PMC6821047_01
Morphology and Gram's staining of P. sputorum. Culture blood samples were transferred to Columbia blood plates and incubated in the 5% CO2 incubator at 37 C for (A) 24 h and (B) 48 h. Gram's staining showed that P. sputorum is (C) a gram-negative bacilli.
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Several tiny, moist and pale colonies appeared on the surface of the plates after 24 h of incubation and transformed into big, moist and ginger colonies over 48 h of incubation (Figure 1).
Phylogenetic analysis. Phylogenetic trees based on 16S rRNA gene sequences indicated the phylogenetic positions of isolates 1 and 2 and of other Pandoraea species. Burkholderia sordidicola S5-BT12826 was used as the outgroup. Bootstrap values (>70%) are shown for appropriate nodes. The scale bars represent the number of nucleotide substitutions per site.
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Phylogenetic analysis based on the 16S rRNA sequences showed that the two isolates were 100% similar to strains of P. sputorum, namely AF139176 and AB480701 (Figure 2).
Relatedness of the two isolates as determined by pulsed-field gel electrophoresis. Chromosomal DNA of the two isolates was digested with restriction endonucleases xbal and spel, followed by pulsed-field gel electrophoresis. (A) Lane 1, Marker; Lane 2, Isolate 1; Lane 3, Isolate 2. (B) Lane 1, Marker; Lane 2, Isolate 1; Lane 3, Isolate 2.
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The two isolates had identical digested DNA patterns (Figure 3), suggesting that they were derived from a common source.