• Journal of fish pathology
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• v.22 no.3
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• pp.229-237
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• 2009

The causative agent for the tunic softness syndrome of the cultured ascidian Halocynthia roretzi from Jan 1999 to Feb 2009 was identified using virus isolation and polymerase chain reaction (PCR). The pathogenicity of the isolated virus MABV UR-1 strain was determined by experimental infection trials. The cytopathic effects was observed in CHSE-214 cell line at a level 5.1% (4/78) in normal ascidian and 1.8% in abnormal ascidian showing tunic softness syndrome signs. MABV gene was detected in 16.8% (18/107) of normal and 13.1% (5/38) of abnormal organisms by PCR. The ratio of MABV isolation and gene detection was similar level in normal and soft tunic diseased ascidian. Based on the VP2/NS junction region sequences, eight strains of virus isolated from ascidian, were included in the same genogroup with MABV which is originally isolated in wide ranges of marine fish and shellfish species. The UR-1 strain caused 60% mortality (36.5% mortality in control group) by immersion infection and 37% mortality (same mortality in control group) in injection infection indicating no significant differences in infected and control groups. These results suggest that ascidian can act as reservoir of the MABV, and this virus is not directly related with the ascidian mortality.

• Journal of fish pathology
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• v.34 no.1
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• pp.63-70
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• 2021

As a part of research to elucidate the pathogenesis of so called Soft Tunic Syndrome(STS), that caused mass mortalities in the cultured sea squirts, Halocynthia roretzi, the epidemiological and pathological analysis were done to both clinically normal and diseased groups of the farms of Tongyoung and Geoje coastal areas in southeast sea from February to July, 2008. In the histological finding of the tunic, most of individuals showed tunic softness syndromes that included the disarrangement and destruction of tunic fiber with the simultaneous presence of flagellates-like cells, recently suspected as main agents of tunic softness syndromes. Simultaneously, the intensive degenerative changes of the skeletal muscle of diseased sea squirts were recognized. The changes were characterized with the hyalinization and condensation of muscle fibril and hemocytic infiltration in the muscle fibers. Those were thought to be a kind of typical Zenker's necrosis as in the skeletal muscle of higher vertebrates. Besides of the diseased sea squirts, Zenker's necrosis of skeletal muscles were seen in the normal ones. Epidemiological inquiry for diseased groups revealed that the higher incidences of tunic softness syndrome were recorded in the fast growing groups and in the sites presuming the organic pollution. And Higher malondialadehyde(MDA) and glutathione peroxidase(GPx) activity were detected in the groups showing STS. Those results suggested that Zenker's necrosis of body muscles was a kind of"nutritional myopathy" by oxidative stress. Conclusively, it was considered that Zenker's necrosis of body muscles gives an important clue for elucidating pathogenesis of STS of cultured squirts. And it seems that the necrosis were caused by the oxidative stress to body muscle during abnormal rapid growth of sea squirts.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.46 no.5
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• pp.575-581
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• 2013

We examined the effect of starvation on the occurrence of tunic softness to determine the cause of mass mortality of cultured Halocynthia roretzi (Drasche) in the Tongyeong region, Korea. In terms of the survival rate of H. roretzi and the occurrence rate of tunic softness, H. roretzi starved for 35 days at water temperatures of 8, 12, and $15^{\circ}C{\pm}0.5^{\circ}C$ (room temperature of $15^{\circ}C{\pm}1^{\circ}C$) did not exhibit tunic softness at water temperatures of either $8^{\circ}C$ or $12^{\circ}C$. for morphological changes, although the tunic of H. roretzi was shrunken and became visibly smaller with a darkening color in all experimental groups, as compared to the state prior to starvation, its tunics bulbs continuously. The ratio of RNA/DNA concentrations and protein contents for each of the tunic sections were lower in the starved group. Our results indicate that tunic softness is not related to feeding deficiency, as no histopathological symptoms were apparent in the digestive gland or tunic of H. roretzi due to starvation.

