISA titers against IHNV between the rIHNV-GΔTM immunized group and the control group suggests that the immunized dose of rIHNV-GΔTM might be too low to induce significant humoral adaptive immune responses in rainbow trout. The involvement of adaptive cellular immunity or innate immunity in the present significantly higher protection by the immunization with rIHNV-GΔTM should be further investigated to know the protection mechanism.