INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY Jan. 1975, p. 50-61 Val. 25, No.

1
Copyright 0 1975 International Association of Microbiological Societies Printed in U S A .

Isolation and Characterization of Staphylococci

from Human Skin
I. Amended Descriptions of Staphylococcus epidermidis and
Staphylococcus saprophyticus and Descriptions of Three New Species:
Staphylococcus cohnii, Staphylococcus haemolyticus, and
Staphylococcus xylosus
KARL H.SCHLEIFER AND WESLEY E. KLOOS
Lehrstuhl fir Mikrobiologie, Universittit Munchen, 8 Munich 19, Germany, and Department of Genetics,
North Carolina State University, Raleigh, North Carolina 27607

Staphylococci were isolated from human skin and subjected to a taxonomic

study. As a result of this study, three new species are being proposed in this
paper: Staphylococcus cohnii, S. haemolyticus, and S. xylosus. The type strains
of these species are DSM (Deutsche Sammlung von Mikroorganismen) 20260,
DSM 20263, and DSM 20266, respectively. Amended descriptions of S. epider-
midis and S. suprophyticus are also given. The main characters for the distinction
of staphylococci and micrococci are mentioned. Staphylococci were classified on
the basis of cell wall composition, lactic acid configuration, and a variety of
morphological and physiological characters. There are some key differential char-
acters of these species which can be determined by simple laboratory procedures.
The failure to ferment trehalose and mannitol is typical for S. epidermidis. The
fermentation of xylose and/or arabinose is a characteristic of S. xylosus. The fail-
ure to ferment sucrose and turanose is typical for s. cohnii. Strains of s. supro-
phyticus do not reduce nitrate, but most of them produce acetylmethylcarbinol
and ferment xylitol. S. haemolyticus is usually hemolysis positive, like S. uureus,
but it does not produce coagulase, does not have strong phosphatase and deoxy-
ribonuclease activities, and does not ferment mannose.

Although gram-positive, catalase-positive compounds in their cell walls. In addition, the

cocci are very common on human skin, the cell wall composition and the type of lactic acid
methods used to classify them are not com- produced from glucose under anaerobic condi-
pletely satifactory. Generally, the scheme de- tions are very valuable in differentiating staphy-
vised by Baird-Parker (2, 3) is employed to lococci (20). This paper reports on the results
divide the family Micrococcaceae. However, of the application of these criteria, as well as of
with this scheme it is impossible to separate morphological and physiological characters, to
weakly anaerobic staphylococci from micro- the classification of staphylococci isolated from
cocci, and such strains are often misidentified human skin.
as micrococci. In applying Baird-Parker’s sys- MATERIALS AND METHODS
tem, Noble (12,13) has found that the predomi-
Bacterial strains. Staphylococci were isolated
nant groups of skin cocci are Staphylococcus from the healthy skins of two groups of people: 20
type 2 (probably S. epidermidis sensu stricto) people living in Raleigh, N.C., who were sampled once
and Micrococcus type 2. The taxonomic status each month for 6 to 13 months, and 20 people living in
of the latter type, which may include some Somerville or New Brunswick, N.J., who were sam-
weakly anaerobic staphylococci, is not clear. pled once during the winter. Samples were taken from
Deoxyribonucleic acid (DNA) base composition two separate sites on the forehead and one site from
and cell wall composition are the characters one cheek, the chin, one external nare, anterior nare,
most; useful in separating Micrococcus from each axilla, each upper and lower arm, and each
upper and lower leg. Representative strains are listed
Stuphylococcus (16, 18, 19). The cell wall pepti- in Tables 2 through 5.
doglycan of the staphylococci contains large Isolation and culture techniques. The procedures
amounts of glycine and is thus clearly different and media used for isolating staphylococci were
from that of the micrococci. Moreover, all identical to those used in previous studies on micro-
staphylococci contain teichoic acids or similar cocci from human skin (10).
50
VOL.25, 1975 STAPHYLOCOCCI FROM HUMAN SKIN. I. 51