• Journal of fish pathology
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• v.24 no.3
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• pp.197-204
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• 2011

The edible ascidian Halocynthia roretzi is a commercially important fisheries resource in Korea. However, for the last several years, outbreaks of mass mortalities of the species have been occurring along the south and east coasts of Korea, where most ascidians are produced. Although it is known that tunic-softness syndrome is associated with these mortality events, the agent causing the syndrome has not yet been confirmed. To determine the agent causing tunic-softness syndrome, healthy and diseased ascidians were collected in March 2011 from Tongyeong, on the south coast of Korea, and were used for biological and pathological investigations. The results showed that diseased ascidians exhibited remarkably reduced body fluid, fatness index, and tunic index compared with healthy specimens. Interestingly, bi-flagellated protozoans were observed specifically in the tissue imprints and tunic cultures of diseased ascidians at an occurrence rate of 97.5%. Histological observation showed that the thickness of the tunics of diseased ascidians was reduced by half, and irregular structure and breakdown of the tunic fiber bundles were observed. In particular, flagellate-like cells were observed in the diseased ascidians. Our study clearly shows that bi-flagellated protists are present only in the softened ascidians, suggesting that the flagellates are partly or entirely associated with soft-tunic syndrome. Accordingly, further investigations to verify the effects of the flagellates found in the present study on soft-tunic syndrome should be conducted.

• Fisheries and Aquatic Sciences
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• v.15 no.2
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• pp.151-155
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• 2012

We used nine microsatellite DNA markers to estimate genetic variation among wild and cultured populations of the sea squirt Halocynthia roretzi. The loci were polymorphic, with 6-32 alleles, and allelic richness ranged from 6.0 to 26.1 in each population. The wild and the cultured populations had similar mean heterozygosities ($H_O$ and $H_E$), allele numbers, and allelic richness. One cultured population with softness syndrome had a lower mean in the observed heterozygosity ($H_O$ = 0.57) and higher mean inbreeding coefficient ($F_{IS}$ = 0.261) than any other populations. This suggests that the loss of genetic variation in the diseased population might be due to increased inbreeding. A neighbor-joining tree and pairwise population estimates of $F_{ST}$ showed moderate genetic differentiation between the wild and the cultured populations. Additionally, the softness syndrome population was genetically divergent from wild populations, but it was genetically close to the cultured populations.

• Proceedings of the Korean Aquaculture Society Conference
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• 2006.05a
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• pp.533-534
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• 2006

• Journal of fish pathology
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• v.23 no.3
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• pp.313-322
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• 2010

Edible tunicate Halocynthia roretzi, one of the most commercially important aquatic organisms in Korea, has been killed by tunic softness syndrome since last decade. The intracellular protistan parasite observed by the transmission electron microscope in hemocytes of the tunicate was considered to be the causative agent of the mass mortality. The goal of the present work is to examine the characteristic features of the parasite by identifying the 18S rDNA sequences of the parasite. The experiments conducted include amplification of presumptive 18S rDNA from diseased tunicate tissues with UNonMet-PCR and sequencing the product. A preliminary phylogenetic analysis was performed on the presumptive parasite rDNA. A digoxigenin labeled DNA probe was designed on the basis of the sequences of rDNA. Dig-ISH assay was conducted to diagnose the protistan parasite. A PCR using UNonMet-PCR primer generated 595 bp SSU rDNA fragment. Subsequently, PCRs with primer pair expended this sequence to 1542 bp. This is the first partial sequences of SSU rDNA gene to be published on the protistan parasite that has presumed causing the mass mortality of tunicate. Since the Dig-ISH technique demonstrated the presence of infection in hemocytes on the all host tissues, the fragment was confirmed to be the intracellular protistan parasite SSU rDNA. A phylogenetic analysis suggested that the protistan parasite may be a unique eukaryote that is closely related to Apicomplexa.

