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Chapter 1 : History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci

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Abstract:

The enterococci are a complex, diverse, and important group of bacteria in terms of their interaction with humans. This chapter presents a variety of techniques and procedures that can be used for isolation and identification of these important microorganisms. Early documentation of microorganisms that are now included in the genus is mainly related to the "streptococci of fecal origin". Enterococci are gram-positive cocci that occur singly, in pairs, or as short chains. Most enterococci, apart from , , , and , hydrolyze pyrrolidonyl-β- naphthylamide (PYR); all strains produce leucine aminopeptidase. The tables depicting the phenotypic characteristics of the species in the chapter are based on correlations between the whole-cell protein (WCP) profiles and the phenotypic tests and, on some occasions, in conjunction with DNA-DNA reassociation experiments. Techniques for isolation of enterococci from inanimate hospital surfaces have been developed. For several years, controversies and confusion surrounded antimicrobial susceptibility testing of enterococcal isolates, particularly with regard to the reliability of phenotypic methods for detection of high-level resistance (HLR) to aminoglycosides and resistance to vancomycin. Some detailing of the specific standard recommendations for detecting HLR to aminoglycosides, as well as resistance to β-lactams and vancomycin, is presented in the chapter as they are considered very helpful for routine detection of these important resistance markers in enterococci. Molecular methods can rapidly detect specific antimicrobial-drug resistance genes and have substantially contributed to the understanding of the spread and genetics of acquired enterococcal resistance.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1

Key Concept Ranking

Antimicrobial Susceptibility Testing
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16s rRNA Sequencing
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Gram-Positive Cocci
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Bacterial Cell Wall
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Enterococcus hirae
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Figures

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Figure 1

Phylogenetic relationships among species of and some related species of other gram-positive cocci, based on the analysis of 16S rRNA gene sequences. The dendrogram was constructed by the CLUSTAL method using the DNASTAR program. The scale units indicate evolutionary values between sequence pairs.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
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Image of Figure 2
Figure 2

Phylogenetic relationships among genera of catalase-negative, gram-positive cocci based on the analysis of 16S rRNA gene sequences. The dendrogram was constructed by the CLUSTAL method using the DNASTAR program. The scale units indicate evolutionary values between sequence pairs.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
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Figure 3

(A and B) SDS-PAGE profiles of whole-cell protein extracts of the different species of . Lanes M, molecular mass markers; A1, ; A2, ; A3, ; A4, ; A5, ; A6, ; A7, ; A8, ; A9, ; A10, ; Bl, ; B2, ; B3, ; B4, ; B5, ; B6, ; B7, ; B8, ; B9, . (C) Dendrogram resulting from the computer-assisted analysis of the protein profiles in panels A and B. The scale represents average percentages of similarity.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
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Image of Figure 4
Figure 4

PFGE profiles of I-digested genomic DNA of strains isolated from patients admitted to hospitals located in Rio de Janeiro, Brazil, showing the clonal relationship of HLR-Ge isolates (A) and the clonal diversity of HLR-St isolates (B). Lanes M, molecular size markers expressed in kilobases. Dendrograms resulted from computer-assisted analysis of the PFGE profiles. The scales represent average percentages of similarity.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
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Figure 5

PFGE profiles of genomic DNA of epidemiologically related and unrelated after digestion with (A) and (B). Lanes M, molecular size markers expressed in kilobases. Dendrograms resulted from computer-assisted analysis of the PFGE profiles. The scales represent average percentages of similarity.

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
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Tables

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Table 1

Phenotypic characteristics of facultatively anaerobic, catalase-negative, gram-positive cocci

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 2

Chronology of entry and references for the application of molecular methods in taxonomic studies of the genus

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 3

Phenotypic characteristics used for identification of species and some related species of other gram-positive cocci

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 4

Phenotypic characteristics used for the differentiation of group III species

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 5

Molecular methods applied for the identification of species

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 6

Major patterns and mechanisms of resistance to antimicrobial agents in enterococci

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 7

Suggested grouping of antimicrobial agents and interpretive criteriaa for disk diffusion and dilution susceptibility testingb for species, according to NCCLS recommendations

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1
Generic image for table
Table 8

Screening methods for detecting vancomycin and high-level aminoglycoside resistance in enterococci

Citation: Facklam R, Carvalho M, Teixeira L. 2002. History, Taxonomy, Biochemical Characteristics, and Antibiotic Susceptibility Testing of Enterococci, p 1-54. In Gilmore M, Clewell D, Courvalin P, Dunny G, Murray B, Rice L (ed), The Enterococci. ASM Press, Washington, DC. doi: 10.1128/9781555817923.ch1