Morphological and colonial characters. Colony examination of the cell wall teichoic acids. In cases
characteristics and cell morphology were determined where both glycerol and ribitol were found in the acid
as described previously for micrococci (10). hydrolysates, the teichoic acids were checked more
Physiological characters. Oxygen and tempera- carefully. The cell walls were extracted with phenol
ture relationships and tolerance to NaCl were deter- (80% [wt/vol] in water) or hot water to remove traces
mined as described previously (10). of membrane teichoic acid. For some representative
Biochemical characters. Catalase and benzidine strains of each group, the teichoic acid composition
tests, DNA base composition, acetylmethylcarbinol was also studied more accurately by gas chromatogra-
production, nitrate reduction, and carbohydrate reac- phy or by enzymatic methods (20).
tions were determined as described previously for Determination of the type of lactic acid pro-
micrococci (10). Bound and free coagulase activities duced. The procedure for determining the type of
were determined on human, rabbit, and bovine plasma lactic acid produced was described previously (20).
(1, 24). Hemolysis was detected on bovine, human, Organisms were grown in yeast extract-peptone-
sheep, and rabbit blood agar plates after 24, 48, and glucose broth under microaerophilic or anaerobic
72 h by using streak cultures. Deoxyribonuclease pro- conditions. They were incubated at 34 C for 5 to 6
duction was determined on deoxyribonuclease test days. The type of lactic acid produced was deter-
agar (Difco) plates after 24 and 48 h by using streak mined enzymatically with L-lactate dehydrogenase
streak cultures. Deoxyribonuclease (DNase) produc- from rabbit muscle (Boehringer, Mannheim) and
tion was determined on deoxyribonuclease test agar D-lactate dehydrogenase obtained from Leuconostoc
(Difco) plates after 24 and 48 h by using streak mesenteroides ATCC 12291 (6).
cultures. Phosphatase activity was determined by a Chemicals. Egg white lysozyme (muramidase) was
modification of the technique of Pennock and Huddy purchased from Sigma Chemical Co., St. Louis. Mo.;
(14), using a 0.005 M phenolphthalein monophos- lysostaphin was from Schwarz/Mann, Orangeburg,
phate sodium salt (Sigma) instead of a 0.01 M N.Y. T h e following carbohydrates were tested:
solution of disodium phenylphosphate. Production of D( + )-glucose, D-D( - )-fructose, D( +)-galactose, D( + ) -
bacteriolytic activity was detected by using freeze- mannose, L(+)-rhamnose, D( +)-xylose, L ( + ) -
dried M . luteus cells (lysozyme substrate, Difco) (26) arabinose, D( - )-ribose, maltose, a-lactose, sucrose,
or living M. luteus (ATCC 27141) cells in an agar D( +)-trehalose, D( +)-turanose, P-gentiobiose, D( + )-
overlay. cellobiose, D( + )-melezitose, raffinose, D-mannitol,
Lysostaphin susceptibility. An agar overlay was glycerol, xylitol. D-sorbitol, rneso-inositol, salicin,
prepared by adding 0.1 ml of a saline cell suspension adonitol (ribitol), dulcitol, earabitol, rneso-erythritol,
(approximately 10' colony-forming units per ml) to a D-erythrose. P-melibiose, a-L-fucose, tagatose, D-lyx-
tube containing 3 ml of fluid, soft peptone-yeast ose, and L-sorbose.
extract-glucose agar (10). The contents were mixed
thoroughly and then poured on the surface of a dry RESULTS AND DISCUSSION
soft peptone-yeast extract-glucose agar plate. A drop Identification of the genus Staphylococcus.
of sterile lysostaphin solution (200 pg/ml) was placed The main characteristics for the differentiation
on the inoculated agar and incubated for 24 to 48 h. of the genera Micrococcus and Staphylococcus
Lysostaphin susceptibility was interpreted according
to the following spot-inhibition scheme: + sensitive are listed in Table 1. Members of the genus
(complete growth inhibition); *, slightly resistant Staphylococcus possess a low guanine plus cyto-
(partial growth inhibition); and -, resistant (no visi- sine ( G + C ) content (30 to 38 mol%) in their
ble growth inhibition). The minimal inhibitory con- DNA, whereas members of the genus
centration of lysostaphin was determined on agar Micrococcus have a rather high G + C content
plates containing different concentrations of lyso- (66 to 73 mol%). These differences in the DNA
staphin. A loopful of a saline cell suspension was base composition are accompanied by differ-
placed on each plate. Plates were incubated for 48 h ences in the chemical composition of the cell
and then examined for growth.
Lysozyme susceptibility. Lysozyme susceptibil-
walls, as discussed above. The difference in the
ity was determined on the same agar plate as used for cell wall peptidoglycan composition is reflected
testing lysostaphin. A drop of a sterile lysozyme by the susceptibility lysostaphin. The main bac-
solution (400 pglml) was placed on the agar plate. teriolytic activity of lysostaphin is due to an
Lysozyme susceptibility was interpreted by using the endopeptidase with a specificity of action
same scheme described for the lysostaphin suscepti- against the pentaglycine interpeptide bridge of
bility test. the peptidoglycan. Thus, all staphylococci
Antibiotic susceptibilities. Procedures for testing tested by the spot test described above showed
susceptibility to penicillin G, streptomycin, erythro- some susceptibility to lysostaphin, whereas
mycin, tetracycline, and- novobiocin were described most micrococci were resistant.
previously ( 10).
Cell wall preparation and analyses. Cell walls Only a few micrococci that were susceptible
were prepared and analyzed as described previously to lysozyme showed a weak susceptibility to ly-
(17, 20). The different peptidoglycan types are abbre- sostaphin. These strains, however, cannot be
viated by the system of Schleifer and Kandler (19).The confused with staphylococci, since all of the
procedure of Wolin et al. (27) was used for qualitative staphylococci were resistant to lysozyme.
52 SCHLEIFER AND KLOOS INT. J. SYST.BACTERIOL.
TABLE
1. Abbreviated scheme for the differentiation of the genera Micrococcus and Staphylococcus
Differentiation test Micrococcus Staphylococcus

G+C content (2mol%) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66-73 30-38

Cell wall teichoic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . - +
Peptidoglycan composition ( > 2 mol of glycindmol
of lysine) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . - +
Lysostaphin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Resistant" Susceptible or
slightly resistant
Anaerobic growth in thio-
glycolate medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -, (h)b +, f
Colony profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Convex (high) Raised to slight convex
Growth a t 45 C . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . - 9 f +, (+, -1
Acid from glycerol (aerobically) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . -, (+)" +
Some strains of M.luteus and a few strains of M.roseus are slightly lysostaphin susceptible. These strains,
however, can be easily distinguished from staphylococci by their susceptibility to lysozyme and their charac-
teristic pigmentation.
M . kristinae produces individual colonies in the anaerobic zone like certain species of staphylococci.
M. kristinue produces acid aerobically from glycerol, and a few strains of M.roseus and M. oarians produce
weak acid.

