• Journal of fish pathology
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• v.21 no.3
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• pp.157-166
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• 2008

Streptococcal diseases are known as serious problems in fresh water and marine fish culture industry worldwide. Recently, the importance of the infection of Streptococcus parauberis has been increased among the streptococcicosis since severe outbreaks in cultured olive flounder (Paralichthys olivaceus) were recorded in Korea. The aims of the present study were to investigate immunomodulatory differences between S. parauberis and S. iniae. In this study, S. parauberis isolated from diseased olive flounder was investigated on the characteristics of morphological and immune responses. Immune response of the fish to the pathogen was characterized by bactericidal activity in the serum, phagocytic activity and reactive oxygen intermediaters (ROIs) production of olive flounder. The capsules of both bacteria, S. parauberis and S. iniae seem to be associated with the ability to resist killing activity of normal sera. Also, capsulated strains could survive in the phagocytes and induce to lower ROIs production.

• Journal of fish pathology
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• v.20 no.1
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• pp.49-60
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• 2007

Non-hemolytic Streptococcus parauberis isolated from diseased olive flounder, Paralichthys olivaceus in the South coast of Korea were identified by physiological, biochemical and genetic analysis in order to define the different characteristics geographically. First, twelve strains of S. parauberis were isolated from catalase-negative gram-positive cocci by multiplex PCR assay. Phenotypic identifications were performed with commercially available kit (API 20 Strep and API ZYM system). Analysis of API profiles of the isolates showed that strains were identified as either of Lactococcus lactis, S. constellatus or S. uberis. Moreover, S. parauberis isolated from olive flounder differed from that of turbot (X89967) to the test of not Voges-Proskauer, arginine, hippurate, alkiline phosphatase and pyrroidonyl arylamidase but β-glucuronidase. All S. parauberis isolates were sensitive to florfenicol, ampicillin, ofloxacin and vancomycin but were resistant to oxolinic acid, flumequine, nalidixic acid and sulfisoxazol. However, the 16S rDNA sequences of the isolates showed 99% similarity to S. parauberis KCTC 3651 (AY584477) and a great homogenecity among the flounder isolates.

• Journal of Life Science
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• v.24 no.4
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• pp.428-436
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• 2014

Loop-mediated isothermal amplification (LAMP) technique relies on autocycling strand displacement DNA sysnthesis without template denaturation steps under isothermal conditions. LAMP has more advantages than modern PCR, as it requires only basic laboratory equipment like an isothermal water bath, oven, and heating cabinet. Hence, in this study, five random primers were designed with Streptococcus parauberis, shikimate kinase Arok gene sequences (Genbank accession number: CP0024711). Two primers were selected based on the ladder pattern. Other optimum reaction conditions like temperature, time, and sensitivity were established and confirmed with conventional SYBR-green PCR. Results confirmed that the designed random primers were species specific, without any non-target DNA amplification under isothermal conditions. Hence, this study suggests that LAMP techniques could be used in the diagnosis of fish pathogen, specifically S. parauberis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.4
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• pp.382-388
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• 2019

Streptococcosis caused by Streptococcus parauberis is a very important disease in farmed olive flounder Paralichthys olivaceus. For most fish pathogens, including S. parauberis, there are no analytical criteria to distinguish antibioticsusceptible strains from antibiotic-resistant strains. In this study, epidemiological cut-off ($CO_{WT}$) values were generated to classify 75 strains of S. parauberis isolated from 1999 to 2018 as wild type (WT) and non-wild type (NWT) using disc diffusion data and normalized resistance interpretation (NRI) analysis. The susceptibility of the isolates to 16 antibiotics was evaluated using CLSI guideline M42-A. The wild-type cut-off values for amoxicillin, erythromycin, oxytetracycline, and florfenicol for S. parauberis were ${\geq}35$, 31, 28, and 27 mm, respectively. The NWT ratios of S. parauberis strains to treatment with GEN, FFC, ENR, SXT, EFT, VAN, and CHL were 17% or less, indicating that these antibiotics may be used to treat streptococcosis caused by S. parauberis. For recent S. parauberis isolates, the NWT ratios for AMX, ERY, OTC and FFC are much higher than for strains isolated from 1999-2007. The $CO_{WT}$ data from this study will assist aquatic animal disease professionals in prescribing appropriate antibiotics for the treatment of streptococcosis caused by S. parauberis, which will help reduce the misuse and abuse of antibiotics in the aquaculture sector.

