JOURNAL OF CLINICAL MICROBIOLOGY, July 1993, p. 1788-1793 0095-1137/93/071788-06$02.00/0 Copyright © 1993, American Society for Microbiology
Vol. 31, No. 7
Evidence of Multiple Taxa within Commercially Available Reference Strains of Corynebacterium xerosis MARIE B. COYLE,l,2* REBECCA B. LEONARD,1t DAVID J. NOWOWIEJSKI,1 ALI MALEKNIAZI,1 AND DONALD J. FINN1*
Department of Laboratory Medicine, Harborview Medical Center, University of Washington, Seattle, Washington 98104,1 and Department of Microbiology, University of Washington, Seattle, Washington 981952 Received 8 February 1993/Accepted 5 April 1993
Attempts to identify coryneform isolates resembling Corynebacterium xerosis can lead clinical microbiologists to identification schemes with conflicting descriptions which result in confusing C. xerosis with Corynebacterium striatum. For the present study we purchased all available American Type Culture Collection and National Collection of Type Cultures reference cultures of C. xerosis (n = 10) and C. striatum (n = 4) and analyzed them as follows: (i) analysis of biochemical reactions in conventional tests and in the Rapid CORYNE system, (iU) whole-cell fatty acid analysis by using the gas-liquid chromatography research software ofMicrobial ID, Inc., and (iii) analysis of DNA homology in dot blot hybridizations. Three C. xerosis strains were indistinguishable from the C. striatum strains in whole-cell fatty acid analyses and DNA hybridizations and shared very similar biochemical reactions. The remaining seven strains of C. xerosis clustered into five groups on the basis of fatty acid patterns, DNA hybridizations, and biochemical tests. No reference strain of C. striatum fit the species description in Bergey's Manual of Systematic Bacteriology. The type strains of both C. striatum and C. xerosis fit their respective descriptions by the Centers for Disease Control and Prevention. This study suggests that the 10 commercially available reference strains of C. xerosis represent six different taxa which should be assigned to new species.
7711 had DNA containing 67 to 68 mol% G+C, which was markedly different from the 57 to 58 mol% found in other reference strains of this species (6, 19, 23, 29). The electrophoretic protein patterns of reference strains of C. xerosis also indicated that they were highly diverse organisms (16). Confusion in the descriptions of C. xerosis and Corynebacterium striatum is found in a comparison of the two major resources available to clinical microbiologists for the identification of coryneform organisms (3, 14). In the CDC system, which includes 34 traits, C xerosis differs from C. striatum only by the former's ability to produce acid from maltose and its lack of hemolysis on sheep blood agar. The maltose reactions listed in Bergey's Manual of Systematic Bacteriology (3) for C. striatum and C xerosis are the opposite of those reported by CDC. The basis for the differences in the two descriptions of C. striatum is the fact that the CDC reactions are based on the C. striatum type strain ATCC 6940 (NCTC 764), whereas the description in Bergey's Manual (3) is from Munch-Petersen's report (22) of bovine strains that were not deposited in either the National Collection of Type Cultures (NCTC) or the American Type Culture Collection (ATCC) (3, 22). The purpose of the present study was to compare all of the commercially available reference strains of C. striatum and C. xerosis in an attempt to resolve the discrepancies in the literature. The results from biochemical tests, whole-cell fatty acid analyses, and DNA-DNA dot blot hybridizations indicate that the 10 reference strains of C. xerosis comprise six different taxa, including one which is indistinguishable from the C. striatum reference strains.
As the survival of severely compromised patients increases with advances in health care, clinical microbiologists more frequently encounter opportunistic coryneforms, commonly called diphtheroids, that cannot be identified by any of the available resources. Unfortunately, the inability of the microbiology laboratory to identify an isolate greatly re-
duces the clinician's recognition of its potential significance. For example, Corynebacterium jeikeium (CDC group JK) was not recognized as an important cause of sepsis in immunocompromised patients until it had been described in a 1979 publication (25). The Special Pathogens Laboratory at the Centers for Disease Control and Prevention (CDC) has addressed the problem of coryneform identification by describing more than 20 biochemically distinct tentative taxa to which they have assigned such names as group JK and group D-2 (14, 15). In spite of major contributions from the CDC, a recent study involving 21 biochemical tests found that only 60% of coryneform clinical isolates could be assigned to any species or CDC group (10). Unfortunately, identification schemes that are limited to validly published species enable reference laboratories to identify only one-third of their coryneform isolates (12, 13). The fact that recognized species within the genus Corynebacterium may be difficult to identify with a high level of confidence can be at least partly attributed to numerous differences within reference strains of a single species (12). Diversity within the reference strains of Corynebacterium xerosis was described in 1970 when Yamada et al. (29) found that the C. xerosis type strain, ATCC 373, and strain ATCC * Corresponding author. Electronic mail address: Coyle@zippy. labmed.washington.edu. t Present address: Associated Regional and University Pathologists, Salt Lake City, UT 84108. * Present address: Department of Pathology, Bay Area Hospital, Coos Bay, OR 97420.
MATERIALS AND METHODS
Strains. All strains used in the present study were freshly purchased from ATCC or NCTC. The following C. striatum strains were used: two cultures of the type strain (ATCC 1788
VOL. 31, 1993
MULTIPLE TAXA WITHIN C. XEROSIS REFERENCE STRAINS
1789
TABLE 1. Conventional biochemical reactions and colony morphologies of C. striatum and C. xerosis reference strains Straina
C. striatum A6940T N764T A43735 A43751
C. xerosis A7094 A9016 N9755 A373T A7711 N7238 N7883 N7929 N8481 N7243
gDNA Catalase NO3 Urease Gelatin Motility Esculin
groupb A A A A
A A A B C D D D E F
Carbohydrate utflization
C Glucose Maltose Sucrose Mannitol Xylose
-
a-1 a-1 a-1
a-i
-
-
a-i
a-i
_
-
+ +
a-1
a-i a-i a-i
+
-
-
-
-
+
-
+
+ +
-
-
-
-
+ +
-
+ +
+
+
-
-
-
-
+
-
+
+ + + + + + +
+e + + + + +
-
-
-
-
-
-
-
-
+ + + + +
-
-
-
-
+ + + + + + +
-
-
+
+
-
-
-
-
+ +
+ -
+ -
-
-
-
-
-
-
+ +e
Morphology
-
+
+ +
-
Serum
stimulation Colonyc Brothd
-
+ -
+9 +B
-
+
of +f o.,f
+
-
+ +
+ +
+ +
-
-
-
+
+ -
a-i
a-2 b-i b-1 d d d d b-2
a-1 a-i
b b -
a-2
a Reference strain numbers preceded by A and N were obtained from ATCC and NCTC, respectively. Superscript T's indicate type strains. b DNA groups are defined in the text, Fig. 2, and Table 4. c Morphology on Trypticase soy blood agar at 48 h. a-1, creamy gray-white colonies, 2 mm in diameter; a-2, colonies resemble a-i colonies, but with tan pigment; b-i, dry, slightly yellow colonies, 0.2 to 1.0 mm in diameter; b-2, colonies resemble b-i colonies, but with white color; d, translucent gray colonies,