Most strains of Staphylococcus demonstrated from the same person, where clonal populations
a good diffuse growth or a t least individual appeared to be rather common. The attempts a t
colonies-in the anaerobic portion of the semi- strain identification were similar to those used
solid thioglycolate medium ( 7 ) . Out of more for micrococci (lo), except that 20 colonies of
than 900 strains of staphylococci, only 46 failed each suspected strain were isolated (when avail-
to grow in the anaerobic portion of this medium. able), and growth on inorganic nitrogen agar
On the other hand, most micrococci, except was not applied to strain analysis.
strains of M. kristinae, did not grow anaerobi- Characterization of coagulase-negative
cally in the thioglycolate medium. As discussed Staphylococcus species. Descriptions of five
previously (lo), the Micrococcus colonies were of the coagulase-negative Staphylococcus spe-
characteristically more convex than those of cies isolated from human skin in this study are
Staphylococcus. Only some strains of M. as follows.
uariuns, which produced colonies with an un- (i) S. epiderrnidis (Winslow and Winslow
usually low convex profile, could be confused 1908) Evans 1916. The following description of
with certain staphylococci. Staphylococci could S. epidermidis is based on a total of 204 strains,
be further distinguished from micrococci by unless noted otherwise.
their growth properties. (i) Freshly isolated The cells were gram-positive cocci, 0.5 to 1.0
staphylococci grew much faster on agar plates pm in diameter, and arranged predominantly in
than did micrococci. (ii) They developed easily pairs and tetrads; only occasionally did they
detectable colonies within 24 h, whereas colo- occur singly.
nies of micrococci were only pin-point or could Colonies were raised but showed depressed,
not be detected before 2 days of growth a t 34 C. dark centers with crowding, elevated tempera-
(iii) Moreover, most strains of staphylococci ture (above 35 C), or age. They were circular,
(90%) did grow a t 45 C. Most micrococci, on the 2.5 to 4.0 mm in diameter, entire, smooth,
other hand, usually did not grow at 45 C; only glistening, and translucent to slightly opaque.
M. luteus strains and some strains of M . lylae Colonies were gray or grayish-white. Only rarely
and M . nishinomiyaensis demonstrated weak did some strains demonstrate a slight yellow or
growth a t this temperature. brownish pigment. Colonies or culture streaks
An additional criterion for the separation of were usually sticky in consistency.
staphylococci from most micrococci is the aero- Growth in the anaerobic portion of the thio-
bic formation of acid from glycerol. In contrast glycolate medium was uniformly dense. All
to the staphylococci, only some of the micro- strains grew well a t NaCl concentrations up to
cocci, like M . kristime and a few strains of M 7.5%. More than 80% of the strains grew only
roseus and M . uarians, could produce acid poorly at 10%and failed to grow a t 15%. All
aerobically from glycerol. strains grew well at 45 C, but grew poorly or
Identification of strains. Strain identifica- failed to grow at 15 C.
tion was necessary for staphylococci isolated None of the strains produced coagulase, and
VOL.25, 1975 STAPHYLOCOCCI FROM HUMAN SKIN. I. 53
about 30% showed weak hemolysis activity. All production, nitrate reduction, growth a t ex-
strains were catalase and benzidine test posi- treme temperature, and susceptibility to novo-
tive. All produced acetylmethylcarbinol. More biocin.
than 80% of the strains reduced nitrate or dem- (ii) S. saprophyticus Fairbrother emend.
onstrated phosphatase activity. About half of Shaw et al. 1951. The following description of S .
the strains showed a weak extracellular deoxy- saprophyticus is based on a total of 83 strains,
ribonuclease activity. Two-thirds of the strains unless noted otherwise.
demonstrated b act eriolyt ic activity . Cells were gram-positive cocci with a mean
All strains produced acid aerobically from diameter of 0.8 to 1.2 pm; they were nonmotile,
glucose, fructose, maltose, sucrose, and glyc- nonsporeforming, and occurred predominantly
erol. Seventy to 90% of the strains produced in pairs or as single cells.
acid from galactose, mannose, lactose, or turan- Colonies were raised to slightly convex, circu-
ose. Less than 20% produced acid for ribose or lar, usually entire, 4.0 to 9.0 mm in diameter,
melezitose. No acid was produced from rham- smooth, glistening, and usually opaque. Colony
nose, xylose, arabinose, trehalose, gentiobiose, pigment was variable; however, most strains
cellobiose, mannitol, xylitol, sorbitol, inositol, were unpigmented or had a slight yellow tint
salicin, adonitol, dulcitol, arabitol, erythritol, which increased in intensity with age.
erythrose, raffinose, melibiose, fucose, tagatose, Growth occurred in both the aerobic and
lyxose, or sorbose. anaerobic portions of thioglycolate medium;
All strains were either susceptible or slightly growth in the anaerobic portion was usually
resistant to lysostaphin and were resistant to dense or showed a gradient from dense to light
lysozyme. They were usually susceptible to growth in the deeper, more anaerobic, portion of
novobiocin. Most strains were susceptible to this medium. Seventeen of the strains isolated
penicillin, erythromycin, and tetracycline. from skin and strains CCM 883 ( = ATCC 15305
Most strains were resistant to streptomycin (5 = NCTC 7292; type strain, reference 22)72204,
pg/ml). and P.O. 3519 (obtained from P. Oeding, Ber-
Twenty-one strains studied previously (group gen, Norway) lowered the pH in yeast extract-
IIAI, reference 20) and 20 strains isolated from glucose broth under anaerobic conditions from
skin contained peptidoglycan of type L-LYS- 6.8 to 5.0-5.4. Low amounts of lactic acid,
Glyr-a,L-Sero.7-l.5, The cell wall teichoic acid of primarily L-lactic acid, were produced by most
these strains was composed of glycerol and strains (Table 2). All strains grew well or at
glucose. least poorly at concentrations of NaCl up to
All strains lowered the pH of yeast extract- 15%. All strains grew well at 15 to 40 C; 36% of
glucose broth under anaerobic conditions from the strains grew poorly and 26% failed to grow a t
6.8 to 4.3-4.6. They produced predominantly 45 c.
L-lactic acid and only small amounts (usually None of the strains produced coagulase or
less than 10%)of D-lactic acid. The type strain, extracellular deoxyribonuclease or showed bac-
ATCC 14990 (8), and 20 strains studied previ- teriolytic or hemolysin activity. All strains were
ously (group IIA1, reference 20) contained an positive for catalase and the benzidine test. All
L-lactate dehydrogenase, which is activated by strains produced acetylmethylcarbinol and,
fructose-176-diphosphate. with the exception of strain SE 11, did not
The G+C content of the DNA, as determined reduce nitrate and did not demonstrate phos-
in six strains, is 33.5 * 0.2 mol%. phatase activity.
Recent studies by Schleifer and Steber (21) All strains produced acid aerobically from
showed t h a t phages isolated from S . glucose, maltose, sucrose, turanose, mannitol,
epidermidis strains by Verhoef et al. (25) ad- and glycerol. At least 80% of the strains pro-
sorbed only to cell walls showing the character- duced acid from fructose, lactose, trehalose, and
istic cell wall composition of s. epidermidis. In xylitol. Strain SE 11 produced acid from galac-
particular, the presence of glucosylated glycerol tose and strain KL 150 produced acid from
teichoic acid was necessary for specific adsorp- gentiobiose. No acid was produced from man-
tion. nose, rhamnose, xylose, arabinose, ribose, cel-
S. epidermidis can be distinguished from lobiose, melezitose, sorbitol, inositol, salicin,
other staphylococci primarily by its cell wall adonitol, dulcitol, arabitol, erythritol, ery-
composition, colony morphology, growth on t h o s e , raffinose, melibiose, fucose, tagatose,
NaCl agar, carbohydrate reaction pattern, and, lyxose, or sorbose.
to a lesser extent, on the type of lactic acid All strains were susceptible to lysostaphin
produced, growth in thioglycolate medium, and resistant to lysozyme. All strains were
ph 0sp h a t as e activity, ac et y lm et h y lcar b inol susceptible to penicillin and tetracyline. Most
54 SCHLEIFER AND KLOOS INT. J. SYST.BACTERIOL.

TABLE
2. Variable characters of 20 strains of S . saprophyticus"
Pept ido-
Anaero- Anaero- Produ c-
Acid (aerobically) glycan
bic obic fer- tion of Nitrate
Strain
molQ Colonyb
thio-
G c C i g me n t glycolate
nentation
L-Lactic Growtt
of glucose acid (%)
at 45 C acetyl-
methyl-
Reduc-
tion -
from
--
1 (mol/mol
ofGlu)
growth' (pH) carbinol
Mtl Xyli sue GlY
-~ - -- -
CCM 883 31.6* + 5.0 87 - + + f + + 0.8 4.3
CCM 2204 30.8d ND 5.0 89 ND + + ND + + 0.8 4.2
P.O. 3519 ND 5.4 72 ND + + ND + + 0.6 5.1
KL 20 35.5 + 5.1 60 f + + f + + 0.7 4.6
Tw 111 36.3 f 5.2 73 +- + + + + + 0.6 4.1
S E 11 + 5.1 82 + + f + + 0.6 4.9
AW 163 f 5.3 100 f + + + + f 0.7 5.1
DM 115 + 5.0 85 f f + + + + 0.6 4.4
KL 251 + 5.2 75 f + + + + + 0.7 4.4
KL 150 + 5.2 80 f + + +- + + 0.7 4.2
KL 172 fC 5.1 91 - f + + 4.5
DM 66 + 5.2 70 f + + f + 4.5
RM 454 f 5.2 67 * f + f + 4.5
DM 100 33.5 + 5.O 66 - + + + + 4.2
GH 196 + 5.0 75 - + + + + 4.1
KL 122 + 5.4 68 - f + + + 4.6
S M 118 f 5.1 80 f f + - + 5.0
SM 295 f 5.4 62 f + + - + 4.5
JRM 27 f 5.0 64 f + + * + 4.2
SM 161 + 5.4 58 - + + -
- +
+ - 4.5
-
Reaction: +, positive; f,weak; -, negative; ND, not determined. Abbreviation of carbohydrates: Ara, arabi-
nose; Fru, fructose; Gal, galactose; Lac, lactose; Man, mannose; Mtl, manitol; Rib, ribose; SUC, sucrose; Tur,
turanose; Xyl, xylose; Xyli, xylitol.
Symbols for pigment: gw, gray-white; gy, gray-yellow; w, white; wg, white to gray; wy, white with yellowish
tint; y, yellow; yo, yellow-orange; yt, yellowish tint.
c Symbols for anaerobic growth in thioglycolate medium: +, uniformly dense; f,gradient of growth from

