Chapter 19 : , , and Other Catalase-Positive Cocci

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Historically, the genera and were placed together with the genera and in the family containing grampositive, catalase-positive cocci. An unrelated species of gram-positive cocci exhibiting positive catalase reaction and occurring in human specimens is , the only species of this genus, which is a member of the family belonging to the order . In this chapter, the term "micrococci" is used to indicate the members of the genus as understood before the emendation, reflecting most of the clinically relevant species. The genus comprises four hoofed-animal-adapted species including , first described as . species can be identified phenotypically on the basis of a variety of conventional characteristics. The diagnosis of staphylococcal toxic shock syndrome (TSS) and staphylococcal scalded skin syndrome (SSSS) is based on clinical signs supplemented by serologic tests and the detection of the toxin production by staphylococcal isolates (, rarely other species). Detection of methicillin-resistant (MRSA) represents the most important task in determining the antimicrobial susceptibilities of staphylococci. Coagulase-negative staphylococci (CoNS) are an important cause of nosocomial bloodstream infections, but they are also the most common contaminants of blood cultures. The considerations of clinical significance discussed for CoNS are also appropriate for members of the and families; however, the criteria used for distinguishing etiologically relevant isolates from contaminants and colonizers, respectively, should be applied much more strictly.

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19
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Image of FIGURE 1

Columbia blood agar plate with 5% sheep blood showing subsp. after 24 h of incubation displaying the typical shapes of the normal phenotype with golden yellow-pigmented colonies surrounded by a hemolysis zone (left) and of the small-colony phenotype characterized by tiny, nonhemolytic, and nonpigmented colonies (right).

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19
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Differentiation of members of the genus from other gram-positive cocci

Symbols and abbreviations: +, 90% or more species or strains positive; ±, 90% or more species or strains weakly positive; –, 90% or more species or strains negative; , 11 to 89% of species or strains positive; ND, not determined. Parentheses indicate a delayed reaction.

Growth on P agar is under aerobic conditions at 35 to 37°C. Positive growth is indicated for detectable formation of colonies of at least 1 mm in diameter; ± indicates detectable formation of colonies of between 0.5 and 1 mm in diameter. Growth on sheep or bovine blood agar is slightly greater but less discriminative between staphylococci and other genera.

Growth is under aerobic conditions at 35 to 37°C for 24 to 48 h. Positive growth is indicated for a number of CFU on selective medium comparable to that on plate count agar and a colony of 0.5 mm in diameter; ± indicates a significant reduction in the number of CFU on the selective medium compared to that on plate count agar, and parentheses indicate a colony of pinpoint size to 0.5 mm in diameter.

Sometimes a weak catalase or pseudocatalase reaction can be observed with certain strains of species designated as catalase negative. In some species, catalase activity may be activated by hemin supplementation.

Detects the presence of cytochromes. Some strains of benzidine test-negative species can synthesize cytochromes on aerobic media supplemented with hemin ( ).

Determined by the modified oxidase test using tetramethyl-p-phenylenediamine dihydrochloride-impregnated disks or strips to detect the presence of cytochrome ( ).

Standard oxidation/fermentation test.

A disk is used. Positive indicates resistance and no zone of inhibition. , , , , and spp. are susceptible and have an inhibition zone of 10 to 25 mm in diameter.

A disk is used. Positive indicates resistance and no zone of inhibition or a zone of up to 9 mm in diameter. Susceptible species have an inhibition zone of 15 to 35 mm in diameter.

Some strains of adhere tenaciously to the surface of agar, and this property is correlated with heavy slime production.

does not demonstrate growth in the anaerobic portion of a thioglycolate medium within 24 h and may produce only very poor growth in this portion following 3 to 5 days of incubation. However, it grows and ferments glucose anaerobically (standard oxidation-fermentation test). Failure to grow anaerobically in thioglycolate may be due in part to inhibition by the ingredients.

species can also be differentiated from species on the basis of their generally larger Gram-stained cell size (≥2 μm) and larger number of chromosome fragments produced by digestion with NotI (12 to 36 fragments).

A few strains demonstrate high-level (MIC, ≥50 μg/ml) erythromycin resistance.

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19
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Differentiation of species

Symbols and abbreviations (unless otherwise indicated): +, 90% or more strains positive; ±, 90% or more strains weakly positive; –, 90% or more strains negative; , 11 to 89% of strains positive; ND, not determined. Parentheses indicate a delayed reaction.

