• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.3
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• pp.357-368
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• 1996

To utilize the ascidian tunic as a natural pigment and dietary sources for rainbow trout (Oncorhynchus mykiss), juvenile were fed on experimental diets containing enzymatic hydrolysates of ascidian tunic treated with three commercial mined enzymes (ultrazyme, cellulase, viscozyme) for 12 weeks. From the results of feeding experiment, similar growth rate was checked in the enzymatic hydrolysis group compared with control, and those were a higher than of ascidian tunic powder group. The total acetone extractable pigment in muscle of the enzymatic hydrolysates group was lower than that of the ascidian extracts group and carophyll pink group until 8 weeks, but the level of those pigment of the enzymatic hydrolysates was similar to the ascidian extracts and carophyll pink group after 12 weeks. The lipid content was increased with the pigment concentration in the all experimental group. But the ascidian tunic pigment did not influence on the composition of the fatty acids in the muscle and liver. From the consideration of results for pigmentation, the enzymatic hydrolysates of ascidian tunic were suitable for both a natural pigment and dietary protein and carbohydrates sources as a substitute synthetic pigment for aquaculture use.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.3
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• pp.393-408
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• 1996

The effect of various levels of ascidian tunic extracts and carophyll pink on the growth rate, pigmentation, lipid and total cholesterol accumulation, and fatty acid compositions were studied in kuruma prawn, Penaeus japonicus. The kuruma prawn was fed the purified diets with or without ascidian tunic extract and carophyll pink at the levels of 100, 200, and 400 ppm for 8 weeks. In the experiment diet with ascidian tunic extracts or carophyll pink, the values of daily growth rate were ranged between $1.065\;to\;1.292%$, compared with control group. The content of astaxanthin in kuruma prawn was not significantly affected by the feeding levels of tunic extracts. Feeding of the tunic extracts, on the other hand, increased the kuruma prawn lipid and total cholesterol content, and pigment deposition in concentration-dependent manners without influencing the free astaxanthin concentration of prawn flesh and heads between two feeding groups(200 and 400 ppm). And it was also demonstrated that the dietary astaxanthin was deposited in kuruma prawn body tissue mainly as astaxanthin esters. The results suggest that the best feeding strategy for pigmentation in kuruma prawns is the diets with ascidian tunic extracts at the level of 4g/kg feed (200 ppm) for 8 weeks.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.6
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• pp.632-637
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• 2001

Components of polysaccharides isolated from ascidian tunic were measuerd by gel filtration, electrophoresis and chemical analyses. The sulfated polysaccharides consisted in sulfate, protein, uronic acid and amino sugars. Hexosamines were composed of arabinose, xylose, glucose, galactose, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine by gas chromatography analysis. The galactose was predominant hexose after autoclave and nutrase digestion followed by DEAE-cellulose ion exchange chromatography and gel-permeation chromatography on Sephadex G-100 and G-25. FT-IR spectra of isolated polysaccharides from ascidian tunic and standard chondroitin sulfates have similar functional groups of the type of vibration and frequency. Molecular weight of isolated polysaccharides by autoclave represented more than 40 kDa by polyacrylamide gel electrophoresis. But the neutrase treatment group divided into three band. The highest molecular band group was shown more than 100 kDa, and the two low molecular band group were shown about 22 kDa and 5 kDa, respectively, compare to standard materials.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.3
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• pp.423-428
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• 1998

The effective extraction methods and chemical components of crude polysaccharides of ascidian tunics were investigated. Tow extraction conditions, autoclaving or enzyme treatment, were applied. The proximate composition of ascidian tunics was not much different between those dried in raw (containing pigments) and those acetone treated and dried (decolorized), showing $50\%$ of carbohydrate and $40\%$ of protein. It was possible to extract up to $10\%$ of crude polysaccharides from ascidian tunics regardless of the extraction methods, autoclaving or enzyme treatment. In case of the latter the extraction yield by neutrase was higher than that with alkalase (Novo co.) or mixture 2000 (Pacific chemical co.). The most effective enzyme concentration and extraction time appeared to be 24 hrs of extraction with $3\%$ neutrase. On the other hand, in autoclave treatment, 6 hrs extraction showed most desirable extraction yield, about $9.7\%$. The compositions of amino acid of decolorized ascidian tunic (acetone treated group) and the crude polysaccharide from the autoclaving (water solubles) or neutrase treatment (enzyme digestibles) were similar to each other. Histidine was the highest both in the neutrase and autoclave treatment group and the yield were $29.2\%,\;20.4\%$, respectively, followed by aspartic acid and glutamic acid. Among the minerals, the content of Ca was significantly high, followed by Mg and Na.

