Utilizing Zebrafish Embryos for Replication of Tulane Virus: A Human Norovirus Surrogate

IF 4.1 2区 农林科学 Q2 ENVIRONMENTAL SCIENCES Food and Environmental Virology Pub Date : 2024-08-23 DOI:10.1007/s12560-024-09610-6
Sahaana Chandran, Kristen E. Gibson
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Abstract

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.

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利用斑马鱼胚胎复制杜兰病毒:人类诺罗病毒代用品
斑马鱼幼虫/胚胎模型已被证明支持七株人类诺瓦克病毒(HuNoV)(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,由于难以从临床来源持续获得 HuNoV 阳性的粪便样本,因此在该模型中评估 HuNoV 代用品非常有价值。本研究评估了斑马鱼胚胎和幼虫作为杜兰病毒(TuV)复制模型的潜力。研究考察了三种感染方法:显微注射、浸泡和喂食。使用液滴数字 PCR 对所有三种感染方法的病毒 RNA 进行量化。将 3 nL 的 TuV 微注射到斑马鱼胚胎 (
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来源期刊
Food and Environmental Virology
Food and Environmental Virology ENVIRONMENTAL SCIENCES-MICROBIOLOGY
CiteScore
6.50
自引率
2.90%
发文量
35
审稿时长
1 months
期刊介绍: Food and Environmental Virology publishes original articles, notes and review articles on any aspect relating to the transmission of pathogenic viruses via the environment (water, air, soil etc.) and foods. This includes epidemiological studies, identification of novel or emerging pathogens, methods of analysis or characterisation, studies on survival and elimination, and development of procedural controls for industrial processes, e.g. HACCP plans. The journal will cover all aspects of this important area, and encompass studies on any human, animal, and plant pathogenic virus which is capable of transmission via the environment or food.
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