• 환경생물
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• 제32권3호
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• pp.225-233
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• 2014

수온조절 (6, 9, 12, 15, 18, 21, 24, $27{\circ}^C$)에 의해 유도된 멍게의 물렁증 진행에 따른 생리학적 특성을 연구하였다. 수온조절에 의한 물렁증 유도율은 수온 $15^{\circ}C$에서 가장 높았으며, $24^{\circ}C$에서 가장 낮았다. 물렁증은 피낭 색깔 및 탄성을 기준으로 S0, S1, S2, S3의 4단계로 구분하였다. 정상개체와 물렁증을 가진 개체들 사이에서 산소소비율과 여수율은 유의한 차이를 보였다. S0에 비해 S3단계에서 혈구 구성비의 가장 뚜렷한 변화는 multi-vacuole cell은 약 1/2 감소하였으며, morula cell은 약 10배 증가하였다. 기관계 구조의 변화는 피낭, 수관, 새낭, 육질부, 소화선에서 상피세포의 변성, 미세섬유의 변성, 혈구의 증가 및 근섬유의 변형이 확인 되었다. 이러한 모든 특징들은 멍게의 정상적인 생리학적 기능에 영향을 미칠 수 있다.

• 한국어병학회지
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• 제34권1호
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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.

• 한국어병학회지
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• 제24권3호
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• pp.197-204
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• 2011

멍게 물렁증 원인규명의 일환으로 2011년 3월에 경남 통영에서 채집된 물렁증 멍게와 건강한 멍게를 대상으로 병리학적 검사를 실시한 결과 물렁증 멍게의 경우 체액량, 비만도 및 피막지수가 현저히 감소하였다. 물렁증 멍게의 피막 imprint 관찰과 배양 시 2개의 편모를 가진 원생동물이 확인되었으며, 물렁증 멍게에 대한 이들 편모충의 감염률은 97.5%인 반면 건강한 멍게에서는 검출되지 않아 본 편모충은 물렁증 멍게에서 특이적으로 검출되는 것으로 조사되었다. 조직학적 검사결과 물렁증 멍게에서 피막섬 유질다발의 붕괴 현상이 관찰되었으며, 편모충으로 추정되는 원생동물들이 다수 관찰되었다. 통영산 물렁증 멍게에서 확인된 편모충은 일본에서 검출된 그것과 매우 유사한 형태적 특징을 갖고 있으며, 향후 공격실험 등 물렁증 유발과 편모충과의 연관성에 대한 추가 연구가 요망된다.

• 한국어병학회지
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• 제22권3호
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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.

• 한국어병학회지
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• 제28권3호
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• pp.117-123
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• 2015

우리나라 멍게 양식의 발전으로 생산량이 점점 증가하였으나 최근 물렁증 (soft tunic syndrome)으로 멍게가 대량 폐사하여 경제적으로 많은 손실을 입고 있다. 이런 물렁증을 일으키는 원인체가 Azumiobodo hoyamushi라고 보고되었다. 하지만 이 원인충을 박멸할 수 있는 치료제 연구는 미비한 실정이다. 저자들에 의한 이전의 연구에서 기존의 산화제, 환원제, 소독제 등에서 물렁증에 대한 치료효과가 있다고 보고하였다. 본 연구에서는 기존의 약제보다 좀 더 친환경적인 유기산을 활용하여 in vitro 및 in vivo 시험을 통하여 연구하였다. In vitro 시험에서 12종의 유기산 중 8종이 $300{\mu}g/ml$에서 100% 원충에 대한 살충효과를 발휘하였다. In vivo에서의 유기산 침투효과를 평가하기 위해 pH의 변화를 측정한 결과, 유기산 처리 후 멍게 피막내 pH는 in vitro 에서 보다는 0.5~1 정도 높고 무처리 피막보다는 2.0 정도가 낮아 부분적으로는 침투가 일어남을 확인하였다. 본 연구를 토대로 친환경적인 유기산을 이용하여 물렁증의 치료에 활용할 수 있을 것이다.

• 한국어병학회지
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• 제26권1호
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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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• 제20권7호
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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.

• Fisheries and Aquatic Sciences
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• 제19권1호
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• pp.1.1-1.6
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• 2016

Although soft tunic syndrome (STS) in the ascidian is a serious disease, helpful measures have yet not been established. It was examined in this study by applying aniti-parasitic drugs to eradicate the causative protozoa Azumiobodo hoyamushi from infected ascidians. Formalin was synergistic in killing parasites in vitro when co-treated with hydrogen peroxide ($H_2O_2$) or bronopol, but not with chloramine-T or povidone-iodine (PVP-I), when tested with in vitro parasite culture. The synergistic effects did not change when $formalin-H_2O_2$ (or bronopol) ratios were changed. It was found that treatment periods less than 60 min achieved a sub-maximal efficacy. Increasing drug concentration while keeping 30 min period improved anti-parasitic effects. Anti-parasitic effects of $formalin(F)+H_2O_2$(H) were also assessed in an in vivo STS model infected with cultured parasites. It was observed that combined 50 (40F + 10H) and 100 (80F +20H) ppm were effective in partially preventing STS-caused mortality. In horizontally transmitted artificial STS model, significant prevention of ascidian mortality was also observed after 50 ppm. Marked reduction of living parasites were noted after drug treatments in vivo. The results provide a highly useful basis to develop a preventive or treatment measure against the currently uncontrollable STS in the ascidian.

• Parasites, Hosts and Diseases
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• 제52권3호
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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.