dense to light in the deeper, more anaerobic, portion of the medium; +C,gradient plus individual colonies; -C,
individual colonies only; - , no growth.
According to Silvestri and Hill (23).

strains were susceptible to streptomycin and other staphylococci primarily on the basis of cell
erythromycin. All strains were resistant to novo- wall composition, carbohydrate reaction pat-
biocin ( 5 pg/ml). tern, and, to lesser extent, on low acid produc-
Seventeen strains isolated from skin and tion under anaerobic conditions, production of
strains CCM 883,2204, and P.O. 3519 contained acetylmethylcarbinol, the failure to reduce ni-
.
peptidoglycan of type ~ - L p - G l y , -,, L-Sero-6-o.8 trate, and resistance to novobiocin.
Their cell wall teichoic acids contained two (iii) S. cohnii sp.n. (c0h'ni.i. M.L. gen. noun
polyols: glycerol and ribitol. In a previous study of Cohn; named for Ferdinand Cohn, a German
(20), only ribitol was determined as a polyol botanist and bacteriologist). The following de-
constituent of the cell wall teichoic acid of scription of S . cohnii is based on a total of 42
strains CCM 883 and 2204, and related strains strains, unless noted otherwise.
(group IIB), but gas chromatographic and en- Cells were gram-positive cocci, 0.5 to 1.2 pm
zymatic analyses showed that glycerol, al- in diameter, occurring predominantly singly or
though usually in much smaller amounts than in pairs (tetrads were found only in a few
ribitol, is also present. strains). Nonmotile and nonsporeforming.
The G + C content of the DNA, as determined Colonies were convex to slightly peaked (a
in five strains, is 33.5 A 1 mol%. few strains had colonies with depressed cen-
Some variable characters and the G + C con- ters), flat edged, circular, 4.0 to 7.5 mm in
tent of the DNA of representative strains of s. diameter, entire, smooth, usually glistening or
suprophyticus are shown in Table 2. glossy, opaque, and usually unpigmented.
S . saprophyticus can be distinguished from Growth occurred in both the aerobic and
VOL.25, 1975 STAPHYLOCOCCI FROM HUMAN SKIN. I. 55
anaerobic portions of thioglycolate medium; in three strains, is 37.2 A 0.5 mol%.
growth in the anaerobic portion was either Variable characters of 20 selected strains and
uniformly dense or showed a gradient from the G + C contents of some representative
dense to light growth in the more anaerobic strains are listed in Table 3. Previously studied
portion of the medium. Twenty-four percent of strains (group IC 3, reference 19) containing a
the strains grew only as single colonies in the similar cell wall composition may also belong to
anaerobic portion. this species.
Twenty-two strains isolated from skin did not Strain DSM 20260 (originally designated GH
produce a pH lower than 5.3 in yeast extract- 137) is the type strain of S . cohnii. A description
glucose broth under anaerobic conditions. A of this strain follows.
small amount of lactic acid was produced, Cells are gram-positive spheres, about 1.2 pm
predominantly L-lactic acid, but some strains in diameter, occurring predominantly in pairs
produced DL-lactic acid. and singly, occasionally in tetrads. Nonmotile
All strains grew well in media containing up and nonsporeforming.
to 10% NaC1, but 64% of the strains grew poorly Agar colonies are circular, entire, about 6.0
and 15%failed to grow in 15% NaC1. All strains mm in diameter, convex to slightly peaked with
grew well a t 15 C, and most grew a t 45 C; only a flat edge, smooth with glossy surface, white (a
7% failed to grow a t 45 C. color photograph of a colony is shown in Fig. lD,
All strains failed to produce coagulase. All reference 9), and opaque.
strains were catalase and benzidine test posi- Chemoorganotrophic: Metabolism is respira-
tive. Thirty-one percent of the strains demon- tory and fermentative.
strated weak hemolysis activity and 55% Catalase and benzidine test positive.
showed no hemolysis activity. More than 80% of Weakly facultatively anaerobic.
the strains produced weak to moderate acetyl- The anaerobic portion of thioglycolate me-
methylcarbinol reactions and failed to reduce dium showed a gradient from dense to light
nitrate. About 20% of the strains demonstrated growth in the deeper, more anaerobic, portion of
weak phosphatase and extracellular deoxyribo- the medium. The pH of yeast extract-glucose
nuclease activities, and 38% exhibited bacteri- broth was lowered under anaerobic conditions
olytic activity. from 6.8 to 5.3. Small amounts of L-lactic acid
All strains produced acid aerobically from were produced.
glucose, fructose, trehalose, and glycerol. About Growth on NaCl agar: Good growth with 10%
80% of the strains produced acid from mannose, NaC1, weak growth a t 15%.
maltose, and mannitol. Some strains produced Temperature relationships: Good growth a t
acid from lactose (31%) or xylitol (38%).Very 15 and 45 C.
few strains produced acid from sucrose (12%) or Coagulase, phosphatase, extracellular deoxy-
galactose (10%). No acid was produced from ribonuclease, and bacteriolytic activities not
rhamnose, xylose, arabinose, ribose, turanose, demonstrated.
gentiobiose, cellobiose, melezitose, sorbitol, Produced partial hemolysis of bovine blood
inositol, salicin, adonitol, dulcitol, arabitol, but not sheep blood.
erythritol, erythrose, raffinose, melibiose, fu- Acetylmethylcarbinol produced.
cose, tagatose, lyxose, or sorbose. Nitrates not reduced.
All strains were susceptible to lysostaphin Acid produced aerobically from glucose, fruc-
and resistant to lysozyme. About one-half of the tose, maltose, trehalose, mannitol, and glycerol.
strains were resistant to streptomycin or eryth- No acid from galactose, mannose, rhamnose,
romycin. More than 90% of the strains were xylose, arabinose, ribose, lactose, sucrose, tu-
susceptible to penicillin or tetracycline. About ranose, gentiobiose, cellobiose, melezitose,
90% of the strains were resistant to novobiocin xylitol, sorbitol, inositol, salicin, adonitol, dul-
(5 pg/ml). citol, arabitol, erythritol, erythrose, raffinose,
Twenty strains isolated from skin contained melibiose, fucose, t agatose, lyxose, or sorbose.
peptidoglycan of type ~-Lys-Gly,.,. Their cell Susceptible to lysostaphin (minimal inhibi-
wall teichoic acid contained glycerol and usu- tory concentration, 50 pg/ml); resistant to lyso-
ally glucose; glucosamine was also present in zyme.
extracted teichoic acids, but generally in low Antibiotic susceptibilities: Susceptible to
amounts. Some strains (KH 201, RM 10, JRM penicillin and tetracycline.
24, JL 143, and BB 3) also contain galactosa- Resistant to streptomycin (5 pg/ml), erythro-
mine in their cell walls. mycin (150 pg/ml), and novobiocin (20 pg/ml).
The G+C content of the DNA, as determined Cell wall peptidoglycan type: ~-Lys-Gly,.
56 SCHLEIFER AND KLOOS INT. J. SYST.BACTERIOL.

