Experimental hepatitis E virus genotype 1 infection in three types of wild rodents

Figures

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  Fig 1. Infection of wild rodents with HEV-1, HEV-3, and HEV-4 strains. (A) Representative photos of three wild rodent species: A. peninsulae, C. rufocanus, and L. brandtii. (B) Experimental design for HEV-1, HEV-3, and HEV-4 intraperitoneal inoculation in the three rodent species, including the number of animals per group and sample collection time points (feces, serum, and tissues). (C–E) Quantification of HEV RNA in fecal samples following inoculation with HEV-1 (C), HEV-3 (D), or HEV-4 (E) at indicated time points. (F) HEV RNA levels in liver tissues of the three rodent species at 7 and 35 dpi with HEV-1. (G) HEV RNA levels in liver tissues at 7 and 35 dpi after inoculation with HEV-3 or HEV-4. (H) Quantification of fecal HEV RNA in A. peninsulae inoculated with cell-derived HEV-1 (Sar55 strain). Viral RNA copy numbers were determined using RT-qPCR. LOQ: limit of quantification. Created in BioRender. He, Z. (2026) https://BioRender.com/wt0uu75.

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  Fig 2. Serial Passage of HEV-1 in A. peninsulae. (A) Schematic diagram of HEV-1 serial passage in A. peninsulae. (B–E) Quantification of fecal HEV RNA by RT-qPCR during passages P1 to P4 in A. peninsulae. (F) Seroconversion to anti-HEV antibodies in P4 animals at baseline and 42 dpi, determined by ELISA (Wantai, Beijing, China). An S/CO ratio >1 was considered positive. (G) Quantification of HEV RNA in liver, spleen, kidney, and intestine at 14 dpi in P4 animals. (H) Layered quantification of HEV RNA in hepatic portal blood of P4 animals after density gradient centrifugation at 14 dpi. Created in BioRender. He, Z. (2026) https://BioRender.com/wt0uu75.

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  Fig 3. Host Responses in A. peninsulae Following HEV-1 Infection. (A) Schematic diagram of the experimental design. (B) Fecal HEV RNA levels following HEV-1 inoculation. (C) HEV RNA levels in liver, spleen, kidney, and intestine at 10 dpi. (D) Serum conversion of anti-HEV antibodies on the 10th dpi after infection. (E) Weekly seroconversion to anti-HEV antibodies in infected animals. (F) Representative H&E-stained liver sections at 10 dpi in HEV-infected (n = 3) and mock-infected (n = 3) animals. Inflammatory cell infiltration is visible in liver tissues from infected animals. Scale bars are shown. (G) Immunohistochemistry for HEV ORF2 in liver tissues from infected and control animals. Arrows indicate positive staining. (H) Volcano plot showing DEGs between HEV-infected (n = 3) and mock-infected (n = 3) liver tissues at 10 dpi. Thresholds: adjusted P < 0.05 and log2FC > 1. (I) GO enrichment analysis of upregulated DEGs. Created in BioRender. He, Z. (2026) https://BioRender.com/wt0uu75.

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  Fig 4. Re-inoculation and Antiviral Treatment Influence HEV-1 Infection in A. peninsulae. (A) Schematic of reinoculation experiment following fecal viral clearance in P1 animals. (B) Fecal HEV RNA levels in reinoculated and positive control groups. (C) Schematic of ribavirin treatment experiment post-inoculation. (D) Fecal HEV RNA levels in ribavirin-treated and untreated groups during infection. *P < 0.05, ****P < 0.0001. Created in BioRender. He, Z. (2026) https://BioRender.com/wt0uu75.

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  Fig 5. Transmission Routes of HEV-1 in A. peninsulae. (A) Schematic of oral inoculation experiment. (B) Fecal HEV RNA levels following oral HEV-1 administration. (C) HEV RNA levels in liver, spleen, kidney, and intestine at 10 dpi. (D) Schematic of co-housing natural transmission experiment. (E) Fecal HEV RNA levels in Donor and Sentinel groups during co-housing. (F) Seroconversion in Donor and Sentinel animals during co-housing. (G) Schematic of vertical transmission experiment during pregnancy. (H) Fecal HEV RNA levels in pregnant A. peninsulae after inoculation. (I) Detection of HEV RNA in the liver tissues of neonates born to HEV-infected dams. Created in BioRender. He, Z. (2026) https://BioRender.com/wt0uu75.