• Journal of fish pathology
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• v.19 no.1
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• pp.83-86
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• 2006

Effect of iron on the extracellular proteolytic activity of live Uronema marium was determined by fluorescence polarization (FP) method. Supplementation of 0.5 and 5.0 μM iron significantly increased caseinolytic activity of live U. marinum. In contrast, supplementation of 50 μM iron showed no significant differences in FP values compared to the control. The present result suggests that iron in cultured water or skin tissue of olive flounder may influence on the penetration and establishment of U. marinum, correlating with modulation of extracellular protease activity of the ciliates.

• 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.

• Fisheries and Aquatic Sciences
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• v.5 no.4
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• pp.311-313
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• 2002

Proteolytic activity of live Uronema manum was analyzed by fluorescence polarization (FP) technique. Protease activity was measured by a decrease in FP value using fluorescein isothiocynate (FITC)-casein as a protein substrate. The results demonstrated an inverse linear relationship between fluorescence polarization (FP) values and live ciliate concentration over the range $1\times10^4\;to\;2\times10^5$ cells/well. However, the FP values of $10-10^3$ live parasites were not different significantly from that of control. Time-dependent decrease in FP value was shown in the wells containing live U. marinum. In the present study, FP assay had the benefit to provide measurements of substrate hydrolysis by live parasites in real-time, and did not require separations, precipitations, or transfers of reaction mixture.

• Fisheries and Aquatic Sciences
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• v.5 no.3
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• pp.145-149
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• 2002

The effects of pH, temperature and various inhibitors on the proteolytic activity of the cell lysate of Uronema marium were investigated using colorimetric and substrate gel electro­phoretic methods. The cell lysate of U. marinum showed proteolytic activity over a wide range of pH, and pH optima ranged from pH 5 to 7. The proteolytic activity was increased according to a rise of temperature but decreased at $40^{\circ}$. The proteolytic activity of the parasite lysate was significantly inhibited by protease inhibitors including trans-epoxysuccinyl -L-leucylamido-(4-guanidino) butane (E-64), pepstatin A, phenyl-methanesulfonyl fluoride(PMSF), and ethylenediamine-tetraacetic acid (EDTA). Preincubation of the lysate with E-64 showed the maximum inhibition of the caseionolytic activity. Four protease bands (152, 97, 67 and 40 kDa) were detected by gelatin SDS-PAGE. Significant inhibition of caseinolytic activity and complete abolition of a 152 kDa band in gelatin SDS-PAGE by EDTA indicated that the cell lysate of U. marinum had a metalloprotease Another three proteolytic bands were inhibited by E64, a cysteine protease inhibitor. Preincubation of the cell lysate with pepstatin or PMSF had no effects on the protease bands.

• Journal of fish pathology
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• v.17 no.2
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• pp.145-149
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• 2004

The definitive identification of ciliate species by morphological characteristics relies on time-consuming and laborious staining techniques. Therefore, in this study, we discriminated 3 scuticociliatosis causing species - Pseudocohnilembus persalinus, Uronema marinum and Philasterides dicentrarchi - in cultured olive flounder by multiplex PCR. The multiplex PCR based on the species-specific amplification of small subunit ribosomal RNA (SS rRNA) gene sequence enabled us to distinguish the 3 scuticociliate species in a simple and rapid manner, even in the sample containing the three species simultaneously. These data suggest that the multiplex PCR strategy would make it possible to avoid the cumbersome and time-consuming procedures of morphological analysis for the definitive identification of scuticociliates.

