Yoganandhan K, Hameed A S Sahul
Centex Shrimp, Faculty of Science, Chalermprakiat Building, Mahidol University, 272 Rama VI Road, Bangkok 10400, Thailand.
Dis Aquat Organ. 2007 Jan 18;73(3):193-9. doi: 10.3354/dao073193.
The freshwater prawn Macrobrachium rosenbergii was experimentally infected with white spot syndrome virus (WSSV) by intramuscular injection. Infection was confirmed by positive, single-step, WSSV polymerase chain reaction (PCR) assays targeting the VP28 gene from Day 2 up to Day 90 post injection (p.i.). Although no mortality of WSSV-infected prawns was observed, bioassays with the black tiger shrimp Penaeus monodon using hemolymph from Day 90 PCR-positive prawns resulted in white spot disease (WSD). Transcriptional analysis of the VP28 gene of WSSV by reverse transcriptase (RT)-PCR assays and Western blot assays revealed transient expression of the VP28-specific transcript in DNase-treated total RNA from hemolymph, gills, head soft tissue and eyestalks at 2 d p.i. By 3 d p.i., the VP28 transcript could no longer be detected in eyestalks and hemolymph but was still lightly detectable in head soft tissue and gills. It became undetectable there from 5 d p.i. onwards, despite the undiminished presence of the virus shown by single-step PCR targeting of the VP28 gene. VP28 was not detected by the less sensitive Western blot in hemolymph at any time during the study period, but it was detectable in all other tested tissues from Days 2 to 4 p.i. Our results demonstrated that M. rosenbergii is tolerant to a relatively constant level of persistent WSSV infection characterized by a low expression of VP28 and, possibly, other virion proteins. Despite this, M. rosenbergii can carry a level of infectious WSSV sufficient to represent a feasible threat to cultivated penaeid shrimp such as P. monodon. It remains to be seen whether a very low virion protein expression relative to the viral copy number may constitute a general decapod characteristic for persistent viral infections that produce no signs of disease.
通过肌肉注射对罗氏沼虾进行实验性感染白斑综合征病毒(WSSV)。在注射后第2天至第90天,通过针对VP28基因的阳性单步WSSV聚合酶链反应(PCR)检测确认感染。尽管未观察到WSSV感染的对虾死亡,但使用第90天PCR阳性对虾的血淋巴对斑节对虾进行生物测定时引发了白斑病(WSD)。通过逆转录酶(RT)-PCR检测和蛋白质免疫印迹分析对WSSV的VP28基因进行转录分析,结果显示在注射后2天,经DNase处理的血淋巴、鳃、头部软组织和眼柄的总RNA中VP28特异性转录本有短暂表达。到注射后3天,眼柄和血淋巴中不再能检测到VP28转录本,但在头部软组织和鳃中仍能轻微检测到。从注射后5天起,尽管通过针对VP28基因的单步PCR显示病毒持续存在,但在这些组织中已无法检测到该转录本。在研究期间的任何时候,蛋白质免疫印迹在血淋巴中均未检测到VP28,但在注射后第2天至第4天,在所有其他测试组织中均可检测到。我们的结果表明,罗氏沼虾对以VP28和可能的其他病毒粒子蛋白低表达为特征的相对恒定水平的持续性WSSV感染具有耐受性。尽管如此,罗氏沼虾携带的传染性WSSV水平足以对养殖对虾如斑节对虾构成切实威胁。相对于病毒拷贝数而言极低的病毒粒子蛋白表达是否可能构成无疾病迹象的持续性病毒感染的十足目动物普遍特征,仍有待观察。