Journal List > J Bacteriol Virol > v.39(3) > 1033930

Ko, Lee, Park, Peck, and Song: Molecular Identification of Clinical Rothia Isolates from Human Patients: Proposal of a Novel Rothia Species, Rothia arfidiae sp. nov.

Abstract

Four Gram-positive cocci were isolated from the cerebrospinal fluid or blood of four different patients, but they could not be identified by an automated conventional identification system, so they were identified using cellular fatty acid (CFA) composition analysis and 16S rRNA gene sequencing analysis. Of these, two strains (SMC-A2662 and SMC-A5889), which were previously supposed to be Rothia dentocariosa according to the API Coryne system, were identified as Rothia aeria by the 16S rRNA gene analysis. SMC-A608, which was unidentified by both the VITEK2 and API Coryne systems, was identified as Rothia mucilaginosa. The one remaining SMC-2244T was distinguished from the other Rothia species by its biochemical profile, its CFA composition and its 16S rRNA gene sequence. Phylogenetic analysis showed that it was closely related to Rothia nasimurium but the 16S rRNA gene sequence dissimilarity of 1.8% was enough to differentiate it from R. nasimurium. Based on both the phenotypic and phylogenetic evidence, we propose a new species name for this bacterium, Rothia arfidiae sp. nov. The results of this study show that several Rothia species were isolated from human and we have identified them using 16S rRNA gene sequences.

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Figure 1.
Phylogenetic relationships of four Rothia isolates based on 16S rRNA gene sequences. This tree was constructed using the Neighbor-joining method and bootstrap values were evaluated from 1,000 replications. The scale bar indicates the estimated number of substitutions per 100 nucleotides. Micrococcus lylae d10 was used as an outlier.
jbv-39-159f1.tif
Table 1.
Rothia isolates identified in this study
Isolates Patient (sex/age) Isolation Specimen Underlying disease Species
SMC-A5889 F / 52 Oct, 2004 Blood Pituitary adenoma Rothia aeria
SMC-A2662 M / 59 Dec, 2004 Blood Chronic renal failure Rothia aeria
SMC-A6087 F / 2 Jun, 2004 Blood Pneumonia Rothia mucilaginosa
SMC-2244T F / 22 Feb, 2004 CSFa Pituitary adenoma Rothia arfidiae sp. nov b

a Cerebrospinal fluid.

b Novel Rothia species proposed in the present study

Table 2.
Biochemical profiles of Rothia isolates a
Isolates Nitrate reduction Pyrrolidonyl arylamidase Alkaline phosphatase α-glucosidase Catalase
SMC-A5889 + + + +
SMC-A2662 + + + +
SMC-A6087 +
SMC-2244T + + + +

a Positive for esculin hydrolysis and fermented glucose, maltose, and sucrose. Negative for β-glucoronidase, β-galactosidase, N-acetyl-β-glucosaminidase, urea and gelatin hydrolysis, and fermented gelatin, ribose, xylose, mannitol, lactose, and glycogen

Table 3.
Fatty acid compositions of four isolates of Rothia species a
Fatty acidcomposition SMC-A5889 SMC-A2662 SMC-A6087 SMC-2244T
anteiso-C13:0 2.0 2.0
iso-C14:0 2.9 3.6 2.2 3.1
C14:0 1.3 1.2 4.9
iso-C15:0 2.8 3.9 1.2 2.4
anteiso-C15:0 47.4 51.5 72.6 68.4
iso-C16:0 15.9 20.5 3.0 6.0
C16:0 6.4 3.3 5.0 5.3
anteiso-C17:0 10.7 13.0 3.7 5.4
C18:1 w9c 5.4 4.4
C18:0 5.2 2.4

a Only fatty acid composition of more than 1.0% was shown

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