TABLE
3. Variable characters of 20 strains of S. cohniia
- -

Peptido-
Produc- glycan
L-
>olon: Growth He- tion of Acid (aerobically)from
mol% Lactic (mol/mol
Strain Pig- on 15% moly- acetyl- of Glu)
G + C ment acid
(%I NaCl sis methyl - - -
carbinol vlan Ma1 Mtl Xyli Ser GlY

- - - - - - -
DSM 20260 37.1 W 100 f + - f f 0 5.0
DSM 20261 36.5 W 100 f + f f f 0 5.3
DSM 20262 38.1 gw 100 f f - f + 0 5.4
DM 224 W f 5.6 100 f f + + 0 5.7
DM272 W 5.3 100 - - - f - 0 5.0
DM 154 gw + 5.3 50 f - + - + 0.2 5 .O
MK247 W + 5.5 57 f - + f + 0 5.5
RM 429 gw *c 5.4 100 f - - f f 0 5.3
CK 27 W + 5.3 55 +- + + + + 0 5.2
LK 478 Yt fC 6.0 73 + + * - 0 5.0
KH 201 Yt - 5.8 100 + - + + f 0 5.1
RM 10 gw *C 5.8 96 f + + + f 0 5.2
PM 244 Yt f 5.4 95 f f f - + 0 4.9
CM 89 W 60 f f + - + 0 5.4
DBM 388 W 50 - + + f + 0 5.0
DBM 247 W f f + * + 0 5.1
JL 143 gw + + + + + 0 5.2
J L 321 W 5.3 + f f f f 0.1 4.9
JRM 24 W f + f - f 0.1 6.0
BB 3 Yt + + -
--
-
-
f
-
0 5.2
-
For symbols and abbreviations, see Table 2.

Cell wall teichoic acid composition: Glycerol unless noted otherwise,

and glucose; glucosamine present in small Cells were gram-positive cocci, 0.8 to 1.3 pm
amounts. in diameter, occurring predominantly in pairs
G+C content of the DNA: 37.1 mol%. and tetrads. Nonmotile and nonsporeforming.
S . cohrzii can be distinguished from all other Colonies were raised to slightly convex, circu-
staphylococci primarily on the basis of carbohy- lar, 4.0 to 9.0 mm in diameter, usually entire,
drate reaction pattern, cell wall composition, smooth, glistening, and usually opaque. Colony
and, to a lesser extent, low acid production pigment was variable; however, most strains
under anaerobic conditions, production of ace- were unpigmented or had a slight yellow tint
tylmethylcarbinol, lack of nitrate reduction, which increased in intensity with age.
and resistance to novobiocin. There are also Growth occurred in both the aerobic and
some differences between S. cohrzii and other anaerobic portions of thioglycolate medium. In
staphylococcal species in colony pigment, cell the anaerobic portion, there was a gradient from
arrangement, growth a t extreme temperatures, dense to light growth or individual colonies in
and coagulase, hemolysin, phosphatase, extra- the deeper, more anaerobic, portion of the
cellular deoxyribonuclease, and bacteriolytic medium.
activities. Twenty-three strains isolated from skin and
(iv) S. huemolyticus sp.n. (hae.mo.ly’ti.cus. strains CCM 1798, 2444, and 2574, P.O. 375, S
M.L.adj. huemolyticus blood-dissolving) . Sev- 1, and S 226 fermented glucose under anaerobic
eral strains of this species were isolated previ- conditions and lowered the pH of yeast extract-
ously but were identified as belonging to Micro- glucose broth from 6.8 to 4.9-5.5. These strains
coccus halodurans, Micrococcus s ~ . ,or s. usually produced D-lactic acid from glucose; two
epidermidis (5, 15; P . Oeding, personal commu- strains also produced 15 to 30% L-lactic acid.
nication). These strains include CCM 1798, All the strains grew well at the NaCl concen-
2444, and 2574; P.O. 375 (P. Oeding, Bergen, trations tested up to 10%; 40% of the strains did
Norway); and S 1and S 226 (S. Schaefler, New not grow with 15%NaCl. All strains grew well at
York, N.Y.). Based on their cell wall composi- 45 C, but 57% of the strains failed to grow a t
tions and lactic acid configurations, these and 15 C.
other strains were placed in staphylococci group All strains were catalase and benzidine test
IIA4 (20). The following description of S. positive. All failed to produce coagulase. All
huemolyticus is based on a total of 117 strains, strains showed moderate or weak hemolysis.
VOL. 25, 1975 STAPHYLOCOCCI FROM HUMAN SKIN.I. 57

