Intertypic reassortment of mammalian orthoreovirus identified in wastewater in Japan

Figures

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  Figure 1. Identification of novel MRV isolates. Electron microscopic observation of novel MRV isolates, (A) THK0325 and (B) THK0617. Negatively stained virions showing icosahedral particles with a diameter of 60–70 nm. Bar = 100 nm.

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  Figure 2. Phylogenetic analysis based on the nucleotide sequences of novel MRV isolates. Phylogenetic trees for the S1 segment were constructed through the maximum-likelihood method and 1000 bootstrap replicates using MEGA software. (A) Novel isolates and prototypes. A baboon orthoreovirus sequence was set as outgroup. (B) MRV sequences obtained from the GenBank database based on the best hits retrieval from the BLAST result. Each ID depicts the MRV genotype, name, host species, country, and accession number. Solid squares indicate the isolates in this study. Scale bars indicate genetic distances (nucleotide substitutions per site).

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  Figure 3. Phylogenetic analysis based on the nucleotide sequences of MRV isolates. Phylogenetic trees for segments aside from S1 were constructed through the maximum-likelihood method and 1000 bootstrap replicates using MEGA software. MRV sequences obtained from the GenBank database based on the best hits retrieval of the BLAST result. Each ID depicts the MRV genotype, name, host species, country, and accession number. Solid square: the isolates in this study. Red: MRV-2 Osaka strains. Green: MRV-1/TS/2011 and MRV-3/TS/2012. Scale bar: genetic distances (nucleotide substitutions per site).

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  Figure 4. Pairwise sequence comparisons of MRV isolates. Nucleotide and amino acid identities between the novel isolates and MRV-2 Osaka strains previously isolated in Japan. Three-color scale formatting was applied to highlight the percentage values.

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  Figure 5. Schematic summary of the genomic compositions of novel MRV isolates and related strains. Based on the nucleotide identity and phylogenetic analysis, segments with close relation were distinguished by colour. Segments of THK0325 and THK0617 showing high nucleotide identity (more than 90% in Fig. 4) with Osaka1994 are in black. In Taiwan strains, S1 of MRV-1 and MRV-2 were closely related to THK0617 (red) and THK0325 (black), respectively (Fig. 2). S2 segments of Osaka2005 and THK0325, and M2, S2, and S3 segments of the THK0617 were related to MRV-1/TS/2011 and MRV-3/TS/2012 (green) (Fig. 3). The sole MRV-3 S1 segment is in light blue, and the unidentified segments of the Taiwan strain are in grey.