• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.671-675
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• 2002

The present study was conducted to elucidate functional properties of sulfated polysaccharides from ascidian tunics, In physical properties of the crude polysaccharides, emulsion ability and foaminess were more excellent compared with chitin and chitosan, particular dye binding capacity was prominent. Anti-blood coagulation of partially purified sulfnted polysaccharides showed with respect to APTT (Activated partial thromboplastin time). Especially, active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed highest activity, which was approximately $20\%$ of the activity of heparin. ACE inhibitory activity also similar to anticoagulant activity. Active fraction $(F_4)$ obtained by DEAE-cellulose ion exchange chromatography showed about $34\%$, in ACE inhibitory activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.4
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• pp.603-608
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• 1998

Lipid oxidation in ascidian was studied when fresh, deshelled and sliced meats were fermented for 50 days at 5$\pm$2$^{\circ}C$ with 8%(w/w) salt and 0.1% papain. Antioxidative effects of butylated hydroxytoluene(BHT) and carotenoid extracts from ascidian tunic on lipid oxidation and oxidationrelated discoloration of ascidian meat during fermentation were investigated. Changes in peroxide value, carbonyl value, thiobarbituric acid value, fatty acids composition, the loss of total carotenoid and sensory evaluation were determined to assess the rancidity. Peroxide and carbonyl values in BHT and carotenoid extract treatments increased less than those of the control during fermentation. TBA value increased until 30 days, hereafter tended to decrease a little in the control during fermentation. TBA value increased until 30 days, hereafter tended to decrease a little in the control but it increased slowly until 40 days in cases of 0.02% BHT or 0.02% BHT with 0.05% carotenoid added. Fatty acids of fresh ascidian composed of polyenoic acid, saturated acid and monoenoic acid of 51.5%, 28.1% and 20.7%, respectively. Saturated fatty acids(C16:0, C14:0, C18:0) and monoenoic acids(C18:1, C16:1) increased while polyenoic acids(C20:5, C22:6) decreased during fermentation. Carotenoid was markedly degraded and discolored in the control during fermentation. But 0.02% BHT and 0.05% carotenoid treatments had bright color like fresh meat during 40 days. The results of sensory evaluation during the fermentation also convinced the retard of discoloration by the addition of BHT and carotenoid.

• 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.

• Parasites, Hosts and Diseases
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• v.52 no.3
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• pp.305-310
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• 2014

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

• Journal of fish pathology
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• v.26 no.1
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• pp.31-38
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• 2013

The edible ascidian, Halocynthia roretzi is a commercially important fisheries resource in Korea. However, there have been outbreaks of mass mortality due to soft tunic syndrome. It was discovered recently that the cause of death is infection by a protozoan parasite Azumiobodo hoyamushi. Alamar blue assay and microscopic counting were used to estimate anti-protozoal effects of 20 drugs having different action mechanisms. Through comparison of alamar blue assay and microscopic counting, 6 drugs were found to be potential in protozoan-killing effects: amphotericin B, formalin, hydrogen peroxide, bithionol, benzalkonium chloride, bronopol (24hr-$EC_{50}{\leq}20{\mu}g/ml$). The preliminary data can be used as a basis to develop anti-protozoal agents against A. hoyamushi.

• Fisheries and Aquatic Sciences
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• v.20 no.7
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• pp.12.1-12.7
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• 2017

Soft tunic syndrome (STS) is a protozoal disease caused by Azumiobodo hoyamushi in the edible ascidian Halocynthia roretzi. Previous studies have proven that combined formalin-hydrogen peroxide ($H_2O_2$) bath is effective in reducing STS progress and mortality. To secure target animal safety for field applications, toxicity of the treatment needs to be evaluated. Healthy ascidians were bathed for 1 week, 1 h a day at various bathing concentrations. Bathing with 5- and 10-fold optimum concentration caused 100% mortality of ascidians, whereas mortality by 0.5- to 2.0-fold solutions was not different from that of control. Of the oxidative damage parameters, MDA levels did not change after 0.5- and 1.0-fold bathing. However, free radical scavenging ability and reducing power were significantly decreased even with the lower-than-optimal 0.5-fold concentration. Glycogen content tended to increase with 1-fold bathing without statistical significance. All changes induced by the 2-fold bathing were completely or partially restored to control levels 48 h post-bathing. Free amino acid analysis revealed a concentration-dependent decline in aspartic acid and cysteine levels. In contrast, alanine and valine levels increased after the 2-fold bath treatment. These data indicate that the currently established effective disinfectant regimen against the parasitic pathogen is generally safe, and the biochemical changes observed are transient, lasting approximately 48 h at most. Low levels of formalin and $H_2O_2$ were detectable 1 h post-bathing; however, the compounds were completely undetectable after 48 h of bathing. Formalin-$H_2O_2$ bathing is effective against STS; however, reasonable care is required in the treatment to avoid unwanted toxicity. Drug residues do not present a concern for consumer safety.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.3
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• pp.447-451
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• 1998

Functional properties such as anti-blood coagulation, angiotensin converting enzyme (ACE) inhibitory activity, fat binding capacity, foaming properties, emulsifying properties and chemical components of sulfated polysaccharides isolated from ascidian tunics were investigated. The sulfated polysaccharide mainly consisted of sulfate, uronic acid, protein, and chondroitin sulfate, among which chondroitin sulfate showed higher concentration while sulfate and uronic acid did lower. Compositional menosaccharides were arabinose, xylose, glucose, galactose, glucuronic acid, N-acetylgalactosamine and N-acetyglucosamine. Especially, galactose content was dominant among them. And emulsifiability and foaminess of the sulfated polysaccharide was higher than the control group. Anti-blood coagulation of sulfated polysaccharide showed with respect to APTT (Activated partial thromboplastin time). ACE inhibitory activity showed about $16.7\%$.

• KSBB Journal
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• v.21 no.3
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• pp.194-198
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• 2006

Dried raw Ascidians(Halocynthia roretzi) shells harvested from fish farms in southern coast area in Korea were used to extract ${\beta}$-carotene using supercritical carbon dioxide($SCO_2$) and with ethanol as a co-solvent at the range of temperatures and pressures, from 25 to $65^{\circ}C$ and 100 to 350 bar respectively. The size of the dried Ascidians shells was around $850{\mu}m$. The system used this study was a semi-batch flow type high pressure unit. The efficiency of ${\beta}$-carotene extraction using $SCO_2$ with and without co-solvent, ethanol, influenced to pressure and temperature changes. The highest solubility of ${\beta}$-carotene in $SCO_2$ was 1.35 mg/g for ${\beta}$-carotene at $35^{\circ}C$ and 350 bar. With addition of 2(v/v%) ethanol the recovery of ${\beta}$-carotene was 93%. As a result of using n-hexane and methanol for rinse, at $35^{\circ}C$ and 350 bar the amount of ${\beta}$-carotene by methanol rinse was 5 times higher than that of n-hexane rinse.