Prokaryotes (2006) 4:5–75 DOI: 10.1007/0-387-30744-3_1 CHAPTER 1.2.1 ehT areneG succocolyhpatS dna succocorcMa The Genera Staphylococcus and Macrococcus FRIEDRICH GÖTZ, TAMMY BANNERMAN AND KARL-HEINZ SCHLEIFER Introduction zolidone (Baker, 1984). Comparative immu- nochemical studies of catalases (Schleifer, 1986), The name Staphylococcus (staphyle, bunch of DNA-DNA hybridization studies, DNA-rRNA grapes) was introduced by Ogston (1883) for the hybridization studies (Schleifer et al., 1979; Kilp- group micrococci causing inflammation and per et al., 1980), and comparative oligonucle- suppuration. He was the first to differentiate otide cataloguing of 16S rRNA (Ludwig et al., two kinds of pyogenic cocci: one arranged in 1981) clearly demonstrated the epigenetic and groups or masses was called “Staphylococcus” genetic difference of staphylococci and micro- and another arranged in chains was named cocci. Members of the genus Staphylococcus “Billroth’s Streptococcus.” A formal description form a coherent and well-defined group of of the genus Staphylococcus was provided by related species that is widely divergent from Rosenbach (1884). He divided the genus into the those of the genus Micrococcus. Until the early two species Staphylococcus aureus and S. albus. 1970s, the genus Staphylococcus consisted of Zopf (1885) placed the mass-forming staphylo- three species: the coagulase-positive species S. cocci and tetrad-forming micrococci in the genus aureus and the coagulase-negative species S. epi- Micrococcus. In 1886, the genus Staphylococcus dermidis and S. saprophyticus, but a deeper look was separated from Micrococcus by Flügge into the chemotaxonomic and genotypic proper- (1886). He differentiated the two genera mainly ties of staphylococci led to the description of on the basis of their action on gelatin and on many new staphylococcal species. Currently, 36 relation to their hosts. Staphylococci liquefied species and several subspecies are recognized in gelatin and were parasitic or pathogenic or both the genus Staphylococcus (Table 1). whereas micrococci were variable in their action The genus Macrococcus has been described on on gelatin and were saprophytic. The genera the basis of comparative 16S rRNA analysis, Staphylococcus, Micrococcus and Planococcus, DNA-DNA hybridization studies, ribotype pat- containing Gram-positive, catalase-positive cocci, terns, cell wall composition, and phenotypic were later placed in the family Micrococcaceae. characteristics (Kloos et al., 1998a). Macrococci Evans et al. (1955) proposed separating staphy- can be separated from staphylococci on the basis lococci from micrococci on the basis of their rela- of a generally higher DNA G+C content (38– tion to oxygen. The facultative anaerobic cocci 45 mol%), absence of cell wall teichoic acids were placed in the genus Staphylococcus and the (with the possible exception of M. caseolyticus), obligate aerobic cocci in the genus Micrococcus. unique ribotype patterns, and generally larger By the mid-1960s, a clear distinction could be cells. Members of the genus Macrococcus are made between staphylococci and micrococci on also oxidase positive whereas most staphylococci the basis of their DNA base composition (Silves- (exceptions: S. lentus, S. sciuri and S. vitulus) are tri and Hill, 1965). Members of the genus Staphy- oxidase negative. There are four species in the lococcus have a DNA G+C content of 33–40 genus Macrococcus (Table 1). mol%, whereas members of the genus Micrococ- cus have a high G+C content of around 70 mol%. Further studies have shown that staphylococci can be distinguished from micrococci and other Isolation Techniques catalase-positive cocci on the basis of their cell Isolation of S. aureus from Foods wall composition (Schleifer and Kandler, 1972; Endl et al., 1983), cytochrome profile (Faller et Staphylococcus aureus has been confirmed to be al., 1980) and menaquinone pattern (Collins and the causative agent of many cases of severe food Jones, 1981), susceptibility to lysostaphin and poisoning; therefore, its presence in foods is of erythromycin (Schleifer and Kloos, 1975b), major concern. Staphylococcus aureus is very bacitracin (Falk and Guering, 1983), and fura- susceptible to heat treatment and most sanitiz- 6F. Götz, T. Bannerman and K.