The hemolysis was best demonstrated with 5.0. Lactic acid was produced solely as the D
bovine blood and is slightly less intense with isomer.
sheep and rabbit blood. More than 80% of the Growth on NaCl agar: Good growth a t lo%,
strains reduced nitrate and produced acetyl- no growth a t 15% NaCl.
methylcarbinol. Most strains failed to demon- Temperature relationships: No growth a t
strate extracellular deoxyribonuclease and 15 C, good growth at 45 C.
phosphatase activity; only 20 to 30% of the Bovine and sheep erythrocytes were hemo-
strains showed weak activities. Bacteriolytic lyzed.
activity was variable. Coagulase, phosphatase, extracellular deoxy-
All strains produced acid aerobically from ribonuclease, and bacteriolytic activities were
glucose, maltose, sucrose, trehalose, and glyc- not demonstrated.
erol. About one-half of the strains produced acid Acetylmethylcarbinol not produced.
from fructose, galactose, lactose, turanose, and Nitrates were reduced.
mannitol. Only a few strains produced acid Acid was produced from glucose, maltose,
from mannose (3%)and ribose (17%).All strains sucrose, trehalose, and glycerol. No acid from
failed to produce acid from rhamnose, xylose, fructose, galactose, mannose, rhamnose, xylose,
arabinose, gentiobiose, cellobiose, melezitose, arabinose, ribose, lactose, turanose, gentiobi-
xylitol, sorbitol, inositol, salicin, adonitol, dul- we, cellobiose, melezitose, mannitol, xylitol,
citol, arabitol, erythritol, erythrose, raffinose, sorbitol, inositol, salicin, adonitol, dulcitol, ara-
melibiose, fucose, tagatose, lyxose, or sorbose. bitol, erythritol, erythrose, raffinose, melibiose,
All strains were slightly resistant or suscepti- fucose, tagatose, lyxose, or sorbose.
ble to lysostaphin and resistant to lysozyme. All Susceptible to lysostaphin (minimal inhibi-
strains, except S 1 and S 226, were susceptible tory concentration, 100 pg/ml); resistant to
to novobiocin and 90% of the strains were lysozyme.
susceptible to erythromycin. Most were suscep- Antibiotic susceptibilities: Susceptible to
tible to penicillin (79%)and tetracycline (91%). penicillin, erythromycin, tetracycline, and no-
Twenty-three strains isolated from skin and vobiocin. Resistant to streptomycin (5 pg/ml).
strains CCM 1798, 2444, and 2574, P.O. 375, S Cell wall peptidoglycan type: L-LYS-G~Y,.,,
1, and S 226 contained the cell wall peptidogly- L-Ser 2 .
can of type L - L ~ S - G ~ ~ ~ . ,L-Sero.,-l.,.
,.,, Their Cell wall teichoic acid composition: Glycerol
cell wall teichoic acid was composed of glycerol and glucosamine.
and glucosamine. G + C content of the DNA: 36.4 mol%.
T h e G + C content of DNA, as determined for S. huemolyticus can be distinguished from all
three strains, was 35.3 f 0.6 mol%. other staphylococci primarily by its hemolytic
Some variable characters of 23 selected activity, anaerobic growth pattern in thioglyco-
strains from skin and the G+C contents of the late medium, formation of D-lactic acid, and
DNA of representative strains are given in cell wall composition. There are also some
Table 4. significant differences between S . huemolyticus
Strain DSM 20263 (originally designated SM and certain other species in colony pigment,
131) is the type strain of S . huemolyticus. A growth a t extreme temperatures, carbohydrate
description of this strain follows. reaction pattern, nitrate reduction, and the lack
Cells were gram-positive spheres, about 1.3 of coagulase, phosphatase, extracellular deoxy-
pm in diameter, occurring predominantly in ri bonuclease, and bacteriolyt ic activities.
tetrads and less frequently in pairs. Nonmotile (v) S. xylosus sp.n. (xy.lo’sus. M.L. adj.
and nonsporeforming. xylosus xylose.) The following description of S.
Agar colonies were circular, entire, 5 to 6 mm xylosus is based on a total of 102 strains, unless
in diameter, slightly convex. White with a noted otherwise.
yellowish tint (see color photograph of colony: Cells were gram-positive cocci, 0.8 to 1.2 pm
Fig. lF, bottom row, reference 9), and smooth in diameter. They were nonmotile and non-
with glistening surface. sporeforming and occurred predominantly in
Chemoorganotrophic: Metabolism is respira- pairs or as single cells, occasionally as tetrads.
tory and fermentative. The colony morphology was highly variable.
Catalase and benzidine tests were positive. Colonies were circular, 5.0 to 10.0 mm in
Facultatively anaerobic. diameter, raised, slightly convex with a flat
Growth in the anaerobic portion of thioglyco- edge or umbonate; the edges were entire, undu-
late medium occurred only in individual colo- late, or crenate; the texture was smooth to
nies. rough; and they were glistening to dull; they
The pH of yeast extract-glucose broth was were usually opaque. Colony pigment was also
lowered under anaerobic conditions from 6.8 to variable. Many strains were orange-yellow or
 1/11
c3
X

TABLE
4. Variable characters of 23 strains of S . haemolyticusa

I
1 Strain mo'%
G+C

DSM 20263 36.4 Yt -C 5.0 100 -

DSM 20264 34.7 W f 5.1 100 f
DSM 20265 34.8 Yt fC 5.1 100 f
KES 23 FP fC 5.2 100 f
SM 102 Yt f 4.9 100 -
KL 109 Yt fC 5.2 1 100 +
MK67 Yt f 5.0 loo -
KL 35 gw + 5.5 100 =t
KL 194 Yt + ' 5.4 100 f
Yt ZtC 5.5 100 f
GH 107 Yt -C 5.4 100 +
MK 50 Yt ItC 5.2 100 -
AW 263 gw *C 5.3 100 -
MK 310 Yt fC 5.0 100 f
AW 174 Yt f 5.3 100 -

