Pathogenicity evaluation and gut microbiota dysbiosis analysis in piglets infected with a novel recombinant mammalian orthoreovirus type 3 strain HN2019-01

doi:10.1016/j.jia.2026.03.016

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Fangfang Han^1,²^{, 3}^*, Jinhui Hou^{1, 2, 3}^*, Yuxin Tang^{1, 2, 3}, Guozheng Li^{1, 2, 3}, Hui Hu^1,^2,³, Jin Yuan^1,^2,³^#, Zhanyong Wei^1,^2,³^#

¹ College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China

² Ministry of Education Key Laboratory for Animal Pathogens and Biosafety, Zhengzhou 450046, China

³ Henan Province Key Laboratory for Animal Food Pathogens Surveillance, Zhengzhou, 450046, China

Highlights:

1. A new recombinant strain of MRV type 3 (MRV HN2019-01) was successfully isolated, and this strain has the capacity to infect multi-species cells.

2. Pathogenicity evaluation showed that MRV HN2019-01 strain caused obvious diarrhea and intestinal villi damage in 5-day-old piglets.

3. MRV HN2019-01 infection disrupted the intestinal barrier, and affected the diversity and composition of the gut microbiota in piglets.

Abstract
References

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摘要

哺乳动物呼肠孤病毒（MRVs）属于呼肠孤病毒科、正呼肠病毒属，是一种无囊膜、分节段的双链RNA病毒，能感染包括人类、猪和蝙蝠在内的多种哺乳动物，引起呼吸道和肠道症状，但MRVs在中国猪群中的流行情况和致病机制尚不清楚。本研究在分离猪流行性腹泻病毒（PEDV）过程中，获得一株MRVs流行毒株，经二代测序、电镜观察等试验，证实该毒株为MRV 3型，将其命名为MRV HN2019-01株。该毒株能在Vero、ST、LLC-PK、IPEC-J2、IPI和Hela细胞中复制，其中在Vero、ST和LLC-PK细胞中复制效率最高。全基因组测序及遗传进化分析结果表明，MRV HN2019-01毒株属于一个新型的重组呼肠孤病毒，与MRV2/117RNA、MRV2/CH/GX/PReoV/2435/2018、MRV3/ZJ2013及MRV3/BM-100株亲缘关系较近。随后对MRV HN2019-01在5日龄仔猪体内的致病性进行评估，结果显示该病毒可引起仔猪腹泻、肠绒毛及肠道屏障损伤。应用16sRNA测序分析仔猪肠道菌群，显示MRV HN2019-01感染可显著改变结肠微生物的结构组成。通过LC-MS/MS代谢组学分析仔猪肠道内容物，结果显示MRV HN2019-01感染仔猪导致肠道内糖类、胆汁酸、3-羟基-L-酪氨酰-AMP、短链脂肪酸这些代谢物质发生显著性改变。KEGG通路富集分析显示，MRV HN2019-01感染组与空白对照组在硫胺素代谢、蛋白质消化吸收以及氨基酸代谢（包括苯丙氨酸、酪氨酸、色氨酸的生物合成）这些通路上有显著差异。本研究为MRV致病机制、疫苗研发提供了重要的理论基础。

Abstract

Mammalian orthoreoviruses (MRVs) have been identified in various mammalian species, including humans. Although there have been numerous reports on the cross-species transmission and genetic reassortment of MRVs among humans, livestock and wildlife, the prevalence and pathogenicity of MRVs on swine herds in China remain largely unknown. In this study, a novel MRV type 3 (MRV3) HN2019-01 strain was isolated on Vero cells from a clinical porcine epidemic diarrhea virus-positive intestinal content of a diarrheic piglet, and was identified using the small RNA deep sequencing, electron microscopic observation and the immunofluorescence assays. This isolated strain was able to replicate in multiple cell lines and showed the best replicative efficiency on Vero cells. Genetic analysis revealed that MRV HN2019-01 was a recombinant strain with gene segments from the swine MRV2 and MRV3, and has a close phylogenetic relationship with the MRV2/117 RNA, MRV2/CH/GX/PReoV/2435/2018, MRV3/ZJ2013 and MRV3/BM-100 strains. Subsequently, the pathogenicity of MRV HN2019-01 was evaluated in 5-day-old piglets. The results showed that the MRV HN2019-01 strain caused diarrhea and intestinal villi damage in piglets. Meanwhile, MRV HN2019-01 infection was shown to affect both the diversity and composition of the colonic microbiota in piglets, with a significant increase in Collinsella and Enterococcus and a notable decrease in Lactobacillus, Desulfovibrio and Ruminococcus. Metabolomic analysis revealed that MRV HN2019-01 infection in piglets induced alterations in multiple intestinal metabolites, including carbohydrates, bile acids, 3-hydroxy-L-tyrosine-AMP and short-chain fatty acids. KEGG pathway enrichment analysis indicated significant differences between the infected group and the control group in pathways such as thiamine metabolism, protein digestion and absorption, tyrosine metabolism and phenylalanine metabolism. This study provided important foundational data for investigating the pathogenic mechanisms and evolutionary characteristics of MRV and for the development of vaccines against MRV.This study provided important foundational data for investigating the pathogenic mechanisms of MRV and for the development of vaccines against MRV.

Keywords: mammalian orthoreovirus isolation genome reassortment pathogenicity gut microbiota

Online: 10 March 2026

Fund: This work was supported by the grants from the Program for Innovative Research Team (in Science and Technology) in University of Henan Province (Grant No. 24IRTSTHN034) and the Henan Province Key Research and Development Special Project (231111113100).

About author: #Correspondence Jin Yuan, E-mail: yuanjin@henau.edu.cn; Zhanyong Wei, Tel: +86-371-55369210, E-mail: weizhanyong@henau.edu.cn * The authors contributed equally to this work.

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Fangfang Han, Jinhui Hou, Yuxin Tang, Guozheng Li, Hui Hu, Jin Yuan, Zhanyong Wei. 2026. Pathogenicity evaluation and gut microbiota dysbiosis analysis in piglets infected with a novel recombinant mammalian orthoreovirus type 3 strain HN2019-01. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.03.016

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