Molecular Analysis of Caprine Enterovirus Circulating in China during 2016–2021: Evolutionary Significance

Figures

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  Figure 1. Geographic locations for specimen collection: The different provinces are marked in different colors with the number of collected samples. Blue: Xinjiang; red: Inner Mongolia; green: Jilin; yellow: Ningxia; purple: Shandong; orange: Henan. N represents the number of samples collected.

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  Figure 2. Immunoreactions of novel caprine/ovine enterovirus isolates to CEV-JL14. Seven representative novel caprine/ovine enterovirus strains were characterized by IPMA. Immunoreactions of the isolated viral strains to a monoclonal antibody against CEV-JL14-VP1 are shown. Strong reactions were revealed in the cells infected with NX-DR26, NMG-F37, SD-S68, JL-LS34, JL-LS127, JL-LS165, JL-LS174, and CEV-JL14 (A–H, respectively), while no reaction was observed for SD-S67 virus and normal Vero cells (I,J). Panels A–I represent the isolates NX-DR26, NMG-F37, SD-S68, JL-LS34, JL-LS127, JL-LS165, JL-LS174, CEV-JL14, and SD-S67, respectively. J represents the non-infected cell control. Bar = 20 μm.

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  Figure 3. Phylogenetic analyses for the novel caprine/ovine enterovirus strains. The reference sequences include the representative sequences of all known EV-G types and representative sequences of all 15 species of the Enterovirus genus as outgroup sequences. The amino acid sequences of 2C+3CD (A) and P1 (B) were used to construct the phylogenetic tree using the NJ method with 1000 bootstrap replications. Bootstrap values of > 50 are shown at the nodes. The scale bar represents 10% nucleotide sequence divergence for NJ methods. Viruses are marked with symbols as follows: ▲ refers to the strains obtained in this study from goats and sheep; ● stands for the CEV-JL14 strain; ○ refers to the HY12 EV-E strain isolated from cattle; △ stands for the SD-S67 strain (an EV-F) isolated from goats.

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  Figure 4. Three novel EV-G types revealed in caprine/ovine enterovirus: Phylogenetic analysis was performed based on the VP1 amino acid sequences. The reference sequences include the representative sequences of all known EV-G types and representative sequences of all 15 species of the Enterovirus genus as outgroup sequences. The phylogenetic tree was generated using the NJ method with 1000 bootstrap replications. Bootstrap values of > 50 are shown at the nodes. The scale bar represents 10% nucleotide sequence divergence for NJ methods. Viruses are marked with symbols as follows: ▲ refers to the strains obtained in this study from goats and sheep; ● stands for the CEV-JL14 strain; ○ refers to the HY12 EV-E strain isolated from cattle; △ stands for the SD-S67 strain (an EV-F) isolated from goats.

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  Figure 5. Recombination analyses of JL-LS34 with EV-G. Each curve is a comparison between the genome being analyzed and a reference genome. The horizontal bar above the curves is an illustration of the enterovirus genome. Each point plotted is the percentage identity within a sliding window 200 nt wide centered on the position plotted, with a step size of 20 nt between points.