Department of Food Science, Center for Food Safety, University of Arkansas System Division of Agriculture, Fayetteville, AR, 72704, USA.
Food Environ Virol. 2024 Dec;16(4):470-478. doi: 10.1007/s12560-024-09610-6. Epub 2024 Aug 23.
The zebrafish larvae/embryo model has been shown to support the replication of seven strains (G1.7[P7], GII.2[P16], GII.3[P16], GII.4[P4], GII.4[P16], GII.6[P7], and GII.17[P13]) of human norovirus (HuNoV). However, due to challenges in consistently obtaining HuNoV-positive stool samples from clinical sources, evaluating HuNoV surrogates in this model is highly valuable. This study assesses the potential of zebrafish embryos and larvae as a model for Tulane virus (TuV) replication. Three infection methods were examined: microinjection, immersion, and feeding. Droplet digital PCR was used to quantify viral RNA across all three infection methods. Microinjection of 3 nL of TuV into zebrafish embryos (< 6-h post-fertilization) resulted in significant replication, with viral RNA levels reaching 6.22 logs at 4-day post-infection. In contrast, the immersion method showed no replication after immersing 4-day post-fertilization (dpf) larvae in TuV suspension for 6 h. Similarly, no replication was observed with the feeding method, where Paramecium caudatum loaded with TuV were fed to 4 dpf larvae. The findings indicate that the zebrafish embryo model supports TuV replication through the microinjection method, suggesting that TuV may serve as a useful surrogate for studying HuNoV pathogenesis. Additionally, TuV can be utilized in place of HuNoV in method optimization studies using the zebrafish embryo model, circumventing the limited availability of HuNoV.
斑马鱼幼虫/胚胎模型已被证明可支持七种人类诺如病毒（HuNoV）株（G1.7[P7]、GII.2[P16]、GII.3[P16]、GII.4[P4]、GII.4[P16]、GII.6[P7]和 GII.17[P13]）的复制。然而，由于从临床来源获得 HuNoV 阳性粪便样本存在挑战，因此评估该模型中的 HuNoV 替代物具有很高的价值。本研究评估了斑马鱼胚胎和幼虫作为 Tulane 病毒（TuV）复制模型的潜力。研究了三种感染方法：显微注射、浸泡和喂养。使用数字滴式 PCR 定量分析了所有三种感染方法下的病毒 RNA。将 3 nL 的 TuV 显微注射到小于 6 小时受精后的斑马鱼胚胎中，导致显著的复制，在感染后 4 天病毒 RNA 水平达到 6.22 对数。相比之下，在将 4 天受精后的幼虫浸泡在 TuV 悬浮液中 6 小时后，浸泡法没有显示出复制。同样，用已加载 TuV 的 Paramecium caudatum 喂养 4 天受精后的幼虫的喂养法也没有观察到复制。研究结果表明，斑马鱼胚胎模型通过显微注射法支持 TuV 复制，表明 TuV 可能作为研究 HuNoV 发病机制的有用替代物。此外，在使用斑马鱼胚胎模型进行方法优化研究时，可以用 TuV 替代 HuNoV，避免 HuNoV 的有限可用性。
