[국내논문]자연산 자주새우(Crangon affinis)에서 흰반점바이러스(WSSV)의 neutralization을 위한 인위감염(experimental infection) Experimental Infection for the Neutralization of White Spot Syndrome Virus (WSSV) in Wild Captured Sand Shrimp, Crangon affinis원문보기
흰반점바이러스(WSSV)는 새우양식산업에 막대한 경제적 손실을 일으키는 가장 치명적인 바이러스성 질병 원인체 중 하나이다. 본 연구는 인위적인 실험모델로서 WSSV에 대한 자주새우(Crangon affinis)의 감수성을 확인하고자 실험을 수행하였다. WSSV가 희석된 수조 내에서 침지법으로 감염된 새우는 감염 후 7일째에 100% 누적폐사율을 보여 자주새우가 WSSV에 대해 매우 높은 감수성을 갖고 있음을 확인하였다. 또한 재조합단백질인 rVP466에 대한 항혈청의 중화효과를 확인하기 위해 항혈청과 반응시킨 바이러스액($1{\times}10^4$ 배로 희석된 WSSV)을 이용하여 침지법으로 자주새우에 대해 공격실험(challenge test)을 수행하였다. 실험 결과, WSSV로 challenge한 감염대조구(positive control)의 새우들은 감염 후 5일째에 100% 폐사하였으며, WSSV와 rVP466 항혈청을 1:0.01, 1:0.1, 1:1로 혼합한 액으로 challenge한 새우들은 감염 후 14일째에 각각 100%, 68.8%, 68.8%의 누적폐사율을 보였다. 따라서 본 연구 결과는 연안서식종인 자주새우가 양식장으로부터 배출된 WSSV에 의해 자연상태에서 감염 될 수 있는 가능성과 함께 항혈청에 대한 중화효과를 나타냄으로써 겨울철 저수온기에 WSSV 감염을 위한 대체 실험생물로서의 유용성을 확인하였다.
흰반점바이러스(WSSV)는 새우양식산업에 막대한 경제적 손실을 일으키는 가장 치명적인 바이러스성 질병 원인체 중 하나이다. 본 연구는 인위적인 실험모델로서 WSSV에 대한 자주새우(Crangon affinis)의 감수성을 확인하고자 실험을 수행하였다. WSSV가 희석된 수조 내에서 침지법으로 감염된 새우는 감염 후 7일째에 100% 누적폐사율을 보여 자주새우가 WSSV에 대해 매우 높은 감수성을 갖고 있음을 확인하였다. 또한 재조합단백질인 rVP466에 대한 항혈청의 중화효과를 확인하기 위해 항혈청과 반응시킨 바이러스액($1{\times}10^4$ 배로 희석된 WSSV)을 이용하여 침지법으로 자주새우에 대해 공격실험(challenge test)을 수행하였다. 실험 결과, WSSV로 challenge한 감염대조구(positive control)의 새우들은 감염 후 5일째에 100% 폐사하였으며, WSSV와 rVP466 항혈청을 1:0.01, 1:0.1, 1:1로 혼합한 액으로 challenge한 새우들은 감염 후 14일째에 각각 100%, 68.8%, 68.8%의 누적폐사율을 보였다. 따라서 본 연구 결과는 연안서식종인 자주새우가 양식장으로부터 배출된 WSSV에 의해 자연상태에서 감염 될 수 있는 가능성과 함께 항혈청에 대한 중화효과를 나타냄으로써 겨울철 저수온기에 WSSV 감염을 위한 대체 실험생물로서의 유용성을 확인하였다.
White spot syndrome virus (WSSV) is one of the most virulent viral agents threatening the penaeid shrimp culture industry. This study was carried out to evaluate the susceptibility of the sand shrimp, Crangon affinis, to WSSV as an alternative experimental model. WSSV caused 100% mortality in C. aff...