-H. Schleifer CHAPTER 1.2.1 Table 1. List of species and subspecies in the genera Macro- ing agents. Hence, when it or its enterotoxins coccus and Staphylococcus. are found in processed foods, poor sanitation is Species name Referencesa usually indicated. Detailed procedures for pre- Macrococcus bovicus Kloos et al., 1998a paring food samples for analysis, isolating and M. carouselicus Kloos et al., 1998a enumerating S. aureus, and detecting staphylo- M. caseolyticus Schleifer et al., 1982 coccal enterotoxins in foods can be found in the Kloos et al., 1998a following texts: Compendium of Methods for the M. equipercicusT Kloos et al., 1998a Microbiological Examination of Foods (Downes Staphylococcus arlettae Schleifer et al., 1984 and Ito, 2001), Official Methods of Analysis of S. auricularis Kloos and Schleifer, 1983a S. aureusT Rosenbach, 1884 AOAC International (Horowitz, 2000), and Bac- S. aureus subsp. anaerobius De la Fuente et al., 1985 teriological Analytical Manual (BAM; United S. aureus subsp. aureus De la Fuente et al., 1985 States Food and Drug Administration, 1995). In S. capitis Kloos and Schleifer, 1975b addition to the AOAC (Association of Official S. capitis subsp. capitis Bannerman and Kloos, 1991 Analytical Chemists) approved microslide test S. capitis subsp. urealyticus Bannerman and Kloos, 1991 method (Horowitz, 2000), it is possible to detect S. caprae Devriese et al., 1983 enterotoxins directly in culture and in contami- S. carnosus Schleifer and Fischer, 1982 S. carnosus subsp. carnosus Probst et al., 1998 nated foods by the following rapid methods: S. carnosus subsp. utilis Probst et al., 1998 radioimmunoassay (RIA) (Miller et al., 1978), S. chromogenes Devriese et al., 1978 enzyme-linked immunosorbent assay (ELISA), S. cohnii Schleifer and Kloos, 1975c and reverse passive latex agglutination (RPLA). S. cohnii subsp. cohnii Kloos and Wolfshohl, 1983b An ELISA kit, available from Tecra Diagnostics S. cohnii subsp. urealyticus Kloos and Wolfshohl, 1983b (Roseville, Australia), is distributed by Interna- S. condimenti Probst et al., 1998 S. delphini Varaldo et al., 1988 tional Bioproducts, Inc. (Redmond, WA, USA), S. epidermidis Winslow and Winslow, 1908 and an RPLA is available from Oxoid (Colum- S. equorum Schleifer et al., 1984 bia, MD, USA). Molecular methods are being S. felis Igimi et al., 1989 investigated for their usefulness in detecting sta- S. fleurettii Vernozy-Rozand et al., 2000 phylococcal enterotoxin. For example, Western S. gallinarum Devriese et al., 1983 immunoblotting has been used to detect staphy- S. haemolyticus Schleifer and Kloos, 1975c S. hominis Kloos and Schleifer, 1975b lococcal enterotoxin A (Rasooly and Rasooly, S. hominis subsp. hominis Kloos et al., 1998b 1998). In this procedure, the staphylococcal S. hominis subsp. Kloos et al., 1998b enterotoxin A (native or heat-denatured) is sep- novobiosepticus arated by size using sodium dodecyl sulfate S. hyicus Devriese et al., 1978 polyacrylamide gel electrophoresis (SDS- S. intermedius Hájek, 1976a PAGE), transferred to a membrane and then S. kloosii Schleifer et al., 1984 the membrane is probed with antibodies. The S. lugdunensis Freney et al., 1988 S. lutrae Foster et al., 1997 polymerase chain reaction (PCR) is an addi- S. muscae Hájek et al., 1992 tional molecular method being studied for its S. pasteuri Chesneau et al., 1993 ability to detect S. aureus and staphylococcal S. piscifermentans Tanasupawat et al., 1992 enterotoxins from food samples (McLauchlin et S. pulverei (= S. vitulinus) Petras, 1998 al., 2000; Atanassova et al., 2001; Tamarapu et Zakrzewska-Czerwinska al., 2001). One potential problem for PCR is the et al., 1995 possibility of false-negative reactions due to S. saccharolyticus Kilpper-Bälz and Schleifer, 1981 PCR inhibitors that might be present in some S. saprophyticus Schleifer and Kloos, 1975c foods. S. saprophyticus subsp. bovis Hájek et al., 1996 Several conventional procedures for isolating S. saprophyticus subsp. Hájek et al., 1996 S. aureus from foods are described below. saprophyticus S. schleiferi Freney et al., 1988 Nonselective Enrichment Procedures It is S. schleiferi subsp. coagulans Igimi et al., 1990 often necessary to use nonselective enrichment S. schleiferi subsp. schleiferi Igimi et al., 1990 S. sciuri Kloos et al., 1976a procedures for the detection of S. aureus in pro- S. sciuri subsp. carnaticus Kloos et al., 1997 cessed foods, especially when it is suspected that S. sciuri subsp. lentus Kloos et al., 1997 the food contains a small number of cells that S. sciuri subsp. rodentium Kloos et al., 1997 may have been injured, e.g., as a result of heat- S. sciuri subsp. sciuri Kloos et al., 1997 ing, freezing, desiccation, or storage, and whose S. simulans Kloos and Schleifer, 1975b growth could be inhibited by toxic components S. succinus Lambert et al., 1998 S. vitulinus Webster et al., 1994 of a selective enrichment media. The following S. warneri Kloos and Schleifer, 1975b nonselective (repair) enrichment procedure is S. xylosus Schleifer and Kloos, 1975c appropriate for this use: T Nonselective Enrichment of S. aureus Abbreviation: , type species. (Downes and Ito, 2001). aFirst description. CHAPTER 1.2.1 The Genera Staphylococcus and Macrococcus 7 Transfer a