• Fisheries and Aquatic Sciences
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• v.26 no.10
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• pp.605-616
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• 2023

Scuticociliatosis, caused by the parasitic pathogen Miamiensis avidus, poses a significant threat to olive flounder farms in South Korea. Infected fish suffer from severe systemic infections affecting various organs, with potential secondary bacterial diseases. This study investigated the emergence of different M. avidus serotypes in 20 olive flounder farms on Jeju island, South Korea, from 2015 to 2020. Additionally, we identified Tenacibaculum maritimum as a co-infecting bacteria. Based on serotyping and monitoring data, we developed a trivalent vaccine targeting two serotypes of M. avidus and one strain of T. maritimum. The efficacy of the vaccine was evaluated under laboratory conditions and demonstrated a relative percentage of survival (RPS) of 75%, 80%, and 93% for M. avidus serotype I, M. avidus serotype II and T. maritimum, respectively. Furthermore, successful field trials conducted on four different olive flounder farms resulted in significantly higher survival rates (52%-76% RPS) and weight gains in vaccinated fish. Overall, this study presents an effective vaccine against M. avidus and T. maritimum infections in farmed olive flounder, making a valuable contribution to sustainable aquaculture in South Korea.

• Journal of fish pathology
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• v.35 no.1
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• pp.113-120
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• 2022

Scuticociliatosis caused by Miamiensis avidus is a very important parasitic disease in olive flounder farming industry. The aim of this study was to determine effect of combined treatment with blue LED (light-emitting diode) illumination and formalin on olive flounder (Paralichthys olivaceus) infected with M. avidus. Different intensity of 405 nm LED (20, 40, and 60 μmol·m^-2·s^-1) was illuminated on 2.2×10⁴ cells/well of M. avidus in a 24 well microplate for 24 h. Also, 2.4×10⁴ cells/well of M. avidus were exposed to varying combinations of 60 μmol·m^-2·s^-1 of 405 nm LED and serial 10-fold dilutions of formalin (from 10 to 100 ppm) for 15, 30, 45, and 60 min. Surviving M. avidus were counted using a hemocytometer. For in vivo test, flounder acclimatized at 11-12 practical salinity unit (psu) were challenged with 2×10⁶ cells/ml of M. avidus by immersion method for 1 h. Then, fish were moved and divided into four groups; "F" group, treated with formalin at 50 ppm; "L" group, treated with 60 μmol·m^-2·s^-1 of 405 nm LED; "C" group, treated with combination of the two methods; and the control group. After treatment for 30 min, fish were transferred to new tanks (salinity = 11-12 psu) and observed for 3 weeks. As a result, illumination of 405 nm LED at 60 μmol·m^-2·s^-1 killed 100% of M. avidus after 12 h, while 67% and 90% of the scuticociliate died at 20 and 40 μmol·m^-2·s^-1, respectively, after 24 h exposure. One hundred percent of M. avidus was killed at 90, 80, 80 and 70 ppm after exposure to formalin for 15, 30, 45 and 60 min, respectively. However, combined method (e.g., 60 μmol·m^-2·s^-1 of 405 nm-LED plus 50 ppm formalin) killed the parasite within 30 min. From in vivo test, similarly, survival rates of fish challenged with M. avidus were 100%, 43%, 29% and 0% in the C, F, L, and control groups, respectively. Results obtained in this study demonstrates that the combined treatment method has clear synergistic effect on scuticociliatosis in fish.

• Journal of fish pathology
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• v.28 no.3
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• pp.171-174
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• 2015

Previously, we had reported that some Miamiensis avidus, a major pathogen of scuticociliatosis in cultured olive flounder, strongly agglutinated by flounder immune sera could escape from the agglutinated mass within a few hours. In the present study, we observed that M. avidus not only escaped from the agglutinated mass but also conducted division(s) before shedding its old covering. Furthermore, ciliates that survived the antibody-dependent complement killing (ADCK) assay were not killed even when re-exposed to a freshly prepared ADCK assay. This result suggests that the liberated ciliates from the ADCK assay might change not only their i-antigen types but also the epitopes of major surface antigens, which debilitate antibody-mediated complement killing ability.

• Journal of fish pathology
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• v.15 no.3
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• pp.111-115
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• 2002

The scuticocilliate, Uronema marinum is a histophagous ciliate and the causative agent of 'scuticociliatosis'in cultured olive flounder Paralichthys olivaceus. In the present study, in vitro efficacy of hydrogen peroxide, formalin and copper sulfate on the scuticocilliate at low salinity was investigated. Each chemical showed synergistic parasiticidal effects with low salinity (salinity in 5 ppt) compared to each chemical alone (salinity in 33 ppt). At low salinity (5‰), ciliates were killed completely within 1.5h by exposure to 50ppm formalin (37% formaldehyde), at 100ppm hydrogen peroxide (30% solution) and at 100ppm copper sulfate (20% solution). The formalin was the most effective chemical against the parasites at low salinity.

• Journal of fish pathology
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• v.15 no.2
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• pp.77-82
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• 2002

Effects of stress-induced suppression of humoral immunity on scuticociliate killing activity of olive flounder plasma were investigated. Changes in glucose level. alternative complement activity and Iysozyme activity of plasma by handling stress were analysed in relation to in vitri parasiticidal activity of plasma. The plasma glucose level was about two times higher in fish after a handling stress than in control fish. Plasma lysozyme activity and natural haemolytic activity were decreased in suessed fish.The scuticociliate killing activity of plasma was significantly lower in stressed fish than in non-stressed control fish. The present results indicated that stress-induced immunodepression could be a cause of scuticociliatosis occurrence in olive flounder.