White spot syndrome virus (WSSV) is one of the most virulent viral agents threatening the penaeid shrimp culture industry. This study was carried out to evaluate the susceptibility of the sand shrimp, Crangon affinis, to WSSV as an alternative experimental model. WSSV caused 100% mortality in C. affinis within 7 days after experimental infection by immersion. Based on challenge studies, it was confirmed that C. affinis could be a potential host in WSSV transmission. Also, the neutralization of WSSV was carried out using an antiserum raised against recombinant envelop protein rVP466 to evaluate the WSSV infection mechanism. A constant amount of WSSV (at $1{\times}10^4$ diluted stocks) was incubated with various amounts of antiserum and then mixed to 20 l reservoir for the immersion challenge of C. affinis for neutralization. At 5 days post challenge, the shrimp in the positive control immersed in the immersion reservoir containing WSSV stock showed 100% mortality. The shrimps challenged with the 3 different mixtures of WSSV and rVP466 antiserum (1:0.1, 1:0.5 and 1:1) showed 100%, 68.8% and 68.8% mortality at 14 days post challenge, respectively. These results indicated that the antiserum raised against rVP466 could block WSSV infection in C. affinis. Therefore, this study confirmed that C. affinis can be naturally infected by WSSV as another potential host and that C. affinis can be used as an alternative experimental animal instead of penaeid shrimps.
White spot syndrome virus (WSSV) is one of the most virulent viral agents threatening the penaeid shrimp culture industry. This study was carried out to evaluate the susceptibility of the sand shrimp, Crangon affinis, to WSSV as an alternative experimental model. WSSV caused 100% mortality in C. affinis within 7 days after experimental infection by immersion. Based on challenge studies, it was confirmed that C. affinis could be a potential host in WSSV transmission. Also, the neutralization of WSSV was carried out using an antiserum raised against recombinant envelop protein rVP466 to evaluate the WSSV infection mechanism. A constant amount of WSSV (at $1{\times}10^4$ diluted stocks) was incubated with various amounts of antiserum and then mixed to 20 l reservoir for the immersion challenge of C. affinis for neutralization. At 5 days post challenge, the shrimp in the positive control immersed in the immersion reservoir containing WSSV stock showed 100% mortality. The shrimps challenged with the 3 different mixtures of WSSV and rVP466 antiserum (1:0.1, 1:0.5 and 1:1) showed 100%, 68.8% and 68.8% mortality at 14 days post challenge, respectively. These results indicated that the antiserum raised against rVP466 could block WSSV infection in C. affinis. Therefore, this study confirmed that C. affinis can be naturally infected by WSSV as another potential host and that C. affinis can be used as an alternative experimental animal instead of penaeid shrimps.
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제안 방법
In this study, DNA fragment encoding WSSV structural protein VP466 was amplified by PCR and cloned into pHCE vector. As shown in Fig.
Therefore, this study was carried out to evaluate the susceptibility of wild captured sand shrimp, Crangon affinis to WSSV. The recombinant protein, rVP466 was used for the production of antiserum and the neutralization was carried out to evaluate the role of VP466 in infection of WSSV.
대상 데이터
Healthy sand shrimp Crangon affinis were purchased from Dadaepo located in Busan, Korea. Shrimps of approximately 3 to 5 g were used in this experiment.
성능/효과
In conclusion, wild captured sand shrimp, Crangon affinis, was susceptible to WSSV infection. It was confirmed that the antiserum raised against rVP466 blocks WSSV infection.
In this study, in vivo neutralization assay with the antiserum against rVP466 showed 31.2% of survival ratio slightly less than that of VP28 [7]. However, the antiserum raised against VP466 could access to the viral antigen by the independent or coordinate manners [15].
affinis can be collected regardless of seasons. Therefore, this study confirmed that C. affinis can be naturally infected by WSSV as another potential host and C. affinis can be used as an alternative experimental animal instead of penaeid shrimps.
후속연구
However, VP28 is not the only virus protein related attachment to the host cell, as it has been reported that many attachment proteins are involved in infection mechanism for the large DNA virus [5]. Therefore, the further studies with other envelop proteins are required for the correct analysis of WSSV attachment, entry and budding in shrimps.
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