低剂量对虾白斑综合征病毒粗提液对斑节对虾体内潜伏期病毒及血细胞的影响  

Influence of low dosage infection on WSSV in latent period and haemocyte of Penaeus monodon

作　　者：张涛[1,2] 郭志勋[1] 黄建华[1] 杨丽诗[1] 温为庚[1] 杨其彬[1] 江世贵[1] 

机构地区：[1]中国水产科学研究院南海水产研究所,广东省渔业生态环境重点实验室,农业部南海渔业资源开发利用重点实验室,广东广州510300 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《水产学报》2012年第10期1544-1553,共10页Journal of Fisheries of China

基　　金：现代农业产业技术体系建设专项(CARS-47);广东省海洋渔业科技推广专项(A201101B03);农业部"九四八"计划资助项目(2010-Z10)

摘　　要：利用含3×103、6×102、2×102copies/mL的对虾白斑综合征病毒(white spot syndromevirus,WSSV)粗提液和PBS液人工注射感染携带病毒量约1×105copies/g的斑节对虾,通过实时定量PCR法、显微镜观察法研究了感染后斑节对虾发病死亡率、肌肉内WSSV含量及总血淋巴细胞数和不同种类血细胞的比例组成变化。结果表明,3种浓度下斑节对虾累积死亡率分别为93.33%±2.89%、56.67%±5.77%和45.00%±5.00%,对照组没有发生死亡。注射3×103copies/mL的浓度组,在注射后30 min时,对虾肌肉内的病毒含量达到最低值(3.54×104copies/g),而后持续上升至48 h时达到最高值(3.12×108copies/g)后再次下降;另两个感染组,在注射后1 h时,肌肉内病毒含量达到最低值(分别为1.11×105、9.54×104copies/g),在感染后6 h时出现一个次高值(分别为1.58×105、1.11×105copies/g),而后又降低,在48 h时达到最高值(分别为1.48×107、5.46×106copies/g)后下降;对照组对虾肌肉内病毒含量没有出现显著变化。不同感染浓度组,斑节对虾总血淋巴细胞数波动幅度和出现峰值的时间不同,特别是注射3×103copies/mL组,对虾总血淋巴细胞数在12和48 h时出现极低值(分别为3.48×106、4.05×106/mL);3个感染组除个别时间点外,对虾总血淋巴细胞数的变化规律与肌肉中WSSV的扩增规律成负相关。4个处理组,半颗粒细胞比例在感染初期呈现明显上升的趋势,后期比例虽有起伏但均维持在较高水平;颗粒细胞和透明细胞比例在感染初期均呈现明显下降的趋势,中期均略有上升,但后期颗粒细胞比例显著低于初期,而透明细胞比例则与初期无显著性差异。The changes in mortalities of Penaeus monodon（with 1 × 10^5 copies/g WSSV） ,WSSV loads in muscle,THC and the percentage of different types of haemocyte were studied by injection with WSSV solution containing 3 × 10^3 ,6 ×10^2 ,2 × 10^2 copies/mL virus and PBS. Real-time PCR, and microscopical examination were used in this experiment. The results showed that the cumulative mortalities were 93.33% ± 2.89% ,56.67% ±5.77% ,45.00% ±5.00% respectively,when the concentration of WSSV solution were 3 × 10^3 ,6 ×10^2 , 2 × 10^2 copies/mL. No death was observed in the PBS group. When the concentration of WSSV solution was 3 × 10^3 copies/mL, the virus load in the muscle reduced to minimum（3.54 ×10^4 copies/ g）at 30 min post-injection, and reached its maximum （3. 12 × 10^8 copies/g）at 48 h post-injection, then decreased again. For the other two experiment groups, the minimal virus load（ 1.11 × 10^5 ,9.54 × 10^4 copies/ g） in the muscle was observed at 1 h post-injection and the highest virus load（ 1.48 × 10^7 ,5.46 ×10^6 copies/ g） occurred at 48 h post-injection,but a second highest virus load（ 1.58 × 10^5 , 1.11 ×10^5 copies/g）appeared at 6 h post-injection. No significant changes were found in the WSSV load of the control group throughout the experiment. The fluctuation range and the peak time of total haemocyte count （THC） were different between different dosage of WSSV solution. Especially THC reduced to minimum（3.48 × 10^6 ,4.05 × 1^06/ mL） at 12 h and 48 h post-injection with 3 × 10^3 copies/mL solution. The variation of THC and WSSV load were negatively correlated except individual time points. Under all groups, the percentage of semi-granular cells showed upward trend obviously early post infection, but later remained at a high level although fluctuated; the percentage of granular and hyalinocyte cells showed downward trend early post-injection significantly, while increased slightly in the middle period, but the percentage of granular cells at the later stage of

关 键 词：斑节对虾 白斑综合征病毒 低剂量感染 死亡率 实时定量PCR 血淋巴细胞 

分 类 号：Q175[生物学—水生生物学] S945.1[生物学—普通生物学]