• Korean Journal of Environmental Biology
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• v.31 no.3
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• pp.204-212
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• 2013

This study analyzed the occurrence of tunic softness, survival rate, metabolic rate and histopathologic changes arising from the effect of hypoxic environment in order to find the causes of occurrences of tunic softness, which manifests as the key phenomenon of mass mortality of Halocynthia roretzi. Regarding the survival of H. roretzi with reduction in dissolved oxygen, all the entities died on the 4th day of exposure to the dissolved oxygen concentration of $2mg\;L^{-1}$ while 50% mortality was observed on the 5th day of exposure to the dissolved oxygen concentration of $3mg\;L^{-1}$. Therefore the 5 days-$LC_{50}$ was found to be $3.55mg\;L^{-1}$ (1.86~$4.96mg\;L^{-1}$). However, occurrence of tunic softness was not observed during the period of exposure to low oxygen concentration. The oxygen consumption rate significantly decreases at the dissolved oxygen concentration of less than $5mg\;L^{-1}$ in comparison to the control group. Therefore, it is presumed that H. roretzi controls the respiration rate for prescribed period of time when exposed to hypoxic environment. Regarding the histopathologic changes in the gill, digestive gland and cyst of H. roretzi due to hypoxic environment, necrosis of epithelial layer, in filtration of blood cells, and condensation of nucleus that compose each of the organs were observed. Regarding morphological changes, the decrease in volume with shrinking of the tunic, discoloration of the internal organs and necrosis of gill and hepatopancreas were observed.

• Journal of Life Science
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• v.23 no.2
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• pp.190-196
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• 2013

The sea squirt, Halocynthia roretzi, has experienced mass mortality due to softness syndrome. The identification of disease-induced genes can provide insights into the development of this syndrome. To identify the genes, we performed differentially expressed gene (DEG) analysis. The expression of the phosphoglycerate kinase (HrPGK) gene was significantly decreased in diseased sea squirts compared to normal ones. We confirmed the result of the DEG analysis through RT-PCR and real-time PCR. In addition, we detected single nucleotide polymorphisms at position -106 (A/T) and -254 (G/T) in the HrPGK gene promoter by genotyping analysis. At the -106 site of the HrPGK gene, the frequency of the AA allele in disease-resistant sea squirts was about two-fold higher than that of sensitive ones, and the frequency of the TT allele in the disease-resistant sea squirts was about six-fold lower. At the -254 site of the HrPGK gene, the frequency of the GT and the GG allele was approximately two-fold higher and two-fold lower, respectively, in the disease-resistant sea squirts compared to the disease-sensitive ones. Analysis of the relationship between the genotypic variation at the -106/-254 promoter and the expression of HrPGK mRNA showed that HrPGK mRNA expression was higher in the -106/-254 AA/GT genotype samples than in the -106/254 TT/GG genotype ones. These results show that sea squirts harboring the AA/GT genotype may have more resistance to mortality than the sea squirts with other genotypes.

• International Journal of Costume and Fashion
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• v.4 no.2
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• pp.18-29
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• 2004

The purpose of this study was to analyze the formative beauty of metro sexual expressed on the 21C men's fashion. in 21C, metro sexual is being accepted as an object to lead the world consumption culture, and it is said to be most appropriate to the definition as a model that can determine the most modern men. Moreover, with well being life style, it shows characteristic that expresses chic sense through fashion. The results of the study were as below. 1. In recent men's wear, sensory feminine that looks young and trendy is being presented through clothes, rather than the image of men who are strong. Therefore, by wearing splendid flower print, bold color and gorgeous accessories, men send off their attractions. 2. The recent body king syndrome gave new aesthetic value on men's body. Therefore, men's body is becoming cultural icon of metro sexual as another face that shows social status and cultural taste. 3. The men's image of metro sexual that is felt from softer skin and hair than women, and well-built body is appealing more through body conscious look. Here, men's body silhouette is being sublimated aesthetically through design with various materials such as see-through and leather etc., and this is body conscious image that can be seen only in metro sexual. Like this, metro sexual fashion sense was expressed by feminine softness sometimes, and beautiful body silhouette sometimes by expressing one's sexiness enough, being away' from conceptual masculine. Therefore, the appearance of metro sexual can be understood that men's body exposure is becoming a social virtue that expresses beauty.