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为探讨转vp28蓝藻(Anabaena sp.PCC7120)口服剂对凡纳滨对虾抗白斑综合征病毒能力及其相应的免疫反应,本研究将此口服剂免疫幼虾7 d,再分别通过投喂攻毒和浸泡攻毒,测定其存活率及相应的免疫指标。投喂攻毒和浸泡攻毒的实验组存活率分别为78.8%和83.19%,表明该口服剂能显著增强对虾抗白斑综合征病毒的能力。蓝藻口服剂免疫对虾的酶活性检测结果显示,超氧化物歧化酶(SOD)、酚氧化酶(PO)、过氧化氢酶(CAT)和碱性磷酸酶(AKP)活性在免疫后2 h均有上升趋势,且在48或96 h达到最高值,这表明该口服剂能引起对虾体内酶活性变化。投喂攻毒的对虾酶活性检测结果显示,实验组攻毒后的对虾肝胰腺SOD活性分别比阳性对照组、野生型组、空载体组显著提高42.10%、32.26%和16.04%,且攻毒后的肌肉SOD活性分别比阴性对照组、阳性对照组、野生型组和空载体组略微提高17.70%、11.50%、15.00%以及10.00%。实验组攻毒后的对虾肝胰腺PO、CAT和AKP活性比阳性对照组分别提高12.17%、88.80%和240.07%,比野生型组分别提高21.49%、30.90%和100%;酸性磷酸酶(ACP)活性比阴性对照组略微提高,而在肌肉中各组ACP活性无显著性差异。同时浸泡攻毒组结果与投喂攻毒组具有类似的趋势。浸泡攻毒的实验组CAT和AKP活性显著高于其余处理组,且CAT活性比投喂攻毒更为显著。浸泡攻毒的实验组肝胰腺PO活性显著高于阳性对照组、野生型组和空载体组,而各组肌肉ACP活性无显著性差异。研究表明,转vp28蓝藻口服剂能够增强凡纳滨对虾抗病能力并延缓对虾死亡。转vp28蓝藻PCC7120本身可作为幼虾饵料直接投喂,无需提取纯化,有望大规模应用于对虾养殖产业。
In order to investigate the anti-white spot syndrome virus (VPV) virulence and its corresponding immune responses of vaba28 (Anabaena sp. PCC7120) to V. vannamei, the oral immunization of this strain was performed on juvenile shrimp for 7 d, And soak attack, determine the survival rate and the corresponding immune indicators. Survival rates of experimental group fed with challenge and immersion challenge were 78.8% and 83.19%, respectively, indicating that the oral agent can significantly enhance the ability of shrimp against white spot syndrome virus. The results showed that the activities of superoxide dismutase (SOD), phenoloxidase (PO), catalase (CAT) and alkaline phosphatase (AKP) Uptrend and reached the highest value at 48 or 96 h, indicating that the oral agent can cause changes in enzyme activity in shrimp. The result of shrimp fed with challenge showed that SOD activity of hepatopancreas increased by 42.10%, 32.26% and 16.04% in the control group, wild type group and empty vector group after challenge, respectively After the muscle SOD activity were slightly increased 17.70%, 11.50%, 15.00% and 10.00% than the negative control group, positive control group, wild type group and empty vector group. The activities of PO, CAT and AKP in liver and pancreas of experimental group were increased by 12.17%, 88.80% and 240.07%, respectively, 21.49%, 30.90% and 100% higher than those in wild type control group respectively. Acid phosphatase ) Activity than the negative control group slightly increased, while in muscle ACP activity of each group no significant difference. At the same time, the result of soaking the challenge group was similar to the challenge group. The activities of CAT and AKP in the experimental group soaked in water were significantly higher than those in the other treatments, and CAT activity was more significant than that in the control group. The activity of PO in hepatopancreas of experimental group was significantly higher than that of positive control group, wild type group and empty vector group, while there was no significant difference in muscle ACP activity in each group. Studies have shown that vp28 cyanobacteria oral agents can enhance the resistance of Litopenaeus vannamei and delay shrimp death. Turn vp28 Cyanobacteria PCC7120 itself can be fed directly as juvenile bait, without extraction and purification, is expected to be widely used in shrimp farming industry.