Romo-Quiñonez Carlos R, Álvarez-Sánchez Ana R, Álvarez-Ruiz Pindaro, Chávez-Sánchez Maria C, Bogdanchikova Nina, Pestryakov Alexey, Mejia-Ruiz Claudio H
Laboratorio Biotecnologia de Organismos Marinos, Programa de Acuicultura, Centro de Investigaciones Biológicas del Noroeste, La Paz, BCS, México.
Universidad Técnica Estatal de Quevedo, Quevedo, Los Rios, Ecuador.
PeerJ. 2020 Feb 27;8:e8446. doi: 10.7717/peerj.8446. eCollection 2020.
In this study, four experimental assays were conducted to evaluate the use of a new silver nanoparticle formulation named Argovit-4, which was prepared with slight modifications to enhance its biological activity against white spot syndrome virus (WSSV) in shrimp culture. The goals of these assays were to (1) determine the protective effect of Argovit-4 against WSSV, (2) determine whether Argovit-4 supplemented in feed exhibits toxicity towards shrimp, (3) determine whether Argovit-4 as antiviral additive in feed can prevent or delay/reduce WSSV-induced shrimp mortality, and (4) determine whether Argovit-4 supplemented in feed alters the early stages of the shrimp immune response. In bioassay 1, several viral inocula calibrated at 7 SID(shrimp infectious doses 50% endpoint) were exposed to 40, 100, 200 and 1,000 ng/SID of Ag and then intramuscularly injected into shrimp for 96 h. In bioassay 2, shrimp were fed Argovit-4 supplemented in feed at different concentrations (10, 100 and 1,000 µg per gram of feed) for 192 h. In bioassay 3, shrimp were treated with Argovit-4 supplemented in feed at different concentrations and then challenged against WSSV for 192 h. In bioassay 4, quantitative real-time RT-qPCR was performed to measure the transcriptional responses of five immune-relevant genes in haemocytes of experimental shrimp treated with Argovit-4 supplemented in feed at 0, 6, 12, 24 and 48 h. The intramuscularly injected Argovit-4 showed a dose-dependent effect ( < 0.05) on the cumulative shrimp mortality from 0-96 h post-infection. In the second bioassay, shrimp fed Argovit-4 supplemented in feed did not show signs of toxicity for the assayed doses over the 192-h experiment. The third and fourth bioassays showed that shrimp challenged with WSSV at 1,000 µg/g feed exhibited reduced mortality without altering the expression of some immune system-related genes according to the observed level of transcriptional. This study is the first show that the new Argovit-4 formulation has potential as an antiviral additive in feed against WSSV and demonstrates a practical therapeutic strategy to control WSSV and possibly other invertebrate pathogens in shrimp aquaculture.
在本研究中，进行了四项实验测定，以评估一种名为Argovit-4的新型银纳米颗粒制剂的用途，该制剂在制备时进行了细微修改，以增强其对虾养殖中白斑综合征病毒（WSSV）的生物活性。这些测定的目的是：（1）确定Argovit-4对WSSV的保护作用；（2）确定饲料中添加的Argovit-4是否对虾具有毒性；（3）确定饲料中作为抗病毒添加剂的Argovit-4是否可以预防或延迟/降低WSSV诱导的虾死亡率；（4）确定饲料中添加的Argovit-4是否会改变虾免疫反应的早期阶段。在生物测定1中，将几种校准为7个虾感染剂量50%终点（SID）的病毒接种物分别暴露于40、100、200和1000 ng/SID的银，然后肌肉注射到虾体内96小时。在生物测定2中，给虾投喂饲料中添加不同浓度（每克饲料10、100和1000 μg）的Argovit-4，持续192小时。在生物测定3中，用饲料中添加不同浓度的Argovit-4处理虾，然后用WSSV攻击192小时。在生物测定4中，进行定量实时RT-qPCR，以测量在0、6、12、24和48小时用饲料中添加Argovit-4处理的实验虾血细胞中五个免疫相关基因的转录反应。肌肉注射的Argovit-4对感染后0至96小时虾的累积死亡率显示出剂量依赖性效应（P<0.05）。在第二个生物测定中，在192小时的实验中，投喂饲料中添加Argovit-4的虾在所测定剂量下未显示出毒性迹象。第三和第四个生物测定表明，以1000 μg/g饲料用WSSV攻击的虾死亡率降低，根据观察到的转录水平，一些免疫系统相关基因的表达未发生改变。本研究首次表明，新型Argovit-4制剂作为饲料中抗WSSV的抗病毒添加剂具有潜力，并展示了一种控制虾养殖中WSSV以及可能其他无脊椎动物病原体的实用治疗策略。