• Journal of Environmental Science International
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• v.18 no.10
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• pp.1171-1179
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• 2009

The solitary ascidian, Halocynthia aurantium, which is commonly called the sea peach because of its coloration and general shape, is a valuable organism of benthic marine population in the northern region of the East Sea, Korea. It is seldom found at a depth of less than 10 meters and the sea peach is frequently observed in large populations between 20 and 100 meters. It appears to prefer attachment to vertical rocks faces and artificial cement blocks exposed to the currents. Mass mortality and reduction of resources in sea peach, H. aurantium, were occurred in the benthic area of the northern region of the East Sea because of the rapid fluctuation of environmental factors such as temperature and salinity due to mass rainfall in summer and going up north of a strong warm current in winter. Therefore, we examined the effects of temperature and salinity on embryonic development of fertilized eggs, tadpole larva to metamorphosis, and attachment to siphon development. Laboratory-raised larvae were studied using a two-factorial experimental design with four levels of temperature(8, 12, 16 and $20^{\circ}C$) and four levels of salinity(20, 25, 30 and 34 psu). The ascidian larvae of H. aurantium survived environmental conditions between temperature of $8{\sim}20^{\circ}C$ and salinity of 25~34 psu and exhibited positive growth at $8{\sim}16^{\circ}C$ and 30~34 psu. Fertilized eggs have not developed at lower salinity of 20 psu irrespective of temperature range tested and have showed an abnormal development at the salinity of 25 psu between higher temperatures of 20 and $24^{\circ}C$. This result suggests that temperature increase and salinity reduction depending on environmental fluctuation may have significant impacts on population variation of H. aurantium in the northern region of the East Sea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.3
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• pp.232-239
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• 1994

In order to determine the utilization of ascidian tunic, which has been blamed for problems of costal environmental pollution when discharged into the sea after being used as a natural dietary pigment sources for rainbow trout(Oncorhynchus mykiss), fingerlings were fed on experimental diets containing acetone-extracts for 6 weeks. The amounts of acetone-extracts were 11,000mg/Kg and contained 50mg/100g wet tissues of carotenoid and $6\%$ of carotenoids were astaxanthin. From the results of feeding experiments, the growth rate in the extract group was a little higher than that of the control and pink groups after 6 weeks. The redness and yellowness of the fish skin and muscle in the extract group were similar to the pink group. Therefore, acetone-extracts of ascidian tunic were judged to be a natural dietary pigment source suitable as a substitute synthetic pigment for aquaculture use.

• 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 Marine Environment & Safety
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• v.18 no.5
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• pp.379-387
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• 2012

In order to see the oceanographic conditions, the observations of aquaculture farm of ascidian in Youngdeuk, the east coast of Korea were conducted through 6 times-23 February, 6 April, 8 June, 19 August, 6 October and 20 December-in 2010. Surveys were conducted in 20 stations bimonthly using SBE 19 CTD instrument. The mixed layer depth (MLD) was deep in winter and shallow in summer. The cold water below $5^{\circ}C$ in temperature was occupied below thermocline through all season. The temperature was high in the southeastern area. The salinity was increased from the coast to the open sea. The halocline was distinct at 20 m depth in August and at 40 m depth in October. The lowest value of salinity was appeared at the depth of 10 m in October. In addition the value of precipitation minus evaporation denoted negative in October. These low saline water seemed to inflow to the coast from the open sea. Therefore the low saline water moved to the east coast of Korea. The EKWC may play an important role to convey the low saline water. It may affect the aquaculture farm along the coast as the mass mortality of ascidian. It needs to clarify the role and pathway of EKWC to transfer the low saline water along the east coast of Korea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.3
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• pp.345-356
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• 1996

Properties of enzymatic hydrolysates from ascidian tunic were assessed on supernatant ratio, solid yields and solid concentration. The concentartion of solid and yields in the extracts were increased as the enzyme concentration raised from $100\;{\mu}l\;to\;1000{\mu}l$ during the extraction period. The optima concentration and reaction time of each enzyme on digestion were $400\;{\mu}l$ 60 minutes, through treated with Duncan's multiple test. The percent of yields of solid, protein and carotenoids for 60 minutes extraction at $400\;{\mu}l$ were $32.32\%,\;1.34\%\;and\;74.60\;mg\%$, respectively, in Viscozyme systems. The extracts were composed with many kinds of carbohydrates such as arabinose, ribose, xylose, galactose, glucose, N-acetyl-D-galactosamine, and N-acetyl-D-glucosamine. Aspartic and glutamic arid were noted as predominant amino acids in all parts. Amino acid profiles of various ascidian tunic part were similiar to each other, but most of essential amino acids content of inter coat was higher than that of root and tunic (body). About sixty six fatty acids components were observed, and their distribution among neutral and polar lipids was compared. The main fatty acids were found to be 14:0, 16:0, 16:1n7, 18:0, 18:1n9, 18:1n7, 18:2n6, 20:5n3, and 22:6n3.