Positive is defined as a colony diameter of ≥6 mm after incubation on P agar at 34 to 35°C for 3 days and at room temperature (ca. 25°C) for an additional 2 days; exceptions are (4 to 6 mm on tryptic soy agar) and (8 to 12 mm on tryptic soy agar).

Positive is defined by the visual detection of carotenoid pigments (e.g., yellow, yellow-orange, or orange) during colony development at normal incubation or room temperatures. Pigments may be enhanced by the addition of milk, fat, glycerol monoacetate, or soaps to P agar.

Growth is in a semisolid thioglycolate medium. Symbols: +, moderate or heavy growth down the tube within 18 to 24 h; ±, heavier growth in the upper portion of the tube and weaker growth in the lower, anaerobic portion of tube; 2, no visible growth within 48 h but very weak diffuse growth or a few scattered, small colonies may be observed in the lower portion of the tube by 72 to 96 h. Parentheses indicate delayed growth appearing within 24 to 72 h, sometimes noted as large, discrete colonies in the lower portion of the tube.

Growth is on P agar or bovine, sheep, or human blood agar at 34 to 37°C. The subspecies of and grow slowly at 35 to 37°C; the optimum growth temperatures of subsp. and subsp. are 30°C and 32°C, respectively, and those of subsp. and of subsp. are 28°C and 32°C, respectively. Anaerobic species and subsp. grow very slowly in the presence of air. subsp. requires the addition of blood, serum, or egg yolk for growth on primary isolation medium. , , and produce just-detectable colonies on P agar in 24 to 36 h, and these colonies remain very small (1 to 2 mm in diameter).

The slide agglutination test using rabbit or human plasma detects the expression of clumping factor. Use human plasma for and . Latex agglutination is less reliable for the detection of clumping factor in .

Hemolysis on bovine blood agar. Symbols and abbreviations: +, wide zone of hemolysis within 24 to 36 h; (+), delayed moderate to wide zone of hemolysis within 48 to 72 h; (), no or delayed hemolysis; –, no or only very narrow (1-mm) zone of hemolysis within 72 h. Some strains designated as negative may produce a slight greening or browning of blood agar. Analysis of hemolysis for both subspecies was performed on Wilkins-Chalgren anaerobe agar plates containing 5% sheep blood.

Catalase and cytochrome synthesis cannot be induced in subsp. by the addition of HO or hemin to the culture medium. Catalase activity can be induced in by hemin supplementation. In this species, cytochromes and are present in small quantities.

Determined by the modified oxidase test to detect the presence of cytochrome ( ).

Determined primarily by commercial rapid identification tests (see the text).

Positive (resistant) is defined by an MIC of ≥1.6 mg/ml or a growth inhibition zone diameter of ≥16 mm with a 5-μg novobiocin disk.

Positive is defined by a growth inhibition zone diameter of <10 mm with a 300-U polymyxin B disk.

Approximately 6 to 15% of strains of are negative for alkaline phosphatase activity, depending on the population sampled. A low but significant number of clinical isolates are phosphatase negative.

All strains tested are also resistant to penicillin G, methicillin, oxacillin, gentamicin, and streptomycin.

Positive with the STAPH-ZYM tests, but negative with the API STAPH tests ( ).

Positive reactions are with the Staph latex agglutination test (Remel) that detects clumping factor and/or protein A.

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19
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Key tests for identification of the most clinically significant species

Symbols and abbreviations (unless otherwise indicated): +, 90% or more strains positive; ±, 90% or more strains weakly positive; –, 90% or more strains negative; , 11 to 89% of strains positive; ND, not determined. Parentheses indicate a delayed reaction.

Descriptions are the same as those in Table 2 .

Alkaline phosphatase reactions tested positive in the STAPH-ZYM gallery but negative in the API STAPH gallery.

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19
Generic image for table

Key tests for the identification of and found in clinical specimens

Symbols and abbreviations: +, 90% or more strains positive; ±, 90% or more strains weakly positive; –, 90% or more strains negative; , 11 to 89% of strains positive; ND, not determined. Parentheses indicate a delayed reaction.

Citation: Becker K, von Eiff C. 2011. , , and Other Catalase-Positive Cocci , p 308-330. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch19

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