• Journal of fish pathology
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• v.31 no.1
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• pp.15-21
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• 2018

Differences in in vivo virulence and in sensitivity to drugs among different isolates of Miamiensis avidus were analyzed. The isolate III showed the highest resistance against the scuticocidal activity of olive flounder (Paralichthys olivaceus) serum, and induced the highest mortalities of olive flounder fingerlings. The isolate II showed significantly higher serum resistance than the isolate I, but in vivo virulence of isolate II was not significantly different from that of isolate I. The secreted proteinases activity of isolate III was significantly higher than that of isolate I and II, and the activity was significantly reduced by the addition of E-64, a cysteine proteinases inhibitor. There were no differences among isolates in the sensitivity to doxycycline, however, there were significant differences in sensitivities to mebendazole and bithionol. These results suggest that the different characteristics of different M. avidus isolates should be taken into consideration for the development of control measures against scuticociliatosis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.1
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• pp.45-51
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• 2014

Scuticociliates are regarded as serious pathogens in marine aquaculture worldwide. In Korea, they cause mass-mortalities in fish such as the commercially important olive flounder Paralichthys olivaceus. In particular, mixed infections of scuticociliates with pathogenic bacteria have been commonly reported. During efforts to identify natural marine-algae derived products that possess anti-bacterial and anti-scuticociliate properties, we found that an 80% methanolic extract of the red alga Polysiphonia morrowii Harvey exhibits both anti-scuticociliate activity against Miamiensis avidus, which is a major causative agent of scuticociliatosis, and anti-bacterial activities against fish pathogenic bacteria. Activity-guided fractionation and isolation of the 80% methanolic extract of P. morrowii yielded three bromophenols, which were identified as 3-bromo-4,5-dihydroxybenzyl methyl ether (1), 3-bromo-4,5-dihydroxybenzaldehyde (2) and urceolatol (3) based on spectroscopic analyses. 3-bromo-4,5-dihydroxybenzyl methyl ether (1) showed the highest anti-bacterial and anti-scuticociliate activities, with a minimal inhibitory concentration (MIC) of $62.5{\mu}g/mL$ (against Vibrio anguillarum) and minimal lethal concentration (MLC) of 62.5 ppm (in seawater). Investigations of the anti-bacterial and anti-scuticociliate activities of seventeen bromophenol derivatives, including the three isolated natural bromophenols, showed that the existence of an electron donating group or atom with a non-covalent electron pair at $C_4$ of the 2-bromophenol structure may be important in anti-scuticociliate activity. These findings suggest that the extract and bromophenol derivatives of P. morrowii may provide useful alternatives in aquaculture anti-scuticociliate therapies.

• Journal of fish pathology
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• v.32 no.1
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• pp.29-35
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• 2019

A monitoring was performed to survey the mortalities that had occurred in the aquaculture farms of olive flounder (Paralichthys olivaceus) in South Korea from 2015 to 2017. The indirect inquiry for entire farms and the sample survey for selected farms were carried out. The aquatic organism disease inspectors, who have a national license for the diagnosis and prevention of aquatic organism diseases and a have close relationship with the farms, investigated the rates and causes of mortalities according to the standard manual. The mortality rate by sample survey of farms in 2015, 2016, and 2017 were 24.78% (Chunnam: 17.86%, Jeju: 28.69), 30.19% (Chunnam: 24.45%, Jeju: 32.65), and 21.59% (Chunnam: 10.57%, Jeju: 26.00%), respectively. The major cause of mortality was scuticociliatosis, and the mortality caused by viral hemorrhagic septicemia and emaciation disease (Jeju) were also high. Our results can contribute to effective establishment prevention of epidemics system and acquired status as a disease-cleansing country.

• Journal of fish pathology
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• v.33 no.2
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• pp.177-184
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• 2020

In an attempt to find a feed additive showing an anti-scuticociliate effect, extracts from Citrus unshiu Peel were tested against virulent scuticociliate infection. The most effective anti-scuticociliate killing activity in vitro was observed in the extract squeezed from homogenizing water-soaked dried tangerine peel (DTP). In addition, we have investigated the effect of DTP as a feed additive on growth rate and anti-parasitic activity of olivaceus flounder. DTP extract added diets (0.1, 0.5, 1, and 5%/feed weight) were fed to flounder for 7 days for checking a growth rate and 14 days for a challenging test. As a result, the feed conversion rate was significantly improved only in 1% DPT extract group compared to the control and 0.5% DTP extract fed group showed 100% of survival rate in the challenge test, all of which indicating that DTP extract would be a potential feed additive against scuticociliatosis.

• Journal of fish pathology
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• v.7 no.2
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• pp.151-160
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• 1994

In order to elucidate the patterns of tissue damage evoked by the scuticociliatids, eighteen fingerlings of Japanese flounder, Paralichthys olivaceus, heavily infected with an unidentified scuticociliatid were histopathologically examined. Skin layer with the underlying musculature were severely necrotized due to the infestation of the ciliates. However in the early lesions, both dermis and myofibres remained relatively intact compared with other surrounding loose connective tissues. Mild damages were found in more dense tissues. One or more scuticociliatids were recognized in the blood and lymph vessels of the loose connective tissues with or without destructive changes. Many of nerve trunks or ganglia were also parasitized with less marked histological damage in the parenchyma. Dura and its adjacent tissues in the spinal cord were severely necrotized with massive accumulation of the ciliates in subdural space. The parasitic invasion in the central nervous system was usually confined to the cortical region. In the gill, variable degenerative changes were occurred due to the invasion of the ciliates recognized in the blood vessels of branchial arches or primary filaments. From these results, it was strongly suggested that the scuticociliatids are very actively penetrated into the deep tissues mainly through the severe destruction of the loose connective tissue components and that the vascular system could play a role in the rapid distribution of the ciliates to the remote tissues or organs.