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. 2023 Feb 4;13:2044. doi: 10.1038/s41598-023-29210-x

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Replication of v2001 in cell culture and in chickens. (a) Plaque areas were measured in CECs infected with vRLORF4mRFP, vUL47eGFP, and v2001. There were no significant differences between parental viruses using ANOVA. (b) Total MD incidence was determined for each group. There were no significant differences in the total number of chickens developing MD in experimentally infected chickens using Fisher’s exact tests. (c) Feathers were plucked from v2001-infected chickens at 35 days pi and expression of RLORF4mRFP and UL47eGFP were visualized using direct fluorescent stereomicroscopy.