멍게, Halocynthia roretzi에서 분리된 해양버나바이러스의 특성과 물렁증과의 관련성
Isolation of marine birnavirus from ascidian Halocynthia roretzi, and its relation with tunic softness syndrome
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초록
양식 멍게의 폐사를 일으키는‘물렁증’의 원인규명을 위해 1999년 1월부터 2009년 2월까지 9 차례에 걸쳐 멍게 샘플링을 실시하여 바이러스를 분리하고 PCR을 이용하여 MABV 유전자를 검출하였다. 또한, 분리된 MABV UR-1주를 이용해 감염실험을 실시하였다. 바이러스 분리 실험에서 정상 개체는 5.1% (4/78), 비정상 개체 는 1.8% (1/56)의 개체에서 바이러스가 분리되었고, PCR에 의한 MABV 검출율은 정상 개체에서는 16.8% (18/107), 비정상 개체에서는 13.1% (5/38)를 나타내었다. MABV분리와 검출율은 정상 개체와 비정상 개체 관계없이 유사한 빈도로 나타났다. 멍게에서 분리된 8개 분리 주는 버나바이러스의 genogroup의 구분에 사용하 는 VP2/NS 경계영역의 염기서열 분석 결과 해산어류에서 분리되는 MABV와 동일한 genogroup에 속하고 있었다. 멍게에서 분리된 UR-1주를 침지와 주사법으로 병원성실험을 한 결과 침지 감염의 대조구는 약 36.5%, 실험구는 약 60%의 폐사율을 보였고, 주사 감염은 실험구와 대조구 모두 약 37%의 누적폐사율을 보여 모든 실험구에서 대조구와 유의적인 차이가 없었다. 그러므로 멍게체내에 MABV가 존재하기는 하지만 병원체를 저장하고 있는 저장소 역할을 하며 멍게 물렁증 폐사와는 연관이 없는 것으로 사료된다.
Abstract
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.
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