Yt fC 5.3 100 f
gw f C 4.9 100 f
SM 254 gw I f 5.3 100 -

RM 357 gw f C 5.0 100 f

MK 104 Yt -C 5.2 100 f
AW 146 gY f 5.1 100 -

g + 5.O 75 +
gw f C 5.3 70 -
1/11
a For symbols and abbreviations, see Table 2.
VOL.25. 1975 STAPHYLOCOCCI FROM HUMAN SKIN. I. 59
yellow-gray. Some strains were highly sectored other related strains studied previously (group
with clones of different pigment intensities or IB, reference 20) contained peptidoglycan of
with pigmented and unpigmented clones. Some type ~-Lys-Gly,-,.The cell wall teichoic acid of
strains were unpigmented a t 35 to 37 C but these strains contained glucosamine and, like
developed intense pigment a t temperatures that of S. suprophyticus, two polyols: glycerol
below 20 C. Approximately 20% of the strains and ribitol. Studies on strains CCM 1400 and
were unpigmented a t all temperatures tested 2606 proved that two distinct cell wall teichoic
from 15 to 45 C. acids, a ribitol and a glycerol teichoic acid,
Growth occurred in both the aerobic and occurred in these strains (Schleifer and van
anaerobic port ions of thioglycolate medium. Schaik, unpublished results).
Growth in the anaerobic portion was often The G + C content of the DNA, as determined
dense; however, 25% of the strains demon- in six strains, is 34.2 f 1.1 mol%.
strated less growth or produced only a few Some variable characters and the G+C con-
colonies in the deeper, more anaerobic, portion tent of DNA of some representative strains are
of this medium. given in Table 5 .
Twenty of the strains isolated from skin in Strains with similar properties were described
this study and strains CCM 1400 and 2210 by Shaw et al. (22) as S. lactis. However, NCTC
lowered the pH of yeast extract-glucose broth 7564 ( = ATCC 15306 = CCM 884) was desig-
under anaerobic conditions from 6.8 to 4.9-5.6. nated by Shaw et al. as the type strain of S.
These strains produced predominantly L-lactic lactis, and this strain, a typical micrococcus
acid. (18), was subsequently designated (11) as the
All strains grew well a t NaCl concentrations neotype strain of Micrococcus uuriuns Migula
up to 10%; 48% of the strains grew poorly and 1900. Thus, S. lactk is a later objective syn-
14% failed to grow a t a concentration of 15%.All onym of M . uarians. Some of the strains previ-
strains grew well at 15 C, but 62% failed to grow ously identified as belonging to S. lactis are now
a t 45 C. placed in S. xylosus.
None of the strains produced coagulase. All Strain DSM 20266 (originally designated KL
strains were positive for catalase and the benzi- 162) is the type strain of S. xylosus. A descrip-
dine test. tion of this strain is presented below.
About 80% of the strains reduced nitrate and Cells were gram-positive cocci, about 1.2 pm
failed to produce or produced only very low in diameter, occurring singly and in pairs. Non-
levels of acetylmet hylcarbinol. All strains failed motile and nonsporeforming.
to demonstrate hemolysis activity, and 80% of Agar colonies were circular, entire, about 4
the strains did not show an extracellular deoxy- mm in diameter, raised to slightly convex,
ribonuclease activity. About 80% of the strains yellow-orange (see color photograph of a colony:
showed phosphatase activity. Bacteriolytic ac- Fig. lC, middle row, reference 9), and opaque.
tivity was usually weak or absent. Chemoorganotrophic: Metabolism is respi-
All strains produced acid aerobically from ratory and fermentative.
glucose, fructose, mannose, maltose, xylose, Catalase and benzidine test positive.
mannitol, and glycerol. About 80% of the strains Faculatively anaerobic.
produced acid from galactose, arabinose, lac- Growth on NaCl agar: Good growth a t 10%
tose, sucrose, trehalose, or turanose. Less than NaC1, poor growth a t 15% NaC1.
30% of the strains produced acid from rham- Temperature relationships: Good growth a t
nose, ribose, gentiobiose, xylitol, sorbitol, or 15 C, but no growth a t 45 C.
inositol. None was produced from cellobiose, Coagulase not produced.
melezitose, salicin, adonitol, dulcitol, arabitol, Nitrates were reduced.
erythritol, erythrose, raffinose, melibiose, fu- Acetylmethylcarbinol not produced.
cose, tagatose, lyxose, or sorbose. Acid produced from glucose, fructose, galac-
All strains were susceptible to lysostaphin tose, mannose, xylose, arabinose, maltose, lac-
and resistant to lysozyme. tose, sucrose, trehalose, turanose, mannitol,
All strains were susceptible to penicillin and and glycerol.
streptomycin. At least 80% of the strains were Acid not produced from rhamnose, ribose,
susceptible to erythromycin and tetracycline. cellobiose, melezitose, xylitol, sorbitol, inositol,
About 70% were resistant to novobiocin (5 salicin, adonitol, dulcitol, arabitol, erythritol,
pdml). erythrose, raffinose, melibiose, fucose, tagatose,
Twenty of the strains isolated from skin in lyxose, or sorbose. Yeast extract-glucose broth is
this study and strains CCM 1400 and 2210 and acidified to pH 5.0 under anaerobic conditions.
60 SCHLEIFER AND KLOOS INT. J. SYST.BACTERIOL.

- TABLE
5. --
Variable characters t 22 stmins of S.-
nYlosusa
Anaer- Peptido-
Anaer- Produc- Acid (aerobically) glycan
obic L-
tion of v'itratc 'has-
2Ofonj obic fermen- ,attic from rnol/mol of'
nol% acetyl- reduc- pha-
Strain P%- .hiogly- tation of acid Glu)
G+C nethyl- tion tase -
ment colate -
glucose (%) :arbinol
growth Mtl Ara XYl Glr
(pH)
- - - - - - -
DSM 20266 36.2 YO + 5.0 95 - + + + + + 5.1
DSM 20267 34.6 YO + 5.1 94 f + + + + + 5.2
DSM 20268 35.2 W + 5.1 90 - + + + + t 5.5
DM37 35.2 YO + 4.9 88 f + + + + + 4.9
DM 30 w + 5.0 85 + - + + + 4.9
SM 212 Y + 4.9 91 f + - + - + 5.5
SM 24'7 w + 5.0 95 f + + + + + 5.0
TW 41 YO + 5.2 91 f + + + + + 5.3
TW 83 YO +C 5.0 82 - + + + + + 5.6
TW 54 Yt + 5.1 100 f + + + + + 5.2
AW 124 35.6 Y + 4.9 88 + + + + + 5.4
KH 168 YO f 5.0 + + + + + + 5.3
GH 30 34.8 W + 4.9 - + + + + + 5.0
SL 114, YO + 5.0 * + + + + + 5.0
SE 4 W + 5.1 + + + + + + 5.6
DM 172 #w + 5.1 f + + + + t 5.6
SM 159 w + 5.1 + + + + + t 5.7
KL 236 W + 5.4 + t + + + 5.2
KL 117 Yt + 5.1 - + + + + f+ i 5.1
AW 244 YO + 5.3 - +
- + + - - 1 5.4
CCM 1400 30.2b gw -C 5.6 - + ND ++ + + j 5.3
CCM 2210 32.W gw -c 5.5 - + -
ND + + + + i 5.2
- - - -- I
For symba and i Ibreviations. see rable 2.
According to BohaEek et a]. (4)
L' According to Silvestri and Hill (23).