• Fisheries and Aquatic Sciences
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• v.6 no.3
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• pp.155-159
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• 2003

Lipid composition and fatty acid composition were characterized in the membrane of a marine ciliate protozoan (Uronema marinum). Phospholipids accounted for 70% of total lipid, and the remainder was neutral lipids. Total phospholipids were separated as phosphatidylcholine $(24.26\%)$, phosphatidylethanolamine $(22.21\%)$, phosphatidylinositol $(6.14\%)$, phosphatidyl­serne $(5.11\%)$, cardiolipin $(3.07\%)$ and unidentified phospholipids $(28.72\%)$ through high performance liquid chromatography (HPLC). Fatty acid composition of neutral lipids and phospholipids was determined by gas chromatography (GC), based solely on comparision of retention times. In neutral lipids, the most abundant fatty acid group was monounsaturated fatty acid $(48.3\% of total fatty acids)$ with oleic acid (18:1) and nervonic acid (24:1). Saturated fatty acids comprised $29.6\%$ of total fatty acids, with palmitic acid (16:0), stearic acid (18:0) ane myristic acid (14:0), and polyunsaturated fatty acid accounted for $33.0\%$ with $Di-homo-\gamma-linolenic$ acid (20:3) and linoleic acid (18:2). Wherease phospholipids predominantly contained the fatty acid group in the following order: polyunsaturated fatty acids $(52.7\%\;of\;total\;fatty\;acids)$ with linoleic acid (18:2) and $\gamma-linolenic$ acid (18:3) > monounsaturated fatty acids $(28.5\%\;of\;total\;fatty\;acids)$ with oleic acid (18:1) and palmitoleic acid (16:1) > saturated fatty acids $(25.5\%\;of\;total\;fatty\;acids)$ with palmitic acid (16:0), stearic acid (18:0) and myristic acid (14:0).

• Journal of fish pathology
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• v.34 no.1
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• pp.47-53
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• 2021

Scuticociliates are one of the serious parasitic threats faced by the marine aquaculturists worldwide. To date, Uronema nigricans, Philasterides dicentrarchi, Miamiensis avidus, Uronema marinum, and Pseudocohnilembus persalinus have been reported as the important culprit species causing scuticociliatosis in fish. The present paper reports the finding of an additional scuticociliate isolate from the gill of diseased olive flounder Paralichthys olivaceus in Korea. Based on the morphological characteristics, a scuticociliate in this study was identified as Paranophrys marina. Phylogenetic analysis placed P. marina as a sister lineage to three species of Pseudocohnilembus and Mesanophrys carcini within the order Philasterida.

• Journal of Species Research
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• v.6 no.spc
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• pp.15-25
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• 2017

Twelve ciliates were collected from freshwater, marine, and brackish water habitats in Korea and taxonomic descriptions are provided based on observations of living cells and protargol-impregnated specimens. These ciliates, recorded for the first time in Korea, were classified into 12 genera, 11 families, 10 orders, and five classes. We provide brief descriptions and remarks, including microphotographs, for the 12 ciliates: Allotricha mollis, Amphileptus eigneri, Didinium gargantua, Holophrya teres, Lacrymaria marina, Novistrombidium apsheronicum, Pelagostrobilidium conicum, Pseudochilodonopsis fluviatilis, Pseudourostyla subtropica, Strombidium conicum, Urocentrum turbo, and Uronema marinum.

• Journal of fish pathology
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• v.21 no.1
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• pp.1-11
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• 2008

We report the existence of new type of phosphatidylcholine-hydrolyzing phospholipase D (PLD), which has been characterized and partially purified in the scuticociliate, Uronema marinum. The enzyme from partial purification showed that it was existed in membrane fraction and was a neutral PLD, which catalyzed both transphosphatidylation and hydrolysis reaction. The activity of partially purified membrane-bound PLD was also found to be optimal at pH 7.0-7.5 for 2 hours at 37℃ and depended strictly on the presence of Ca2+ (2.5 mM) and Mg2+ (1.6 mM). Immunoblot analysis indicated that the enzyme was distinct from hPLD1 (human PLD1) and hPLD2 (human PLD2) because it was not recognized by a polyclonal antibody raised to the 12 terminal amino acid of these enzymes. We also found that the membrane-bound PLD is a PIP2-dependent PLD and that GTP-binding proteins are not implicated in the regulation of this enzyme: This enzyme activity is markedly stimulated by phosphatidylinositol 4,5-bisphosphate (PIP2) but not by the small G-protein Arf and GTPrS. In addition, this enzyme was capable of hydrolyzing phosphatidylcholine (PC) but not phosphatidylethanolamine (PE), implying that PC was a preferred substrate.