• Journal of fish pathology
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• v.33 no.2
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• pp.119-125
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• 2020

During the period from 2003 to 2020, a total of 470 Streptococcus species were isolated from farmed flounder in Jeju. Out of 470 isolates, 92 (19.6%) isolates and 378 isolates (80.4%) were identified as Streptococcus iniae and S. parauberis, respectively by multiplex PCR assay. During that period, the percentage of S. iniae decreased from 56.9% in 2003 to 0.0% in 2020 whereas that of S. parauberis increased from 43.1% in 2003 to 100% in 2020. In the PCR assay for serotyping, the isolated S. parauberis showed 3 subserotypes, Ia (34.9%), Ib/Ic (46.3%) and II (18.8%). In 2003 and 2004, serotype II was dominant at 59.1% and 50.0% of isolation rates, however between 2005 and 2009, subserotype Ib/Ic was dominant (57.6%, 86.0%, 84.6%, 57.9%, and 83.3%). After 2010, except for 2015, subserotype Ia was the most dominant one. In the last 3 years (2018 to 2020), subserotype Ia was most abundant (70%), followed by subserotype Ib/Ic (16-30%). Serotype II was not isolated in 2018, but in 2019 and 2020, it showed an increased tendency to 3.4% and 16.7%, respectively. It is believed that continuous monitoring is necessary for research on counter-measures against streptococcosis of flounder.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.899-903
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• 2005

Two infectious species of Streptococcosis pathogens were detected by multiplex PCR assay. Detection rates of Streptococcus iniae and S. parauberis could reach 44.9% and 55.1% respectively for one year during 2004 to 2005 in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeju island. These findings showed that S. parauberis strains were important pathogen with streptococcosis of olive flounder in Jeiu island. In the present study we have investigated the interspecific relationship of all Jeju area of S. parauberis by RAPD analysis. Represent strains divided to four groups by RAPD fingerprints. The important differences observed between the olive flounder isolates suggest that they could constitute a well-differentiated group or a separate clonal line within this bacterial species. Though, serological research of S. parauberis strains in Jeju island not exist yet. These strains doing the serological evolution.

• Journal of fish pathology
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• v.23 no.1
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• pp.37-45
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• 2010

In this study, we carried out research on the level of single and multi-drug resistance of bacteria isolated from cultured flounder Paralichthys olivaceus. One hundred sixty one bacteria were isolated from cultured olive flounder Paralichthys olivaceus in Korea and the isolates consisted of Edwardsiella tarda (n=32), Vibrio ichthyoenteri (n=37), Vibrio spp. (n=54), Streptococcus parauberis (n=28) and Streptococcus spp. (n=10). These E. tarda isolates were highly resistant in the order of tetracycline (84.4%) and oxolinic acid (71.9%). V. ichthyoenteri and Vibrio spp. showed resistance ampicillin (94.6% and 81.5%) and tetracycline (56.8% and 42.6%). S. parauberis isolates were resistant ampicillin (57.1%), tetracycline (57.1%) and erythromycin (35.7%). Of the isolates, 84.4% of E. tarda, 73.0% of V. ichthyoenteri, 57.4% of Vibrio spp., 42.8% of S. parauberis and 70.0% of Streptococcus spp. isolates exhibited multi-drug resistance against more than two antibiotics.

• Journal of fish pathology
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• v.21 no.3
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• pp.209-217
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• 2008

A disease with mass mortality characterized exophthalmia and abdominal distension by ascites occurred in farmed starry flounder, Platichthys stellatus in Pohang since October of 2007. Severe necrosis and infiltration of inflammatory cells were observed in tissue histopathological sections from liver, kidney, spleen and heart of the diseased fish. The catalase-negative and gram positive cocci with -hemolysis were isolated from moribund fish. Analysis of the API profiles and the multiplex PCR of the isolates showed that strains were identified as Streptococcus parauberis. In API profiles, S. parauberis isolated from starry flounder differed from those of olive flounder, Paralichthys olivaceus to the test of arginine dihydrolase, D-mannitol, D-sorbitol and α-lactose. And the strains, PH0710 and PH0711, were strongly lethal for starry flounder (LD50 ranging from 4.2×103~4.2×102 cfu/fish at 7 days after intraperitoneally inoculation). This is the first report on a S. parauberis infection in starry flounder.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.3
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• pp.247-254
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• 2014

To investigate the acquisition of quinolone resistance, we examined mutations in the quinolone resistance-determining region (QRDR) of type II topoisomerase genes in ciprofloxacin (CIP)-resistant clinical isolates and in vitro mutants of Streptococcus parauberis. The CIP-resistant clinical isolates had one base change responsible for a Ser-79${\rightarrow}$Thr in the QRDR of parC. However, the CIP-resistant in vitro mutants had an altered QRDR of parC (Ser-79${\rightarrow}$Ile) that differed from that of the isolates. None of the CIP-resistant S. parauberis clinical isolates or in vitro mutants exhibited amino acid changes in gyrA or gyrB. However, even though involvement in the increased resistance was not clear, an Arg-449${\rightarrow}$Ser mutation outside of the QRDR of parE was detected in CIP-resistant mutant 2P1. These results suggest that the topoisomerase IV gene, parC (and possibly parE, as well), is the primary ciprofloxacin target in S. parauberis. Additionally we established a high-resolution melting (HRM) assay capable of detecting the dominant mutation in four type II topoisomerase genes conferring ciprofloxacin resistance. These rapid and reliable assays may provide a convenient method of surveillance for genetic mutations conferring antibiotic resistance.

• Journal of fish pathology
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• v.22 no.3
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• pp.263-273
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• 2009

There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.