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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 (&lt;\u20096-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\u00a0h. Similarly, no replication was observed with the feeding method, where <jats:italic>Paramecium caudatum<\/jats:italic> 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.<\/jats:p>","DOI":"10.1007\/s12560-024-09610-6","type":"journal-article","created":{"date-parts":[[2024,8,23]],"date-time":"2024-08-23T18:03:34Z","timestamp":1724436214000},"page":"470-478","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Utilizing Zebrafish Embryos for Replication of Tulane Virus: A Human Norovirus Surrogate"],"prefix":"10.1007","volume":"16","author":[{"given":"Sahaana","family":"Chandran","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristen E.","family":"Gibson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,8,23]]},"reference":[{"issue":"8","key":"9610_CR1","doi-asserted-by":"publisher","first-page":"725","DOI":"10.1016\/s1473-3099(14)70767-4","volume":"14","author":"SM Ahmed","year":"2014","unstructured":"Ahmed, S. 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