L-Lactic acid is predominantly produced. temperature range than most other staphylo-
Susceptible to lysostaphin (minimal inhibi- cocci. The common properties of the novobi-
tory concentration, 50 pg/ml) and resistant to ocin-resistant species may have been the reason
lysozyme. why the strains of these species were classified
Antibiotic susceptibilities: Susceptible to in the new edition of Bergey's Manual (Baird-
penicillin, erythromycin, streptomycin, and tet- Parker, personal communication) as belonging
racycline. Resistant to novobiocin ( 5 pg/ml). to S . saprophyticus. Our studies, however, have
Cell wall peptidoglycan type: L - L Y S - G ~Cell
Y~. clearly demonstrated that a separation of these
wall teichoic acid composition: glycerol, ribi- novobiocin-resistant strains into three distinct
tol, and glucosamine. species is feasible. The cell wall peptidoglycan
G+C content of the DNA: 36.2 mol%. type differentiates S. saprophyticus from S .
S . xylosus can be distinguished from all the cohnii and S. xylosus, and the chemical compo-
other staphylococci primarily on the basis of sition of the cell wall teichoic acids separates S .
its carbohydrate fermentation pattern, cell cohnii from S . xylosus and S . saprophyticus. In
wall composition, and growth a t extreme tem- addition, the failure to reduce nitrate and the
peratures and, to a lesser extent, on its reduc- lack of phosphatase activity distinguish S.
tion of nitrate, production of phosphatase, re- saprophyticus and S . cohnii from most strains
sistance to novobiocin, and low acid production of S. xylosus. The carbohydrate reaction pat-
under anaerobic conditions. tern is also a valuable character for the separa-
Several of the properties overlap with those of tion of these three species. Thus, S . xylosus
S . cohnii and S. saprophyticus, suggesting a strains usually ferment xylose, arabinose, and
relatively close relationship among these three 8-gentiobiose in contrast to most other staphy-
species. With respect to cell wall composition, lococci. Most strains of S . saprophyticus are
S. xylosus and S . cohnii contain identical pepti- distinguished by their ability to ferment xylitol,
doglycan types, whereas with S. xylosus and S . whereas S. cohnii strains usually differ from
saprophyticus the chemical compositions of the other staphylococci by their inability to ferment
cell wall teichoic acids are identical. Low acid sucrose or turanose.
production from glucose under anaerobic condi-
tions and resistance to novobiocin are properties ACKNOWLEDGMENTS
common to all three species. Moreover, the We are greatly indebted to E. Hagner, M. Musselwhite, I.
three species have a lower optimal growth Pomper, and B. Popp for their excellent technical assistance
Va. 25, 1975 STAF'HnOCOCCI FROM HUMAN SKIN. I. 61
throughout this study. J . Clin. Pathol. 22:249-253.
This research was supported by a grant of the Deutsche 13. Noble, W. C. 1969.Distribution of the Micrococcaceae.
Forschungsgemeinschaft and by Public Health Service re- Br. J. Dermatol. 81(Suppl. 1):27-32.
search grant AI 08255 from the National Institute of Allergy 14. Pennock, C. A., and R. B. Huddy. 1967. Phosphatase
and Infectious Diseases. reaction of coagulase-negative staphylococci and mi-
crococci. J. Pathol. Bacteriol. 93:M-W.
REPRINT REQUESTS 15. Schaefler, S. 1971.Staphylococcus epidermidis BV: anti-
biotic resistance patterns, physiological characteris-
Address requests for reprints to: Dr. K. H. Schleifer, tics, and bacteriophage susceptibility. Appl. Microbiol,
Lehrstuhl Mikrobiologie, Universitat Munchen, 8 Miinchen 22:693-699.
19,Menzinger Str. 67,BRD, Germany. 16. Schleifer, K. H.1973.Chemical composition of staphylo-
coccal cell walls, p. 13-32.In J. Jeljaszewicz and W.
LITERATURE CITED Hryniewicz (ed.), Contributions to microbiology and
1. Bailey, W. R., and E. G. Scott. 1966.Diagnostic microbi- immunology, vol. 1. Staphylococci and staphylococcal
ology. The c. V. Mmby Co., St. Louis. infections. S. Karger, Basel.
2. Baird-Parker, A. C. 1963.A classification of micrococci 17. Schleifer, K. H., and 0. Kandler. 1967.Zur chemischen
and staphylococci based on physiological and biochem- Zusammensetzung der Zellwand der Streptokokken. I.
ical tests. J. Gen. Microbiol. 30:409-427. Die Aminostiuresequenz des Mureins von Streptococ-
3. Baird-Parker, A. C. 1965.The classification of staphylo- cus thermophilus und Streptococcus faecalis. Arch.
cocci and micrococci from world-wide sources. J. Gen. Mikrobiol. 57:335-364.
Microbiol. 38:363-387. 18. Schleifer, K. H. and 0.Kandler. 1970. Amino acid
4. Bohacek, J., M. Kocur, and T. Martinec. 1965.Deoxyri- sequence of the murein of P~ROCOCCUS and other
bonucleic acid base composition and taxonomy of the Micrococcaceae. J. Bacteriol. 103:387-392.
genus Micrococcus. Publ. Fac. Sci. Univ. J.E. PurkynB, 19. Schleifer, K. H.,and 0. Kandler. 1972.The peptidogly-
Bmo. K35:318-322. can types of bacterial cell walls and their taxonomic
5. Buck, J. D. 1965.Micrococcus halodumna nsp. Int. Bull. implications. Bacteriol. Rev. 36:407-477.
Bacteriol. Nomencl. Taxon. 15:181-184. 20. Schleifer, K. H.,and M. Kocur. 1973. Classification of
6. Dennis, D. 1962. Lactic acid racemase from Clostridium staphylococci based on the chemical and biochemical
butylicum, p. 430-432. In S. P. Colowick and N. 0. properties. Arch. Mikrobiol. 93:65-85.
Kaplan (ed.),Methods in enzymology, vol. 5. Aca- 21. Schleifer, K. H.,and J. Steber. 1974.Chemische Untersu-
demic Press Inc., New York. chungen am Phagenrezeptor von Staphylococcus
7. Evans, J. B., and W. E. Kloos. 1972.Use of shake cultures epidermidis. Arch. Mikrobiol. 98:251-270.
in a semisolid thioglycolate medium for differentiating 22. bhaw, C.,J. M. Stitt, and S. T. Cowan. 1951.Staphylo-
staphylococci from micrococci. Appl. Microbiol. cocci and their classification. J. Gen. Microbiol.
23~326-331. 5:1010-1023.
8. Hugh, R., and Me. A. Ellis. 1968.The neotype strain for 23. Silvestri, L. G., and L. R. Hill. 1965.Agreement between
Staphylococcus epidermidis (Winslow and Winslow deoxyribonucleic acid base composition and taxometric
1808) Evans 1916. Int. J. Syst. Bacteriol. 18:231-239. classification of gram-positive cocci. J. Bacteriol,
9. Klooe. W. E.. and K. H. Schleifer. 1975. Isolation and 90 ~136-140.
characterization of staphylococci from human skin. 11. 24. Subcommittee on Taxonomy of Staphylococci and Micro-
Deecription of four new species: Staphylococcus cocci. 1965. Recommendations. Int. Bull. Bacteriol.
warnerr, Staphylococcus capitia, Staphylococcus Nomencl. Taxon. 15:109-110.
hominis, and Staphylococcus simuhns. Int. J. Syst. 25. Verhoef, J., C. P. A. van Boven, and K. C. Winkler. 1971.
Bacteriol. 25:62-79. Lysogeny in coagulase-negative staphylococci. J. Med.
10. K l m , W. E., T. G. Tornabene, andK. H. Schieifer. 1974. Microbiol. 4:405-412.
Isolation and characterization of micrococci from 26. Wadstrom, T., and K. Hisatsune. 1970. Bacteriolytic
human skin, including two new species: Micrococcus enzymes from Staphylococcus aureua. Purification of
1yloe.and Micrococcus kristinae. Int. J. Syst. Bacteriol. an endo-B-N-acetylglucosaminidase. Biochem. J.
24:79-101. 120:725-734.
11. Kocur, M., and T. Martinec. 1972.Taxonomic status of 27. Wolin, M. J., A. R. Archibald. and J. Baddiley. 1966.
Micrococcus varians Migula 1900 and designation of Changes in wall teichoic acid resulting from mutations
the neotype strain. Int. J. Syst. Bacteriol. 22:228-232. of Staphylococcus aureus. Nature (London)
12. Noble, W. C. 1969.Skin carriage of the Micrococcaceae. 209:484-486.